Patent Publication Number: US-2023151341-A1

Title: Method for specifically editing genomic dna and application thereof

Description:
FIELD OF THE INVENTION 
     The present invention relates to the field of bioengineering technology, and in particular relates to a method for specifically modulating the methylation/demethylation status of genomic DNA and use thereof. 
     BACKGROUND OF THE INVENTION 
     DNA methylation is one of the important modifications in epigenetic modulation and is called the “fifth base” in mammalian DNA except for the four bases of ATCG. As a covalent modification, DNA methylation plays an important role in normal differentiation and disease development and can be stably inherited in cell differentiation of higher eukaryotic organs, and it is found in zebrafish that DNA methylation can be passed on to the next generation through sperm. Under the influence of cell differentiation, disease and environment, the methylation status of DNA will change greatly. 
     Studies have shown that DNA methylation is closely related to the occurrence and development of tumors. Changes in DNA methylation status include hypermethylation and hypomethylation. In general, DNA hypermethylation in the promoter region of the gene has the effect of silencing gene expression, while hypomethylation activates gene expression. DNA analysis of different tumor cells showed that the probability of genetic mutations in cancerous cells was much lower than expected. In the transcriptome range, gene expression inhibition by promoter hypermethylation in colorectal cancer was detected, and it was found that up to 5% of known genes have abnormal promoter hypermethylation in tumor cells. Therefore, it can be speculated that DNA methylation changes may play a greater role in cell malignant transformation than genetic mutations. 
     Target-specific nucleic acid editing techniques, especially the specific editing of genomic DNA, have always been an important technical basis for gene therapy. With the deepening of epigenetics research, more and more studies have shown that the methylation of the genome is directly involved in transcriptional modulation and other modulation of the genome, while the promotor and enhancer regions of an active expression gene are usually hypomethylated. Therefore, a nucleotide editing technique capable of specific demethylation is very important for the transcriptional activation of silenced genes. 
     Currently, site-specific and region-specific demethylation processes have been reported. For example, genomic remodeling of germ cells is often accompanied by large-scale demethylation. In addition, 5mC can be oxidized by certain enzymes (such as Tet) to 5hmC, followed by NER or BER process to be finally demethylated. Xu Guoliang, et al., have reported and filed a patent application for demethylation by reagents such as Tet dioxygenase and thymidine DNA glycosylase in 2015, but this method has not been able to accurately edit a certain site, being an important bottleneck for use in gene therapy or experimental technology tools. 
     Certain members of the Apobec protein family have the ability to deaminate 5mC into T in single-stranded DNA. With such characteristics and the precise positioning ability of the CRISPR protein family, it has become possible to develop a system that can accurately edit methylation at a specific site in the genome. 
     SUMMARY OF THE INVENTION 
     In order to solve the above problems, the present invention provides a method for editing a target nucleic acid molecule, comprising the steps of:
     (1) obtaining a recombinant vector encoding a fusion protein (A) and a small guide RNA (sgRNA) (B), wherein the fusion protein (A) comprises an Apobec family protein domain at N-terminal and a Cas9 family or a Cpf1 family protein domain whose nuclease activity is inactivated at C-terminal, and the small guide RNA has a complementary region to a target editing region of the target nucleic acid molecule, wherein the target editing region of the target nucleic acid molecule includes at least one methylated cytosine nucleotide;   (2) contacting the recombinant vector encoding the fusion protein (A) and the small guide RNA (sgRNA) (B) obtained in the step (1) with the target nucleic acid molecule.   

     The recombinant vector in the above steps may be a recombinant vector in which two vectors respectively encode the fusion protein (A) and the small guide RNA (sgRNA) (B), or a recombinant vector in which a recombinant vector encodes both the fusion protein (A) and the small guide RNA (sgRNA) (B). 
     In a preferred embodiment, the Apobec family protein at N-terminal of the fusion protein is selected from the group consisting of human Apobec3A or Apobec3H, or a protein having deamination activity with 95% or more homology to human Apobec3A or Apobec3H. More preferably, the Apobec protein is Apobec3H or Apobec3A. 
     In another preferred embodiment, the Cas9 family protein whose nuclease activity is inactivated at C-terminal of the fusion protein is the one obtained by mutating aspartic acid at position 10 and histidine at position 840 in the wild-type Cas9 protein to alanine and alanine, or the Cpf1 protein whose nuclease activity is inactivated at C-terminal of the fusion protein is the one obtained by mutating aspartic acid to alanine at position 908 in the wide-type Cpf1 protein. 
     In order to provide better spatial structural flexibility for the two protein domains of the fusion protein, a linker consisting of 3-14 motifs can be added between the two domains of the fusion protein. The motif is selected from (GGS). The longer the linker is, the higher the spatial flexibility of the protein is and the larger the editable target area is. 
     To facilitate expression and purification of the fusion protein, a purification tag sequence can also be included. A commonly used purification tag is 6xHis. 
     In a more preferred embodiment, the fusion protein is selected from any of the sequences of SEQ ID NOs. 201-207. 
     The present invention also provides a gene sequence encoding the above fusion protein sequence, which is preferably selected from the group consisting of SEQ ID NOs. 301-307. 
     The present invention also provides a recombinant vector comprising any of the above gene sequences, which may be a prokaryotic expression vector or a eukaryotic expression vector, including but not limited to a plasmid vector, a viral vector, and the like, for the purpose of subsequent experiments. 
     Another aspect of the invention provides a small guide RNA molecule. In a preferred embodiment, the small guide RNA is 60 to 80 bp in length. In another preferred embodiment, the complementary region of the small guide RNA to the target nucleic acid molecule is 18 to 25 bp in length, preferably 20 bp. 
     A method for editing a target nucleic acid molecule in vitro, comprising the steps of: (1) obtaining a recombinant vector encoding a fusion protein (A) and a small guide RNA (sgRNA) (B), wherein the fusion protein (A) comprises an Apobec family protein domain at N-terminal and a Cas9 family or a Cpf1 family protein domain whose nuclease activity is inactivated at C-terminal, and the small guide RNA has a complementary region to a target editing region of the target nucleic acid molecule, wherein the target editing region of the target nucleic acid molecule includes at least one methylated cytosine nucleotide;
     (2) contacting the fusion protein (A) and the small guide RNA (sgRNA) (B) with the target nucleic acid molecule;   (3) after a high temperature termination reaction, adding an effective amount of TDG, and carrying out a reaction at 42° C. for 6 to 8 hours; and   (4) adding an effective amount of EDTA, formamide and NaOH, and carrying out a reaction at 90 to 95° C. for 5 to 10 minutes.   

     The present invention also provides use of the method for editing a target nucleic acid molecule for specifically modulating genomic DNA methylation/demethylation status. 
     In the method for editing a target nucleic acid molecule according to the present invention, the target nucleic acid molecule contains at least one methylated cytosine nucleotide, the methylated cytidine nucleotide is associated with diseases such as cancer, genetic disorders, developmental errors and the like. The method for editing a target nucleic acid molecule can be used for the treatment of a disease associated with cytosine nucleotide methylation, including but not limited to diseases associated with abnormal cell differentiation. 
     The Beneficial Effects of the Present Invention 
     In the present invention, the Apobec protein having deamination activity is guided to the methylated cytosine position of the target nucleic acid molecule to modify the methylated cytosine by the guidance of sgRNA and the specific binding function of the mutant Cas9 or Cpf1. Further, the methylated cytosine is removed by an in vivo DNA repair mechanism to achieve specific editing of the target nucleic acid molecule. The gene editing method of the present invention has high specificity and has no dependence on the upstream and downstream sequences of the target site, and thus has universal applicability. Moreover, the gene editing method of the present invention only edits the target, does not produce off-target effects, and does not introduce insertion or deletion mutations during editing, thus has low toxic side effects. 
    
    
     
       BRIEF DESCRIPTION OF THE DRAWINGS 
         FIG.  1    shows a schematic diagram of extracellular editing of fusion protein. 
         FIG.  2    shows a schematic diagram of intracellular editing of fusion protein. 
         FIG.  3    shows tests for active intensities and ranges of several fusion proteins in vitro. 
         FIG.  4    shows effect of the base located adjacent to upstream of the editing target site on editing efficiency. 
         FIG.  5    shows editing results in two groups of HEK293 cell lines. 
         FIG.  6    shows editing results of the two fusion proteins in the same region of the PC3 cell line. 
     
    
    
     DETAILED DESCRIPTION OF THE INVENTION 
     The Cas9 or Cpf1 protein is a double-stranded DNA nuclease that binds to a targeting sequence and cleaves double-stranded DNA under the action of a small guide RNA (sgRNA). The Cas9 protein whose nuclease activity is inactivated retains the activity of binding to the targeting sequence, but does not cleave the target site. In the present invention, the methylated cytosine in the targeted sequence region is deaminated by fusing the Cas9 or Cpf1 protein whose nuclease activity is inactivated with the Apobec protein having deamination activity and guiding the Apobec protein to the target sequence region of the target nucleic acid molecule by the mutated Cas9 protein or Cpf1 protein, so that the target Met-C becomes T under deamination and does not pair with G on the complementary chain to form a protrusion. The addition of an effective amount of TDG after termination of the reaction by high temperature (the main effect is to inactivate the fusion protein by high temperature, usually at a temperature of 90 to 95° C.) removes the mismatched T base, thereby forming a deletion at the editing target site of the substrate. The dsDNA then changes back to ssDNA and cleaves at the base deletion site by the combined action of an effective amount of EDTA, formamide and NaOH. 
     Based on the above experiments, the applicant has found that the fusion protein Apobec-dCas9 or Apobec-dCpf1 enables site-specifically editing of methylated cytosine site in the target sequence region, which does not rely on the upstream and downstream sequences of the methylated cytosine site, has universal applicability, does not cause off-target effects, and does not introduce other insertion or deletion mutations, so there are no other toxic side effects. 
     The details will be further described below by way of specific examples. However, it should be understood that the specific embodiments are only used to explain the present invention and are not intended to limit the scope of the present invention. The instruments, devices, reagents, methods and the like used in the present application are all instruments, devices, reagents and methods commonly used in the art unless otherwise specified. 
     Examples 
     Example 1. Recombinant Protein Expression and Purification 
     Invitrogen was commissioned to synthesize 6His-NLS-Apobec3H-linker (GGS-GGS-GGS) dCas9(Asp10Ala/His840A1a), 6His-NLS-Apobec3H-linker (GGS-GGS-GGS-GGSGGS-GGS-GGS), 6His-NLS-Apobec3H-linker (GGS-GGS-GGS-GGS-GGS-GGSGGS-GGS-GGS-GGS-GGS-GGS-GGS-GGS)-dCas9(Asp10Ala/His840A1a), 6HisNLS-Apobec3A-linker (GGS-GGS-GGS)-dCas9(Asp10Ala/Hi s840A1a) dCas9(Asp10Ala/His840A1a), 6His-NLS-Apobec3 A-linker (GGS-GGS-GGS-GGSGGS-GGS-GGS)-dCas9(Asp10Ala/His840A1a), 6His-NLS-Apobec3A-linker (GGS-GGS-GGS-GGS-GGS-GGS-GGS-GGS-GGS-GGS-GGS-GGS-GGS-GGS) dCas9(Asp10Ala/His840A1a), 6His-NLS-Apobec3H-linker (GGS-GGS-GGS-GGSGGS-GGS-GGS)-dCpf1(Asp908A1a) gene sequences, respectively SEQ ID NO. 301, NO. 302, NO. 303, NO. 304, NO. 305, NO. 306 and NO. 307, and a Nco I endonuclease site was introduced at the 5′ end of the gene fragment, and a Hind III endonuclease site was introduced at the 3′ end. The synthesized gene fragment and the pET28a (+) vector were respectively double digested with Nco I and Hind III, and the gene fragment and the vector fragment were ligated with T4 DNA ligase, and DH5a competent cells (Tiangen Biochemical Technology (Beijing) Co., Ltd.) were routinely transformed, and positive clones were selected according to kanamycin resistance, then the plasmids were extracted. The recombinant plasmid was identified by Nco I and Hind III double digestion and agarose gel electrophoresis. Meanwhile, Invitrogen was commissioned to sequence the recombinant plasmid, and the results of the sequencing were analyzed using BioEdit software. The results were identical to the designed sequence, indicating that the recombinant plasmid was successfully constructed. 
     The obtained positive clone plasmid was transformed into  E. coli . BL21 (DE3) competent cells (Tiangen Biotechnology (Beijing) Co., Ltd.), and cultured overnight at 37° C. in LB medium containing 100 μg/mlkanamycin, and then transferred to 1 L of the same LB medium and cultured at 37° C. to OD=0.6 about. The medium was then cooled to 4° C. and induced to express for approximately 16 hours by the addition of 0.5 mM IPTG. The cells were collected by centrifugation at 4000 g and resuspended in lysis buffer (50 mM Tris pH=7.0, 1 M NaCl, 20% glycerol, 10 mM TCEP). The cells were lysed by ultrasonic method (6W output for 8 minutes, on for 20 seconds and off for 20 seconds), and the supernatant was separated by centrifugation at 25,000 g. The supernatant was incubated with Nickel resin (ThermoFisher) at 4° C. for 1 hour, then passed through a gravity column and washed with 40 ml of lysis buffer. The recombinant protein was eluted with a 285 mM lysis buffer, diluted to 0.1 M NaCl and concentrated to the appropriate concentration with a centrifuge tube. The quality and concentration of the recombinant protein were determined by SDS Page. 
     The recombinant protein sequences were SEQ ID NO. 201-207. 
     Example 2. sgRNA In Vitro Transcription 
     Based on the 34 dsDNA substrate sequences to be tested (SEQ ID NO. 39-54 and their complementary strands 55-70, 71-85 and their complementary strands 86-100, 101-104 and their complementary strands 105-108) and the pFYF320 vector sequence providing the sgRNA universal sequence, the sgRNA forward primer (SEQ ID NO. 2-17, 18-34, and 35-38) and the reverse primer (SEQ ID NO. 1) were respectively designed. The sgRNA was obtained from a linear DNA fragment containing the T7 promoter by TranscriptAid T7 High Yield Transcription Kit (ThermoFisher Scientific), using DpnI to remove the template DNA, and then purified using a MEGAclear Kit (ThermoFisher Scientific), and the mass was detected by UV absorption. 
     Example 3. Substrate Preparation 
     Invitrogen was commissioned to synthesize the forward and reverse oligonucleic acid strand sequences of the substrate sequence, wherein the 5′ end of the positive strand sequence was labeled with FAM fluorescent labeling. 2 OD single-stranded oligonucleic acid strands were separately dissolved in 500 μl of water, and an equal amount of the positive and negative chain solutions were mixed and allowed to stand for 5 minutes to obtain a double-stranded substrate (dsDNA). 
     Fifteen sequences as SEQ ID NO. 39-54 were used for the dCas9 fusion protein demethylation range test. 
     Fifteen sequences as SEQ ID NO. 71-85 were used for the dCas9 fusion protein demethylation range test. 
     Four sequences as SEQ ID NO. 101-104 were used to test the effect of the base located adjacent to upstream of the target site on activity. 
     Example 4. In Vitro Activity Test 
     The recombinant fusion protein obtained in Example 1 was separately mixed with the sgRNA obtained in Example 2 in a molar ratio of 1:1, and allowed to stand at room temperature for 5 minutes. The corresponding dsDNA substrate was added to a final concentration of 125 nM and reacted at 37° C. for 2 hours. After the obtained dsDNA was purified using EconoSpin micro spin column (Epoch Life Science), 1 unit of TDG (NEB) was added and reacted at 37° C. for 1 hour. After the reaction, 10 μl of formamide, 1 μl of 0.5 M EDTA, and 0.5 μl of 5 M NaOH were added, and the mixture was reacted at 95° C. for 5 minutes. The product was isolated on 10% TBE-urea gel. 
     The target DNA strand contained the target Met-C and the 3′ end was labeled with the fluorophore FAM. Under the action of the recombinant protein, Met-C was converted to T and thus could not be paired with G of the complementary strand. Under the action of TDG, the mismatched T was going to be excised, leaving a base deletion site. Under the action of formamide and NaOH, the double strand became a single strand and was further cleaved at the base deletion site, thereby forming a short strand labeled with a fluorescent group FAM. The long and short chain marked DNAs were separated in urea gel. If a long and a short band appeared on the gel, it indicated that the recombinant protein was active. 
     Example 5. Preparation of dsDNA Substrate for Pyrosequencing 
     Invitrogen was commissioned to synthesize the forward and reverse oligonucleic acid strand sequences of the substrate sequence, wherein the 5′ end of the positive strand sequence was labeled with FAM fluorescent labeling. 2 OD single-stranded oligonucleic acid strands were separately dissolved in 500 μl of water, and an equal amount of the positive and negative chain solutions were mixed and allowed to stand for 5 minutes to obtain a double-stranded substrate (dsDNA). The recombinant fusion protein obtained in Example 1 was separately mixed with the sgRNA obtained in Example 2 in a molar ratio of 1:1, and allowed to stand at room temperature for 5 minutes. The corresponding dsDNA substrate was added to a final concentration of 125 nM and reacted at 37° C. for 2 hours. The reacted dsDNA was purified using EconoSpin micro spin column (Epoch Life Science) and submitted to BGI for pyrosequencing after sulfite treatment and amplication with designed primers. 
     Example 6. In Vivo Activity Assay 
     (1) Cell culture 
     The HEK293 cell line or PC3 cell line was maintained in Dulbecco&#39;s Modified Eagle&#39;s Medium plus under an environment of 37° C. and 5% carbon dioxide. 
     (2) Construction of PX330 recombinant protein expression vector 
     Invitrogen was commissioned to synthesize 6His-NLS-Apobec3H-linker (GGS-GGS-GGS) dCas9(Asp10A1a/His840Ala), 6His-NLS-Apobec3H-linker (GGS-GGS-GGSGGS-GGS-GGS-GGS), 6His-NLS-Apobec3H-linker (GGS-GGS-GGS-GGSGGS-GGS-GGS-GGS-GGS-GGS-GGS-GGS-GGS-GGS) dCas9(Asp10Ala/His840A1a), 6His-NLS-Apobec3A-linker (GGS-GGS-GGS) dCas9(Asp10Ala/His840A1a)-dCas9(Asp10Ala/His840Ala), 6His-NLSApobec3A-linker (GGS-GGS-GGS-GGS-GGS-GGS-GGS) dCas9(Asp10Ala/His840A1a), 6His-NLS-Apobec3A-linker (GGS-GGS-GGSGGS-GGS-GGS-GGS-GGS-GGS-GGS-GGS-GGS-GGS-GGS) dCas9(Asp10A1a/His840Ala), 6His-NLS-Apobec3H-linker (GGS-GGS-GGSGGS-GGS-GGS-GGS)-dCpf1(Asp908A1a) gene sequences, respectively SEQ ID NO. 301, NO. 302, NO. 303, NO. 304, NO. 305, NO. 306 and NO. 307, and a BamHI endonuclease site was introduced at the 5′ end of the gene fragment, and an AgeI endonuclease site was introduced at the 3′ end. The synthesized gene fragment and the pX330 vector (Addgene) were respectively double digested with BamHI and AgeI, and the gene fragment and the vector fragment were ligated with T4 DNA ligase. It was confirmed by sequencing that the recombinant vector was constructed correctly. The sgRNA vectors corresponding to the five intracellular experiments inserted the corresponding PCR products (obtained by PCR from forward primers 121, 123, 125, 127, 129 and reverse primers 1, 122, 124, 126, 128, 130) through MluI and SpeI double digestion. 
     (3) Transfection 
     A. One day before transfection, HEK293 cells or PC3 cells were inoculated in a medium that did not contain antibiotics, and the confluence of the cells at the time of transfection was 30-50%. 
     B. Preparation of transfection samples: 
     1 μl of 20 μM pX330 recombinant vector and 1.5 μl of cell transfection reagent Lipofectamine™ 2000 (Invitrogen) were diluted in 0.05 ml Opti-MEM (Invitogen), gently mixed and incubated for 5 minutes. The control group was a blank pX330 vector that did not clone any foreign gene. 
     The diluted pX330 recombinant vector and Lipofectamine™ 2000 (Invitrogen) were incubated at room temperature for 20 minutes to form a recombinant vector-Lipofectamine™ 2000 (Invitrogen) complex and a blank vector-Lipofectamine 2000 (Invitrogen) complex. The incubation time should not exceed 30 minutes, and a longer incubation time may reduce activity. 
     The vector-Lipofectamine™ 2000 complex was added to each well containing cells and medium, and the plate was gently shaken back and forth, and incubated at 37° C. in a CO 2  incubator for 72 hours. 
     The transfected cells were harvested 3 days later and the genomic DNA was purified by Agencourt DNA dvance Genomic DNA Isolation Kit (Beckman Coulter). Sample preparation was carried out by the method of Example 5, and the obtained sample was subjected to pyrosequencing by BGI Shenzhen. 
     Example 7. Determination of Demethylation Site Range 
     According to Example 2, the inventor synthesized 30 ssDNA (15 fusion proteins for dCas9, 15 fusion proteins for dCpf1) of 59 bases in length as reaction substrates, their complementary ssDNA, and corresponding sgRNA primers. The 5′ end of the reaction substrate ssDNA was modified by the fluorophore FAM with a methylated C (Met-C) in between, which is the target of editing. After the ssDNA formed a dsDNA substrate with its complementary strand, the Cas9 region of the recombinant protein bound to the corresponding region in the middle of the dsDNA under the guidance of the corresponding sgRNA, and melted about 20 bases in the region, that was, formed a single-stranded region in the middle of the dsDNA. The target Met-C was in this region and was named as substrate 4-20 based on its distance to the 5′-end double-stranded region (4-20 bases). When the recombinant protein bound to different sgRNAs and then interacted with the corresponding dsDNA substrates for a certain period of time, some of the target Met-C became T under deamination and did not pair with G on the complementary strand to form a protrusion. The addition of 1 Unit of TDG after termination of the reaction at high temperature removed the mismatched T base, resulting in a deletion at the editing target of the substrate. The dsDNA then changed back to ssDNA and was cleaved at the base deletion site by the combined action of EDTA (0.5 μl at a concentration of 0.5 M), formamide (10 μl) and NaOH (1 μl at 5 M). Since both the cleaved 5′-end short-chain ssDNA and the unacting ssDNA substrate had a specific FAM fluorophore label at the 5′ end, the relative ratio of the two could be accurately estimated, and the efficiency of the recombinant protein to change Met-C to T at this site could be inferred. 
     As shown in  FIG.  3   , by experimental results on 15 different substrates, it can be seen that for the dCas9 fusion protein with a linker of (GGS) 3, Met-C within a range of 7-10 bases from the first base at the 5′ end of the single-stranded region after melting the double-strand in the target region can be changed to T, but not outside the range; for fusion proteins with a linker (GGS) 7 and (GGS) 14, the distances of the editing interval are 6-11 bases and 5-13 bases. This range will be slightly wider due to the length of the linker becoming longer. This range will be an important basis for our subsequent experimental design and future gene therapy design sgRNA. 
     It can also be seen from the results that A3H was slightly more active than A3A. 
     As can be seen from the results, the dCpf1 fusion protein with a linker of (GGS) 7 in length had similar activity, and the distance of the action range was 7-12 bases. 
     In the control group, the synthesized T was used as a positive control, and the wrong sgRNA and Cas-9 or Cpf1 without sgRNA were used as negative controls. 
     The control experiment was mainly to prove two problems: first, our method is feasible. One of the groups in which the formation of short-chain DNA were clearly seen was chosen, the same ssDNA substrate was synthesized but the Met-C therein was changed to T, that was, the function of the recombinant protein was artificially completed. The same operations were employed. As a result, the formation of short-chain DNA was also observed. It was proved that the short-chain DNA in the experimental results was actually produced by the action of the recombinant protein on the target DNA. Second, by continuing the next experimental procedure by allowing the recombinant protein not to bind to sgRNA or to bind to unpaired sgRNA, no short-chain DNA was produced, demonstrating that such editing was directed. 
     Example 8. Effect of Bases Upstream and Downstream of the Action Site 
     A recombinant protein (a linker of GGS*7, and Apobec protein of A3H) was used as a subject for the study on effect of the base located adjacent to upstream of the editing target site on demethylation activity. 
     Based on previous studies of the Apobec protein family, the base located adjacent to upstream of the editing target site has a direct effect on their activities. The substrate with Met-C at position 7 was selected and the previous base was changed to A, T, C and G, respectively. As shown in  FIG.  4   , the test results show that the sequence of the previous base has no effect on the editing efficiency, which proves the versatility of the technology. 
     Example 9. Efficiency of Intracellular Demethylation 
     When it had been demonstrated that the recombinant protein had an ideal ability to change Met-C to T outside the cell, it was desirable to further verify whether such activity remains in the cell, the intensity of the activity, and whether T is repaired into a normal C by the cell&#39;s own DNA repair mechanism after the reaction, thereby achieving the effect of site-specific demethylation. The applicant designed three sets of intracellular experiments, and the promoter regions of three different genes were selected for demethylation testing. 
     The first intracellular editing target was the two methylated C of the U.S. Pat. Nos. 17,741,472 and 17,741,474 loci on chromosome 11 in the HEK293 cell line, located in the promoter region of the gene MYOD1. As shown in  FIG.  5   , this experiment demonstrated that the system could accurately edit the chosen one in two methylation modifications that were close to each other. 
     The second editing target was a methylated C of the 31138558 locus on chromosome 6 in the HEK293 cell line, located in the promoter region of the gene POUF1. As shown in  FIG.  5   , this experiment also achieved the desired editing effect. 
     The third editing target was a methylated C of the 113875226 locus on chromosome 2 in the PC3 cell line, located in the promoter region of the gene IL1RN. As shown in  FIG.  6   , the system can edit one or two of the two adjacent methylated sites by a reasonable sgRNA design. 
     Recombinant vectors were separately constructed and transfected into cells using the method described in Example 6, and the editing results were evaluated by pyrosequencing. 
     Example 10. Proportion of Indel (Insertion and Deletion) in Cells after Editing 
     Based on the sequencing results of the above experiments, the cases of base insertion and deletion occurring near the target site throughout the process were also counted. From the sequencing results, there was no phenomenon of insertion and deletion of bases around. 
     The nucleic acid sequences used in the examples are specifically shown in the following table. 
     
