Abstract:
Host cells that exhibit regulated expression of a test gene, are rapidly identifyed by providing a plurality of host cells, each host cell comprising a polynucleotide, the polynucleotide comprising in order a regulatable promoter; a test gene; an IRES sequence; and a surface marker coding sequence, where the surface marker comprises a secretion signal sequence, a detectable label protein, and a membrane anchor; wherein expression of said test gene also results in expression of said surface marker; inducing said promoter; and selecting a host cell that displays said surface marker on its surface.

Description:
FIELD OF THE INVENTION  
         [0001]    This invention is related generally to the fields of molecular biology and genomics. More particularly, the invention relates to nucleic acid vectors and methods for rapidly identifying transformed cells with regulated expression of a heterologous polynucleotide.  
         BACKGROUND OF THE INVENTION  
         [0002]    Identification of clonal eukaryotic cells lines that have regulated expression of target cDNAs or genes has been a time consuming and expensive process. During the course of cell biological and functional genomic investigations it is often desirable to create cell lines in which a cDNA is expressed in a regulated manner. For this purpose many types of regulons (combinations of transactivators and regulated promoters) have been invented. These include the tetracycline regulon (U. Baron, et al.,  Nuc Acids Res  (1995) 23(17):3605-06), the ecdysone regulon (D. No, et al.,  Proc Natl Acad Sci USA  (1996) 93(8):3346-51), regulons controlled by a transplanted  E. coli  Lac/La repressor system, the heat shock regulon, and the metalothionine regulon. These systems provide, with some effort, clonal cell lines in which the cDNA is regulated by application of an exogenous stimulator: tetracycline, ecdysone, isopropylthiogal-actopyranoside (ITGP), heat or heavy metals, respectively. The time and expense of creating these cells lines arises from the need to isolate and expand numerous single cell clones and analyze each clone for appropriate regulation of the query gene. If a way could be devised to use a rapid cell sorting technique such as flow cytometry, cell panning, colorimetric overlays or magnetic selection to identify candidate regulated clones, significant time and effort could be saved.  
         SUMMARY OF THE INVENTION  
         [0003]    We have now invented a construct and method for rapidly labeling host cells, and for determining which cells provide regulated expression of a test gene (and surface label).  
           [0004]    One aspect of the invention is a polynucleotide, comprising a regulatable promoter; a test gene; an IRES sequence; a surface marker coding sequence, said surface marker comprising a secretion signal sequence, a detectable label protein, and a membrane anchor; wherein expression of said test gene also results in expression of said surface marker.  
           [0005]    Another aspect of the invention is a host cell, comprising: a polynucleotide, said polynucleotide comprising a regulatable promoter, a test gene, an IRES sequence, a surface marker coding sequence, said surface marker comprising a secretion signal sequence, a detectable label protein, and a membrane anchor; wherein expression of said test gene also results in expression of said surface marker.  
           [0006]    Another aspect of the invention is a method of identifying a host cell that exhibits regulated expression of a test gene, comprising: providing a plurality of host cells, each host cell comprising a polynucleotide, said polynucleotide comprising in order a regulatable promoter; a test gene; an IRES sequence; and a surface marker coding sequence, said surface marker comprising a secretion signal sequence, a detectable label protein, and a membrane anchor; wherein expression of said test gene also results in expression of said surface marker; inducing said promoter; and selecting a host cell that displays said surface marker on its surface.  
           [0007]    Another aspect of the invention is a method of identifying a host cell that exhibits regulated expression of a test gene, comprising: providing a plurality of host cells, each host cell comprising a polynucleotide, said polynucleotide comprising in order a regulatable promoter; a test gene; an IRES sequence; and a surface marker coding sequence, said surface marker comprising a secretion signal sequence, a detectable label protein, and a membrane anchor; wherein expression of said test gene also results in expression of said surface marker; selecting a plurality of host cells that do not exhibit said surface marker in the absence of induction of said promoter; inducing said promoter; and selecting a host cell that exhibits expression of said surface marker. 
       
    
    
     BRIEF DESCRIPTION OF THE FIGURES  
       [0008]    [0008]FIG. 1 is a diagram of an embodiment of the invention (the pFastFind vector, SEQ ID NO:5).  
         [0009]    [0009]FIG. 2 is a diagram of the pFastFind-delta vector (SEQ ID NO:6), a version having a reduced size surface epitope marker.  
         [0010]    [0010]FIG. 3 is a diagram of the pFastFind-point vector (SEQ ID NO:7), a version with a point mutation that eliminates the alkaline phosphatase enzymatic activity from the surface epitope marker while maintaining the size and epitope characteristics.  
         [0011]    [0011]FIG. 4 is a summary protocol for isolation of regulated expression clones using vectors of the invention.  
         [0012]    [0012]FIG. 5 presents data demonstrating that pFastFind results in the preparation of cell lines with regulated expression that persists for 36 hours after ponasterone-A treatment, and is induced by micromolar concentrations of ponasterone-A.  
         [0013]    [0013]FIG. 6 presents data demonstrating that the surrogate marker expression correlates with the amount of mRNA encoding the query gene. 
     
    
     DETAILED DESCRIPTION  
       [0014]    Definitions:  
         [0015]    The terms “test gene” and “query gene” refer to a polynucleotide to be examined, whether its function is known or unknown, regardless of whether it is synthetic or identical to a known sequence.  
         [0016]    The term “IRES” refers to an internal ribosome binding site, or other sequence capable of serving as a translational initiation point when transcribed into mRNA.  
         [0017]    The term “surface marker” refers to a protein which is associated with the surface of a host cell following translation. In general, the surface marker must be detectable either directly or through binding a labeled binding partner.  
         [0018]    The term “regulatable promoter” refers to a polynucleotide sequence capable of controlling the transcription of an adjacent polynucleotide, and which can be controlled by altering or adjusting the host cell&#39;s environment. The environment can be adjusted by addition or subtraction of various factors or compounds, by altering the temperature, pressure, concentration of media components, radiation, and the like.  
         [0019]    The term “FACS” refers to fluorescence-activated cell sorting, and includes any method for separating cells on the basis of a visible or fluorescent label. The label can be attached directly to the cell (for example, it can be expressed as a cell surface protein), or can be bound to the cell surface (for example, by allowing a labeled antibody to recognize and bind to a cell surface antigen).  
         [0020]    General Method:  
         [0021]    The vector of the invention and its method of use allows rapid isolation of candidate eukaryotic cell clones in which a query gene is regulated by exogenous application of an appropriate stimulus. The vector is arranged such that the query gene can be cloned immediately downstream of the regulated promoter by means of a multiple cloning site (MCS). Downstream of the multiple cloning site is placed an internal ribosome entry site (IRES); and downstream of the IRES is placed a cell membrane-localized protein for which an epitope recognized by a convenient antibody is available (a surrogate surface marker). Thus, since the surface epitope is co-cistronic with the query gene, both the query gene and the surface epitope will be elevated in response to the exogenous stimulator.  
         [0022]    The use of a surrogate surface marker for the query gene allows isolation of clonal cell lines with stimulator-induced expression by means of flow cytometry, magnetic cell sorting, cell panning, cell enrichment by column chromatography, by use of colorimetric cell overlay methods, and other cell enrichment techniques. The use of a surrogate surface marker for the query gene circumvents any need for a specific antibody to the query gene&#39;s encoded protein, it circumvents any need for a biochemical assay for the query gene&#39;s product. The surrogate surface marker allows rapid reconfirmation of the regulation and expression of the query gene by use of the above mentioned techniques.  
         [0023]    Suitable surrogate surface markers include, without limitation, placental alkaline phosphatase, β-lactamase, β2-microglobulin, and the like. If desired, one can select or construct any distinct surface protein, and prepare antibodies capable of recognizing the protein by conventional methods. The polynucleotide encoding the surface marker preferably further includes a secretion signal sequence (or other sequence that provides for export of the protein to the outer surface of the cell), and a transmembrane anchor (or other sequence that insures that the protein will remain associated with the cell surface). The surface marker is preferably relatively non-toxic to the cell. The surface marker can exhibit enzymatic activity, which can be used as a label (for example, alkaline phosphatase, β-galactosidase, and the like), or can have rely solely on binding (for example, as an epitope or ligand-binding partner), or can include both enzymatic and ligand-binding features.  
         [0024]    The presence of a surface marker permits one to quickly separate host cells that express the test gene (and thus the surface marker) from those that do not. Such separation can be effected by means of FACS (fluorescence-activated cell sorting), affinity panning, affinity column separation, and the like. Thus, one can identify host cells that express the test gene without the need to identify another phenotype or altered characteristic that results from the test gene expression. Further, one can separate host cells in which expression is regulated from cells in which expression is either constitutive or non-existent, by selecting cells that do not express the surface marker when the promoter is repressed or not induced, and from that pool selecting cells that express the marker following induction of the promoter. These cells can also be removed by using an antibody specific for the surface marker in combination with complement. It is also possible to perform the selection steps in reverse order, or to repeat the steps several times, although one may need to wait a sufficient period of time for marker present on the host cell surface to be cleared. Additionally, one can select several different pools of cells by using different methods for inducing the promoters, for example, where the vector is cloned into position adjacent to a plurality of different promoters, or next to promoters randomly. For example, one can select a pool of cells that do not express the surface marker constitutively, and from this pool select a subset of cells that express the surface marker in response to a change in temperature. The cells that were not selected can be subjected to other conditions, for example the presence or absence of a nutrient, and any cells that respond to such conditions are then selected.  
         [0025]    The table below illustrates the advantages of the method of the invention over conventional procedures.  
                                           Conventional   pFastFind       Cell Cloning   Technology   Method                   Time Elapsed   35 days   30 days       Manpower   14 days    5 days       Screening method on clonal cell lines   RT-PCR   FACS       Number of clones analyzed   21   23       Number of regulated clones   1   10       Cloning efficiency   4.8%   43.5%                  
 