       
         
           
               
               
               
             
               
                   
               
               
                 Seq 
                   
                   
               
               
                 ID 
                   
                   
               
               
                 no. 
                 Name 
                 Sequence (5′-3′) 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
            
               
                 1 
                 Rev_sgRNA_T7 
                 AAAAAAAGCACCGACTCGGTG 
               
               
                   
               
               
                 2 
                 Fwd_sgRNA_T7_ds 
                 TAATACGACTCACTATAGGTATCGGATTTATTTATTTAAGTTT 
               
               
                   
                 DNA_4 
                 TAGAGCTAGAAATAGC 
               
               
                   
               
               
                 3 
                 Fwd_sgRNA_T7_ds 
                 TAATACGACTCACTATAGGTTATCGGATTTATTTATTTAGTTT 
               
               
                   
                 DNA_5 
                 TAGAGCTAGAAATAGC 
               
               
                   
               
               
                 4 
                 Fwd_sgRNA_T7_ds 
                 TAATACGACTCACTATAGGTTTATCGGATTTATTTATTAGTTT 
               
               
                   
                 DNA6 
                 TAGAGCTAGAAATAGC 
               
               
                   
               
               
                 5 
                 Fwd_sgRNA_T7_ds 
                 TAATACGACTCACTATAGGATTTATCGGATTTATTTATTGTTT 
               
               
                   
                 DNA_7 
                 TAGAGCTAGAAATAGC 
               
               
                   
               
               
                 6 
                 Fwd_sgRNA_T7_ds 
                 TAATACGACTCACTATAGGTATTTATCGGATTTATTTATGTTT 
               
               
                   
                 DNA_8 
                 TAGAGCTAGAAATAGC 
               
               
                   
               
               
                 7 
                 Fwd_sgRNA_T7_ds 
                 TAATACGACTCACTATAGGTTATTTATCGGATTTATTTAGTTT 
               
               
                   
                 DNA_9 
                 TAGAGCTAGAAATAGC 
               
               
                   
               
               
                 8 
                 Fwd_sgRNA_T7_ds 
                 TAATACGACTCACTATAGGATTATTTATCGGATTTATTTGTTT 
               
               
                   
                 DNA_10 
                 TAGAGCTAGAAATAGC 
               
               
                   
               
               
                 9 
                 Fwd_sgRNA_T7_ds 
                 TAATACGACTCACTATAGGTATTATTTATCGGATTTATTGTTT 
               
               
                   
                 DNA_11 
                 TAGAGCTAGAAATAGC 
               
               
                   
               
               
                 10 
                 Fwd_sgRNA_T7_ds 
                 TAATACGACTCACTATAGGATTATTATTATCGGATTTATGTTT 
               
               
                   
                 DNA_12 
                 TAGAGCTAGAAATAGC 
               
               
                   
               
               
                 11 
                 Fwd_sgRNA_T7_ds 
                 TAATACGACTCACTATAGGTATTATATTTATCGGATTTAGTTT 
               
               
                   
                 DNA_13 
                 TAGAGCTAGAAATAGC 
               
               
                   
               
               
                 12 
                 Fwd_sgRNA_T7_ds 
                 TAATACGACTCACTATAGGTTATTATATTTATCGGATTTGTTT 
               
               
                   
                 DNA_14 
                 TAGAGCTAGAAATAGC 
               
               
                   
               
               
                 13 
                 Fwd_sgRNA_T7_ds 
                 TAATACGACTCACTATAGGATTATTATATTTATCGGATTGTTT 
               
               
                   
                 DNA_15 
                 TAGAGCTAGAAATAGC 
               
               
                   
               
               
                 14 
                 Fwd_sgRNA_T7_ds 
                 TAATACGACTCACTATAGGTATTATTATATTTATCGGATGTTT 
               
               
                   
                 DNA_16 
                 TAGAGCTAGAAATAGC 
               
               
                   
               
               
                 15 
                 Fwd_sgRNA_T7_ds 
                 TAATACGACTCACTATAGGATTATTATTATTATATCGGAGTTT 
               
               
                   
                 DNA_17 
                 TAGAGCTAGAAATAGC 
               
               
                   
               
               
                 16 
                 Fwd_sgRNA_T7_ds 
                 TAATACGACTCACTATAGGATTATTATTATTATTATATCGTTT 
               
               
                   
                 DNA_20 
                 TAGAGCTAGAAATAGC 
               
               
                   
               
               
                 17 
                 Fwd_sgRNA_T7_ds 
                 TAATACGACTCACTATAGGTATAGGATTTATTTATTTAAGTTT 
               
               
                   
                 DNA_noC 
                 TAGAGCTAGAAATAGC 
               
               
                   
               
               
                 18 
                 Fwd_crRNA_T7 
                 TAATACGACTCACTATAGGAATTTCTACTGTTGTAGATG 
               
               
                   
               
               
                 19 
                 Rev_crRNA_T7_dsD 
                 TTAAATAAATAAATCCGATACATCTACAACAGTAGAAATTCC 
               
               
                   
                 NA_4 
                 TATAGTGAGTCGTATTA 
               
               
                   
               
               
                 20 
                 Rev_crRNA_T7_dsD 
                 TAAATAAATAAATCCGATAACATCTACAACAGTAGAAATTCC 
               
               
                   
                 NA_5 
                 TATAGTGAGTCGTATTA 
               
               
                   
               
               
                 21 
                 Rev_crRNA_T7_dsD 
                 TAATAAATAAATCCGATAAACATCTACAACAGTAGAAATTCC 
               
               
                   
                 NA_6 
                 TATAGTGAGTCGTATTA 
               
               
                   
               
               
                 22 
                 Rev_crRNA_T7_dsD 
                 AATAAATAAATCCGATAAATCATCTACAACAGTAGAAATTCC 
               
               
                   
                 NA_7 
                 TATAGTGAGTCGTATTA 
               
               
                   
               
               
                 23 
                 Rev_crRNA_T7_dsD 
                 ATAAATAAATCCGATAAATACATCTACAACAGTAGAAATTCC 
               
               
                   
                 NA_8 
                 TATAGTGAGTCGTATTA 
               
               
                   
               
               
                 24 
                 Rev_crRNA_T7_dsD 
                 TAAATAAATCCGATAAATAACATCTACAACAGTAGAAATTCC 
               
               
                   
                 NA_9 
                 TATAGTGAGTCGTATTA 
               
               
                   
               
               
                 25 
                 Rev_crRNA_T7_dsD 
                 AAATAAATCCGATAAATAATCATCTACAACAGTAGAAATTCC 
               
               
                   
                 NA_10 
                 TATAGTGAGTCGTATTA 
               
               
                   
               
               
                 26 
                 Rev_crRNA_T7_dsD 
                 AATAAATCCGATAAATAATACATCTACAACAGTAGAAATTCC 
               
               
                   
                 NA_11 
                 TATAGTGAGTCGTATTA 
               
               
                   
               
               
                 27 
                 Rev_crRNA_T7_dsD 
                 ATAAATCCGATAATAATAATCATCTACAACAGTAGAAATTCC 
               
               
                   
                 NA_12 
                 TATAGTGAGTCGTATTA 
               
               
                   
               
               
                 28 
                 Rev_crRNA_T7_dsD 
                 TAAATCCGATAAATATAATACATCTACAACAGTAGAAATTCC 
               
               
                   
                 NA_13 
                 TATAGTGAGTCGTATTA 
               
               
                   
               
               
                 29 
                 Rev_crRNA_T7_dsD 
                 AAATCCGATAAATATAATAACATCTACAACAGTAGAAATTCC 
               
               
                   
                 NA_14 
                 TATAGTGAGTCGTATTA 
               
               
                   
               
               
                 30 
                 Rev_crRNA_T7_dsD 
                 AATCCGATAAATATAATAATCATCTACAACAGTAGAAATTCC 
               
               
                   
                 NA_15 
                 TATAGTGAGTCGTATTA 
               
               
                   
               
               
                 31 
                 Rev_crRNA_T7_dsD 
                 ATCCGATAAATATAATAATACATCTACAACAGTAGAAATTCC 
               
               
                   
                 NA_16 
                 TATAGTGAGTCGTATTA 
               
               
                   
               
               
                 32 
                 Rev_crRNA_T7_dsD 
                 TCCGATATAATAATAATAATCATCTACAACAGTAGAAATTCC 
               
               
                   
                 NA_17 
                 TATAGTGAGTCGTATTA 
               
               
                   
               
               
                 33 
                 Rev_crRNA_T7_dsD 
                 GATATAATAATAATAATAATCATCTACAACAGTAGAAATTCC 
               
               
                   
                 NA_20 
                 TATAGTGAGTCGTATTA 
               
               
                   
               
               
                 34 
                 Rev_crRNA_T7_dsD 
                 TTAAATAAATAAATCCTATACATCTACAACAGTAGAAATTCC 
               
               
                   
                 NA_noC 
                 TATAGTGAGTCGTATTA 
               
               
                   
               
               
                 35 
                 Fwd_sgRNA_6T 
                 TAATACGACTCACTATAGGTTATTTCGTGGATTTATTTAGTTT 
               
               
                   
                   
                 TAGAGCTAGAAATAGC 
               
               
                   
               
               
                 36 
                 Fwd_sgRNA_6A 
                 TAATACGACTCACTATAGGTTATTTCGTGGATTTATTTAGTTT 
               
               
                   
                   
                 TAGAGCTAGAAATAGC 
               
               
                   
               
               
                 37 
                 Fwd_sgRNA_6C 
                 TAATACGACTCACTATAGGTTATTTCGTGGATTTATTTAGTTT 
               
               
                   
                   
                 TAGAGCTAGAAATAGC 
               
               
                   
               
               
                 38 
                 Fwd_sgRNA_6G 
                 TAATACGACTCACTATAGGTTATTTCGTGGATTTATTTAGTTT 
               
               
                   
                   
                 TAGAGCTAGAAATAGC 
               
               
                   
               
               
                 39 
                 dCas9_ds_4 
                 FAM- 
               
               
                   
                   
                 GGTAGTTAGGATGAATGGAAGGTTGGTATAGCCTATmet- 
               
               
                   
                   
                 CGGATTTATTTATTTAAT 
               
               
                   
                   
                 GGATGACCTCTGGATCCATG 
               
               
                   
               
               
                 40 
                 dCas9_ds_5 
                 FAM- 
               
               
                   
                   
                 GGTAGTTAGGATGAATGGAAGGTTGGTATAGCCTTATmet- 
               
               
                   
                   
                 CGGATTTATTTATTTAT 
               
               
                   
                   
                 GGATGACCTCTGGATCCATG 
               
               
                   
               
               
                 41 
                 dCas9_ds_6 
                 FAM- 
               
               
                   
                   
                 GGTAGTTAGGATGAATGGAAGGTTGGTATAGCCTTTATmet- 
               
               
                   
                   
                 CGGATTTATTTATTAT 
               
               
                   
                   
                 GGATGACCTCTGGATCCATG 
               
               
                   
               
               
                 42 
                 dCas9_ds_7 
                 FAM- 
               
               
                   
                   
                 GGTAGTTAGGATGAATGGAAGGTTGGTATAGCCATTTATmet- 
               
               
                   
                   
                 CGGATTTATTTATTT 
               
               
                   
                   
                 GGATGACCTCTGGATCCATG 
               
               
                   
               
               
                 43 
                 dCas9_ds_8 
                 FAM- 
               
               
                   
                   
                 GGTAGTTAGGATGAATGGAAGGTTGGTATAGCCTATTTATmet 
               
               
                   
                   
                 -CGGATTTATTTATT 
               
               
                   
                   
                 GGATGACCTCTGGATCCATG 
               
               
                   
               
               
                 44 
                 dCas9_ds_9 
                 FAM- 
               
               
                   
                   
                 GGTAGTTAGGATGAATGGAAGGTTGGTATAGCCTTATTTATm 
               
               
                   
                   
                 et-CGGATTTATTTAT 
               
               
                   
                   
                 GGATGACCTCTGGATCCATG 
               
               
                   
               
               
                 45 
                 dCas9_ds_10 
                 FAM- 
               
               
                   
                   
                 GGTAGTTAGGATGAATGGAAGGTTGGTATAGCCATTATTTAT 
               
               
                   
                   
                 met-CGGATTTATTTT 
               
               
                   
                   
                 GGATGACCTCTGGATCCATG 
               
               
                   
               
               
                 46 
                 dCas9_ds_11 
                 FAM- 
               
               
                   
                   
                 GGTAGTTAGGATGAATGGAAGGTTGGTATAGCCTATTATTTA 
               
               
                   
                   
                 Tmet-CGGATTTATTT 
               
               
                   
                   
                 GGATGACCTCTGGATCCATG 
               
               
                   
               
               
                 47 
                 dCas9_ds_12 
                 FAM- 
               
               
                   
                   
                 GGTAGTTAGGATGAATGGAAGGTTGGTATAGCCATTATTATT 
               
               
                   
                   
                 ATmet-CGGATTTATT 
               
               
                   
                   
                 GGATGACCTCTGGATCCATG 
               
               
                   
               
               
                 48 
                 dCas9_ds_13 
                 FAM- 
               
               
                   
                   
                 GGTAGTTAGGATGAATGGAAGGTTGGTATAGCCTATTATATT 
               
               
                   
                   
                 TATmet-CGGATTTAT 
               
               
                   
                   
                 GGATGACCTCTGGATCCATG 
               
               
                   
               
               
                 49 
                 dCas9_ds_14 
                 FAM- 
               
               
                   
                   
                 GGTAGTTAGGATGAATGGAAGGTTGGTATAGCCTTATTATAT 
               
               
                   
                   
                 TTATmet-CGGATTTT 
               
               
                   
                   
                 GGATGACCTCTGGATCCATG 
               
               
                   
               
               
                 50 
                 dCas9_ds_15 
                 FAM- 
               
               
                   
                   
                 GGTAGTTAGGATGAATGGAAGGTTGGTATAGCCATTATTATA 
               
               
                   
                   
                 TTTATmet-CGGATTT 
               
               
                   
                   
                 GGATGACCTCTGGATCCATG 
               
               
                   
               
               
                 51 
                 dCas9_ds_16 
                 FAM- 
               
               
                   
                   
                 GGTAGTTAGGATGAATGGAAGGTTGGTATAGCCTATTATTAT 
               
               
                   
                   
                 ATTTATmet-CGGATT 
               
               
                   
                   
                 GGATGACCTCTGGATCCATG 
               
               
                   
               
               
                 52 
                 dCas9_ds_17 
                 FAM- 
               
               
                   
                   
                 GGTAGTTAGGATGAATGGAAGGTTGGTATAGCCATTATTATT 
               
               
                   
                   
                 ATTATATmet-CGGAT 
               
               
                   
                   
                 GGATGACCTCTGGATCCATG 
               
               
                   
               
               
                 53 
                 dCas9_ds_20 
                 FAM- 
               
               
                   
                   
                 GGTAGTTAGGATGAATGGAAGGTTGGTATAGCCATTATTATT 
               
               
                   
                   
                 ATTATTATATmet-CT 
               
               
                   
                   
                 GGATGACCTCTGGATCCATG 
               
               
                   
               
               
                 54 
                 dCas9_ds_noC 
                 FAM- 
               
               
                   
                   
                 GGTAGTTAGGATGAATGGAAGGTTGGTATAGCCTATAGGATT 
               
               
                   
                   
                 TATTTATTTAAT 
               
               
                   
                   
                 GGATGACCTCTGGATCCATG 
               
               
                   
               
               
                 55 
                 dCas9_ds_com_4 
                 CATGGATCCAGAGGTCATCCATTAAATAAATAAATCCGATAG 
               
               
                   
                   
                 GCTATACCAACCTTCC 
               
               
                   
                   
                 ATTCATCCTAACTACC 
               
               
                   
               
               
                 56 
                 dCas9_ds_com_5 
                 CATGGATCCAGAGGTCATCCATAAATAAATAAATCCGATAA 
               
               
                   
                   
                 GGCTATACCAACCTTCC 
               
               
                   
                   
                 ATTCATCCTAACTACC 
               
               
                   
               
               
                 57 
                 dCas9_ds_com_6 
                 CATGGATCCAGAGGTCATCCATAATAAATAAATCCGATAAA 
               
               
                   
                   
                 GGCTATACCAACCTTCC 
               
               
                   
                   
                 ATTCATCCTAACTACC 
               
               
                   
               
               
                 58 
                 dCas9_ds_com_7 
                 CATGGATCCAGAGGTCATCCAAATAAATAAATCCGATAAAT 
               
               
                   
                   
                 GGCTATACCAACCTTCC 
               
               
                   
                   
                 ATTCATCCTAACTACC 
               
               
                   
               
               
                 59 
                 dCas9_ds_com_8 
                 CATGGATCCAGAGGTCATCCAATAAATAAATCCGATAAATA 
               
               
                   
                   
                 GGCTATACCAACCTTCC 
               
               
                   
                   
                 ATTCATCCTAACTACC 
               
               
                   
               
               
                 60 
                 dCas9_ds_com_9 
                 CATGGATCCAGAGGTCATCCATAAATAAATCCGATAAATAA 
               
               
                   
                   
                 GGCTATACCAACCTTCC 
               
               
                   
                   
                 ATTCATCCTAACTACC 
               
               
                   
               
               
                 61 
                 dCas9_ds_com_10 
                 CATGGATCCAGAGGTCATCCAAAATAAATCCGATAAATAAT 
               
               
                   
                   
                 GGCTATACCAACCTTCC 
               
               
                   
                   
                 ATTCATCCTAACTACC 
               
               
                   
               
               
                 62 
                 dCas9_ds_com_11 
                 CATGGATCCAGAGGTCATCCAAATAAATCCGATAAATAATA 
               
               
                   
                   
                 GGCTATACCAACCTTCC 
               
               
                   
                   
                 ATTCATCCTAACTACC 
               
               
                   
               
               
                 63 
                 dCas9_ds_com_12 
                 CATGGATCCAGAGGTCATCCAATAAATCCGATAATAATAATG 
               
               
                   
                   
                 GCTATACCAACCTTCC 
               
               
                   
                   
                 ATTCATCCTAACTACC 
               
               
                   
               
               
                 64 
                 dCas9_ds_com_13 
                 CATGGATCCAGAGGTCATCCATAAATCCGATAAATATAATAG 
               
               
                   
                   
                 GCTATACCAACCTTCC 
               
               
                   
                   
                 ATTCATCCTAACTACC 
               
               
                   
               
               
                 65 
                 dCas9_ds_com_14 
                 CATGGATCCAGAGGTCATCCAAAATCCGATAAATATAATAA 
               
               
                   
                   
                 GGCTATACCAACCTTCC 
               
               
                   
                   
                 ATTCATCCTAACTACC 
               
               
                   
               
               
                 66 
                 dCas9_ds_com_15 
                 CATGGATCCAGAGGTCATCCAAATCCGATAAATATAATAATG 
               
               
                   
                   
                 GCTATACCAACCTTCC 
               
               
                   
                   
                 ATTCATCCTAACTACC 
               
               
                   
               
               
                 67 
                 dCas9_ds_com_16 
                 CATGGATCCAGAGGTCATCCAATCCGATAAATATAATAATAG 
               
               
                   
                   
                 GCTATACCAACCTTCC 
               
               
                   
                   
                 ATTCATCCTAACTACC 
               
               
                   
               
               
                 68 
                 dCas9_ds_com_17 
                 CATGGATCCAGAGGTCATCCATCCGATATAATAATAATAATG 
               
               
                   
                   
                 GCTATACCAACCTTCC 
               
               
                   
                   
                 ATTCATCCTAACTACC 
               
               
                   
               
               
                 69 
                 dCas9_ds_com_20 
                 CATGGATCCAGAGGTCATCCAGATATAATAATAATAATAATG 
               
               
                   
                   
                 GCTATACCAACCTTCC 
               
               
                   
                   
                 ATTCATCCTAACTACC 
               
               
                   
               
               
                 70 
                 dCas9_ds_com_noC 
                 CATGGATCCAGAGGTCATCCATTAAATAAATAAATCCTATAG 
               
               
                   
                   
                 GCTATACCAACCTTCC 
               
               
                   
                   
                 ATTCATCCTAACTACC 
               
               
                   
               
               
                 71 
                 dCpf1_ds_4 
                 FAM-GGTACCCGGGGATCCTTTATATmet- 
               
               
                   
                   
                 CGGATTTATTTATTTAAGTTAAAAAGCTTGGCGTAAT 
               
               
                   
               
               
                 72 
                 dCpf1_ds_5 
                 FAM-GGTACCCGGGGATCCTTTATTATmet- 
               
               
                   
                   
                 CGGATTTATTTATTTAGTTAAAAAGCTTGGCGTAAT 
               
               
                   
               
               
                 73 
                 dCpf1_ds_6 
                 FAM-GGTACCCGGGGATCCTTTATTTATmet- 
               
               
                   
                   
                 CGGATTTATTTATTAGTTAAAAAGCTTGGCGTAAT 
               
               
                   
               
               
                 74 
                 dCpf1_ds_7 
                 FAM-GGTACCCGGGGATCCTTTAATTTATmet- 
               
               
                   
                   
                 CGGATTTATTTATTGTTAAAAAGCTTGGCGTAAT 
               
               
                   
               
               
                 75 
                 dCpf1_ds_8 
                 FAM-GGTACCCGGGGATCCTTTATATTTATmet- 
               
               
                   
                   
                 CGGATTTATTTATGTTAAAAAGCTTGGCGTAAT 
               
               
                   
               
               
                 76 
                 dCpf1_ds_9 
                 FAM-GGTACCCGGGGATCCTTTATTATTTATmet- 
               
               
                   
                   
                 CGGATTTATTTAGTTAAAAAGCTTGGCGTAAT 
               
               
                   
               
               
                 77 
                 dCpf1_ds_10 
                 FAM-GGTACCCGGGGATCCTTTAATTATTTATmet- 
               
               
                   
                   
                 CGGATTTATTTGTTAAAAAGCTTGGCGTAAT 
               
               
                   
               
               
                 78 
                 dCpf1_ds_11 
                 FAM-GGTACCCGGGGATCCTTTATATTATTTATmet- 
               
               
                   
                   
                 CGGATTTATTGTTAAAAAGCTTGGCGTAAT 
               
               
                   
               
               
                 79 
                 dCpf1_ds_12 
                 FAM-GGTACCCGGGGATCCTTTAATTATTATTATmet- 
               
               
                   
                   
                 CGGATTTATGTTAAAAAGCTTGGCGTAAT 
               
               
                   
               
               
                 80 
                 dCpf1_ds_13 
                 FAM-GGTACCCGGGGATCCTTTATATTATATTTATmet- 
               
               
                   
                   
                 CGGATTTAGTTAAAAAGCTTGGCGTAAT 
               
               
                   
               
               
                 81 
                 dCpf1_ds_14 
                 FAM-GGTACCCGGGGATCCTTTATTATTATATTTATmet- 
               
               
                   
                   
                 CGGATTTGTTAAAAAGCTTGGCGTAAT 
               
               
                   
               
               
                 82 
                 dCpf1_ds_15 
                 FAM-GGTACCCGGGGATCCTTTAATTATTATATTTATmet- 
               
               
                   
                   
                 CGGATTGTTAAAAAGCTTGGCGTAAT 
               
               
                   
               
               
                 83 
                 dCpf1_ds_16 
                 FAM-GGTACCCGGGGATCCTTTATATTATTATATTTATmet- 
               
               
                   
                   
                 CGGATGTTAAAAAGCTTGGCGTAAT 
               
               
                   
               
               
                 84 
                 dCpf1_ds_17 
                 FAM-GGTACCCGGGGATCCTTTAATTATTATTATTATATmet- 
               
               
                   
                   
                 CGGAGTTAAAAAGCTTGGCGTAAT 
               
               
                   
               
               
                 85 
                 dCpf1_ds_20 
                 FAM- 
               
               
                   
                   
                 GGTACCCGGGGATCCTTTAATTATTATTATTATTATATmet- 
               
               
                   
                   
                 CGTTAAAAAGCTTGGCGTAAT 
               
               
                   
               
               
                 86 
                 dCpf1_ds_com_4 
                 ATTACGCCAAGCTTTTTAACTTAAATAAATAAATCCGATATA 
               
               
                   
                   
                 AAGGATCCCCGGGTACC 
               
               
                   
               
               
                 87 
                 dCpf1_ds_com_5 
                 ATTACGCCAAGCTTTTTAACTAAATAAATAAATCCGATAATA 
               
               
                   
                   
                 AAGGATCCCCGGGTACC 
               
               
                   
               
               
                 88 
                 dCpf1_ds_com_6 
                 ATTACGCCAAGCTTTTTAACTAATAAATAAATCCGATAAATA 
               
               
                   
                   
                 AAGGATCCCCGGGTACC 
               
               
                   
               
               
                 89 
                 dCpf1_ds_com_7 
                 ATTACGCCAAGCTTTTTAACAATAAATAAATCCGATAAATTA 
               
               
                   
                   
                 AAGGATCCCCGGGTACC 
               
               
                   
               
               
                 90 
                 dCpf1_ds_com_8 
                 ATTACGCCAAGCTTTTTAACATAAATAAATCCGATAAATATA 
               
               
                   
                   
                 AAGGATCCCCGGGTACC 
               
               
                   
               
               
                 91 
                 dCpf1_ds_com_9 
                 ATTACGCCAAGCTTTTTAACTAAATAAATCCGATAAATAATA 
               
               
                   
                   
                 AAGGATCCCCGGGTACC 
               
               
                   
               
               
                 92 
                 dCpf1_ds_com_10 
                 ATTACGCCAAGCTTTTTAACAAATAAATCCGATAAATAATTA 
               
               
                   
                   
                 AAGGATCCCCGGGTACC 
               
               
                   
               
               
                 93 
                 dCpf1_ds_com_11 
                 ATTACGCCAAGCTTTTTAACAATAAATCCGATAAATAATATA 
               
               
                   
                   
                 AAGGATCCCCGGGTACC 
               
               
                   
               
               
                 94 
                 dCpf1_ds_com_12 
                 ATTACGCCAAGCTTTTTAACATAAATCCGATAATAATAATTA 
               
               
                   
                   
                 AAGGATCCCCGGGTACC 
               
               
                   
               
               
                 95 
                 dCpf1_ds_com_13 
                 ATTACGCCAAGCTTTTTAACTAAATCCGATAAATATAATATA 
               
               
                   
                   
                 AAGGATCCCCGGGTACC 
               
               
                   
               
               
                 96 
                 dCpf1_ds_com_14 
                 ATTACGCCAAGCTTTTTAACAAATCCGATAAATATAATAATA 
               
               
                   
                   
                 AAGGATCCCCGGGTACC 
               
               
                   
               
               
                 97 
                 dCpf1_ds_com_15 
                 ATTACGCCAAGCTTTTTAACAATCCGATAAATATAATAATTA 
               
               
                   
                   
                 AAGGATCCCCGGGTACC 
               
               
                   
               
               
                 98 
                 dCpf1_ds_com_16 
                 ATTACGCCAAGCTTTTTAACATCCGATAAATATAATAATATA 
               
               
                   
                   
                 AAGGATCCCCGGGTACC 
               
               
                   
               
               
                 99 
                 dCpf1_ds_com_17 
                 ATTACGCCAAGCTTTTTAACTCCGATATAATAATAATAATTA 
               
               
                   
                   
                 AAGGATCCCCGGGTACC 
               
               
                   
               
               
                 100 
                 dCpf1_ds_com_20 
                 ATTACGCCAAGCTTTTTAACGATATAATAATAATAATAATTA 
               
               
                   
                   
                 AAGGATCCCCGGGTACC 
               
               
                   
               
               
                 101 
                 dCas9_ds_6T 
                 ACGTAAACGGCCACAAGTTCTTATTTmet- 
               
               
                   
                   
                 CGTGGATTTATTTATGGCATCTTCTTCAAGGAC 
               
               
                   
               
               
                 102 
                 dCas9_ds_6A 
                 ACGTAAACGGCCACAAGTTCTTATTAmet- 
               
               
                   
                   
                 CGTGGATTTATTTATGGCATCTTCTTCAAGGAC 
               
               
                   
               
               
                 103 
                 dCas9_ds_6C 
                 ACGTAAACGGCCACAAGTTCTTATTCmet- 
               
               
                   
                   
                 CGTGGATTTATTTATGGCATCTTCTTCAAGGAC 
               
               
                   
               
               
                 104 
                 dCas9_ds_6G 
                 ACGTAAACGGCCACAAGTTCTTATTGmet- 
               
               
                   
                   
                 CGTGGATTTATTTATGGCATCTTCTTCAAGGAC 
               
               
                   
               
               
                 105 
                 dCas9_ds_com_6T 
                 GTCCTTGAAGAAGATGCCATAAATAAATCCACGAAATAAGA 
               
               
                   
                   
                 ACTTGTGGCCGTTTACGT 
               
               
                   
               
               
                 106 
                 dCas9_ds_com_6A 
                 GTCCTTGAAGAAGATGCCATAAATAAATCCACGTAATAAGA 
               
               
                   
                   
                 ACTTGTGGCCGTTTACGT 
               
               
                   
               
               
                 107 
                 dCas9_ds_com_6C 
                 GTCCTTGAAGAAGATGCCATAAATAAATCCACGGAATAAGA 
               
               
                   
                   
                 ACTTGTGGCCGTTTACGT 
               
               
                   
               
               
                 108 
                 dCas9_ds_com_6G 
                 GTCCTTGAAGAAGATGCCATAAATAAATCCACGCAATAAGA 
               
               
                   
                   
                 ACTTGTGGCCGTTTACGT 
               
               
                   
               
               
                 109 
                 ds_6_F 
                 CGTAAACGGCCACAAGTTCTTAT 
               
               
                   
               
               
                 110 
                 ds_6_R 
                 GTCCTTGAAGAAGATGCCATAAA 
               
               
                   
               
               
                 111 
                 ds_6_S 
                 CGGCCACAAGTTCTTAT 
               
               
                   
               
               
                 112 
                 HEK293T-T1-F 
                 GGATTTGYGTTTTTTYGAAGATTTGG 
               
               
                   
               
               
                 113 
                 HEK293T-T1-R 
                 AAATACRAATACTCTTCRAATTTCAAAAAC 
               
               
                   
               
               
                 114 
                 HEK293T-T1-S 
                 GTTTTTTAGAAGATTTGGAT 
               
               
                   
               
               
                 115 
                 HEK293T-T2-F 
                 GTTTTGAATGAATGTGTGTATATATGTATG 
               
               
                   
               
               
                 116 
                 HEK293T-T2-R 
                 CTAACAAAAACCAAACTAATTCTTATCTAC 
               
               
                   
               
               
                 117 
                 HEK293T-T2-S 
                 ATGAATGTGTGTATATATGTATGAG 
               
               
                   
               
               
                 118 
                 PC3-F 
                 TAAGGGTTTTYGGAAYGGGGT 
               
               
                   
               
               
                 119 
                 PC3-R 
                 CCAAACAAAACATCCCTCAAC 
               
               
                   
               
               
                 120 
                 PC3-S 
                 GGGTTGTGTGAGTGGG 
               
               
                   
               
               
                 121 
                 HEK293T-gRNA1-F 
                 CACCG GGACCCGCGCCTGATGCACG 
               
               
                   
               
               
                 122 
                 HEK293T-gRNA1-R 
                 AAAC CGTGCATCAGGCGCGGGTCC C 
               
               
                   
               
               
                 123 
                 HEK293T-gRNA2-F 
                 CACCG GAGCTGGCGGCAGTCGGGGT 
               
               
                   
               
               
                 124 
                 HEK293T-gRNA2-R 
                 AAAC ACCCCGACTGCCGCCAGCTC C 
               
               
                   
               
               
                 125 
                 Gfap-gRNA-F 
                 CACCG TTCCGAGAAGTCTATTGAGC 
               
               
                   
               
               
                 126 
                 Gfap-gRNA-R 
                 AAAC GCTCAATAGACTTCTCGGAA C 
               
               
                   
               
               
                 127 
                 PMP24-gRNA-F 
                 CACCG TGGGGCCGTCGGGCCGGGCT 
               
               
                   
               
               
                 128 
                 PMP24-gRNA-R 
                 AAAC AGCCCGGCCCGACGGCCCCA C 
               
               
                   
               
               
                 129 
                 C/EBPδ-gRNA-F 
                 CACCG TCAGCCGGGGCTAGAAAAGG 
               
               
                   
               
            
           
         
       
     
     The sequences of protein domains are as follows: 
     
       
         
           
               
               
            
               
                   
                 APOBEC3A 
               
               
                   
                 MEASPASGPRHLMDPHIFTSNFNNGIGRHKTYLCYEVERL 
               
               
                   
                   
               
               
                   
                 DNGTSVKMDQHRGFLHNQAKNLLCGFYGRHAELRFLDLVP 
               
               
                   
                   
               
               
                   
                 SLQLDPAQIYRVTWFISWSPCFSWGCAGEVRAFLQENTHV 
               
               
                   
                   
               
               
                   
                 RLRIFAARIYDYDPLYKEALQMLRDAGAQVSIMTYDEFKH 
               
               
                   
                   
               
               
                   
                 CWDTFVDHQGCPFQPWDGLDEHSQALSGRLRAILQNQGN 
               
               
                   
                   
               
               
                   
                 &gt;AP0BEC3H Hyplotype II 
               
               
                   
                 MALLTAETFRLQFNNKRRLRRPYYPRKALLCYQLTPQNGS 
               
               
                   
                   
               
               
                   
                 TPTRGYFENKKKCHAEICFINEIKSMGLDETQCYQVTCYL 
               
               
                   
                   
               
               
                   
                 TWSPCSSCAWELVDFIKAHDHLNLRIFASRLYYHWCKPQQ 
               
               
                   
                   
               
               
                   
                 DGLRLLCGSQVPVEVMGFPEFADCWENFVDHEKPLSFNPY 
               
               
                   
                   
               
               
                   
                 KMLEELDKNSRAIKRRLDRIKS 
               
               
                   
                   
               
               
                   
                 &gt;Cas9 
               
               
                   
                 MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDR 
               
               
                   
                   
               
               
                   
                 HSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRIC 
               
               
                   
                   
               
               
                   
                 YLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFG 
               
               
                   
                   
               
               
                   
                 NIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAH 
               
               
                   
                   
               
               
                   
                 MIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENP 
               
               
                   
                   
               
               
                   
                 INASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGN 
               
               
                   
                   
               
               
                   
                 LIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLA 
               
               
                   
                   
               
               
                   
                 QIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSAS 
               
               
                   
                   
               
               
                   
                 MIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYA 
               
               
                   
                   
               
               
                   
                 GYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR 
               
               
                   
                   
               
               
                   
                 KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKI 
               
               
                   
                   
               
               
                   
                 EKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEE 
               
               
                   
                   
               
               
                   
                 VVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTV 
               
               
                   
                   
               
               
                   
                 YNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVT 
               
               
                   
                   
               
               
                   
                 VKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKI 
               
               
                   
                   
               
               
                   
                 IKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYA 
               
               
                   
                   
               
               
                   
                 HLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTIL 
               
               
                   
                   
               
               
                   
                 DFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSL 
               
               
                   
                   
               
               
                   
                 HEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIV 
               
               
                   
                   
               
               
                   
                 IEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHP 
               
               
                   
                   
               
               
                   
                 VENTQLQNEKLYEYYLQNGRDMYVDQELDINRLSDYDVDH 
               
               
                   
                   
               
               
                   
                 IVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMK 
               
               
                   
                   
               
               
                   
                 NYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQ 
               
               
                   
                   
               
               
                   
                 LVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKS 
               
               
                   
                   
               
               
                   
                 KLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKK 
               
               
                   
                   
               
               
                   
                 YPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYS 
               
               
                   
                   
               
               
                   
                 NIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDF 
               
               
                   
                   
               
               
                   
                 ATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLI 
               
               
                   
                   
               
               
                   
                 ARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSV 
               
               
                   
                   
               
               
                   
                 KELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPK 
               
               
                   
                   
               
               
                   
                 YSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLAS 
               
               
                   
                   
               
               
                   
                 HYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRV 
               
               
                   
                   
               
               
                   
                 ILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGA 
               
               
                   
                   
               
               
                   
                 PAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRI 
               
               
                   
                   
               
               
                   
                 DLSQLGGDPPKKKRKV 
               
               
                   
                   
               
               
                   
                 &gt;Cpf1 
               
               
                   
                 MTQFEGFTNLYQVSKTLRFELIPQGKTLKHIQEQGFIEED 
               
               
                   
                   
               
               
                   
                 KARNDHYKELKPIIDRIYKTYADQCLQLVQLDWENLSAAI 
               
               
                   
                   
               
               
                   
                 DSYRKEKTEETRNALIEEQATYRNAIHDYFIGRTDNLTDA 
               
               
                   
                   
               
               
                   
                 INKRHAEIYKGLFKAELFNGKVLKQLGTVTTTEHENALLR 
               
               
                   
                   
               
               
                   
                 SFDKFTTYFSGFYENRKNVFSAEDISTAIPHRIVQDNFPK 
               
               
                   
                   
               
               
                   
                 FKENCHIFTRLITAVPSLREHFENVKKAIGIFVSTSIEEV 
               
               
                   
                   
               
               
                   
                 FSFPFYNQLLTQTQIDLYNQLLGGISREAGTEKIKGLNEV 
               
               
                   
                   
               
               
                   
                 LNLAIQKNDETAHIIASLPHRFIPLFKQILSDRNTLSFIL 
               
               
                   
                   
               
               
                   
                 EEFKSDEEVIQSFCKYKTLLRNENVLETAEALFNELNSID 
               
               
                   
                   
               
               
                   
                 LTHIFISHKKLETISSALCDHWDTLRNALYERRISELTGK 
               
               
                   
                   
               
               
                   
                 ITKSAKEKVQRSLKHEDINLQEIISAAGKELSEAFKQKTS 
               
               
                   
                   
               
               
                   
                 EILSHAHAALDQPLPTTLKKQEEKEILKSQLDSLLGLYHL 
               
               
                   
                   
               
               
                   
                 LDWFAVDESNEVDPEFSARLTGIKLEMEPSLSFYNKARNY 
               
               
                   
                   
               
               
                   
                 ATKKPYSVEKFKLNFQMPTLASGWDVNKEKNNGAILFVKN 
               
               
                   
                   
               
               
                   
                 GLYYLGIMPKQKGRYKALSFEPTEKTSEGFDKMYYDYFPD 
               
               
                   
                   
               
               
                   
                 AAKMIPKCSTQLKAVTAHFQTHTTPILLSNNFIEPLEITK 
               
               
                   
                   
               
               
                   
                 EIYDLNNPEKEPKKFQTAYAKKTGDQKGYREALCKWIDFT 
               
               
                   
                   
               
               
                   
                 RDFLSKYTKTTSIDLSSLRPSSQYKDLGEYYAELNPLLYH 
               
               
                   
                   
               
               
                   
                 ISFQRIAEKEIMDAVETGKLYLFQIYNKDFAKGHHGKPNL 
               
               
                   
                   
               
               
                   
                 HTLYWTGLFSPENLAKTSIKLNGQAELFYRPKSRMKRMAH 
               
               
                   
                   
               
               
                   
                 RLGEKMLNKKLKDQKTPIPDTLYQELYDYVNHRLSHDLSD 
               
               
                   
                   
               
               
                   
                 EARALLPNVITKEVSHEIIKDRRFTSDKFFFHVPITLNYQ 
               
               
                   
                   
               
               
                   
                 AANSPSKFNQRVNAYLKEHPETPIIGIDRGERNLIYITVI 
               
               
                   
                   
               
               
                   
                 DSTGKILEQRSLNTIQQFDYQKKLDNREKERVAARQAWSV 
               
               
                   
                   
               
               
                   
                 VGTIKDLKQGYLSQVIHEIVDLMIHYQAVVVLENLNFGFK 
               
               
                   
                   
               
               
                   
                 SKRTGIAEKAVYQQFEKMLIDKLNCLVLKDYPAEKVGGVL 
               
               
                   
                   
               
               
                   
                 NPYQLTDQFTSFAKMGTQSGFLFYVPAPYTSKIDPLTGFV 
               
               
                   
                   
               
               
                   
                 DPFVWKTIKNHESRKHFLEGFDFLHYDVKTGDFILHFKMN 
               
               
                   
                   
               
               
                   
                 RNLSFQRGLPGFMPAWDIVFEKNETQFDAKGTPFIAGKRI 
               
               
                   
                   
               
               
                   
                 VPVIENHRFTGRYRDLYPANELIALLEEKGIVFRDGSNIL 
               
               
                   
                   
               
               
                   
                 PKLLENDDSHAIDTMVALIRSVLQMRNSNAATGEDYINSP 
               
               
                   
                   
               
               
                   
                 VRDLNGVCFDSRFQNPEWPMDADANGAYHIALKGQLLLNH 
               
               
                   
                   
               
               
                   
                 LKESKDLKLQNGISNQDWLAYIQELRN 
               
               
                   
                   
               
               
                   
                 Seq ID NO 201: 
               
               
                   
                 &gt;6his-NLS-A3A-GGS3-dCas9 
               
               
                   
                 HHHHHH-SSGLVPRGSHM-PKKKRKV-MEASPASGPRHLM 
               
               
                   
                   
               
               
                   
                 DPHIFTSNFNNGIGRHKTYLCYEVERLDNGTSVKMDQHRG 
               
               
                   
                   
               
               
                   
                 FLHNQAKNLLCGFYGRHAELRFLDLVPSLQLDPAQIYRVT 
               
               
                   
                   
               
               
                   
                 WFISWSPCFSWGCAGEVRAFLQENTHVRLRIFAARIYDYD 
               
               
                   
                   
               
               
                   
                 PLYKEALQMLRDAGAQVSIMTYDEFKHCWDTFVDHQGCPF 
               
               
                   
                   
               
               
                   
                 QPWDGLDEHSQALSGRLRAILQNQGN-GGSGGSGGS-MDK 
               
               
                   
                   
               
               
                   
                 KYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSI 
               
               
                   
                   
               
               
                   
                 KKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQ 
               
               
                   
                   
               
               
                   
                 EIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIV 
               
               
                   
                   
               
               
                   
                 DEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK 
               
               
                   
                   
               
               
                   
                 FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINA 
               
               
                   
                   
               
               
                   
                 SGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIA 
               
               
                   
                   
               
               
                   
                 LSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIG 
               
               
                   
                   
               
               
                   
                 DQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIK 
               
               
                   
                   
               
               
                   
                 RYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYI 
               
               
                   
                   
               
               
                   
                 DGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQR 
               
               
                   
                   
               
               
                   
                 TFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKI 
               
               
                   
                   
               
               
                   
                 LTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVD 
               
               
                   
                   
               
               
                   
                 KGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNE 
               
               
                   
                   
               
               
                   
                 LTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQ 
               
               
                   
                   
               
               
                   
                 LKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKD 
               
               
                   
                   
               
               
                   
                 KDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLF 
               
               
                   
                   
               
               
                   
                 DDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFL 
               
               
                   
                   
               
               
                   
                 KSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEH 
               
               
                   
                   
               
               
                   
                 IANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEM 
               
               
                   
                   
               
               
                   
                 ARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVEN 
               
               
                   
                   
               
               
                   
                 TQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDAIVP 
               
               
                   
                   
               
               
                   
                 QSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYW 
               
               
                   
                   
               
               
                   
                 RQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVE 
               
               
                   
                   
               
               
                   
                 TRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLV 
               
               
                   
                   
               
               
                   
                 SDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPK 
               
               
                   
                   
               
               
                   
                 LESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIM 
               
               
                   
                   
               
               
                   
                 NFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATV 
               
               
                   
                   
               
               
                   
                 RKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARK 
               
               
                   
                   
               
               
                   
                 KDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKEL 
               
               
                   
                   
               
               
                   
                 LGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSL 
               
               
                   
                   
               
               
                   
                 FELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYE 
               
               
                   
                   
               
               
                   
                 KLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILA 
               
               
                   
                   
               
               
                   
                 DANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAA 
               
               
                   
                   
               
               
                   
                 FKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLS 
               
               
                   
                   
               
               
                   
                 QLGGDPPKKKRKV 
               
               
                   
                   
               
               
                   
                 Seq ID NO 202: 
               
               
                   
                 &gt;6his-NLS-A3A-GGS7-dCas9 
               
               
                   
                 HHHHHH-SSGLVPRGSHM-PKKKRKV-EASPASGPRHLMD 
               
               
                   
                   
               
               
                   
                 PHIFTSNFNNGIGRHKTYLCYEVERLDNGTSVKMDQHRGF 
               
               
                   
                   
               
               
                   
                 LHNQAKNLLCGFYGRHAELRFLDLVPSLQLDPAQIYRVTW 
               
               
                   
                   
               
               
                   
                 FISWSPCFSWGCAGEVRAFLQENTHVRLRIFAARIYDYDP 
               
               
                   
                   
               
               
                   
                 LYKEALQMLRDAGAQVSIMTYDEFKHCWDTFVDHQGCPFQ 
               
               
                   
                   
               
               
                   
                 PWDGLDEHSQALSGRLRAILQNQGN-GGSGGSGGSGGSGG 
               
               
                   
                   
               
               
                   
                 SGGSGGS-MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKF 
               
               
                   
                   
               
               
                   
                 KVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRY 
               
               
                   
                   
               
               
                   
                 TRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKK 
               
               
                   
                   
               
               
                   
                 HERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLR 
               
               
                   
                   
               
               
                   
                 LIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTY 
               
               
                   
                   
               
               
                   
                 NQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGE 
               
               
                   
                   
               
               
                   
                 KKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYD 
               
               
                   
                   
               
               
                   
                 DDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEI 
               
               
                   
                   
               
               
                   
                 TKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFF 
               
               
                   
                   
               
               
                   
                 DQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVK 
               
               
                   
                   
               
               
                   
                 LNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPF 
               
               
                   
                   
               
               
                   
                 LKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEET 
               
               
                   
                   
               
               
                   
                 ITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHS 
               
               
                   
                   
               
               
                   
                 LLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLL 
               
               
                   
                   
               
               
                   
                 FKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLG 
               
               
                   
                   
               
               
                   
                 TYHDLLKlIKDKDFLDNEENEDILEDIVLTLTLFEDREMI 
               
               
                   
                   
               
               
                   
                 EERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRD 
               
               
                   
                   
               
               
                   
                 KQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQ 
               
               
                   
                   
               
               
                   
                 VSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMG 
               
               
                   
                   
               
               
                   
                 RHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELG 
               
               
                   
                   
               
               
                   
                 SQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINR 
               
               
                   
                   
               
               
                   
                 LSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS 
               
               
                   
                   
               
               
                   
                 EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELD 
               
               
                   
                   
               
               
                   
                 KAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIRE 
               
               
                   
                   
               
               
                   
                 VKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAV 
               
               
                   
                   
               
               
                   
                 VGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKA 
               
               
                   
                   
               
               
                   
                 TAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEI 
               
               
                   
                   
               
               
                   
                 VWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILP 
               
               
                   