         [0026]    [0026]FIG. 1 is a schematic view of an embodiment of the invention (the pFastFind vector, SEQ ID NO:5). It highlights the neomycin resistance gene (herring-bone), the ecdysone inducible promoter (vertical hatching), the multiple cloning site (“MCS”) (grid), internal ribosome initiation sequence (IRES) (diagonal hatching), polyadenylation sequence (check hatching), secretion signal sequence (stippled hatching), surface marker alkaline phosphatase (horizontal hatching), and the transmembrane region from the PDEF receptor (brick pattern). The “M” indicates a methionine start site for translation of the surface marker, and the stop sign indicates the stop sequence for translation of the surface marker. the target cDNA or query gene is inserted in the MCS.  
         [0027]    [0027]FIG. 2 is a schematic view of another embodiment of the invention (the pFastFind-delta vector, SEQ ID NO:6). Its highlighted features are depicted as in FIG. 1. The deletion of residues 98-260 of the surface marker is shown as a white box.  
         [0028]    [0028]FIG. 3 is a schematic view of another embodiment of the invention (the pFastFind-point vector, SEQ ID NO:7). Its highlighted features are depicted as in FIG. 1. S→A indicates the point mutation that eliminates the catalytic activity of the alkaline phosphatase surface marker.  
       EXAMPLES  
       [0029]    The following examples are provided as a guide for the practitioner of ordinary skill in the art. Nothing in the examples is intended to limit the claimed invention. Unless otherwise specified, all reagents are used in accordance with the manufacturer&#39;s recommendations, and all reactions are performed at standard temperature and pressure.  
       Example 1  
     pFastFind-JNK3  
       [0030]    (A) Construction of Vector pFastFind-JNK3  
         [0031]    1. Building pIND-IRES: The following oligonucleotides were used to PCR amplify the IRES sequence from pIRES-EYFP (Clontech):  
         [0032]    5′TCATCATCATCTCGAGCCAATTCCGCCCCTCTCCCTC (SEQ ID NO:1);  
         [0033]    5′TCTCTCTAGACCGGGTTGTGGCAAGCTTATCATC (SEQ ID NO:2).  
         [0034]    The resulting IRES cDNA was digested with XhoI and XbaI and ligated to pIND (Invitrogen) treated with XhoI and XbaI and calf intestine phosphatase (CIP). The ligated plasmid was transformed and pIND-IRES was isolated and verified by restriction digests.  
         [0035]    2. Building pIND-IRES-SEAP: Full length secreted alkaline phosphatase (SEAP) cDNA was amplified from pSEAPBasic2 (Clontech) using primers containing NheI at the 5′ end and XbaI at the 3′ end. The resulting SEAP cDNA was digested with NheI and XbaI and ligated to pIND-IRES treated with XbaI and CIP. The ligated plasmid was transformed and pIND-IRES-SEAP was isolated and verified by restriction digests.  
         [0036]    3. Building pIND-IRES/SEAP-TM (pFastFind): The following oligos were used to amplify the TM fragment from pDisplay (Invitrogen):  
         [0037]    5′CTCTAGTCTAGAGTCGGGGCGGCCGGCCGCTTCGAGCAGACATCTCCCGGGAATCGCGGCTGCAG (SEQ ID NO:3);  
         [0038]    5′TAGCTAGCTGATCTCGAGCGGCCGCCTGAACGT (SEQ ID NO:4).  
         [0039]    The resulting TM PCR product was topo cloned into pcDNA3.1 and isolated by digestion with XbaI and NheI. The XbaI-TM-NheI fragment was ligated to pIND-IRES/SEAP treated with XbaI and CIP. The ligated plasmid was transformed and pIND-IRES/SEAP-TM was isolated and verified by restriction digests.  
         [0040]    4. Building pFastFind-JNK3: pFastFind-JNK3 (SEQ ID NO:8) was constructed by inserting the protein coding sequence of JNK3 into pFastFind. The JNK3 sequence was isolated using polymerase chain reactions (Stratagene) from a human brain cDNA library using primers that encode a mammalian translation consensus sequence and the coding sequence surrounding the initiator methionine and stop codons of JNK3 (Genbank Accession number U07620). The PCR product was first subcloned into pIND, amplified by PCR, inserted into pcDNA2.1-TOPO and then subcloned in between SpeI and EcoRV restriction sites of pFastFind. The DNA sequence of the pFastFind-JNK3 vector was determined throughout the JNK3 coding region. The protein sequence of the JNK3 clone agreed with the consensus alignment of JNK3 clones present in Genbank with the exception of the following changes: Tyr to Cys at residue 240.  
         [0041]    (B) Use of pFastFind-JNK3 to Isolate Clonal Cell Lines with Regulated Expression of JNK3  
         [0042]    JNK3 is a member of the MAP kinase family of protein kinases. JNK3 shares sequence homology and common biochemical substrates with its close relative JNK1 (an important regulator of early response genes); however, its role in cell physiology is unclear. An understanding of JNK3 role in cell physiology is of immediate interest. To study this gene, we constructed a cell line in which JNK3&#39;s expression and activity could be regulated by the application of an exogenous agent that is inert to the engineered cell line.  
         [0043]    Cell lines were isolated using either of two protocols. Protocol A (48-58 days) comprised: (a) transfecting cells with pFastFind; (b) selecting the transformed cells with neomycin for 15 days; (c) adding ponasterone A for 1 day; (d) isolating single cells displaying the surface marker by FACS; (e) allowing the single cells to multiply for 30-40 days; and (f) analyzing single clones treated with and without ponasterone A by FACS. Protocol B (59-79 days) comprised: (a) transfecting cells with pFastFind; (b) selecting the transformed cells with neomycin for 15 days; (c) adding ponasterone A for 1 day; (d) isolating an enriched pool of single cells displaying the surface marker by FACS; (e) allowing the single cells to multiply for 10-20 days; (f) adding ponasterone A for 1 day; (g) isolating single surface-marker positive cells using FACS; (h) allowing the single cells to multiply for 30-40 days; and (i) analyzing single clones treated with and without ponasterone A by FACS.  
         [0044]    The pFastFind-JNK3 vector was transfected into ECR-293 cells (Invitrogen), and treated with neomycin for 15 days. Surviving cells were treated with 5 μM ponasterone A and rendered into single cell suspension by trypsinization. The cell suspension was incubated with anti-SEAP monoclonal antibody for 1 hour at room temperature and subsequently incubated with fluorescein-conjugated anti-mouse IgG for 30 minutes at room temperature in dark. The labeled cell suspension was resuspended into phosphate buffered saline containing 5 μg/ml of propidium iodide and analyzed for fluorescein fluorescence using CellQuest software on FACSVantageSE (Becton Dickinson). Cells with fluorescein fluorescence&gt;20-30 and propidium iodide fluorescence&lt;100 were gated and collected as population or single cell clones using CloneCyt software.  
         [0045]    Single cells were expanded and then re-analyzed using FACS for their expression of the surface marker after treatment with ponasterone A for 24-48 h. Cells showing increased expression of the surface marker in the presence of ponasterone A as compared to unstained controls were analyzed at varying times (0-48 h) after the start of ponasterone A treatment and were subjected to increasing doses (0-30 μM) of ponasterone A. FIG. 6 shows one clone as an example where the expression of the surface marker is significantly increased with time after the addition of ponasterone A as well as with increasing concentrations of ponasterone A. Cell clones showing increased expression of the surfaces marker were also analyzed using semi-quantitative RT-PCR with primers specific for the query gene (JNK 3). FIG. 7 shows an example where the expression of the query gene increases in a similar fashion as the expression of the surface marker after the addition of ponasterone A. This shows that the methods described here can lead to a rapid selection of cell clones regulating the expression of the query gene.  
     