                   
               
               
                   
                 KRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKG 
               
               
                   
                   
               
               
                   
                 KSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKK 
               
               
                   
                   
               
               
                   
                 DLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKY 
               
               
                   
                   
               
               
                   
                 VNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQ 
               
               
                   
                   
               
               
                   
                 ISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHL 
               
               
                   
                   
               
               
                   
                 FTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSI 
               
               
                   
                   
               
               
                   
                 TGLYETRIDLSQLGGDPPKKKRKV 
               
               
                   
                   
               
               
                   
                 Seq ID NO 203: 
               
               
                   
                 &gt;6his-NLS-A3A-GGS14-dCas9 
               
               
                   
                 HHHHHH-SSGLVPRGSHM-PKKKRKV-EASPASGPRHLMD 
               
               
                   
                   
               
               
                   
                 PHIFTSNFNNGIGRHKTYLCYEVERLDNGTSVKMDQHRGF 
               
               
                   
                   
               
               
                   
                 LHNQAKNLLCGFYGRHAELRFLDLVPSLQLDPAQIYRVTW 
               
               
                   
                   
               
               
                   
                 FISWSPCFSWGCAGEVRAFLQENTHVRLRIFAARIYDYDP 
               
               
                   
                   
               
               
                   
                 LYKEALQMLRDAGAQVSIMTYDEFKHCWDTFVDHQGCPFQ 
               
               
                   
                   
               
               
                   
                 PWDGLDEHSQALSGRLRAILQNQGN-GGSGGSGGSGGSGG 
               
               
                   
                   
               
               
                   
                 SGGSGGSGGSGGSGGSGGSGGSGGSGGS-MDKKYSIGLAI 
               
               
                   
                   
               
               
                   
                 GTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGAL 
               
               
                   
                   
               
               
                   
                 LFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMA 
               
               
                   
                   
               
               
                   
                 KVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEK 
               
               
                   
                   
               
               
                   
                 YPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIE 
               
               
                   
                   
               
               
                   
                 GDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAI 
               
               
                   
                   
               
               
                   
                 LSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPN 
               
               
                   
                   
               
               
                   
                 FKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFL 
               
               
                   
                   
               
               
                   
                 AAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQD 
               
               
                   
                   
               
               
                   
                 LTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEE 
               
               
                   
                   
               
               
                   
                 FYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIP 
               
               
                   
                   
               
               
                   
                 HQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYY 
               
               
                   
                   
               
               
                   
                 VGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSF 
               
               
                   
                   
               
               
                   
                 IERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVT 
               
               
                   
                   
               
               
                   
                 EGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKK 
               
               
                   
                   
               
               
                   
                 IECFDSVEISGVEDRFNASLGTYHDLLKlIKDKDFLDNEE 
               
               
                   
                   
               
               
                   
                 NEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQL 
               
               
                   
                   
               
               
                   
                 KRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANR 
               
               
                   
                   
               
               
                   
                 NFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSP 
               
               
                   
                   
               
               
                   
                 AIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQ 
               
               
                   
                   
               
               
                   
                 KGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKL 
               
               
                   
                   
               
               
                   
                 YLYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDS 
               
               
                   
                   
               
               
                   
                 IDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKL 
               
               
                   
                   
               
               
                   
                 ITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV 
               
               
                   
                   
               
               
                   
                 AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQ 
               
               
                   
                   
               
               
                   
                 FYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYG 
               
               
                   
                   
               
               
                   
                 DYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEIT 
               
               
                   
                   
               
               
                   
                 LANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQ 
               
               
                   
                   
               
               
                   
                 VNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKY 
               
               
                   
                   
               
               
                   
                 GGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMER 
               
               
                   
                   
               
               
                   
                 SSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRK 
               
               
                   
                   
               
               
                   
                 RMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPED 
               
               
                   
                   
               
               
                   
                 NEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVL 
               
               
                   
                   
               
               
                   
                 SAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTI 
               
               
                   
                   
               
               
                   
                 DRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGDPPK 
               
               
                   
                   
               
               
                   
                 KKRKV 
               
               
                   
                   
               
               
                   
                 Seq ID NO 204: 
               
               
                   
                 &gt;6his-NLS-A3H-GGS3-dCas9 
               
               
                   
                 HHHHHH-SSGLVPRGSHM-PKKKRKV-MALLTAETFRLQF 
               
               
                   
                   
               
               
                   
                 NNKRRLRRPYYPRKALLCYQLTPQNGSTPTRGYFENKKKC 
               
               
                   
                   
               
               
                   
                 HAEICFINEIKSMGLDETQCYQVTCYLTWSPCSSCAWELV 
               
               
                   
                   
               
               
                   
                 DFIKAHDHLNLRIFASRLYYHWCKPQQDGLRLLCGSQVPV 
               
               
                   
                   
               
               
                   
                 EVMGFPEFADCWENFVDHEKPLSFNPYKMLEELDKNSRAI 
               
               
                   
                   
               
               
                   
                 KRRLDRIKS-GGSGGSGGS-MDKKYSIGLAIGTNSVGWAV 
               
               
                   
                   
               
               
                   
                 ITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAE 
               
               
                   
                   
               
               
                   
                 ATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHR 
               
               
                   
                   
               
               
                   
                 LEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRK 
               
               
                   
                   
               
               
                   
                 KLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSD 
               
               
                   
                   
               
               
                   
                 VDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSR 
               
               
                   
                   
               
               
                   
                 RLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAE 
               
               
                   
                   
               
               
                   
                 DAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAI 
               
               
                   
                   
               
               
                   
                 LLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR 
               
               
                   
                   
               
               
                   
                 QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPIL 
               
               
                   
                   
               
               
                   
                 EKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELH 
               
               
                   
                   
               
               
                   
                 AILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNS 
               
               
                   
                   
               
               
                   
                 RFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDK 
               
               
                   
                   
               
               
                   
                 NLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFL 
               
               
                   
                   
               
               
                   
                 SGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEI 
               
               
                   
                   
               
               
                   
                 SGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIV 
               
               
                   
                   
               
               
                   
                 LTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWG 
               
               
                   
                   
               
               
                   
                 RLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDD 
               
               
                   
                   
               
               
                   
                 SLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQT 
               
               
                   
                   
               
               
                   
                 VKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRER 
               
               
                   
                   
               
               
                   
                 MKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGR 
               
               
                   
                   
               
               
                   
                 DMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRS 
               
               
                   
                   
               
               
                   
                 DKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNL 
               
               
                   
                   
               
               
                   
                 TKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMN 
               
               
                   
                   
               
               
                   
                 TKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINN 
               
               
                   
                   
               
               
                   
                 YHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRK 
               
               
                   
                   
               
               
                   
                 MIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKR 
               
               
                   
                   
               
               
                   
                 PLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEV 
               
               
                   
                   
               
               
                   
                 QTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVA 
               
               
                   
                   
               
               
                   
                 YSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPID 
               
               
                   
                   
               
               
                   
                 FLEAKGYKEVKKDLIKLPKYSLFELENGRKRMLASAGELQ 
               
               
                   
                   
               
               
                   
                 KGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQ 
               
               
                   
                   
               
               
                   
                 HKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKP 
               
               
                   
                   
               
               
                   
                 IREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKE 
               
               
                   
                   
               
               
                   
                 VLDATLIHQSITGLYETRIDLSQLGGDPPKKKRKV 
               
               
                   
                   
               
               
                   
                 Seq ID NO 205: 
               
               
                   
                 &gt;6his-NLS-A3H-GGS7-dCas9 
               
               
                   
                 HHHHHH-SSGLVPRGSHM-PKKKRKV-MALLTAETFRLQF 
               
               
                   
                   
               
               
                   
                 NNKRRLRRPYYPRKALLCYQLTPQNGSTPTRGYFENKKKC 
               
               
                   
                   
               
               
                   
                 HAEICFINEIKSMGLDETQCYQVTCYLTWSPCSSCAWELV 
               
               
                   
                   
               
               
                   
                 DFIKAHDHLNLRIFASRLYYHWCKPQQDGLRLLCGSQVPV 
               
               
                   
                   
               
               
                   
                 EVMGFPEFADCWENFVDHEKPLSFNPYKMLEELDKNSRAI 
               
               
                   
                   
               
               
                   
                 KRRLDRIKS-GGSGGSGGSGGSGGSGGSGGS-MDKKYSIG 
               
               
                   
                   
               
               
                   
                 LAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLI 
               
               
                   
                   
               
               
                   
                 GALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSN 
               
               
                   
                   
               
               
                   
                 EMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAY 
               
               
                   
                   
               
               
                   
                 HEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHF 
               
               
                   
                   
               
               
                   
                 LIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDA 
               
               
                   
                   
               
               
                   
                 KAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGL 
               
               
                   
                   
               
               
                   
                 TPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYAD 
               
               
                   
                   
               
               
                   
                 LFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEH 
               
               
                   
                   
               
               
                   
                 HQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGAS 
               
               
                   
                   
               
               
                   
                 QEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNG 
               
               
                   
                   
               
               
                   
                 SIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRI 
               
               
                   
                   
               
               
                   
                 PYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASA 
               
               
                   
                   
               
               
                   
                 QSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVK 
               
               
                   
                   
               
               
                   
                 YVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDY 
               
               
                   
                   
               
               
                   
                 FKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLD 
               
               
                   
                   
               
               
                   
                 NEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVM 
               
               
                   
                   
               
               
                   
                 KQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGF 
               
               
                   
                   
               
               
                   
                 ANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLA 
               
               
                   
                   
               
               
                   
                 GSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQ 
               
               
                   
                   
               
               
                   
                 TTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQN 
               
               
                   
                   
               
               
                   
                 EKLYLYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLK 
               
               
                   
                   
               
               
                   
                 DDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLN 
               
               
                   
                   
               
               
                   
                 AKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQIT 
               
               
                   
                   
               
               
                   
                 KHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRK 
               
               
                   
                   
               
               
                   
                 DFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEF 
               
               
                   
                   
               
               
                   
                 VYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT 
               
               
                   
                   
               
               
                   
                 EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLS 
               
               
                   
                   
               
               
                   
                 MPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDP 
               
               
                   
                   
               
               
                   
                 KKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITI 
               
               
                   
                   
               
               
                   
                 MERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELEN 
               
               
                   
                   
               
               
                   
                 GRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGS 
               
               
                   
                   
               
               
                   
                 PEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLD 
               
               
                   
                   
               
               
                   
                 KVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFD 
               
               
                   
                   
               
               
                   
                 TTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGD 
               
               
                   
                   
               
               
                   
                 PPKKKRKV 
               
               
                   
                   
               
               
                   
                 Seq ID NO 206: 
               
               
                   
                 &gt;6his-NLS-A3H-GGS14-dCas9 
               
               
                   
                 HHHHHH-SSGLVPRGSHM-PKKKRKV-MALLTAETFRLQF 
               
               
                   
                   
               
               
                   
                 NNKRRLRRPYYPRKALLCYQLTPQNGSTPTRGYFENKKKC 
               
               
                   
                   
               
               
                   
                 HAEICFINEIKSMGLDETQCYQVTCYLTWSPCSSCAWELV 
               
               
                   
                   
               
               
                   
                 DFIKAHDHLNLRIFASRLYYHWCKPQQDGLRLLCGSQVPV 
               
               
                   
                   
               
               
                   
                 EVMGFPEFADCWENFVDHEKPLSFNPYKMLEELDKNSRAI 
               
               
                   
                   
               
               
                   
                 KRRLDRIKS-GGSGGSGGSGGSGGSGGSGGSGGSGGSGGS 
               
               
                   
                   
               
               
                   
                 GGSGGSGGSGGS-MDKKYSIGLAIGTNSVGWAVITDEYKV 
               
               
                   
                   
               
               
                   
                 PSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRT 
               
               
                   
                   
               
               
                   
                 ARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLV 
               
               
                   
                   
               
               
                   
                 EEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTD 
               
               
                   
                   
               
               
                   
                 KADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQ 
               
               
                   
                   
               
               
                   
                 LVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIA 
               
               
                   
                   
               
               
                   
                 QLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLS 
               
               
                   
                   
               
               
                   
                 KDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILR 
               
               
                   
                   
               
               
                   
                 VNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKY 
               
               
                   
                   
               
               
                   
                 KEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTE 
               
               
                   
                   
               
               
                   
                 ELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQE 
               
               
                   
                   
               
               
                   
                 DFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTR 
               
               
                   
                   
               
               
                   
                 KSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKV 
               
               
                   
                   
               
               
                   
                 LPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKA 
               
               
                   
                   
               
               
                   
                 IVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRF 
               
               
                   
                   
               
               
                   
                 NASLGTYHDLLKlIKDKDFLDNEENEDILEDIVLTLTLFE 
               
               
                   
                   
               
               
                   
                 DREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLI 
               
               
                   
                   
               
               
                   
                 NGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKED 
               
               
                   
                   
               
               
                   
                 IQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDEL 
               
               
                   
                   
               
               
                   
                 VKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEG 
               
               
                   
                   
               
               
                   
                 IKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQE 
               
               
                   
                   
               
               
                   
                 LDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKS 
               
               
                   
                   
               
               
                   
                 DNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGG 
               
               
                   
                   
               
               
                   
                 LSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDEND 
               
               
                   
                   
               
               
                   
                 KLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDA 
               
               
                   
                   
               
               
                   
                 YLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQ 
               
               
                   
                   
               
               
                   
                 EIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNG 
               
               
                   
                   
               
               
                   
                 ETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK 
               
               
                   
                   
               
               
                   
                 ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVA 
               
               
                   
                   
               
               
                   
                 KVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGY 
               
               
                   
                   
               
               
                   
                 KEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELA 
               
               
                   
                   
               
               
                   
                 LPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLD 
               
               
                   
                   
               
               
                   
                 EIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAE 
               
               
                   
                   
               
               
                   
                 NIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATL 
               
               
                   
                   
               
               
                   
                 IHQSITGLYETRIDLSQLGGDPPKKKRKV 
               
               
                   
                   
               
               
                   
                 Seq ID NO 207: 
               
               
                   
                 &gt;6his-NLS-A3H-GGS7-dCpf1 gene sequence 
               
               
                   
                 HHHHHH-SSGLVPRGSHM-PKKKRKV-MALLTAETFRLQF 
               
               
                   
                   
               
               
                   
                 NNKRRLRRPYYPRKALLCYQLTPQNGSTPTRGYFENKKKC 
               
               
                   
                   
               
               
                   
                 HAEICFINEIKSMGLDETQCYQVTCYLTWSPCSSCAWELV 
               
               
                   
                   
               
               
                   
                 DFIKAHDHLNLRIFASRLYYHWCKPQQDGLRLLCGSQVPV 
               
               
                   
                   
               
               
                   
                 EVMGFPEFADCWENFVDHEKPLSFNPYKMLEELDKNSRAI 
               
               
                   
                   
               
               
                   
                 KRRLDRIKS-GGSGGSGGSGGSGGSGGSGGS-KLTQFEGF 
               
               
                   
                   
               
               
                   
                 TNLYQVSKTLRFELIPQGKTLKHIQEQGFIEEDKARNDHY 
               
               
                   
                   
               
               
                   
                 KELKPIIDRIYKTYADQCLQLVQLDWENLSAAIDSYRKEK 
               
               
                   
                   
               
               
                   
                 TEETRNALIEEQATYRNAIHDYFIGRTDNLTDAINKRHAE 
               
               
                   
                   
               
               
                   
                 IYKGLFKAELFNGKVLKQLGTVTTTEHENALLRSFDKFTT 
               
               
                   
                   
               
               
                   
                 YFSGFYENRKNVFSAEDISTAIPHRIVQDNFPKFKENCHI 
               
               
                   
                   
               
               
                   
                 FTRLITAVPSLREHFENVKKAIGIFVSTSIEEVFsFPFYN 
               
               
                   
                   
               
               
                   
                 QLLTQTQIDLYNQLLGGISREAGTEKIKGLNEVLNLAIQK 
               
               
                   
                   
               
               
                   
                 NDETAHIIASLPHRFIPLFKQILSDRNTLSFILEEFKSDE 
               
               
                   
                   
               
               
                   
                 EVIQSFCKYKTLLRNENVLETAEALFNELNSIDLTHIFIS 
               
               
                   
                   
               
               
                   
                 HKKLETISSALCDHWDTLRNALYERRISELTGKITKSAKE 
               
               
                   
                   
               
               
                   
                 KVQRSLKHEDINLQEIISAAGKELSEAFKQKTSEILSHAH 
               
               
                   
                   
               
               
                   
                 AALDQPLPTTLKKQEEKEILKSQLDSLLGLYHLLDWFAVD 
               
               
                   
                   
               
               
                   
                 ESNEVDPEFSARLTGIKLEMEPSLSFYNKARNYATKKPYS 
               
               
                   
                   
               
               
                   
                 VEKFKLNFQMPTLASGWDVNKEKNNGAILFVKNGLYYLGI 
               
               
                   
                   
               
               
                   
                 MPKQKGRYKALSFEPTEKTSEGFDKMYYDYFPDAAKMIPK 
               
               
                   
                   
               
               
                   
                 CSTQLKAVTAHFQTHTTPILLSNNFIEPLEITKEIYDLNN 
               
               
                   
                   
               
               
                   
                 PEKEPKKFQTAYAKKTGDQKGYREALCKWIDFTRDFLSKY 
               
               
                   
                   
               
               
                   
                 TKTTSIDLSSLRPSSQYKDLGEYYAELNPLLYHISFQRIA 
               
               
                   
                   
               
               
                   
                 EKEIMDAVETGKLYLFQIYNKDFAKGHHGKPNLHTLYWTG 
               
               
                   
                   
               
               
                   
                 LFSPENLAKTSIKLNGQAELFYRPKSRMKRMAHRLGEKML 
               
               
                   
                   
               
               
                   
                 NKKLKDQKTPIPDTLYQELYDYVNHRLSHDLSDEARALLP 
               
               
                   
                   
               
               
                   
                 NVITKEVSHEIIKDRRFTSDKFFFHVPITLNYQAANSPSK 
               
               
                   
                   
               
               
                   
                 FNQRVNAYLKEHPETPIIGIARGERNLIYITVIDSTGKIL 
               
               
                   
                   
               
               
                   
                 EQRSLNTIQQFDYQKKLDNREKERVAARQAWSVVGTIKDL 
               
               
                   
                   
               
               
                   
                 KQGYLSQVIHEIVDLMIHYQAVVVLENLNFGFKSKRTGIA 
               
               
                   
                   
               
               
                   
                 EKAVYQQFEKMLIDKLNCLVLKDYPAEKVGGVLNPYQLTD 
               
               
                   
                   
               
               
                   
                 QFTSFAKMGTQSGFLFYVPAPYTSKIDPLTGFVDPFVWKT 
               
               
                   
                   
               
               
                   
                 IKNHESRKHFLEGFDFLHYDVKTGDFILHFKMNRNLSFQR 
               
               
                   
                   
               
               
                   
                 GLPGFMPAWDIVFEKNETQFDAKGTPFIAGKRIVPVIENH 
               
               
                   
                   
               
               
                   
                 RFTGRYRDLYPANELIALLEEKGIVFRDGSNILPKLLEND 
               
               
                   
                   
               
               
                   
                 DSHAIDTMVALIRSVLQMRNSNAATGEDYINSPVRDLNGV 
               
               
                   
                   
               
               
                   
                 CFDSRFQNPEWPMDADANGAYHIALKGQLLLNHLKESKDL 
               
               
                   
                   
               
               
                   
                 KLQNGISNQDWLAYIQELRN 
               
               
                   
                   
               
               
                   
                 Seq ID NO 301: 
               
               
                   
                 &gt;6his-NLS-A3A-GGS3-dCas9 gene sequence 
               
               
                   
                 ATGggcagcagccatcatcatcatcatcacagcagcggcc 
               
               
                   
                   
               
               
                   
                 tggtgccgcgcggcagccatatgccaaagaagaagcggaa 
               
               
                   
                   
               
               
                   
                 ggtcGAAGCCAGCCCAGCATCCGGGCCCAGACACTTGATG 
               
               
                   
                   
               
               
                   
                 GATCCACACATATTCACTTCCAACTTTAACAATGGCATTG 
               
               
                   
                   
               
               
                   
                 GAAGGCATAAGACCTACCTGTGCTACGAAGTGGAGCGCCT 
               
               
                   
                   
               
               
                   
                 GGACAATGGCACCTCGGTCAAGATGGACCAGCACAGGGGC 
               
               
                   
                   
               
               
                   
                 TTTCTACACAACCAGGCTAAGAATCTTCTCTGTGGCTTTT 
               
               
                   
                   
               
               
                   
                 ACGGCCGCCATGCGCAGCTGCGCTTCTTGGACCTGGTTCC 
               
               
                   
                   
               
               
                   
                 TTCTTTGCAGTTGGACCCGGCCCAGATCTACAGGGTCACT 
               
               
                   
                   
               
               
                   
                 TGGTTCATCTCCTGGAGCCCCTGCTTCTCCTGGGGCTGTG 
               
               
                   
                   
               
               
                   
                 CCGGGGAAGTGCGTGCGTTCCTTCAGGAGAACACACACGT 
               
               
                   
                   
               
               
                   
                 GAGACTGCGTATCTTCGCTGCCCGCATCTATGATTACGAC 
               
               
                   
                   
               
               
                   
                 CCCCTATATAAGGAGGCACTGCAAATGCTGCGGGATGCTG 
               
               
                   
                   
               
               
                   
                 GGGCCCAAGTCTCCATCATGACCTACGATGAATTTAAGCA 
               
               
                   
                   
               
               
                   
                 CTGCTGGGACACCTTTGTGGACCACCAGGGATGTCCCTTC 
               
               
                   
                   
               
               
                   
                 CAGCCCTGGGATGGACTAGATGAGCACAGCCAAGCCCTGA 
               
               
                   
                   
               
               
                   
                 GTGGGAGGCTGCGGGCCATTCTCCAGAATCAGGGAAACGG 
               
               
                   
                   
               
               
                   
                 AGGAAGTGGAGGAAGTGGAGGAAGTaagcttgacaagaag 
               
               
                   
                   
               
               
                   
                 tacagcatcggcctggccatcggcaccaactctgtgggct 
               
               
                   
                   
               
               
                   
                 gggccgtgatcaccgacgagtacaaggtgcccagcaagaa 
               
               
                   
                   
               
               
                   
                 attcaaggtgctgggcaacaccgaccggcacagcatcaag 
               
               
                   
                   
               
               
                   
                 aagaacctgatcggagccctgctgttcgacagcggcgaaa 
               
               
                   
                   
               
               
                   
                 cagccgaggccacccggctgaagagaaccgccagaagaag 
               
               
                   
                   
               
               
                   
                 atacaccagacggaagaaccggatctgctatctgcaagag 
               
               
                   
                   
               
               
                   
                 atcttcagcaacgagatggccaaggtggacgacagcttct 
               
               
                   
                   
               
               
                   
                 tccacagactggaagagtccttcctggtggaagaggataa 
               
               
                   
                   
               
               
                   
                 gaagcacgagcggcaccccatcttcggcaacatcgtggac 
               
               
                   
                   
               
               
                   
                 gaggtggcctaccacgagaagtaccccaccatctaccacc 
               
               
                   
                   
               
               
                   
                 tgagaaagaaactggtggacagcaccgacaaggccgacct 
               
               
                   
                   
               
               
                   
                 gcggctgatctatctggccctggcccacatgatcaagttc 
               
               
                   
                   
               
               
                   
                 cggggccacttcctgatcgagggcgacctgaaccccgaca 
               
               
                   
                   
               
               
                   
                 acagcgacgtggacaagctgttcatccagctggtgcagac 
               
               
                   
                   
               
               
                   
                 ctacaaccagctgttcgaggaaaaccccatcaacgccagc 
               
               
                   
                   
               
               
                   
                 ggcgtggacgccaaggccatcctgtctgccagactgagca 
               
               
                   
                   
               
               
                   
                 agagcagacggctggaaaatctgatcgcccagctgcccgg 
               
               
                   
                   
               
               
                   
                 cgagaagaagaatggcctgttcggaaacctgattgccctg 
               
               
                   
                   
               
               
                   
                 agcctgggcctgacccccaacttcaagagcaacttcgacc 
               
               
                   
                   
               
               
                   
                 tggccgaggatgccaaactgcagctgagcaaggacaccta 
               
               
                   
                   
               
               
                   
                 cgacgacgacctggacaacctgctggcccagatcggcgac 
               
               
                   