       
       
         1 
         
           
             8  
           
           
             1  
             37  
             DNA  
             PCR primer  
           
            1 

tcatcatcat ctcgagccaa ttccgcccct ctccctc                              37 

 
           
             2  
             34  
             DNA  
             PCR primer  
           
            2 

tctctctaga ccgggttgtg gcaagcttat catc                                 34 

 
           
             3  
             65  
             DNA  
             PCR primer  
           
            3 

ctctagtcta gagtcggggc ggccggccgc ttcgagcaga catctcccgg gaatcgcggc     60 

tgcag                                                                 65 

 
           
             4  
             33  
             DNA  
             PCR primer  
           
            4 

tagctagctg atctcgagcg gccgcctgaa cgt                                  33 

 
           
             5  
             7563  
             DNA  
             pFastFind  
           
            5 

agatctcggc cgcatattaa gtgcattgtt ctcgataccg ctaagtgcat tgttctcgtt     60 

agctcgatgg acaagtgcat tgttctcttg ctgaaagctc gatggacaag tgcattgttc    120 

tcttgctgaa agctcgatgg acaagtgcat tgttctcttg ctgaaagctc agtacccggg    180 

agtaccctcg accgccggag tataaataga ggcgcttcgt ctacggagcg acaattcaat    240 

tcaaacaagc aaagtgaaca cgtcgctaag cgaaagctaa gcaaataaac aagcgcagct    300 

gaacaagcta aacaatctgc agtaaagtgc aagttaaagt gaatcaatta aaagtaacca    360 

gcaaccaagt aaatcaactg caactactga aatctgccaa gaagtaatta ttgaatacaa    420 

gaagagaact ctgaatactt tcaacaagtt accgagaaag aagaactcac acacagctag    480 

cgtttaaact taagcttggt accgagctcg gatccactag tccagtgtgg tggaattctg    540 

cagatatcca gcacagtggc ggccgctcga gccaattccg cccctctccc tccccccccc    600 

ctaacgttac tggccgaagc cgcttggaat aaggccggtg tgcgtttgtc tatatgtgat    660 

tttccaccat attgccgtct tttggcaatg tgagggcccg gaaacctggc cctgtcttct    720 

tgacgagcat tcctaggggt ctttcccctc tcgccaaagg aatgcaaggt ctgttgaatg    780 

tcgtgaagga agcagttcct ctggaagctt cttgaagaca aacaacgtct gtagcgaccc    840 

tttgcaggca gcggaacccc ccacctggcg acaggtgcct ctgcggccaa aagccacgtg    900 

tataagatac acctgcaaag gcggcacaac cccagtgcca cgttgtgagt tggatagttg    960 

tggaaagagt caaatggctc tcctcaagcg tattcaacaa ggggctgaag gatgcccaga   1020 

aggtacccca ttgtatggga tctgatctgg ggcctcggtg cacatgcttt acatgtgttt   1080 

agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg ggacgtggtt ttcctttgaa   1140 

aaacacgatg ataagcttgc cacaacccgg tctagcccgg gctcgagatc tgcgatctaa   1200 

gtaagcttcg aatcgcgaat tcgcccacca tgctgctgct gctgctgctg ctgggcctga   1260 

ggctacagct ctccctgggc atcatcccag ttgaggagga gaacccggac ttctggaacc   1320 

gcgaggcagc cgaggccctg ggtgccgcca agaagctgca gcctgcacag acagccgcca   1380 

agaacctcat catcttcctg ggcgatggga tgggggtgtc tacggtgaca gctgccagga   1440 

tcctaaaagg gcagaagaag gacaaactgg ggcctgagat acccctggcc atggaccgct   1500 

tcccatatgt ggctctgtcc aagacataca atgtagacaa acatgtgcca gacagtggag   1560 

ccacagccac ggcctacctg tgcggggtca agggcaactt ccagaccatt ggcttgagtg   1620 

cagccgcccg ctttaaccag tgcaacacga cacgcggcaa cgaggtcatc tccgtgatga   1680 

atcgggccaa gaaagcaggg aagtcagtgg gagtggtaac caccacacga gtgcagcacg   1740 

cctcgccagc cggcacctac gcccacacgg tgaaccgcaa ctggtactcg gacgccgacg   1800 

tgcctgcctc ggcccgccag gaggggtgcc aggacatcgc tacgcagctc atctccaaca   1860 

tggacattga cgtgatccta ggtggaggcc gaaagtacat gtttcgcatg ggaaccccag   1920 

accctgagta cccagatgac tacagccaag gtgggaccag gctggacggg aagaatctgg   1980 

tgcaggaatg gctggcgaag cgccagggtg cccggtatgt gtggaaccgc actgagctca   2040 

tgcaggcttc cctggacccg tctgtgaccc atctcatggg tctctttgag cctggagaca   2100 

tgaaatacga gatccaccga gactccacac tggacccctc cctgatggag atgacagagg   2160 

ctgccctgcg cctgctgagc aggaaccccc gcggcttctt cctcttcgtg gagggtggtc   2220 

gcatcgacca tggtcatcat gaaagcaggg cttaccgggc actgactgag acgatcatgt   2280 

tcgacgacgc cattgagagg gcgggccagc tcaccagcga ggaggacacg ctgagcctcg   2340 

tcactgccga ccactcccac gtcttctcct tcggaggcta ccccctgcga gggagctcca   2400 

tcttcgggct ggcccctggc aaggcccggg acaggaaggc ctacacggtc ctcctatacg   2460 

gaaacggtcc aggctatgtg ctcaaggacg gcgcccggcc ggatgttacc gagagcgaga   2520 

gcgggagccc cgagtatcgg cagcagtcag cagtgcccct ggacgaagag acccacgcag   2580 

gcgaggacgt ggcggtgttc gcgcgcggcc cgcaggcgca cctggttcac ggcgtgcagg   2640 

agcagacctt catagcgcac gtcatggcct tcgccgcctg cctggagccc tacaccgcct   2700 

gcgacctggc gccccccgcc ggcaccaccg acgccgcgca cccgggttac tctagagtcg   2760 

gggcggccgg ccgcttcgag cagacatctc ccgggaatcc gcggctgcag gtcgacgaac   2820 

aaaaactcat ctcagaagag gatctgaatg ctgtgggcca ggacacgcag gaggtcatcg   2880 

tggtgccaca ctccttgccc tttaaggtgg tggtgatctc agccatcctg gccctggtgg   2940 

tgctcaccat catctccctt atcatcctca tcatgctttg gcagaagaag ccacgttagg   3000 

cggccgctcg agatcagcta gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc   3060 

ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg   3120 

tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag   3180 

gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga   3240 

caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag   3300 

ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt   3360 

ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc   3420 

tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg   3480 

catcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta   3540 

gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt   3600 

ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat   3660 

ctcggtctat tcttttgatt tataagggat tttggggatt tcggcctatt ggttaaaaaa   3720 

tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg   3780 

tgtggaaagt ccccaggctc cccaggcagg cagaagtatg caaagcatgc atctcaatta   3840 

gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat   3900 

gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac   3960 

tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga   4020 

ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg   4080 

cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga   4140 

gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc   4200 

cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga   4260 

tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct   4320 

gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac   4380 

gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct   4440 

attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt   4500 

atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt   4560 

cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt   4620 

cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag   4680 

gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt   4740 

gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg   4800 

tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg   4860 

cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg   4920 

catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg   4980 

accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat   5040 

gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg   5100 

gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac   5160 

aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt   5220 

tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc   5280 

tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca   5340 

attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg   5400 

agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg   5460 

tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc   5520 

tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta   5580 

tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag   5640 

aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg   5700 

tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg   5760 

tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg   5820 

cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga   5880 

agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc   5940 

tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt   6000 

aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact   6060 

ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg   6120 

cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt   6180 

accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt   6240 

ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct   6300 

ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg   6360 

gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt   6420 

aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt   6480 

gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc   6540 

gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg   6600 

cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc   6660 

gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg   6720 

gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca   6780 

ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga   6840 

tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct   6900 

ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg   6960 

cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca   7020 

accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata   7080 

cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct   7140 

tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact   7200 

cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa   7260 

acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc   7320 

atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga   7380 

tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga   7440 

aaagtgccac ctgacgtcga cggatcgggc gcctatgtat aaacttacat aaatcttttt   7500 

atttgtttat ccccaaggcg cgtgtaaagg ggcttttcac ggtggactgc agctgcctag   7560 

ccc                                                                 7563 

 
           
             6  
             7469  
             DNA  
             pFastFind-delta  
           
            6 

agatctcggc cgcatattaa gtgcattgtt ctcgataccg ctaagtgcat tgttctcgtt     60 

agctcgatgg acaagtgcat tgttctcttg ctgaaagctc gatggacaag tgcattgttc    120 

tcttgctgaa agctcgatgg acaagtgcat tgttctcttg ctgaaagctc agtacccggg    180 

agtaccctcg accgccggag tataaataga ggcgcttcgt ctacggagcg acaattcaat    240 

tcaaacaagc aaagtgaaca cgtcgctaag cgaaagctaa gcaaataaac aagcgcagga    300 

cttgttcgat ttgttagacg tcatttcacg ttcaatttca cttagttaat tttcattggt    360 

cgttggttca tttagttgac gttgatgact ttagacggtt cttcattaat aacttatgtt    420 

cttctcttga gacttatgaa agttgttgtt accgagaaag aagaactcac acacagctag    480 

cgtttaaact taagcttggt accgagctcg gatccactag tccagtgtgg tggaattctg    540 

cagatatcca gcacagtggc ggccgctcga gccaattccg cccctctccc tccccccccc    600 

ctaacgttac tggccgaagc cgcttggaat aaggccggtg tgcgtttgtc tatatgtgat    660 

tttccaccat attgccgtct tttggcaatg tgagggcccg gaaacctggc cctgtcttct    720 

tgacgagcat tcctaggggt ctttcccctc tcgccaaagg aatgcaaggt ctgttgaatg    780 

tcgtgaagga agcagttcct ctggaagctt cttgaagaca aacaacgtct gtagcgaccc    840 

tttgcaggca gcggaacccc ccacctggcg acaggtgcct ctgcggccaa aagccacgtg    900 

tataagatac acctgcaaag gcggcacaac cccagtgcca cgttgtgagt tggatagttg    960 

tggaaagagt caaatggctc tcctcaagcg tattcaacaa ggggctgaag gatgcccaga   1020 

aggtacccca ttgtatggga tctgatctgg ggcctcggtg cacatgcttt acatgtgttt   1080 

agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg ggacgtggtt ttcctttgaa   1140 