                   
               
               
                   
                 cagtacgccgacctgtttctggccgccaagaacctgtccg 
               
               
                   
                   
               
               
                   
                 acgccatcctgctgagcgacatcctgagagtgaacaccga 
               
               
                   
                   
               
               
                   
                 gatcaccaaggcccccctgagcgcctctatgatcaagaga 
               
               
                   
                   
               
               
                   
                 tacgacgagcaccaccaggacctgaccctgctgaaagctc 
               
               
                   
                   
               
               
                   
                 tcgtgcggcagcagctgcctgagaagtacaaagagatttt 
               
               
                   
                   
               
               
                   
                 cttcgaccagagcaagaacggctacgccggctacattgac 
               
               
                   
                   
               
               
                   
                 ggcggagccagccaggaagagttctacaagttcatcaagc 
               
               
                   
                   
               
               
                   
                 ccatcctggaaaagatggacggcaccgaggaactgctcgt 
               
               
                   
                   
               
               
                   
                 gaagctgaacagagaggacctgctgcggaagcagcggacc 
               
               
                   
                   
               
               
                   
                 ttcgacaacggcagcatcccccaccagatccacctgggag 
               
               
                   
                   
               
               
                   
                 agctgcacgccattctgcggcggcaggaagatttttaccc 
               
               
                   
                   
               
               
                   
                 attcctgaaggacaaccgggaaaagatcgagaagatcctg 
               
               
                   
                   
               
               
                   
                 accttccgcatcccctactacgtgggccctctggccaggg 
               
               
                   
                   
               
               
                   
                 gaaacagcagattcgcctggatgaccagaaagagcgagga 
               
               
                   
                   
               
               
                   
                 aaccatcaccccctggaacttcgaggaagtggtggacaag 
               
               
                   
                   
               
               
                   
                 ggcgcttccgcccagagcttcatcgagcggatgaccaact 
               
               
                   
                   
               
               
                   
                 tcgataagaacctgcccaacgagaaggtgctgcccaagca 
               
               
                   
                   
               
               
                   
                 cagcctgctgtacgagtacttcaccgtgtataacgagctg 
               
               
                   
                   
               
               
                   
                 accaaagtgaaatacgtgaccgagggaatgagaaagcccg 
               
               
                   
                   
               
               
                   
                 ccttcctgagcggcgagcagaaaaaggccatcgtggacct 
               
               
                   
                   
               
               
                   
                 gctgttcaagaccaaccggaaagtgaccgtgaagcagctg 
               
               
                   
                   
               
               
                   
                 aaagaggactacttcaagaaaatcgagtgcttcgactccg 
               
               
                   
                   
               
               
                   
                 tggaaatctccggcgtggaagatcggttcaacgcctccct 
               
               
                   
                   
               
               
                   
                 gggcacataccacgatctgctgaaaattatcaaggacaag 
               
               
                   
                   
               
               
                   
                 gacttcctggacaatgaggaaaacgaggacattctggaag 
               
               
                   
                   
               
               
                   
                 atatcgtgctgaccctgacactgtttgaggacagagagat 
               
               
                   
                   
               
               
                   
                 gatcgaggaacggctgaaaacctatgcccacctgttcgac 
               
               
                   
                   
               
               
                   
                 gacaaagtgatgaagcagctgaagcggcggagatacaccg 
               
               
                   
                   
               
               
                   
                 gctggggcaggctgagccggaagctgatcaacggcatccg 
               
               
                   
                   
               
               
                   
                 ggacaagcagtccggcaagacaatcctggatttcctgaag 
               
               
                   
                   
               
               
                   
                 tccgacggcttcgccaacagaaacttcatgcagctgatcc 
               
               
                   
                   
               
               
                   
                 acgacgacagcctgacctttaaagaggacatccagaaagc 
               
               
                   
                   
               
               
                   
                 ccaggtgtccggccagggcgatagcctgcacgagcacatt 
               
               
                   
                   
               
               
                   
                 gccaatctggccggcagccccgccattaagaagggcatcc 
               
               
                   
                   
               
               
                   
                 tgcagacagtgaaggtggtggacgagctcgtgaaagtgat 
               
               
                   
                   
               
               
                   
                 gggccggcacaagcccgagaacatcgtgatcgaaatggcc 
               
               
                   
                   
               
               
                   
                 agagagaaccagaccacccagaagggacagaagaacagcc 
               
               
                   
                   
               
               
                   
                 gcgagagaatgaagcggatcgaagagggcatcaaagagct 
               
               
                   
                   
               
               
                   
                 gggcagccagatcctgaaagaacaccccgtggaaaacacc 
               
               
                   
                   
               
               
                   
                 cagctgcagaacgagaagctgtacctgtactacctgcaga 
               
               
                   
                   
               
               
                   
                 atgggcgggatatgtacgtggaccaggaactggacatcaa 
               
               
                   
                   
               
               
                   
                 ccggctgtccgactacgatgtggacgctatcgtgcctcag 
               
               
                   
                   
               
               
                   
                 agctttctgaaggacgactccatcgacaacaaggtgctga 
               
               
                   
                   
               
               
                   
                 ccagaagcgacaagaaccggggcaagagcgacaacgtgcc 
               
               
                   
                   
               
               
                   
                 ctccgaagaggtcgtgaagaagatgaagaactactggcgg 
               
               
                   
                   
               
               
                   
                 cagctgctgaacgccaagctgattacccagagaaagttcg 
               
               
                   
                   
               
               
                   
                 acaatctgaccaaggccgagagaggcggcctgagcgaact 
               
               
                   
                   
               
               
                   
                 ggataaggccggcttcatcaagagacagctggtggaaacc 
               
               
                   
                   
               
               
                   
                 cggcagatcacaaagcacgtggcacagatcctggactccc 
               
               
                   
                   
               
               
                   
                 ggatgaacactaagtacgacgagaatgacaagctgatccg 
               
               
                   
                   
               
               
                   
                 ggaagtgaaagtgatcaccctgaagtccaagctggtgtcc 
               
               
                   
                   
               
               
                   
                 gatttccggaaggatttccagttttacaaagtgcgcgaga 
               
               
                   
                   
               
               
                   
                 tcaacaactaccaccacgcccacgacgcctacctgaacgc 
               
               
                   
                   
               
               
                   
                 cgtcgtgggaaccgccctgatcaaaaagtaccctaagctg 
               
               
                   
                   
               
               
                   
                 gaaagcgagttcgtgtacggcgactacaaggtgtacgacg 
               
               
                   
                   
               
               
                   
                 tgcggaagatgatcgccaagagcgagcaggaaatcggcaa 
               
               
                   
                   
               
               
                   
                 ggctaccgccaagtacttcttctacagcaacatcatgaac 
               
               
                   
                   
               
               
                   
                 tttttcaagaccgagattaccctggccaacggcgagatcc 
               
               
                   
                   
               
               
                   
                 ggaagcggcctctgatcgagacaaacggcgaaaccgggga 
               
               
                   
                   
               
               
                   
                 gatcgtgtgggataagggccgggattttgccaccgtgcgg 
               
               
                   
                   
               
               
                   
                 aaagtgctgagcatgccccaagtgaatatcgtgaaaaaga 
               
               
                   
                   
               
               
                   
                 ccgaggtgcagacaggcggcttcagcaaagagtctatcct 
               
               
                   
                   
               
               
                   
                 gcccaagaggaacagcgataagctgatcgccagaaagaag 
               
               
                   
                   
               
               
                   
                 gactgggaccctaagaagtacggcggcttcgacagcccca 
               
               
                   
                   
               
               
                   
                 ccgtggcctattctgtgctggtggtggccaaagtggaaaa 
               
               
                   
                   
               
               
                   
                 gggcaagtccaagaaactgaagagtgtgaaagagctgctg 
               
               
                   
                   
               
               
                   
                 gggatcaccatcatggaaagaagcagcttcgagaagaatc 
               
               
                   
                   
               
               
                   
                 ccatcgactttctggaagccaagggctacaaagaagtgaa 
               
               
                   
                   
               
               
                   
                 aaaggacctgatcatcaagctgcctaagtactccctgttc 
               
               
                   
                   
               
               
                   
                 gagctggaaaacggccggaagagaatgctggcctctgccg 
               
               
                   
                   
               
               
                   
                 gcgaactgcagaagggaaacgaactggccctgccctccaa 
               
               
                   
                   
               
               
                   
                 atatgtgaacttcctgtacctggccagccactatgagaag 
               
               
                   
                   
               
               
                   
                 ctgaagggctcccccgaggataatgagcagaaacagctgt 
               
               
                   
                   
               
               
                   
                 ttgtggaacagcacaagcactacctggacgagatcatcga 
               
               
                   
                   
               
               
                   
                 gcagatcagcgagttctccaagagagtgatcctggccgac 
               
               
                   
                   
               
               
                   
                 gctaatctggacaaagtgctgtccgcctacaacaagcacc 
               
               
                   
                   
               
               
                   
                 gggataagcccatcagagagcaggccgagaatatcatcca 
               
               
                   
                   
               
               
                   
                 cctgtttaccctgaccaatctgggagcccctgccgccttc 
               
               
                   
                   
               
               
                   
                 aagtactttgacaccaccatcgaccggaagaggtacacca 
               
               
                   
                   
               
               
                   
                 gcaccaaagaggtgctggacgccaccctgatccaccagag 
               
               
                   
                   
               
               
                   
                 catcaccggcctgtacgagacacggatcgacctgtctcag 
               
               
                   
                   
               
               
                   
                 ctgggaggcgactaactcgag 
               
               
                   
                   
               
               
                   
                 Seq ID NO 302: 
               
               
                   
                 &gt;6his-NLS-A3A-GGS7-dCas9 gene sequence 
               
               
                   
                 ATGggcagcagccatcatcatcatcatcacagcagcggcc 
               
               
                   
                   
               
               
                   
                 tggtgccgcgcggcagccatatgccaaagaagaagcggaa 
               
               
                   
                   
               
               
                   
                 ggtcGAAGCCAGCCCAGCATCCGGGCCCAGACACTTGATG 
               
               
                   
                   
               
               
                   
                 GATCCACACATATTCACTTCCAACTTTAACAATGGCATTG 
               
               
                   
                   
               
               
                   
                 GAAGGCATAAGACCTACCTGTGCTACGAAGTGGAGCGCCT 
               
               
                   
                   
               
               
                   
                 GGACAATGGCACCTCGGTCAAGATGGACCAGCACAGGGGC 
               
               
                   
                   
               
               
                   
                 TTTCTACACAACCAGGCTAAGAATCTTCTCTGTGGCTTTT 
               
               
                   
                   
               
               
                   
                 ACGGCCGCCATGCGCAGCTGCGCTTCTTGGACCTGGTTCC 
               
               
                   
                   
               
               
                   
                 TTCTTTGCAGTTGGACCCGGCCCAGATCTACAGGGTCACT 
               
               
                   
                   
               
               
                   
                 TGGTTCATCTCCTGGAGCCCCTGCTTCTCCTGGGGCTGTG 
               
               
                   
                   
               
               
                   
                 CCGGGGAAGTGCGTGCGTTCCTTCAGGAGAACACACACGT 
               
               
                   
                   
               
               
                   
                 GAGACTGCGTATCTTCGCTGCCCGCATCTATGATTACGAC 
               
               
                   
                   
               
               
                   
                 CCCCTATATAAGGAGGCACTGCAAATGCTGCGGGATGCTG 
               
               
                   
                   
               
               
                   
                 GGGCCCAAGTCTCCATCATGACCTACGATGAATTTAAGCA 
               
               
                   
                   
               
               
                   
                 CTGCTGGGACACCTTTGTGGACCACCAGGGATGTCCCTTC 
               
               
                   
                   
               
               
                   
                 CAGCCCTGGGATGGACTAGATGAGCACAGCCAAGCCCTGA 
               
               
                   
                   
               
               
                   
                 GTGGGAGGCTGCGGGCCATTCTCCAGAATCAGGGAAACGG 
               
               
                   
                   
               
               
                   
                 AGGAAGTGGAGGAAGTGGAGGAAGTGGAGGAAGTGGAGGA 
               
               
                   
                   
               
               
                   
                 AGTGGAGGAAGTGGAGGAAGTaagcttgacaagaagtaca 
               
               
                   
                   
               
               
                   
                 gcatcggcctggccatcggcaccaactctgtgggctgggc 
               
               
                   
                   
               
               
                   
                 cgtgatcaccgacgagtacaaggtgcccagcaagaaattc 
               
               
                   
                   
               
               
                   
                 aaggtgctgggcaacaccgaccggcacagcatcaagaaga 
               
               
                   
                   
               
               
                   
                 acctgatcggagccctgctgttcgacagcggcgaaacagc 
               
               
                   
                   
               
               
                   
                 cgaggccacccggctgaagagaaccgccagaagaagatac 
               
               
                   
                   
               
               
                   
                 accagacggaagaaccggatctgctatctgcaagagatct 
               
               
                   
                   
               
               
                   
                 tcagcaacgagatggccaaggtggacgacagcttcttcca 
               
               
                   
                   
               
               
                   
                 cagactggaagagtccttcctggtggaagaggataagaag 
               
               
                   
                   
               
               
                   
                 cacgagcggcaccccatcttcggcaacatcgtggacgagg 
               
               
                   
                   
               
               
                   
                 tggcctaccacgagaagtaccccaccatctaccacctgag 
               
               
                   
                   
               
               
                   
                 aaagaaactggtggacagcaccgacaaggccgacctgcgg 
               
               
                   
                   
               
               
                   
                 ctgatctatctggccctggcccacatgatcaagttccggg 
               
               
                   
                   
               
               
                   
                 gccacttcctgatcgagggcgacctgaaccccgacaacag 
               
               
                   
                   
               
               
                   
                 cgacgtggacaagctgttcatccagctggtgcagacctac 
               
               
                   
                   
               
               
                   
                 aaccagctgttcgaggaaaaccccatcaacgccagcggcg 
               
               
                   
                   
               
               
                   
                 tggacgccaaggccatcctgtctgccagactgagcaagag 
               
               
                   
                   
               
               
                   
                 cagacggctggaaaatctgatcgcccagctgcccggcgag 
               
               
                   
                   
               
               
                   
                 aagaagaatggcctgttcggaaacctgattgccctgagcc 
               
               
                   
                   
               
               
                   
                 tgggcctgacccccaacttcaagagcaacttcgacctggc 
               
               
                   
                   
               
               
                   
                 cgaggatgccaaactgcagctgagcaaggacacctacgac 
               
               
                   
                   
               
               
                   
                 gacgacctggacaacctgctggcccagatcggcgaccagt 
               
               
                   
                   
               
               
                   
                 acgccgacctgtttctggccgccaagaacctgtccgacgc 
               
               
                   
                   
               
               
                   
                 catcctgctgagcgacatcctgagagtgaacaccgagatc 
               
               
                   
                   
               
               
                   
                 accaaggcccccctgagcgcctctatgatcaagagatacg 
               
               
                   
                   
               
               
                   
                 acgagcaccaccaggacctgaccctgctgaaagctctcgt 
               
               
                   
                   
               
               
                   
                 gcggcagcagctgcctgagaagtacaaagagattttcttc 
               
               
                   
                   
               
               
                   
                 gaccagagcaagaacggctacgccggctacattgacggcg 
               
               
                   
                   
               
               
                   
                 gagccagccaggaagagttctacaagttcatcaagcccat 
               
               
                   
                   
               
               
                   
                 cctggaaaagatggacggcaccgaggaactgctcgtgaag 
               
               
                   
                   
               
               
                   
                 ctgaacagagaggacctgctgcggaagcagcggaccttcg 
               
               
                   
                   
               
               
                   
                 acaacggcagcatcccccaccagatccacctgggagagct 
               
               
                   
                   
               
               
                   
                 gcacgccattctgcggcggcaggaagatttttacccattc 
               
               
                   
                   
               
               
                   
                 ctgaaggacaaccgggaaaagatcgagaagatcctgacct 
               
               
                   
                   
               
               
                   
                 tccgcatcccctactacgtgggccctctggccaggggaaa 
               
               
                   
                   
               
               
                   
                 cagcagattcgcctggatgaccagaaagagcgaggaaacc 
               
               
                   
                   
               
               
                   
                 atcaccccctggaacttcgaggaagtggtggacaagggcg 
               
               
                   
                   
               
               
                   
                 cttccgcccagagcttcatcgagcggatgaccaacttcga 
               
               
                   
                   
               
               
                   
                 taagaacctgcccaacgagaaggtgctgcccaagcacagc 
               
               
                   
                   
               
               
                   
                 ctgctgtacgagtacttcaccgtgtataacgagctgacca 
               
               
                   
                   
               
               
                   
                 aagtgaaatacgtgaccgagggaatgagaaagcccgcctt 
               
               
                   
                   
               
               
                   
                 cctgagcggcgagcagaaaaaggccatcgtggacctgctg 
               
               
                   
                   
               
               
                   
                 ttcaagaccaaccggaaagtgaccgtgaagcagctgaaag 
               
               
                   
                   
               
               
                   
                 aggactacttcaagaaaatcgagtgcttcgactccgtgga 
               
               
                   
                   
               
               
                   
                 aatctccggcgtggaagatcggttcaacgcctccctgggc 
               
               
                   
                   
               
               
                   
                 acataccacgatctgctgaaaattatcaaggacaaggact 
               
               
                   
                   
               
               
                   
                 tcctggacaatgaggaaaacgaggacattctggaagatat 
               
               
                   
                   
               
               
                   
                 cgtgctgaccctgacactgtttgaggacagagagatgatc 
               
               
                   
                   
               
               
                   
                 gaggaacggctgaaaacctatgcccacctgttcgacgaca 
               
               
                   
                   
               
               
                   
                 aagtgatgaagcagctgaagcggcggagatacaccggctg 
               
               
                   
                   
               
               
                   
                 gggcaggctgagccggaagctgatcaacggcatccgggac 
               
               
                   
                   
               
               
                   
                 aagcagtccggcaagacaatcctggatttcctgaagtccg 
               
               
                   
                   
               
               
                   
                 acggcttcgccaacagaaacttcatgcagctgatccacga 
               
               
                   
                   
               
               
                   
                 cgacagcctgacctttaaagaggacatccagaaagcccag 
               
               
                   
                   
               
               
                   
                 gtgtccggccagggcgatagcctgcacgagcacattgcca 
               
               
                   
                   
               
               
                   
                 atctggccggcagccccgccattaagaagggcatcctgca 
               
               
                   
                   
               
               
                   
                 gacagtgaaggtggtggacgagctcgtgaaagtgatgggc 
               
               
                   
                   
               
               
                   
                 cggcacaagcccgagaacatcgtgatcgaaatggccagag 
               
               
                   
                   
               
               
                   
                 agaaccagaccacccagaagggacagaagaacagccgcga 
               
               
                   
                   
               
               
                   
                 gagaatgaagcggatcgaagagggcatcaaagagctgggc 
               
               
                   
                   
               
               
                   
                 agccagatcctgaaagaacaccccgtggaaaacacccagc 
               
               
                   
                   
               
               
                   
                 tgcagaacgagaagctgtacctgtactacctgcagaatgg 
               
               
                   
                   
               
               
                   
                 gcgggatatgtacgtggaccaggaactggacatcaaccgg 
               
               
                   
                   
               
               
                   
                 ctgtccgactacgatgtggacgctatcgtgcctcagagct 
               
               
                   
                   
               
               
                   
                 ttctgaaggacgactccatcgacaacaaggtgctgaccag 
               
               
                   
                   
               
               
                   
                 aagcgacaagaaccggggcaagagcgacaacgtgccctcc 
               
               
                   
                   
               
               
                   
                 gaagaggtcgtgaagaagatgaagaactactggcggcagc 
               
               
                   
                   
               
               
                   
                 tgctgaacgccaagctgattacccagagaaagttcgacaa 
               
               
                   
                   
               
               
                   
                 tctgaccaaggccgagagaggcggcctgagcgaactggat 
               
               
                   
                   
               
               
                   
                 aaggccggcttcatcaagagacagctggtggaaacccggc 
               
               
                   
                   
               
               
                   
                 agatcacaaagcacgtggcacagatcctggactcccggat 
               
               
                   
                   
               
               
                   
                 gaacactaagtacgacgagaatgacaagctgatccgggaa 
               
               
                   
                   
               
               
                   
                 gtgaaagtgatcaccctgaagtccaagctggtgtccgatt 
               
               
                   
                   
               
               
                   
                 tccggaaggatttccagttttacaaagtgcgcgagatcaa 
               
               
                   
                   
               
               
                   
                 caactaccaccacgcccacgacgcctacctgaacgccgtc 
               
               
                   
                   
               
               
                   
                 gtgggaaccgccctgatcaaaaagtaccctaagctggaaa 
               
               
                   
                   
               
               
                   
                 gcgagttcgtgtacggcgactacaaggtgtacgacgtgcg 
               
               
                   
                   
               
               
                   
                 gaagatgatcgccaagagcgagcaggaaatcggcaaggct 
               
               
                   
                   
               
               
                   
                 accgccaagtacttcttctacagcaacatcatgaactttt 
               
               
                   
                   
               
               
                   
                 tcaagaccgagattaccctggccaacggcgagatccggaa 
               
               
                   
                   
               
               
                   
                 gcggcctctgatcgagacaaacggcgaaaccggggagatc 
               
               
                   
                   
               
               
                   
                 gtgtgggataagggccgggattttgccaccgtgcggaaag 
               
               
                   
                   
               
               
                   
                 tgctgagcatgccccaagtgaatatcgtgaaaaagaccga 
               
               
                   
                   
               
               
                   
                 ggtgcagacaggcggcttcagcaaagagtctatcctgccc 
               
               
                   
                   
               
               
                   
                 aagaggaacagcgataagctgatcgccagaaagaaggact 
               
               
                   
                   
               
               
                   
                 gggaccctaagaagtacggcggcttcgacagccccaccgt 
               
               
                   
                   
               
               
                   
                 ggcctattctgtgctggtggtggccaaagtggaaaagggc 
               
               
                   
                   
               
               
                   
                 aagtccaagaaactgaagagtgtgaaagagctgctgggga 
               
               
                   
                   
               
               
                   
                 tcaccatcatggaaagaagcagcttcgagaagaatcccat 
               
               
                   
                   
               
               
                   
                 cgactttctggaagccaagggctacaaagaagtgaaaaag 
               
               
                   
                   
               
               
                   
                 gacctgatcatcaagctgcctaagtactccctgttcgagc 
               
               
                   
                   
               
               
                   
                 tggaaaacggccggaagagaatgctggcctctgccggcga 
               
               
                   
                   
               
               
                   
                 actgcagaagggaaacgaactggccctgccctccaaatat 
               
               
                   
                   
               
               
                   
                 gtgaacttcctgtacctggccagccactatgagaagctga 
               
               
                   
                   
               
               
                   
                 agggctcccccgaggataatgagcagaaacagctgtttgt 
               
               
                   
                   
               
               
                   
                 ggaacagcacaagcactacctggacgagatcatcgagcag 
               
               
                   
                   
               
               
                   
                 atcagcgagttctccaagagagtgatcctggccgacgcta 
               
               
                   
                   
               
               
                   
                 atctggacaaagtgctgtccgcctacaacaagcaccggga 
               
               
                   
                   
               
               
                   
                 taagcccatcagagagcaggccgagaatatcatccacctg 
               
               
                   
                   
               
               
                   
                 tttaccctgaccaatctgggagcccctgccgccttcaagt 
               
               
                   
                   
               
               
                   
                 actttgacaccaccatcgaccggaagaggtacaccagcac 
               
               
                   
                   
               
               
                   
                 caaagaggtgctggacgccaccctgatccaccagagcatc 
               
               
                   
                   
               
               
                   
                 accggcctgtacgagacacggatcgacctgtctcagctgg 
               
               
                   
                   
               
               
                   
                 gaggcgactaactcgag 
               
               
                   
                   
               
               
                   
                 Seq ID NO 303: 
               
               
                   
                 &gt;6his-NLS-A3A-GGS14-dCas9 gene sequence 
               
               
                   
                 ATGggcagcagccatcatcatcatcatcacagcagcggcc 
               
               
                   
                   
               
               
                   
                 tggtgccgcgcggcagccatatgccaaagaagaagcggaa 
               
               
                   
                   
               
               
                   
                 ggtcGAAGCCAGCCCAGCATCCGGGCCCAGACACTTGATG 
               
               
                   
                   
               
               
                   
                 GATCCACACATATTCACTTCCAACTTTAACAATGGCATTG 
               
               
                   
                   
               
               
                   