aaacacgatg ataagcttgc cacaacccgg tctagcccgg gctcgagatc tgcgatctaa   1200 

gtaagcttcg aatcgcgaat tcgcccacca tgctgctgct gctgctgctg ctgggcctga   1260 

ggctacagct ctccctgggc atcatcccag ttgaggagga gaacccggac ttctggaacc   1320 

gcgaggcagc cgaggccctg ggtgccgcca agaagctgca gcctgcacag acagccgcca   1380 

agaacctcat catcttcctg ggcgatggga tgggggtgtc tacggtgaca gctgccagga   1440 

tcctaaaagg gcagaagaag gacaaactgg ggcctgagat acccctggcc atggaccgct   1500 

tcccatatgt ggctctgtcc aagacataca atgtagacaa acatgtgcca gacgctggag   1560 

ccacagccac ggcctacctg tgcggggtca agggcaactt ccagaccatt ggcttgagtg   1620 

cagccgcccg ctttaaccag tgcaacacga cacgcggcaa cgaggtcatc tccgtgatga   1680 

atcgggccaa gaaagcaggg aagtcagtgg gagtggtaac caccacacga gtgcagcacg   1740 

cctcgccagc cggcacctac gcccacacgg tgaaccgcaa ctggtactcg gacgccgacg   1800 

tgcctgcctc ggcccgccag gaggggtgcc aggacatcgc tacgcagctc atctccaaca   1860 

tggacattga cgtgatccta ggtggaggcc gaaagtacat gtttcgcatg ggaaccccag   1920 

accctgagta cccagatgac tacagccaag gtgggaccag gctggacggg aagaatctgg   1980 

tgcaggaatg gctggcgaag cgccagggtg cccggtatgt gtggaaccgc actgagctca   2040 

tgcaggcttc cctggacccg tctgtgaccc atctcatggg tctctttgag cctggagaca   2100 

tgaaatacga gatccaccga gactccacac tggacccctc cctgatggag atgacagagg   2160 

ctgccctgcg cctgctgagc aggaaccccc gcggcttctt cctcttcgtg gagggtggtc   2220 

gcatcgacca tggtcatcat gaaagcaggg cttaccgggc actgactgag acgatcatgt   2280 

tcgacgacgc cattgagagg gcgggccagc tcaccagcga ggaggacacg ctgagcctcg   2340 

tcactgccga ccactcccac gtcttctcct tcggaggcta ccccctgcga gggagctcca   2400 

tcttcgggct ggcccctggc aaggcccggg acaggaaggc ctacacggtc ctcctatacg   2460 

gaaacggtcc aggctatgtg ctcaaggacg gcgcccggcc ggatgttacc gagagcgaga   2520 

gcgggagccc cgagtatcgg cagcagtcag cagtgcccct ggacgaagag acccacgcag   2580 

gcgaggacgt ggcggtgttc gcgcgcggcc cgcaggcgca cctggttcac ggcgtgcagg   2640 

agcagacctt catagcgcac gtcatggcct tcgccgcctg cctggagccc tacaccgcct   2700 

gcgacctggc gccccccgcc ggcaccaccg acgccgcgca cccgggttac tctagagtcg   2760 

gggcggccgg ccgcttcgag cagacatctc ccgggaatcc gcggctgcag gtcgacgaac   2820 

aaaaactcat ctcagaagag gatctgaatg ctgtgggcca ggacacgcag gaggtcatcg   2880 

tggtgccaca ctccttgccc tttaaggtgg tggtgatctc agccatcctg gccctggtgg   2940 

tgctcaccat catctccctt atcatcctca tcatgctttg gcagaagaag ccacgttagg   3000 

cggccgctcg agatcagcta gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc   3060 

ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg   3120 

tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag   3180 

gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga   3240 

caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag   3300 

ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt   3360 

ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc   3420 

tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg   3480 

catcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta   3540 

gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt   3600 

ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat   3660 

ctcggtctat tcttttgatt tataagggat tttggggatt tcggcctatt ggttaaaaaa   3720 

tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg   3780 

tgtggaaagt ccccaggctc cccaggcagg cagaagtatg caaagcatgc atctcaatta   3840 

gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat   3900 

gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac   3960 

tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga   4020 

ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg   4080 

cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga   4140 

gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc   4200 

cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga   4260 

tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct   4320 

gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac   4380 

gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct   4440 

attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt   4500 

atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt   4560 

cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt   4620 

cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag   4680 

gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt   4740 

gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg   4800 

tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg   4860 

cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg   4920 

catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg   4980 

accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat   5040 

gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg   5100 

gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac   5160 

aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt   5220 

tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc   5280 

tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca   5340 

attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg   5400 

agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg   5460 

tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc   5520 

tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta   5580 

tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag   5640 

aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg   5700 

tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg   5760 

tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg   5820 

cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga   5880 

agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc   5940 

tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt   6000 

aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact   6060 

ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg   6120 

cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt   6180 

accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt   6240 

ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct   6300 

ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg   6360 

gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt   6420 

aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt   6480 

gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc   6540 

gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg   6600 

cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc   6660 

gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg   6720 

gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca   6780 

ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga   6840 

tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct   6900 

ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg   6960 

cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca   7020 

accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata   7080 

cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct   7140 

tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact   7200 

cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa   7260 

acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc   7320 

atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga   7380 

tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga   7440 

aaagtgccac ctgacgtcga cggatcggg                                     7469 

 
           
             7  
             6980  
             DNA  
             pFastFind-point  
           
            7 

agatctcggc cgcatattaa gtgcattgtt ctcgataccg ctaagtgcat tgttctcgtt     60 

agctcgatgg acaagtgcat tgttctcttg ctgaaagctc gatggacaag tgcattgttc    120 

tcttgctgaa agctcgatgg acaagtgcat tgttctcttg ctgaaagctc agtacccggg    180 

agtaccctcg accgccggag tataaataga ggcgcttcgt ctacggagcg acaattcaat    240 

tcaaacaagc aaagtgaaca cgtcgctaag cgaaagctaa gcaaataaac aagcgcagct    300 

gaacaagcta aacaatctgc agtaaagtgc aagttaaagt gaatcaatta aaagtaacca    360 

gcaaccaagt aaatcaactg caactactga aatctgccaa gaagtaatta ttgaatacaa    420 

gaagagaact ctgaatactt tcaacaagtt accgagaaag aagaactcac acacagctag    480 

cgtttaaact taagcttggt accgagctcg gatccactag tccagtgtgg tggaattctg    540 

cagatatcca gcacagtggc ggccgctcga gccaattccg cccctctccc tccccccccc    600 

ctaacgttac tggccgaagc cgcttggaat aaggccggtg tgcgtttgtc tatatgtgat    660 

tttccaccat attgccgtct tttggcaatg tgagggcccg gaaacctggc cctgtcttct    720 

tgacgagcat tcctaggggt ctttcccctc tcgccaaagg aatgcaaggt ctgttgaatg    780 

tcgtgaagga agcagttcct ctggaagctt cttgaagaca aacaacgtct gtagcgaccc    840 

tttgcaggca gcggaacccc ccacctggcg acaggtgcct ctgcggccaa aagccacgtg    900 

tataagatac acctgcaaag gcggcacaac cccagtgcca cgttgtgagt tggatagttg    960 

tggaaagagt caaatggctc tcctcaagcg tattcaacaa ggggctgaag gatgcccaga   1020 

aggtacccca ttgtatggga tctgatctgg ggcctcggtg cacatgcttt acatgtgttt   1080 

agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg ggacgtggtt ttcctttgaa   1140 

aaacacgatg ataagcttgc cacaacccgg tctagcccgg gctcgagatc tgcgatctaa   1200 

gtaagcttcg aatcgcgaat tcgcccacca tgctgctgct gctgctgctg ctgggcctga   1260 

ggctacagct ctccctgggc atcatcccag ttgaggagga gaacccggac ttctggaacc   1320 

gcgaggcagc cgaggccctg ggtgccgcca agaagctgca gcctgcacag acagccgcca   1380 

agaacctcat catcttcctg ggcgatggga tgggggtgtc tacggtgaca gctgccagga   1440 

tcctaaaagg gcagaagaag gacaaactgg ggcctgagat acccctggcc atggaccgct   1500 

tcccatatgt ggctctgtcc aagacataca atgtagacaa acatgtgcca gacagtggag   1560 

ccacagccac ggcctacctg tgcggggtca agggcaactt ccagaccatt ggcttgagtg   1620 

cagccgcccg ctttaaccag tgcaacacga cacgcggcaa cgaggtcatc tccgtgatga   1680 

atcgggccaa gaaagcaggg gcgggcttct tcctcttcgt ggagggtggt cgcatcgacc   1740 

atggtcatca tgaaagcagg gcttaccggg cactgactga gacgatcatg ttcgacgacg   1800 

ccattgagag ggcgggccag ctcaccagcg aggaggacac gctgagcctc gtcactgccg   1860 

accactccca cgtcttctcc ttcggaggct accccctgcg agggagctcc atcttcgggc   1920 

tggcccctgg caaggcccgg gacaggaagg cctacacggt cctcctatac ggaaacggtc   1980 

caggctatgt gctcaaggac ggcgcccggc cggatgttac cgagagcgag agcgggagcc   2040 

ccgagtatcg gcagcagtca gcagtgcccc tggacgaaga gacccacgca ggcgaggacg   2100 

tggcggtgtt cgcgcgcggc ccgcaggcgc acctggttca cggcgtgcag gagcagacct   2160 

tcatagcgca cgtcatggcc ttcgccgcct gcctggagcc ctacaccgcc tgcgacctgg   2220 

cgccccccgc cggcaccacc gacgccgcgc acccgggtta ctctagagtc ggggcggccg   2280 

gccgcttcga gcagacatct cccgggaatc cgcggctgca ggtcgacgaa caaaaactca   2340 

tctcagaaga ggatctgaat gctgtgggcc aggacacgca ggaggtcatc gtggtgccac   2400 

actccttgcc ctttaaggtg gtggtgatct cagccatcct ggccctggtg gtgctcacca   2460 

tcatctccct tatcatcctc atcatgcttt ggcagaagaa gccacgttag gcggccgctc   2520 

gagatcagct agagggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg   2580 

ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc   2640 

cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc   2700 

tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag   2760 

gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc   2820 

tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac   2880 

gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc   2940 

ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg gcatcccttt   3000 

agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg   3060 

ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac   3120 

gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta   3180 

ttcttttgat ttataaggga ttttggggat ttcggcctat tggttaaaaa atgagctgat   3240 

ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag   3300 

tccccaggct ccccaggcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac   3360 

caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa   3420 

ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag   3480 

ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc   3540 

cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt   3600 

ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg   3660 

aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt   3720 

ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt   3780 

gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc   3840 

cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc   3900 

ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga   3960 

agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat   4020 

ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca   4080 

agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga   4140 

tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc   4200 

gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat   4260 

catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga   4320 

ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg   4380 

ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt   4440 

ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa   4500 

gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg   4560 

ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg   4620 

ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc   4680 

aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg   4740 

tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg   4800 

gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac   4860 

aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc   4920 

acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg   4980 

cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct   5040 

tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac   5100 

tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga   5160 

gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat   5220 

aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac   5280 

ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct   5340 

gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg   5400 

ctttctcaat gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg   5460 

ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt   5520 

cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg   5580 

attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac   5640 

ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga   5700 

aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt   5760 

gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt   5820 

tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga   5880 

ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc   5940 

taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct   6000 

atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata   6060 

actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca   6120 

cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga   6180 

agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga   6240 

gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg   6300 

gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga   6360 

gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt   6420 

gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct   6480 

cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca   6540 

ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat   6600 

accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga   6660 

aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc   6720 

aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg   6780 

caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc   6840 

ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt   6900 

gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca   6960 

cctgacgtcg acggatcggg                                               6980 

 
           