                 GAAGGCATAAGACCTACCTGTGCTACGAAGTGGAGCGCCT 
               
               
                   
                   
               
               
                   
                 GGACAATGGCACCTCGGTCAAGATGGACCAGCACAGGGGC 
               
               
                   
                   
               
               
                   
                 TTTCTACACAACCAGGCTAAGAATCTTCTCTGTGGCTTTT 
               
               
                   
                   
               
               
                   
                 ACGGCCGCCATGCGCAGCTGCGCTTCTTGGACCTGGTTCC 
               
               
                   
                   
               
               
                   
                 TTCTTTGCAGTTGGACCCGGCCCAGATCTACAGGGTCACT 
               
               
                   
                   
               
               
                   
                 TGGTTCATCTCCTGGAGCCCCTGCTTCTCCTGGGGCTGTG 
               
               
                   
                   
               
               
                   
                 CCGGGGAAGTGCGTGCGTTCCTTCAGGAGAACACACACGT 
               
               
                   
                   
               
               
                   
                 GAGACTGCGTATCTTCGCTGCCCGCATCTATGATTACGAC 
               
               
                   
                   
               
               
                   
                 CCCCTATATAAGGAGGCACTGCAAATGCTGCGGGATGCTG 
               
               
                   
                   
               
               
                   
                 GGGCCCAAGTCTCCATCATGACCTACGATGAATTTAAGCA 
               
               
                   
                   
               
               
                   
                 CTGCTGGGACACCTTTGTGGACCACCAGGGATGTCCCTTC 
               
               
                   
                   
               
               
                   
                 CAGCCCTGGGATGGACTAGATGAGCACAGCCAAGCCCTGA 
               
               
                   
                   
               
               
                   
                 GTGGGAGGCTGCGGGCCATTCTCCAGAATCAGGGAAACGG 
               
               
                   
                   
               
               
                   
                 AGGAAGTGGAGGAAGTGGAGGAAGTGGAGGAAGTGGAGGA 
               
               
                   
                   
               
               
                   
                 AGTGGAGGAAGTGGAGGAAGTGGAGGAAGTGGAGGAAGTG 
               
               
                   
                   
               
               
                   
                 GAGGAAGTGGAGGAAGTGGAGGAAGTGGAGGAAGTGGAGG 
               
               
                   
                   
               
               
                   
                 AAGTaagcttgacaagaagtacagcatcggcctggccatc 
               
               
                   
                   
               
               
                   
                 ggcaccaactctgtgggctgggccgtgatcaccgacgagt 
               
               
                   
                   
               
               
                   
                 acaaggtgcccagcaagaaattcaaggtgctgggcaacac 
               
               
                   
                   
               
               
                   
                 cgaccggcacagcatcaagaagaacctgatcggagccctg 
               
               
                   
                   
               
               
                   
                 ctgttcgacagcggcgaaacagccgaggccacccggctga 
               
               
                   
                   
               
               
                   
                 agagaaccgccagaagaagatacaccagacggaagaaccg 
               
               
                   
                   
               
               
                   
                 gatctgctatctgcaagagatcttcagcaacgagatggcc 
               
               
                   
                   
               
               
                   
                 aaggtggacgacagcttcttccacagactggaagagtcct 
               
               
                   
                   
               
               
                   
                 tcctggtggaagaggataagaagcacgagcggcaccccat 
               
               
                   
                   
               
               
                   
                 cttcggcaacatcgtggacgaggtggcctaccacgagaag 
               
               
                   
                   
               
               
                   
                 taccccaccatctaccacctgagaaagaaactggtggaca 
               
               
                   
                   
               
               
                   
                 gcaccgacaaggccgacctgcggctgatctatctggccct 
               
               
                   
                   
               
               
                   
                 ggcccacatgatcaagttccggggccacttcctgatcgag 
               
               
                   
                   
               
               
                   
                 ggcgacctgaaccccgacaacagcgacgtggacaagctgt 
               
               
                   
                   
               
               
                   
                 tcatccagctggtgcagacctacaaccagctgttcgagga 
               
               
                   
                   
               
               
                   
                 aaaccccatcaacgccagcggcgtggacgccaaggccatc 
               
               
                   
                   
               
               
                   
                 ctgtctgccagactgagcaagagcagacggctggaaaatc 
               
               
                   
                   
               
               
                   
                 tgatcgcccagctgcccggcgagaagaagaatggcctgtt 
               
               
                   
                   
               
               
                   
                 cggaaacctgattgccctgagcctgggcctgacccccaac 
               
               
                   
                   
               
               
                   
                 ttcaagagcaacttcgacctggccgaggatgccaaactgc 
               
               
                   
                   
               
               
                   
                 agctgagcaaggacacctacgacgacgacctggacaacct 
               
               
                   
                   
               
               
                   
                 gctggcccagatcggcgaccagtacgccgacctgtttctg 
               
               
                   
                   
               
               
                   
                 gccgccaagaacctgtccgacgccatcctgctgagcgaca 
               
               
                   
                   
               
               
                   
                 tcctgagagtgaacaccgagatcaccaaggcccccctgag 
               
               
                   
                   
               
               
                   
                 cgcctctatgatcaagagatacgacgagcaccaccaggac 
               
               
                   
                   
               
               
                   
                 ctgaccctgctgaaagctctcgtgcggcagcagctgcctg 
               
               
                   
                   
               
               
                   
                 agaagtacaaagagattttcttcgaccagagcaagaacgg 
               
               
                   
                   
               
               
                   
                 ctacgccggctacattgacggcggagccagccaggaagag 
               
               
                   
                   
               
               
                   
                 ttctacaagttcatcaagcccatcctggaaaagatggacg 
               
               
                   
                   
               
               
                   
                 gcaccgaggaactgctcgtgaagctgaacagagaggacct 
               
               
                   
                   
               
               
                   
                 gctgcggaagcagcggaccttcgacaacggcagcatcccc 
               
               
                   
                   
               
               
                   
                 caccagatccacctgggagagctgcacgccattctgcggc 
               
               
                   
                   
               
               
                   
                 ggcaggaagatttttacccattcctgaaggacaaccggga 
               
               
                   
                   
               
               
                   
                 aaagatcgagaagatcctgaccttccgcatcccctactac 
               
               
                   
                   
               
               
                   
                 gtgggccctctggccaggggaaacagcagattcgcctgga 
               
               
                   
                   
               
               
                   
                 tgaccagaaagagcgaggaaaccatcaccccctggaactt 
               
               
                   
                   
               
               
                   
                 cgaggaagtggtggacaagggcgcttccgcccagagcttc 
               
               
                   
                   
               
               
                   
                 atcgagcggatgaccaacttcgataagaacctgcccaacg 
               
               
                   
                   
               
               
                   
                 agaaggtgctgcccaagcacagcctgctgtacgagtactt 
               
               
                   
                   
               
               
                   
                 caccgtgtataacgagctgaccaaagtgaaatacgtgacc 
               
               
                   
                   
               
               
                   
                 gagggaatgagaaagcccgccttcctgagcggcgagcaga 
               
               
                   
                   
               
               
                   
                 aaaaggccatcgtggacctgctgttcaagaccaaccggaa 
               
               
                   
                   
               
               
                   
                 agtgaccgtgaagcagctgaaagaggactacttcaagaaa 
               
               
                   
                   
               
               
                   
                 atcgagtgcttcgactccgtggaaatctccggcgtggaag 
               
               
                   
                   
               
               
                   
                 atcggttcaacgcctccctgggcacataccacgatctgct 
               
               
                   
                   
               
               
                   
                 gaaaattatcaaggacaaggacttcctggacaatgaggaa 
               
               
                   
                   
               
               
                   
                 aacgaggacattctggaagatatcgtgctgaccctgacac 
               
               
                   
                   
               
               
                   
                 tgtttgaggacagagagatgatcgaggaacggctgaaaac 
               
               
                   
                   
               
               
                   
                 ctatgcccacctgttcgacgacaaagtgatgaagcagctg 
               
               
                   
                   
               
               
                   
                 aagcggcggagatacaccggctggggcaggctgagccgga 
               
               
                   
                   
               
               
                   
                 agctgatcaacggcatccgggacaagcagtccggcaagac 
               
               
                   
                   
               
               
                   
                 aatcctggatttcctgaagtccgacggcttcgccaacaga 
               
               
                   
                   
               
               
                   
                 aacttcatgcagctgatccacgacgacagcctgaccttta 
               
               
                   
                   
               
               
                   
                 aagaggacatccagaaagcccaggtgtccggccagggcga 
               
               
                   
                   
               
               
                   
                 tagcctgcacgagcacattgccaatctggccggcagcccc 
               
               
                   
                   
               
               
                   
                 gccattaagaagggcatcctgcagacagtgaaggtggtgg 
               
               
                   
                   
               
               
                   
                 acgagctcgtgaaagtgatgggccggcacaagcccgagaa 
               
               
                   
                   
               
               
                   
                 catcgtgatcgaaatggccagagagaaccagaccacccag 
               
               
                   
                   
               
               
                   
                 aagggacagaagaacagccgcgagagaatgaagcggatcg 
               
               
                   
                   
               
               
                   
                 aagagggcatcaaagagctgggcagccagatcctgaaaga 
               
               
                   
                   
               
               
                   
                 acaccccgtggaaaacacccagctgcagaacgagaagctg 
               
               
                   
                   
               
               
                   
                 tacctgtactacctgcagaatgggcgggatatgtacgtgg 
               
               
                   
                   
               
               
                   
                 accaggaactggacatcaaccggctgtccgactacgatgt 
               
               
                   
                   
               
               
                   
                 ggacgctatcgtgcctcagagctttctgaaggacgactcc 
               
               
                   
                   
               
               
                   
                 atcgacaacaaggtgctgaccagaagcgacaagaaccggg 
               
               
                   
                   
               
               
                   
                 gcaagagcgacaacgtgccctccgaagaggtcgtgaagaa 
               
               
                   
                   
               
               
                   
                 gatgaagaactactggcggcagctgctgaacgccaagctg 
               
               
                   
                   
               
               
                   
                 attacccagagaaagttcgacaatctgaccaaggccgaga 
               
               
                   
                   
               
               
                   
                 gaggcggcctgagcgaactggataaggccggcttcatcaa 
               
               
                   
                   
               
               
                   
                 gagacagctggtggaaacccggcagatcacaaagcacgtg 
               
               
                   
                   
               
               
                   
                 gcacagatcctggactcccggatgaacactaagtacgacg 
               
               
                   
                   
               
               
                   
                 agaatgacaagctgatccgggaagtgaaagtgatcaccct 
               
               
                   
                   
               
               
                   
                 gaagtccaagctggtgtccgatttccggaaggatttccag 
               
               
                   
                   
               
               
                   
                 ttttacaaagtgcgcgagatcaacaactaccaccacgccc 
               
               
                   
                   
               
               
                   
                 acgacgcctacctgaacgccgtcgtgggaaccgccctgat 
               
               
                   
                   
               
               
                   
                 caaaaagtaccctaagctggaaagcgagttcgtgtacggc 
               
               
                   
                   
               
               
                   
                 gactacaaggtgtacgacgtgcggaagatgatcgccaaga 
               
               
                   
                   
               
               
                   
                 gcgagcaggaaatcggcaaggctaccgccaagtacttctt 
               
               
                   
                   
               
               
                   
                 ctacagcaacatcatgaactttttcaagaccgagattacc 
               
               
                   
                   
               
               
                   
                 ctggccaacggcgagatccggaagcggcctctgatcgaga 
               
               
                   
                   
               
               
                   
                 caaacggcgaaaccggggagatcgtgtgggataagggccg 
               
               
                   
                   
               
               
                   
                 ggattttgccaccgtgcggaaagtgctgagcatgccccaa 
               
               
                   
                   
               
               
                   
                 gtgaatatcgtgaaaaagaccgaggtgcagacaggcggct 
               
               
                   
                   
               
               
                   
                 tcagcaaagagtctatcctgcccaagaggaacagcgataa 
               
               
                   
                   
               
               
                   
                 gctgatcgccagaaagaaggactgggaccctaagaagtac 
               
               
                   
                   
               
               
                   
                 ggcggcttcgacagccccaccgtggcctattctgtgctgg 
               
               
                   
                   
               
               
                   
                 tggtggccaaagtggaaaagggcaagtccaagaaactgaa 
               
               
                   
                   
               
               
                   
                 gagtgtgaaagagctgctggggatcaccatcatggaaaga 
               
               
                   
                   
               
               
                   
                 agcagcttcgagaagaatcccatcgactttctggaagcca 
               
               
                   
                   
               
               
                   
                 agggctacaaagaagtgaaaaaggacctgatcatcaagct 
               
               
                   
                   
               
               
                   
                 gcctaagtactccctgttcgagctggaaaacggccggaag 
               
               
                   
                   
               
               
                   
                 agaatgctggcctctgccggcgaactgcagaagggaaacg 
               
               
                   
                   
               
               
                   
                 aactggccctgccctccaaatatgtgaacttcctgtacct 
               
               
                   
                   
               
               
                   
                 ggccagccactatgagaagctgaagggctcccccgaggat 
               
               
                   
                   
               
               
                   
                 aatgagcagaaacagctgtttgtggaacagcacaagcact 
               
               
                   
                   
               
               
                   
                 acctggacgagatcatcgagcagatcagcgagttctccaa 
               
               
                   
                   
               
               
                   
                 gagagtgatcctggccgacgctaatctggacaaagtgctg 
               
               
                   
                   
               
               
                   
                 tccgcctacaacaagcaccgggataagcccatcagagagc 
               
               
                   
                   
               
               
                   
                 aggccgagaatatcatccacctgtttaccctgaccaatct 
               
               
                   
                   
               
               
                   
                 gggagcccctgccgccttcaagtactttgacaccaccatc 
               
               
                   
                   
               
               
                   
                 gaccggaagaggtacaccagcaccaaagaggtgctggacg 
               
               
                   
                   
               
               
                   
                 ccaccctgatccaccagagcatcaccggcctgtacgagac 
               
               
                   
                   
               
               
                   
                 acggatcgacctgtctcagctgggaggcgactaactcgag 
               
               
                   
                   
               
               
                   
                   
               
               
                   
                 Seq ID NO 304: 
               
               
                   
                 &gt;6his-NLS-A3H-GGS3-dCas9 gene sequence 
               
               
                   
                 ATGggcagcagccatcatcatcatcatcacagcagcggcc 
               
               
                   
                   
               
               
                   
                 tggtgccgcgcggcagccatatgccaaagaagaagcggaa 
               
               
                   
                   
               
               
                   
                 ggtcGCTCTTCTTACTGCTGAAACTTTTCGTCTCCAATTT 
               
               
                   
                   
               
               
                   
                 AATAATAAACGCCGTCTGCGTCGCCCGTATTACCCGCGCA 
               
               
                   
                   
               
               
                   
                 AGGCGCTGCTGTGTTACCAACTGACCCCACAAAACGGTTC 
               
               
                   
                   
               
               
                   
                 CACCCCGACTCGCGGTTACTTTGAGAATAAGAAAAAATGT 
               
               
                   
                   
               
               
                   
                 CACGCTGAGATCTGTTTCATTAACGAAATCAAATCTATGG 
               
               
                   
                   
               
               
                   
                 GCCTGGATGAAACTCAGTGCTACCAGGTCACCTGCTACCT 
               
               
                   
                   
               
               
                   
                 GACCTGGAGCCCGTGTAGCTCTTGCGCGTGGGAACTGGTT 
               
               
                   
                   
               
               
                   
                 GACTTCATCAAAGCGCACGACCATCTGAACCTGCGTATCT 
               
               
                   
                   
               
               
                   
                 TCGCTTCCCGCCTGTACTATCACTGGTGCAAGCCGCAACA 
               
               
                   
                   
               
               
                   
                 GGATGGCCTGCGCCTGCTGTGTGGTTCTCAGGTTCCGGTT 
               
               
                   
                   
               
               
                   
                 GAAGTTATGGGTTTCCCGGAGTTTGCGGACTGCTGGGAAA 
               
               
                   
                   
               
               
                   
                 ACTTTGTTGACCATGAGAAGCCACTGTCCTTTAACCCGTA 
               
               
                   
                   
               
               
                   
                 TAAAATGCTGGAAGAGCTGGACAAAAACTCTCGTGCTATC 
               
               
                   
                   
               
               
                   
                 AAGCGCCGTCTGGATCGTATCAAGTCTGGAGGAAGTGGAG 
               
               
                   
                   
               
               
                   
                 GAAGTGGAGGAAGTagcttgacaagaagtacagcatcggc 
               
               
                   
                   
               
               
                   
                 ctggccatcggcaccaactctgtgggctgggccgtgatca 
               
               
                   
                   
               
               
                   
                 ccgacgagtacaaggtgcccagcaagaaattcaaggtgct 
               
               
                   
                   
               
               
                   
                 gggcaacaccgaccggcacagcatcaagaagaacctgatc 
               
               
                   
                   
               
               
                   
                 ggagccctgctgttcgacagcggcgaaacagccgaggcca 
               
               
                   
                   
               
               
                   
                 cccggctgaagagaaccgccagaagaagatacaccagacg 
               
               
                   
                   
               
               
                   
                 gaagaaccggatctgctatctgcaagagatcttcagcaac 
               
               
                   
                   
               
               
                   
                 gagatggccaaggtggacgacagcttcttccacagactgg 
               
               
                   
                   
               
               
                   
                 aagagtccttcctggtggaagaggataagaagcacgagcg 
               
               
                   
                   
               
               
                   
                 gcaccccatcttcggcaacatcgtggacgaggtggcctac 
               
               
                   
                   
               
               
                   
                 cacgagaagtaccccaccatctaccacctgagaaagaaac 
               
               
                   
                   
               
               
                   
                 tggtggacagcaccgacaaggccgacctgcggctgatcta 
               
               
                   
                   
               
               
                   
                 tctggccctggcccacatgatcaagttccggggccacttc 
               
               
                   
                   
               
               
                   
                 ctgatcgagggcgacctgaaccccgacaacagcgacgtgg 
               
               
                   
                   
               
               
                   
                 acaagctgttcatccagctggtgcagacctacaaccagct 
               
               
                   
                   
               
               
                   
                 gttcgaggaaaaccccatcaacgccagcggcgtggacgcc 
               
               
                   
                   
               
               
                   
                 aaggccatcctgtctgccagactgagcaagagcagacggc 
               
               
                   
                   
               
               
                   
                 tggaaaatctgatcgcccagctgcccggcgagaagaagaa 
               
               
                   
                   
               
               
                   
                 tggcctgttcggaaacctgattgccctgagcctgggcctg 
               
               
                   
                   
               
               
                   
                 acccccaacttcaagagcaacttcgacctggccgaggatg 
               
               
                   
                   
               
               
                   
                 ccaaactgcagctgagcaaggacacctacgacgacgacct 
               
               
                   
                   
               
               
                   
                 ggacaacctgctggcccagatcggcgaccagtacgccgac 
               
               
                   
                   
               
               
                   
                 ctgtttctggccgccaagaacctgtccgacgccatcctgc 
               
               
                   
                   
               
               
                   
                 tgagcgacatcctgagagtgaacaccgagatcaccaaggc 
               
               
                   
                   
               
               
                   
                 ccccctgagcgcctctatgatcaagagatacgacgagcac 
               
               
                   
                   
               
               
                   
                 caccaggacctgaccctgctgaaagctctcgtgcggcagc 
               
               
                   
                   
               
               
                   
                 agctgcctgagaagtacaaagagattttcttcgaccagag 
               
               
                   
                   
               
               
                   
                 caagaacggctacgccggctacattgacggcggagccagc 
               
               
                   
                   
               
               
                   
                 caggaagagttctacaagttcatcaagcccatcctggaaa 
               
               
                   
                   
               
               
                   
                 agatggacggcaccgaggaactgctcgtgaagctgaacag 
               
               
                   
                   
               
               
                   
                 agaggacctgctgcggaagcagcggaccttcgacaacggc 
               
               
                   
                   
               
               
                   
                 agcatcccccaccagatccacctgggagagctgcacgcca 
               
               
                   
                   
               
               
                   
                 ttctgcggcggcaggaagatttttacccattcctgaagga 
               
               
                   
                   
               
               
                   
                 caaccgggaaaagatcgagaagatcctgaccttccgcatc 
               
               
                   
                   
               
               
                   
                 ccctactacgtgggccctctggccaggggaaacagcagat 
               
               
                   
                   
               
               
                   
                 tcgcctggatgaccagaaagagcgaggaaaccatcacccc 
               
               
                   
                   
               
               
                   
                 ctggaacttcgaggaagtggtggacaagggcgcttccgcc 
               
               
                   
                   
               
               
                   
                 cagagcttcatcgagcggatgaccaacttcgataagaacc 
               
               
                   
                   
               
               
                   
                 tgcccaacgagaaggtgctgcccaagcacagcctgctgta 
               
               
                   
                   
               
               
                   
                 cgagtacttcaccgtgtataacgagctgaccaaagtgaaa 
               
               
                   
                   
               
               
                   
                 tacgtgaccgagggaatgagaaagcccgccttcctgagcg 
               
               
                   
                   
               
               
                   
                 gcgagcagaaaaaggccatcgtggacctgctgttcaagac 
               
               
                   
                   
               
               
                   
                 caaccggaaagtgaccgtgaagcagctgaaagaggactac 
               
               
                   
                   
               
               
                   
                 ttcaagaaaatcgagtgcttcgactccgtggaaatctccg 
               
               
                   
                   
               
               
                   
                 gcgtggaagatcggttcaacgcctccctgggcacatacca 
               
               
                   
                   
               
               
                   
                 cgatctgctgaaaattatcaaggacaaggacttcctggac 
               
               
                   
                   
               
               
                   
                 aatgaggaaaacgaggacattctggaagatatcgtgctga 
               
               
                   
                   
               
               
                   
                 ccctgacactgtttgaggacagagagatgatcgaggaacg 
               
               
                   
                   
               
               
                   
                 gctgaaaacctatgcccacctgttcgacgacaaagtgatg 
               
               
                   
                   
               
               
                   
                 aagcagctgaagcggcggagatacaccggctggggcaggc 
               
               
                   
                   
               
               
                   
                 tgagccggaagctgatcaacggcatccgggacaagcagtc 
               
               
                   
                   
               
               
                   
                 cggcaagacaatcctggatttcctgaagtccgacggcttc 
               
               
                   
                   
               
               
                   
                 gccaacagaaacttcatgcagctgatccacgacgacagcc 
               
               
                   
                   
               
               
                   
                 tgacctttaaagaggacatccagaaagcccaggtgtccgg 
               
               
                   
                   
               
               
                   
                 ccagggcgatagcctgcacgagcacattgccaatctggcc 
               
               
                   
                   
               
               
                   
                 ggcagccccgccattaagaagggcatcctgcagacagtga 
               
               
                   
                   
               
               
                   
                 aggtggtggacgagctcgtgaaagtgatgggccggcacaa 
               
               
                   
                   
               
               
                   
                 gcccgagaacatcgtgatcgaaatggccagagagaaccag 
               
               
                   
                   
               
               
                   
                 accacccagaagggacagaagaacagccgcgagagaatga 
               
               
                   
                   
               
               
                   
                 agcggatcgaagagggcatcaaagagctgggcagccagat 
               
               
                   
                   
               
               
                   
                 cctgaaagaacaccccgtggaaaacacccagctgcagaac 
               
               
                   
                   
               
               
                   
                 gagaagctgtacctgtactacctgcagaatgggcgggata 
               
               
                   
                   
               
               
                   
                 tgtacgtggaccaggaactggacatcaaccggctgtccga 
               
               
                   
                   
               
               
                   
                 ctacgatgtggacgctatcgtgcctcagagctttctgaag 
               
               
                   
                   
               
               
                   
                 gacgactccatcgacaacaaggtgctgaccagaagcgaca 
               
               
                   
                   
               
               
                   
                 agaaccggggcaagagcgacaacgtgccctccgaagaggt 
               
               
                   
                   
               
               
                   
                 cgtgaagaagatgaagaactactggcggcagctgctgaac 
               
               
                   
                   
               
               
                   
                 gccaagctgattacccagagaaagttcgacaatctgacca 
               
               
                   
                   
               
               
                   
                 aggccgagagaggcggcctgagcgaactggataaggccgg 
               
               
                   
                   
               
               
                   
                 cttcatcaagagacagctggtggaaacccggcagatcaca 
               
               
                   
                   
               
               
                   
                 aagcacgtggcacagatcctggactcccggatgaacacta 
               
               
                   
                   
               
               
                   
                 agtacgacgagaatgacaagctgatccgggaagtgaaagt 
               
               
                   
                   
               
               
                   
                 gatcaccctgaagtccaagctggtgtccgatttccggaag 
               
               
                   
                   
               
               
                   
                 gatttccagttttacaaagtgcgcgagatcaacaactacc 
               
               
                   
                   
               
               
                   
                 accacgcccacgacgcctacctgaacgccgtcgtgggaac 
               
               
                   
                   
               
               
                   
                 cgccctgatcaaaaagtaccctaagctggaaagcgagttc 
               
               
                   
                   
               
               
                   
                 gtgtacggcgactacaaggtgtacgacgtgcggaagatga 
               
               
                   
                   
               
               
                   
                 tcgccaagagcgagcaggaaatcggcaaggctaccgccaa 
               
               
                   
                   
               
               
                   
                 gtacttcttctacagcaacatcatgaactttttcaagacc 
               
               
                   
                   
               
               
                   
                 gagattaccctggccaacggcgagatccggaagcggcctc 
               
               
                   
                   
               
               
                   
                 tgatcgagacaaacggcgaaaccggggagatcgtgtggga 
               
               
                   
                   
               
               
                   
                 taagggccgggattttgccaccgtgcggaaagtgctgagc 
               
               
                   
                   
               
               
                   
                 atgccccaagtgaatatcgtgaaaaagaccgaggtgcaga 
               
               
                   
                   
               
               
                   
                 caggcggcttcagcaaagagtctatcctgcccaagaggaa 
               
               
                   
                   
               
               
                   
                 cagcgataagctgatcgccagaaagaaggactgggaccct 
               
               
                   
                   
               
               
                   
                 aagaagtacggcggcttcgacagccccaccgtggcctatt 
               
               
                   
                   
               
               
                   
                 ctgtgctggtggtggccaaagtggaaaagggcaagtccaa 
               
               
                   
                   
               
               
                   
                 gaaactgaagagtgtgaaagagctgctggggatcaccatc 
               
               
                   
                   
               
               
                   
                 atggaaagaagcagcttcgagaagaatcccatcgactttc 
               
               
                   
                   
               
               
                   
                 tggaagccaagggctacaaagaagtgaaaaaggacctgat 
               
               
                   
                   
               
               
                   
                 catcaagctgcctaagtactccctgttcgagctggaaaac 
               
               
                   
                   
               
               
                   
                 ggccggaagagaatgctggcctctgccggcgaactgcaga 
               
               
                   
                   
               
               
                   
                 agggaaacgaactggccctgccctccaaatatgtgaactt 
               
               
                   
                   
               
               
                   
                 cctgtacctggccagccactatgagaagctgaagggctcc 
               
               
                   
                   
               
               
                   
                 cccgaggataatgagcagaaacagctgtttgtggaacagc 
               
               
                   
                   
               
               
                   
                 acaagcactacctggacgagatcatcgagcagatcagcga 
               
               
                   
                   
               
               
                   
                 gttctccaagagagtgatcctggccgacgctaatctggac 
               
               
                   
                   
               
               
                   
                 aaagtgctgtccgcctacaacaagcaccgggataagccca 
               
               
                   
                   
               
               
                   
                 tcagagagcaggccgagaatatcatccacctgtttaccct 
               
               
                   
                   
               
               
                   
                 gaccaatctgggagcccctgccgccttcaagtactttgac 
               
               
                   
                   
               
               
                   
                 accaccatcgaccggaagaggtacaccagcaccaaagagg 
               
               
                   
                   
               
               
                   
                 tgctggacgccaccctgatccaccagagcatcaccggcct 
               
               
                   
                   
               
               
                   
                 gtacgagacacggatcgacctgtctcagctgggaggcgac 
               
               
                   
                   
               
               
                   
                 taactcgag 
               
               
                   
                   
               
               
                   
                 Seq ID NO 305: 
               
               
                   
                 &gt;6his-NLS-A3H-GGS7-dCas9 gene sequence 
               
               
                   
                 ATGggcagcagccatcatcatcatcatcacagcagcggcc 
               
               
                   
                   
               
               
                   
                 tggtgccgcgcggcagccatatgccaaagaagaagcggaa 
               
               
                   
                   
               
               
                   
                 ggtcGCTCTTCTTACTGCTGAAACTTTTCGTCTCCAATTT 
               
               
                   
                   
               
               
                   
                 AATAATAAACGCCGTCTGCGTCGCCCGTATTACCCGCGCA 
               
               
                   
                   
               
               
                   
                 AGGCGCTGCTGTGTTACCAACTGACCCCACAAAACGGTTC 
               
               
                   
                   
               
               
                   
                 CACCCCGACTCGCGGTTACTTTGAGAATAAGAAAAAATGT 
               
               
                   
                   
               
               
                   
                 CACGCTGAGATCTGTTTCATTAACGAAATCAAATCTATGG 
               
               
                   
                   
               
               
                   
                 GCCTGGATGAAACTCAGTGCTACCAGGTCACCTGCTACCT 
               
               
                   
                   
               
               
                   
                 GACCTGGAGCCCGTGTAGCTCTTGCGCGTGGGAACTGGTT 
               
               
                   
                   
               
               
                   
                 GACTTCATCAAAGCGCACGACCATCTGAACCTGCGTATCT 
               
               
                   
                   
               
               
                   
                 TCGCTTCCCGCCTGTACTATCACTGGTGCAAGCCGCAACA 
               
               
                   
                   
               
               
                   
                 GGATGGCCTGCGCCTGCTGTGTGGTTCTCAGGTTCCGGTT 
               
               
                   
                   
               
               
                   
                 GAAGTTATGGGTTTCCCGGAGTTTGCGGACTGCTGGGAAA 
               
               
                   
                   
               
               
                   
                 ACTTTGTTGACCATGAGAAGCCACTGTCCTTTAACCCGTA 
               
               
                   
                   
               
               
                   
                 TAAAATGCTGGAAGAGCTGGACAAAAACTCTCGTGCTATC 
               
               
                   
                   
               
               
                   
                 AAGCGCCGTCTGGATCGTATCAAGTCTGGAGGAAGTGGAG 
               
               
                   
                   
               
               
                   
                 GAAGTGGAGGAAGTGGAGGAAGTGGAGGAAGTGGAGGAAG 
               
               
                   
                   
               
               
                   
                 TGGAGGAAGTaagcttgacaagaagtacagcatcggcctg 
               
               
                   
                   
               
               
                   
                 gccatcggcaccaactctgtgggctgggccgtgatcaccg 
               
               
                   
                   
               
               
                   
                 acgagtacaaggtgcccagcaagaaattcaaggtgctggg 
               
               
                   
                   
               
               
                   
                 caacaccgaccggcacagcatcaagaagaacctgatcgga 
               
               
                   
                   
               
               
                   
                 gccctgctgttcgacagcggcgaaacagccgaggccaccc 
               
               
                   
                   
               
               
                   
                 ggctgaagagaaccgccagaagaagatacaccagacggaa 
               
               
                   
                   
               
               
                   
                 gaaccggatctgctatctgcaagagatcttcagcaacgag 
               
               
                   
                   
               
               
                   
                 atggccaaggtggacgacagcttcttccacagactggaag 
               
               
                   
                   
               
               
                   
                 agtccttcctggtggaagaggataagaagcacgagcggca 
               
               
                   
                   
               
               
                   
                 ccccatcttcggcaacatcgtggacgaggtggcctaccac 
               
               
                   
                   
               
               
                   
                 gagaagtaccccaccatctaccacctgagaaagaaactgg 
               
               
                   
                   
               
               
                   
                 tggacagcaccgacaaggccgacctgcggctgatctatct 
               
               
                   
                   
               
               
                   
                 ggccctggcccacatgatcaagttccggggccacttcctg 
               
               
                   
                   
               
               
                   
                 atcgagggcgacctgaaccccgacaacagcgacgtggaca 
               
               
                   
                   
               
               
                   
                 agctgttcatccagctggtgcagacctacaaccagctgtt 
               
               
                   
                   
               
               
                   
                 cgaggaaaaccccatcaacgccagcggcgtggacgccaag 
               
               
                   
                   
               
               
                   
                 gccatcctgtctgccagactgagcaagagcagacggctgg 
               
               
                   
                   
               
               
                   
                 aaaatctgatcgcccagctgcccggcgagaagaagaatgg 
               
               
                   
                   
               
               
                   
                 cctgttcggaaacctgattgccctgagcctgggcctgacc 
               
               
                   
                   
               
               
                   
                 cccaacttcaagagcaacttcgacctggccgaggatgcca 
               
               
                   
                   
               
               
                   
                 aactgcagctgagcaaggacacctacgacgacgacctgga 
               
               
                   
                   
               
               
                   
                 caacctgctggcccagatcggcgaccagtacgccgacctg 
               
               
                   
                   
               
               
                   
                 tttctggccgccaagaacctgtccgacgccatcctgctga 
               
               
                   
                   
               
               
                   
                 gcgacatcctgagagtgaacaccgagatcaccaaggcccc 
               
               
                   
                   
               
               
                   
                 cctgagcgcctctatgatcaagagatacgacgagcaccac 
               
               
                   
                   
               
               
                   
                 caggacctgaccctgctgaaagctctcgtgcggcagcagc 
               
               
                   
                   
               
               
                   
                 tgcctgagaagtacaaagagattttcttcgaccagagcaa 
               
               
                   
                   
               
               
                   
                 gaacggctacgccggctacattgacggcggagccagccag 
               
               
                   
                   
               
               
                   
                 gaagagttctacaagttcatcaagcccatcctggaaaaga 
               
               
                   
                   
               
               
                   
                 tggacggcaccgaggaactgctcgtgaagctgaacagaga 
               
               
                   
                   
               
               
                   
                 ggacctgctgcggaagcagcggaccttcgacaacggcagc 
               
               
                   
                   
               
               
                   
                 atcccccaccagatccacctgggagagctgcacgccattc 
               
               
                   
                   
               
               
                   
                 tgcggcggcaggaagatttttacccattcctgaaggacaa 
               
               
                   
                   
               
               
                   
                 ccgggaaaagatcgagaagatcctgaccttccgcatcccc 
               
               
                   
                   
               
               
                   
                 tactacgtgggccctctggccaggggaaacagcagattcg 
               
               
                   
                   
               
               
                   
                 cctggatgaccagaaagagcgaggaaaccatcaccccctg 
               
               
                   
                   
               
               
                   
                 gaacttcgaggaagtggtggacaagggcgcttccgcccag 
               
               
                   
                   
               
               
                   
                 agcttcatcgagcggatgaccaacttcgataagaacctgc 
               
               
                   
                   
               
               
                   
                 ccaacgagaaggtgctgcccaagcacagcctgctgtacga 
               
               
                   
                   
               
               
                   
                 gtacttcaccgtgtataacgagctgaccaaagtgaaatac 
               
               
                   
                   
               
               
                   
                 gtgaccgagggaatgagaaagcccgccttcctgagcggcg 
               
               
                   
                   
               
               
                   
                 agcagaaaaaggccatcgtggacctgctgttcaagaccaa 
               
               
                   
                   
               
               
                   
                 ccggaaagtgaccgtgaagcagctgaaagaggactacttc 
               
               
                   
                   
               
               
                   
                 aagaaaatcgagtgcttcgactccgtggaaatctccggcg 
               
               
                   
                   
               
               
                   
                 tggaagatcggttcaacgcctccctgggcacataccacga 
               
               
                   
                   
               
               
                   
                 tctgctgaaaattatcaaggacaaggacttcctggacaat 
               
               
                   
                   
               
               
                   
                 gaggaaaacgaggacattctggaagatatcgtgctgaccc 
               
               
                   
                   
               
               
                   
                 tgacactgtttgaggacagagagatgatcgaggaacggct 
               
               
                   
                   
               
               
                   
                 gaaaacctatgcccacctgttcgacgacaaagtgatgaag 
               
               
                   
                   
               
               
                   
                 cagctgaagcggcggagatacaccggctggggcaggctga 
               
               
                   
                   
               
               
                   
                 gccggaagctgatcaacggcatccgggacaagcagtccgg 
               
               
                   
                   
               
               
                   
                 caagacaatcctggatttcctgaagtccgacggcttcgcc 
               
               
                   
                   
               
               
                   
                 aacagaaacttcatgcagctgatccacgacgacagcctga 
               
               
                   
                   
               
               
                   
                 cctttaaagaggacatccagaaagcccaggtgtccggcca 
               
               
                   
                   
               
               
                   
                 gggcgatagcctgcacgagcacattgccaatctggccggc 
               
               
                   
                   
               
               
                   
                 agccccgccattaagaagggcatcctgcagacagtgaagg 
               
               
                   
                   
               
               
                   
                 tggtggacgagctcgtgaaagtgatgggccggcacaagcc 
               
               
                   
                   
               
               
                   
                 cgagaacatcgtgatcgaaatggccagagagaaccagacc 
               
               
                   
                   
               
               
                   
                 acccagaagggacagaagaacagccgcgagagaatgaagc 
               
               
                   
                   
               
               
                   
                 ggatcgaagagggcatcaaagagctgggcagccagatcct 
               
               
                   
                   
               
               
                   
                 gaaagaacaccccgtggaaaacacccagctgcagaacgag 
               
               
                   
                   
               
               
                   
                 aagctgtacctgtactacctgcagaatgggcgggatatgt 
               
               
                   
                   
               
               
                   
                 acgtggaccaggaactggacatcaaccggctgtccgacta 
               
               
                   
                   
               
               
                   
                 cgatgtggacgctatcgtgcctcagagctttctgaaggac 
               
               
                   
                   
               
               
                   
                 gactccatcgacaacaaggtgctgaccagaagcgacaaga 
               
               
                   
                   
               
               
                   
                 accggggcaagagcgacaacgtgccctccgaagaggtcgt 
               
               
                   
                   
               
               
                   
                 gaagaagatgaagaactactggcggcagctgctgaacgcc 
               
               
                   
                   
               
               
                   
                 aagctgattacccagagaaagttcgacaatctgaccaagg 
               
               
                   
                   
               
               
                   
                 ccgagagaggcggcctgagcgaactggataaggccggctt 
               
               
                   
                   
               
               
                   
                 catcaagagacagctggtggaaacccggcagatcacaaag 
               
               
                   
                   
               
               
                   
                 cacgtggcacagatcctggactcccggatgaacactaagt 
               
               
                   
                   
               
               
                   
                 acgacgagaatgacaagctgatccgggaagtgaaagtgat 
               
               
                   
                   
               
               
                   
                 caccctgaagtccaagctggtgtccgatttccggaaggat 
               
               
                   
                   
               
               
                   
                 ttccagttttacaaagtgcgcgagatcaacaactaccacc 
               
               
                   
                   
               
               
                   
                 acgcccacgacgcctacctgaacgccgtcgtgggaaccgc 
               
               
                   
                   
               
               
                   
                 cctgatcaaaaagtaccctaagctggaaagcgagttcgtg 
               
               
                   
                   
               
               
                   
                 tacggcgactacaaggtgtacgacgtgcggaagatgatcg 
               
               
                   
                   
               
               
                   
                 ccaagagcgagcaggaaatcggcaaggctaccgccaagta 
               
               
                   
                   
               
               
                   
                 cttcttctacagcaacatcatgaactttttcaagaccgag 
               
               
                   
                   
               
               
                   
                 attaccctggccaacggcgagatccggaagcggcctctga 
               
               
                   
                   
               
               
                   
                 tcgagacaaacggcgaaaccggggagatcgtgtgggataa 
               
               
                   
                   
               
               
                   
                 gggccgggattttgccaccgtgcggaaagtgctgagcatg 
               
               
                   
                   
               
               
                   
                 ccccaagtgaatatcgtgaaaaagaccgaggtgcagacag 
               
               
                   
                   
               
               
                   
                 gcggcttcagcaaagagtctatcctgcccaagaggaacag 
               
               
                   
                   
               
               
                   
                 cgataagctgatcgccagaaagaaggactgggaccctaag 
               
               
                   
                   
               
               
                   
                 aagtacggcggcttcgacagccccaccgtggcctattctg 
               
               
                   
                   
               
               
                   
                 tgctggtggtggccaaagtggaaaagggcaagtccaagaa 
               
               
                   
                   
               
               
                   
                 actgaagagtgtgaaagagctgctggggatcaccatcatg 
               
               
                   
                   
               
               
                   
                 gaaagaagcagcttcgagaagaatcccatcgactttctgg 
               
               
                   
                   
               
               
                   
                 aagccaagggctacaaagaagtgaaaaaggacctgatcat 
               
               
                   
                   
               
               
                   
                 caagctgcctaagtactccctgttcgagctggaaaacggc 
               
               
                   
                   
               
               
                   
                 cggaagagaatgctggcctctgccggcgaactgcagaagg 
               
               
                   
                   
               
               
                   
                 gaaacgaactggccctgccctccaaatatgtgaacttcct 
               
               
                   
                   
               
               
                   
                 gtacctggccagccactatgagaagctgaagggctccccc 
               
               
                   
                   
               
               
                   
                 gaggataatgagcagaaacagctgtttgtggaacagcaca 
               
               
                   
                   
               
               
                   
                 agcactacctggacgagatcatcgagcagatcagcgagtt 
               
               
                   
                   
               
               
                   
                 ctccaagagagtgatcctggccgacgctaatctggacaaa 
               
               
                   
                   
               
               
                   
                 gtgctgtccgcctacaacaagcaccgggataagcccatca 
               
               
                   
                   
               
               
                   
                 gagagcaggccgagaatatcatccacctgtttaccctgac 
               
               
                   
                   
               
               
                   
                 caatctgggagcccctgccgccttcaagtactttgacacc 
               
               
                   
                   
               
               
                   
                 accatcgaccggaagaggtacaccagcaccaaagaggtgc 
               
               
                   
                   
               
               
                   
                 tggacgccaccctgatccaccagagcatcaccggcctgta 
               
               
                   
                   
               
               
                   
                 cgagacacggatcgacctgtctcagctgggaggcgactaa 
               
               
                   
                   
               
               
                   
                 ctcgag 
               
               
                   
                   
               
               
                   
                 Seq ID NO 306: 
               
               
                   
                 &gt;6his-NLS-A3H-GGS14-dCas9 gene sequence 
               
               
                   
                 ATGggcagcagccatcatcatcatcatcacagcagcggcc 
               
               
                   
                   
               
               
                   
                 tggtgccgcgcggcagccatatgccaaagaagaagcggaa 
               
               
                   
                   
               
               
                   
                 ggtcGCTCTTCTTACTGCTGAAACTTTTCGTCTCCAATTT 
               
               
                   
                   
               
               
                   
                 AATAATAAACGCCGTCTGCGTCGCCCGTATTACCCGCGCA 
               
               
                   
                   
               
               
                   
                 AGGCGCTGCTGTGTTACCAACTGACCCCACAAAACGGTTC 
               
               
                   
                   
               
               
                   
                 CACCCCGACTCGCGGTTACTTTGAGAATAAGAAAAAATGT 
               
               
                   
                   
               
               
                   
                 CACGCTGAGATCTGTTTCATTAACGAAATCAAATCTATGG 
               
               
                   
                   
               
               
                   
                 GCCTGGATGAAACTCAGTGCTACCAGGTCACCTGCTACCT 
               
               
                   
                   
               
               
                   
                 GACCTGGAGCCCGTGTAGCTCTTGCGCGTGGGAACTGGTT 
               
               
                   
                   
               
               
                   
                 GACTTCATCAAAGCGCACGACCATCTGAACCTGCGTATCT 
               
               
                   
                   
               
               
                   
                 TCGCTTCCCGCCTGTACTATCACTGGTGCAAGCCGCAACA 
               
               
                   
                   
               
               
                   
                 GGATGGCCTGCGCCTGCTGTGTGGTTCTCAGGTTCCGGTT 
               
               
                   
                   
               
               
                   
                 GAAGTTATGGGTTTCCCGGAGTTTGCGGACTGCTGGGAAA 
               
               
                   
                   
               
               
                   
                 ACTTTGTTGACCATGAGAAGCCACTGTCCTTTAACCCGTA 
               
               
                   
                   
               
               
                   
                 TAAAATGCTGGAAGAGCTGGACAAAAACTCTCGTGCTATC 
               
               
                   
                   
               
               
                   
                 AAGCGCCGTCTGGATCGTATCAAGTCTGGAGGAAGTGGAG 
               
               
                   
                   
               
               
                   
                 GAAGTGGAGGAAGTGGAGGAAGTGGAGGAAGTGGAGGAAG 
               
               
                   
                   
               
               
                   
                 TGGAGGAAGTGGAGGAAGTGGAGGAAGTGGAGGAAGTGGA 
               
               
                   
                   
               
               
                   
                 GGAAGTGGAGGAAGTGGAGGAAGTGGAGGAAGTaagcttg 
               
               
                   
                   
               
               
                   
                 acaagaagtacagcatcggcctggccatcggcaccaactc 
               
               
                   
                   
               
               
                   
                 tgtgggctgggccgtgatcaccgacgagtacaaggtgccc 
               
               
                   
                   
               
               
                   
                 agcaagaaattcaaggtgctgggcaacaccgaccggcaca 
               
               
                   
                   
               
               
                   
                 gcatcaagaagaacctgatcggagccctgctgttcgacag 
               
               
                   
                   
               
               
                   
                 cggcgaaacagccgaggccacccggctgaagagaaccgcc 
               
               
                   
                   
               
               
                   
                 agaagaagatacaccagacggaagaaccggatctgctatc 
               
               
                   
                   
               
               
                   
                 tgcaagagatcttcagcaacgagatggccaaggtggacga 
               
               
                   
                   
               
               
                   
                 cagcttcttccacagactggaagagtccttcctggtggaa 
               
               
                   
                   
               
               
                   
                 gaggataagaagcacgagcggcaccccatcttcggcaaca 
               
               
                   
                   
               
               
                   
                 tcgtggacgaggtggcctaccacgagaagtaccccaccat 
               
               
                   
                   
               
               
                   
                 ctaccacctgagaaagaaactggtggacagcaccgacaag 
               
               
                   
                   
               
               
                   
                 gccgacctgcggctgatctatctggccctggcccacatga 
               
               
                   
                   
               
               
                   
                 tcaagttccggggccacttcctgatcgagggcgacctgaa 
               
               
                   
                   
               
               
                   
                 ccccgacaacagcgacgtggacaagctgttcatccagctg 
               
               
                   
                   
               
               
                   
                 gtgcagacctacaaccagctgttcgaggaaaaccccatca 
               
               
                   
                   
               
               
                   
                 acgccagcggcgtggacgccaaggccatcctgtctgccag 
               
               
                   
                   
               
               
                   
                 actgagcaagagcagacggctggaaaatctgatcgcccag 
               
               
                   
                   
               
               
                   
                 ctgcccggcgagaagaagaatggcctgttcggaaacctga 
               
               
                   
                   
               
               
                   
                 ttgccctgagcctgggcctgacccccaacttcaagagcaa 
               
               
                   
                   
               
               
                   
                 cttcgacctggccgaggatgccaaactgcagctgagcaag 
               
               
                   
                   
               
               
                   
                 gacacctacgacgacgacctggacaacctgctggcccaga 
               
               
                   
                   
               
               
                   
                 tcggcgaccagtacgccgacctgtttctggccgccaagaa 
               
               
                   
                   
               
               
                   
                 cctgtccgacgccatcctgctgagcgacatcctgagagtg 
               
               
                   
                   
               
               
                   
                 aacaccgagatcaccaaggcccccctgagcgcctctatga 
               
               
                   
                   
               
               
                   
                 tcaagagatacgacgagcaccaccaggacctgaccctgct 
               
               
                   
                   
               
               
                   
                 gaaagctctcgtgcggcagcagctgcctgagaagtacaaa 
               
               
                   
                   
               
               
                   
                 gagattttcttcgaccagagcaagaacggctacgccggct 
               
               
                   
                   
               
               
                   
                 acattgacggcggagccagccaggaagagttctacaagtt 
               
               
                   
                   
               
               
                   
                 catcaagcccatcctggaaaagatggacggcaccgaggaa 
               
               
                   
                   
               
               
                   
                 ctgctcgtgaagctgaacagagaggacctgctgcggaagc 
               
               
                   
                   
               
               
                   
                 agcggaccttcgacaacggcagcatcccccaccagatcca 
               
               
                   
                   
               
               
                   
                 cctgggagagctgcacgccattctgcggcggcaggaagat 
               
               
                   
                   
               
               
                   
                 ttttacccattcctgaaggacaaccgggaaaagatcgaga 
               
               
                   
                   
               
               
                   
                 agatcctgaccttccgcatcccctactacgtgggccctct 
               
               
                   
                   
               
               
                   
                 ggccaggggaaacagcagattcgcctggatgaccagaaag 
               
               
                   
                   
               
               
                   
                 agcgaggaaaccatcaccccctggaacttcgaggaagtgg 
               
               
                   
                   
               
               
                   
                 tggacaagggcgcttccgcccagagcttcatcgagcggat 
               
               
                   
                   
               
               
                   
                 gaccaacttcgataagaacctgcccaacgagaaggtgctg 
               
               
                   
                   
               
               
                   
                 cccaagcacagcctgctgtacgagtacttcaccgtgtata 
               
               
                   
                   
               
               
                   
                 acgagctgaccaaagtgaaatacgtgaccgagggaatgag 
               
               
                   
                   
               
               
                   
                 aaagcccgccttcctgagcggcgagcagaaaaaggccatc 
               
               
                   
                   
               
               
                   
                 gtggacctgctgttcaagaccaaccggaaagtgaccgtga 
               
               
                   
                   
               
               
                   
                 agcagctgaaagaggactacttcaagaaaatcgagtgctt 
               
               
                   
                   
               
               
                   
                 cgactccgtggaaatctccggcgtggaagatcggttcaac 
               
               
                   
                   
               
               
                   
                 gcctccctgggcacataccacgatctgctgaaaattatca 
               
               
                   
                   
               
               
                   
                 aggacaaggacttcctggacaatgaggaaaacgaggacat 
               
               
                   
                   
               
               
                   
                 tctggaagatatcgtgctgaccctgacactgtttgaggac 
               
               
                   
                   
               
               
                   
                 agagagatgatcgaggaacggctgaaaacctatgcccacc 
               
               
                   
                   
               
               
                   
                 tgttcgacgacaaagtgatgaagcagctgaagcggcggag 
               
               
                   
                   
               
               
                   
                 atacaccggctggggcaggctgagccggaagctgatcaac 
               
               
                   
                   
               
               
                   
                 ggcatccgggacaagcagtccggcaagacaatcctggatt 
               
               
                   
                   
               
               
                   
                 tcctgaagtccgacggcttcgccaacagaaacttcatgca 
               
               
                   
                   
               
               
                   
                 gctgatccacgacgacagcctgacctttaaagaggacatc 
               
               
                   
                   
               
               
                   
                 cagaaagcccaggtgtccggccagggcgatagcctgcacg 
               
               
                   
                   
               
               
                   
                 agcacattgccaatctggccggcagccccgccattaagaa 
               
               
                   
                   
               
               
                   
                 gggcatcctgcagacagtgaaggtggtggacgagctcgtg 
               
               
                   
                   
               
               
                   
                 aaagtgatgggccggcacaagcccgagaacatcgtgatcg 
               
               
                   
                   
               
               
                   
                 aaatggccagagagaaccagaccacccagaagggacagaa 
               
               
                   
                   
               
               
                   
                 gaacagccgcgagagaatgaagcggatcgaagagggcatc 
               
               
                   
                   
               
               
                   
                 aaagagctgggcagccagatcctgaaagaacaccccgtgg 
               
               
                   
                   
               
               
                   
                 aaaacacccagctgcagaacgagaagctgtacctgtacta 
               
               
                   
                   
               
               
                   
                 cctgcagaatgggcgggatatgtacgtggaccaggaactg 
               
               
                   
                   
               
               
                   
                 gacatcaaccggctgtccgactacgatgtggacgctatcg 
               
               
                   
                   
               
               
                   
                 tgcctcagagctttctgaaggacgactccatcgacaacaa 
               
               
                   
                   
               
               
                   
                 ggtgctgaccagaagcgacaagaaccggggcaagagcgac 
               
               
                   
                   
               
               
                   
                 aacgtgccctccgaagaggtcgtgaagaagatgaagaact 
               
               
                   
                   
               
               
                   
                 actggcggcagctgctgaacgccaagctgattacccagag 
               
               
                   
                   
               
               
                   
                 aaagttcgacaatctgaccaaggccgagagaggcggcctg 
               
               
                   
                   
               
               
                   
                 agcgaactggataaggccggcttcatcaagagacagctgg 
               
               
                   
                   
               
               
                   
                 tggaaacccggcagatcacaaagcacgtggcacagatcct 
               
               
                   
                   
               
               
                   
                 ggactcccggatgaacactaagtacgacgagaatgacaag 
               
               
                   
                   
               
               
                   
                 ctgatccgggaagtgaaagtgatcaccctgaagtccaagc 
               
               
                   
                   
               
               
                   
                 tggtgtccgatttccggaaggatttccagttttacaaagt 
               
               
                   
                   
               
               
                   
                 gcgcgagatcaacaactaccaccacgcccacgacgcctac 
               
               
                   
                   
               
               
                   
                 ctgaacgccgtcgtgggaaccgccctgatcaaaaagtacc 
               
               
                   
                   
               
               
                   
                 ctaagctggaaagcgagttcgtgtacggcgactacaaggt 
               
               
                   
                   
               
               
                   
                 gtacgacgtgcggaagatgatcgccaagagcgagcaggaa 
               
               
                   
                   
               
               
                   
                 atcggcaaggctaccgccaagtacttcttctacagcaaca 
               
               
                   
                   
               
               
                   
                 tcatgaactttttcaagaccgagattaccctggccaacgg 
               
               
                   
                   
               
               
                   
                 cgagatccggaagcggcctctgatcgagacaaacggcgaa 
               
               
                   
                   
               
               
                   
                 accggggagatcgtgtgggataagggccgggattttgcca 
               
               
                   
                   
               
               
                   
                 ccgtgcggaaagtgctgagcatgccccaagtgaatatcgt 
               
               
                   
                   
               
               
                   
                 gaaaaagaccgaggtgcagacaggcggcttcagcaaagag 
               
               
                   
                   
               
               
                   
                 tctatcctgcccaagaggaacagcgataagctgatcgcca 
               
               
                   
                   
               
               
                   
                 gaaagaaggactgggaccctaagaagtacggcggcttcga 
               
               
                   
                   
               
               
                   
                 cagccccaccgtggcctattctgtgctggtggtggccaaa 
               
               
                   
                   
               
               
                   
                 gtggaaaagggcaagtccaagaaactgaagagtgtgaaag 
               
               
                   
                   
               
               
                   
                 agctgctggggatcaccatcatggaaagaagcagcttcga 
               
               
                   
                   
               
               
                   
                 gaagaatcccatcgactttctggaagccaagggctacaaa 
               
               
                   
                   
               
               
                   
                 gaagtgaaaaaggacctgatcatcaagctgcctaagtact 
               
               
                   
                   
               
               
                   
                 ccctgttcgagctggaaaacggccggaagagaatgctggc 
               
               
                   
                   
               
               
                   
                 ctctgccggcgaactgcagaagggaaacgaactggccctg 
               
               
                   
                   
               
               
                   
                 ccctccaaatatgtgaacttcctgtacctggccagccact 
               
               
                   
                   
               
               
                   
                 atgagaagctgaagggctcccccgaggataatgagcagaa 
               
               
                   
                   
               
               
                   
                 acagctgtttgtggaacagcacaagcactacctggacgag 
               
               
                   
                   
               
               
                   
                 atcatcgagcagatcagcgagttctccaagagagtgatcc 
               
               
                   
                   
               
               
                   
                 tggccgacgctaatctggacaaagtgctgtccgcctacaa 
               
               
                   
                   
               
               
                   
                 caagcaccgggataagcccatcagagagcaggccgagaat 
               
               
                   
                   
               
               
                   
                 atcatccacctgtttaccctgaccaatctgggagcccctg 
               
               
                   
                   
               
               
                   
                 ccgccttcaagtactttgacaccaccatcgaccggaagag 
               
               
                   
                   
               
               
                   
                 gtacaccagcaccaaagaggtgctggacgccaccctgatc 
               
               
                   
                   
               
               
                   
                 caccagagcatcaccggcctgtacgagacacggatcgacc 
               
               
                   
                   
               
               
                   
                 tgtctcagctgggaggcgactaactcgag 
               
               
                   
                   
               
               
                   
                 Seq ID NO 307: 
               
               
                   
                 &gt;6his-NLS-A3H-GGS7-dCpf1 gene sequence 
               
               
                   
                 ATGggcagcagccatcatcatcatcatcacagcagcggcc 
               
               
                   
                   
               
               
                   
                 tggtgccgcgcggcagccatatgccaaagaagaagcggaa 
               
               
                   
                   
               
               
                   
                 ggtcGCTCTTCTTACTGCTGAAACTTTTCGTCTCCAATTT 
               
               
                   
                   
               
               
                   
                 AATAATAAACGCCGTCTGCGTCGCCCGTATTACCCGCGCA 
               
               
                   
                   
               
               
                   
                 AGGCGCTGCTGTGTTACCAACTGACCCCACAAAACGGTTC 
               
               
                   
                   
               
               
                   
                 CACCCCGACTCGCGGTTACTTTGAGAATAAGAAAAAATGT 
               
               
                   
                   
               
               
                   
                 CACGCTGAGATCTGTTTCATTAACGAAATCAAATCTATGG 
               
               
                   
                   
               
               
                   
                 GCCTGGATGAAACTCAGTGCTACCAGGTCACCTGCTACCT 
               
               
                   
                   
               
               
                   
                 GACCTGGAGCCCGTGTAGCTCTTGCGCGTGGGAACTGGTT 
               
               
                   
                   
               
               
                   
                 GACTTCATCAAAGCGCACGACCATCTGAACCTGCGTATCT 
               
               
                   
                   
               
               
                   
                 TCGCTTCCCGCCTGTACTATCACTGGTGCAAGCCGCAACA 
               
               
                   
                   
               
               
                   
                 GGATGGCCTGCGCCTGCTGTGTGGTTCTCAGGTTCCGGTT 
               
               
                   
                   
               
               
                   
                 GAAGTTATGGGTTTCCCGGAGTTTGCGGACTGCTGGGAAA 
               
               
                   
                   
               
               
                   
                 ACTTTGTTGACCATGAGAAGCCACTGTCCTTTAACCCGTA 
               
               
                   
                   
               
               
                   
                 TAAAATGCTGGAAGAGCTGGACAAAAACTCTCGTGCTATC 
               
               
                   
                   
               
               
                   
                 AAGCGCCGTCTGGATCGTATCAAGTCTGGAGGAAGTGGAG 
               
               
                   
                   
               
               
                   
                 GAAGTGGAGGAAGTGGAGGAAGTGGAGGAAGTGGAGGAAG 
               
               
                   
                   
               
               
                   
                 TGGAGGAAGTATGACACAGTTCGAGGGCTTTACCAACCTG 
               
               
                   
                   
               
               
                   
                 TATCAGGTGAGCAAGACACTGCGGTTTGAGCTGATCCCAC 
               
               
                   
                   
               
               
                   
                 AGGGCAAGACCCTGAAGCACATCCAGGAGCAGGGCTTCAT 
               
               
                   
                   
               
               
                   
                 CGAGGAGGACAAGGCCCGCAATGATCACTACAAGGAGCTG 
               
               
                   
                   
               
               
                   
                 AAGCCCATCATCGATCGGATCTACAAGACCTATGCCGACC 
               
               
                   
                   
               
               
                   
                 AGTGCCTGCAGCTGGTGCAGCTGGATTGGGAGAACCTGAG 
               
               
                   
                   
               
               
                   
                 CGCCGCCATCGACTCCTATAGAAAGGAGAAAACCGAGGAG 
               
               
                   
                   
               
               
                   
                 ACAAGGAACGCCCTGATCGAGGAGCAGGCCACATATCGCA 
               
               
                   
                   
               
               
                   
                 ATGCCATCCACGACTACTTCATCGGCCGGACAGACAACCT 
               
               
                   
                   
               
               
                   
                 GACCGATGCCATCAATAAGAGACACGCCGAGATCTACAAG 
               
               
                   
                   
               
               
                   
                 GGCCTGTTCAAGGCCGAGCTGTTTAATGGCAAGGTGCTGA 
               
               
                   
                   
               
               
                   
                 AGCAGCTGGGCACCGTGACCACAACCGAGCACGAGAACGC 
               
               
                   
                   
               
               
                   
                 CCTGCTGCGGAGCTTCGACAAGTTTACAACCTACTTCTCC 
               
               
                   
                   
               
               
                   
                 GGCTTTTATGAGAACAGGAAGAACGTGTTCAGCGCCGAGG 
               
               
                   
                   
               
               
                   
                 ATATCAGCACAGCCATCCCACACCGCATCGTGCAGGACAA 
               
               
                   
                   
               
               
                   
                 CTTCCCCAAGTTTAAGGAGAATTGTCACATCTTCACACGC 
               
               
                   
                   
               
               
                   
                 CTGATCACCGCCGTGCCCAGCCTGCGGGAGCACTTTGAGA 
               
               
                   
                   
               
               
                   
                 ACGTGAAGAAGGCCATCGGCATCTTCGTGAGCACCTCCAT 
               
               
                   
                   
               
               
                   
                 CGAGGAGGTGTTTTCCTTCCCTTTTTATAACCAGCTGCTG 
               
               
                   
                   
               
               
                   
                 ACACAGACCCAGATCGACCTGTATAACCAGCTGCTGGGAG 
               
               
                   
                   
               
               
                   
                 GAATCTCTCGGGAGGCAGGCACCGAGAAGATCAAGGGCCT 
               
               
                   
                   
               
               
                   
                 GAACGAGGTGCTGAATCTGGCCATCCAGAAGAATGATGAG 
               
               
                   
                   
               
               
                   
                 ACAGCCCACATCATCGCCTCCCTGCCACACAGATTCATCC 
               
               
                   
                   
               
               
                   
                 CCCTGTTTAAGCAGATCCTGTCCGATAGGAACACCCTGTC 
               
               
                   
                   
               
               
                   
                 TTTCATCCTGGAGGAGTTTAAGAGCGACGAGGAAGTGATC 
               
               
                   
                   
               
               
                   
                 CAGTCCTTCTGCAAGTACAAGACACTGCTGAGAAACGAGA 
               
               
                   
                   
               
               
                   
                 ACGTGCTGGAGACAGCCGAGGCCCTGTTTAACGAGCTGAA 
               
               
                   
                   
               
               
                   
                 CAGCATCGACCTGACACACATCTTCATCAGCCACAAGAAG 
               
               
                   
                   
               
               
                   
                 CTGGAGACAATCAGCAGCGCCCTGTGCGACCACTGGGATA 
               
               
                   
                   
               
               
                   
                 CACTGAGGAATGCCCTGTATGAGCGGAGAATCTCCGAGCT 
               
               
                   
                   
               
               
                   
                 GACAGGCAAGATCACCAAGTCTGCCAAGGAGAAGGTGCAG 
               
               
                   
                   
               
               
                   
                 CGCAGCCTGAAGCACGAGGATATCAACCTGCAGGAGATCA 
               
               
                   
                   
               
               
                   
                 TCTCTGCCGCAGGCAAGGAGCTGAGCGAGGCCTTCAAGCA 
               
               
                   
                   
               
               
                   
                 GAAAACCAGCGAGATCCTGTCCCACGCACACGCCGCCCTG 
               
               
                   
                   
               
               
                   
                 GATCAGCCACTGCCTACAACCCTGAAGAAGCAGGAGGAGA 
               
               
                   
                   
               
               
                   
                 AGGAGATCCTGAAGTCTCAGCTGGACAGCCTGCTGGGCCT 
               
               
                   
                   
               
               
                   
                 GTACCACCTGCTGGACTGGTTTGCCGTGGATGAGTCCAAC 
               
               
                   
                   
               
               
                   
                 GAGGTGGACCCCGAGTTCTCTGCCCGGCTGACCGGCATCA 
               
               
                   
                   
               
               
                   
                 AGCTGGAGATGGAGCCTTCTCTGAGCTTCTACAACAAGGC 
               
               
                   
                   
               
               
                   
                 CAGAAATTATGCCACCAAGAAGCCCTACTCCGTGGAGAAG 
               
               
                   
                   
               
               
                   
                 TTCAAGCTGAACTTTCAGATGCCTACACTGGCCTCTGGCT 
               
               
                   
                   
               
               
                   
                 GGGACGTGAATAAGGAGAAGAACAATGGCGCCATCCTGTT 
               
               
                   
                   
               
               
                   
                 TGTGAAGAACGGCCTGTACTATCTGGGCATCATGCCAAAG 
               
               
                   
                   
               
               
                   
                 CAGAAGGGCAGGTATAAGGCCCTGAGCTTCGAGCCCACAG 
               
               
                   
                   
               
               
                   
                 AGAAAACCAGCGAGGGCTTTGATAAGATGTACTATGACTA 
               
               
                   
                   
               
               
                   
                 CTTCCCTGATGCCGCCAAGATGATCCCAAAGTGCAGCACC 
               
               
                   
                   
               
               
                   
                 CAGCTGAAGGCCGTGACAGCCCACTTTCAGACCCACACAA 
               
               
                   
                   
               
               
                   
                 CCCCCATCCTGCTGTCCAACAATTTCATCGAGCCTCTGGA 
               
               
                   
                   
               
               
                   
                 GATCACAAAGGAGATCTACGACCTGAACAATCCTGAGAAG 
               
               
                   
                   
               
               
                   
                 GAGCCAAAGAAGTTTCAGACAGCCTACGCCAAGAAAACCG 
               
               
                   
                   
               
               
                   
                 GCGACCAGAAGGGCTACAGAGAGGCCCTGTGCAAGTGGAT 
               
               
                   
                   
               
               
                   
                 CGACTTCACAAGGGATTTTCTGTCCAAGTATACCAAGACA 
               
               
                   
                   
               
               
                   
                 ACCTCTATCGATCTGTCTAGCCTGCGGCCATCCTCTCAGT 
               
               
                   
                   
               
               
                   
                 ATAAGGACCTGGGCGAGTACTATGCCGAGCTGAATCCCCT 
               
               
                   
                   
               
               
                   
                 GCTGTACCACATCAGCTTCCAGAGAATCGCCGAGAAGGAG 
               
               
                   
                   
               
               
                   
                 ATCATGGATGCCGTGGAGACAGGCAAGCTGTACCTGTTCC 
               
               
                   
                   
               
               
                   
                 AGATCTATAACAAGGACTTTGCCAAGGGCCACCACGGCAA 
               
               
                   
                   
               
               
                   
                 GCCTAATCTGCACACACTGTATTGGACCGGCCTGTTTTCT 
               
               
                   
                   
               
               
                   
                 CCAGAGAACCTGGCCAAGACAAGCATCAAGCTGAATGGCC 
               
               
                   
                   
               
               
                   
                 AGGCCGAGCTGTTCTACCGCCCTAAGTCCAGGATGAAGAG 
               
               
                   
                   
               
               
                   
                 GATGGCACACCGGCTGGGAGAGAAGATGCTGAACAAGAAG 
               
               
                   
                   
               
               
                   
                 CTGAAGGATCAGAAAACCCCAATCCCCGACACCCTGTACC 
               
               
                   
                   
               
               
                   
                 AGGAGCTGTACGACTATGTGAATCACAGACTGTCCCACGA 
               
               
                   
                   
               
               
                   
                 CCTGTCTGATGAGGCCAGGGCCCTGCTGCCCAACGTGATC 
               
               
                   
                   
               
               
                   
                 ACCAAGGAGGTGTCTCACGAGATCATCAAGGATAGGCGCT 
               
               
                   
                   
               
               
                   
                 TTACCAGCGACAAGTTCTTTTTCCACGTGCCTATCACACT 
               
               
                   
                   
               
               
                   
                 GAACTATCAGGCCGCCAATTCCCCATCTAAGTTCAACCAG 
               
               
                   
                   
               
               
                   
                 AGGGTGAATGCCTACCTGAAGGAGCACCCCGAGACACCTA 
               
               
                   
                   
               
               
                   
                 TCATCGGCATCGATCGGGGCGAGAGAAACCTGATCTATAT 
               
               
                   
                   
               
               
                   
                 CACAGTGATCGACTCCACCGGCAAGATCCTGGAGCAGCGG 
               
               
                   
                   
               
               
                   
                 AGCCTGAACACCATCCAGCAGTTTGATTACCAGAAGAAGC 
               
               
                   
                   
               
               
                   
                 TGGACAACAGGGAGAAGGAGAGGGTGGCAGCAAGGCAGGC 
               
               
                   
                   
               
               
                   
                 CTGGTCTGTGGTGGGCACAATCAAGGATCTGAAGCAGGGC 
               
               
                   
                   
               
               
                   
                 TATCTGAGCCAGGTCATCCACGAGATCGTGGACCTGATGA 
               
               
                   
                   
               
               
                   
                 TCCACTACCAGGCCGTGGTGGTGCTGGAGAACCTGAATTT 
               
               
                   
                   
               
               
                   
                 CGGCTTTAAGAGCAAGAGGACCGGCATCGCCGAGAAGGCC 
               
               
                   
                   
               
               
                   
                 GTGTACCAGCAGTTCGAGAAGATGCTGATCGATAAGCTGA 
               
               
                   
                   
               
               
                   
                 ATTGCCTGGTGCTGAAGGACTATCCAGCAGAGAAAGTGGG 
               
               
                   
                   
               
               
                   
                 AGGCGTGCTGAACCCATACCAGCTGACAGACCAGTTCACC 
               
               
                   
                   
               
               
                   
                 TCCTTTGCCAAGATGGGCACCCAGTCTGGCTTCCTGTTTT 
               
               
                   
                   
               
               
                   
                 ACGTGCCTGCCCCATATACATCTAAGATCGATCCCCTGAC 
               
               
                   
                   
               
               
                   
                 CGGCTTCGTGGACCCCTTCGTGTGGAAAACCATCAAGAAT 
               
               
                   
                   
               
               
                   
                 CACGAGAGCCGCAAGCACTTCCTGGAGGGCTTCGACTTTC 
               
               
                   
                   
               
               
                   
                 TGCACTACGACGTGAAAACCGGCGACTTCATCCTGCACTT 
               
               
                   
                   
               
               
                   
                 TAAGATGAACAGAAATCTGTCCTTCCAGAGGGGCCTGCCC 
               
               
                   
                   
               
               
                   
                 GGCTTTATGCCTGCATGGGATATCGTGTTCGAGAAGAACG 
               
               
                   
                   
               
               
                   
                 AGACACAGTTTGACGCCAAGGGCACCCCTTTCATCGCCGG 
               
               
                   
                   
               
               
                   
                 CAAGAGAATCGTGCCAGTGATCGAGAATCACAGATTCACC 
               
               
                   
                   
               
               
                   
                 GGCAGATACCGGGACCTGTATCCTGCCAACGAGCTGATCG 
               
               
                   
                   
               
               
                   
                 CCCTGCTGGAGGAGAAGGGCATCGTGTTCAGGGATGGCTC 
               
               
                   
                   
               
               
                   
                 CAACATCCTGCCAAAGCTGCTGGAGAATGACGATTCTCAC 
               
               
                   
                   
               
               
                   
                 GCCATCGACACCATGGTGGCCCTGATCCGCAGCGTGCTGC 
               
               
                   
                   
               
               
                   
                 AGATGCGGAACTCCAATGCCGCCACAGGCGAGGACTATAT 
               
               
                   
                   
               
               
                   
                 CAACAGCCCCGTGCGCGATCTGAATGGCGTGTGCTTCGAC 
               
               
                   
                   
               
               
                   
                 TCCCGGTTTCAGAACCCAGAGTGGCCCATGGACGCCGATG 
               
               
                   
                   
               
               
                   
                 CCAATGGCGCCTACCACATCGCCCTGAAGGGCCAGCTGCT 
               
               
                   
                   
               
               
                   
                 GCTGAATCACCTGAAGGAGAGCAAGGATCTGAAGCTGCAG 
               
               
                   
                   
               
               
                   
                 AACGGCATCTCCAATCAGGACTGGCTGGCCTACATCCAGG 
               
               
                   
                   
               
               
                   
                 AGCTGCGCAACAAAAGGCCGGCGGCCACGAAAAAGGCCGG 
               
               
                   
                   
               
               
                   
                 CCAGGCAAAAAAGAAAAAGGGATCCTACCCATACGATGTT 
               
               
                   
                   
               
               
                   
                 CCAGATTACGCTTATCCCTACGACGTGCCTGATTATGCAT 
               
               
                   
                   
               
               
                   
                 ACCCATATGATGTCCCCGACTATGCCTAAG