             8  
             8750  
             DNA  
             pFastFind-JNK3  
           
            8 

agatctcggc cgcatattaa gtgcattgtt ctcgataccg ctaagtgcat tgttctcgtt     60 

agctcgatgg acaagtgcat tgttctcttg ctgaaagctc gatggacaag tgcattgttc    120 

tcttgctgaa agctcgatgg acaagtgcat tgttctcttg ctgaaagctc agtacccggg    180 

agtaccctcg accgccggag tataaataga ggcgcttcgt ctacggagcg acaattcaat    240 

tcaaacaagc aaagtgaaca cgtcgctaag cgaaagctaa gcaaataaac aagcgcagct    300 

gaacaagcta aacaatctgc agtaaagtgc aagttaaagt gaatcaatta aaagtaacca    360 

gcaaccaagt aaatcaactg caactactga aatctgccaa gaagtaatta ttgaatacaa    420 

gaagagaact ctgaatactt tcaacaagtt accgagaaag aagaactcac acacagctag    480 

cgtttaaact taagcttggt accgagctcg gatccactag tacaaccatg gtgagcctcc    540 

atttcttata ctactgcagt gaaccaacat tggatgtgaa aattgccttt tgtcagggat    600 

tcgataaaca agtggatgtg tcatatattg ccaaacatta caacatgagc aaaagcaaag    660 

ttgacaacca gttctacagt gtggaagtgg gagactcaac cttcacagtt ctcaagcgct    720 

accagaatct aaagcctatt ggctctgggg ctcagggcat agtttgtgcc gcgtatgatg    780 

ctgtccttga cagaaatgtg gccattaaga agctcagcag accctttcag aaccaaacac    840 

atgccaagag agcgtaccgg gagctggtcc tcatgaagtg tgtgaaccat aaaaacatta    900 

ttagtttatt aaatgtcttc acaccccaga aaacgctgga ggagttccaa gatgtttact    960 

tagtaatgga actgatggat gccaacttat gtcaagtgat tcagatggaa ttagaccatg   1020 

agcgaatgtc ttacctgctg taccaaatgt tgtgtggcat taagcacctc cattctgctg   1080 

gaattattca cagggattta aaaccaagta acattgtagt caagtctgat tgcacattga   1140 

aaatcctgga ctttggactg gccaggacag caggcacaag cttcatgatg actccatatg   1200 

tggtgacacg ttattacaga gcccctgagg tcatcctggg gatgggctgc aaggagaacg   1260 

tggatatatg gtctgtggga tgcattatgg gagaaatggt tcgccacaaa atcctctttc   1320 

caggaaggga ctatattgac cagtggaata aggtaattga acaactagga acaccatgtc   1380 

cagaattcat gaagaaattg caacccacag taagaaacta tgtggagaat cggcccaagt   1440 

atgcgggact caccttcccc aaactcttcc cagattccct cttcccagcg gactccgagc   1500 

acaataaact caaagccagc caagccaggg acttgttgtc aaagatgcta gtgattgacc   1560 

cagcaaaaag aatatcagtg gacgacgcct tacagcatcc ctacatcaac gtctggtatg   1620 

acccagccga agtggaggcg cctccacctc agatatatga caagcagttg gatgaaagag   1680 

aacacacaat tgaagaatgg aaagaactta tctacaagga agtaatgaat tcagaagaaa   1740 

agactaaaaa tggtgtagta aaaggacagc cttctccttc agcacaggtg cagcagtgag   1800 

actacaagga cgatgatgac gccgatatcc agcacagtgg cggccgctcg agccaattcc   1860 

gcccctctcc ctcccccccc cctaacgtta ctggccgaag ccgcttggaa taaggccggt   1920 

gtgcgtttgt ctatatgtga ttttccacca tattgccgtc ttttggcaat gtgagggccc   1980 

ggaaacctgg ccctgtcttc ttgacgagca ttcctagggg tctttcccct ctcgccaaag   2040 

gaatgcaagg tctgttgaat gtcgtgaagg aagcagttcc tctggaagct tcttgaagac   2100 

aaacaacgtc tgtagcgacc ctttgcaggc agcggaaccc cccacctggc gacaggtgcc   2160 

tctgcggcca aaagccacgt gtataagata cacctgcaaa ggcggcacaa ccccagtgcc   2220 

acgttgtgag ttggatagtt gtggaaagag tcaaatggct ctcctcaagc gtattcaaca   2280 

aggggctgaa ggatgcccag aaggtacccc attgtatggg atctgatctg gggcctcggt   2340 

gcacatgctt tacatgtgtt tagtcgaggt taaaaaaacg tctaggcccc ccgaaccacg   2400 

gggacgtggt tttcctttga aaaacacgat gataagcttg ccacaacccg gtctagcccg   2460 

ggctcgagat ctgcgatcta agtaagcttc gaatcgcgaa ttcgcccacc atgctgctgc   2520 

tgctgctgct gctgggcctg aggctacagc tctccctggg catcatccca gttgaggagg   2580 

agaacccgga cttctggaac cgcgaggcag ccgaggccct gggtgccgcc aagaagctgc   2640 

agcctgcaca gacagccgcc aagaacctca tcatcttcct gggcgatggg atgggggtgt   2700 

ctacggtgac agctgccagg atcctaaaag ggcagaagaa ggacaaactg gggcctgaga   2760 

tacccctggc catggaccgc ttcccatatg tggctctgtc caagacatac aatgtagaca   2820 

aacatgtgcc agacagtgga gccacagcca cggcctacct gtgcggggtc aagggcaact   2880 

tccagaccat tggcttgagt gcagccgccc gctttaacca gtgcaacacg acacgcggca   2940 

acgaggtcat ctccgtgatg aatcgggcca agaaagcagg gaagtcagtg ggagtggtaa   3000 

ccaccacacg agtgcagcac gcctcgccag ccggcaccta cgcccacacg gtgaaccgca   3060 

actggtactc ggacgccgac gtgcctgcct cggcccgcca ggaggggtgc caggacatcg   3120 

ctacgcagct catctccaac atggacattg acgtgatcct aggtggaggc cgaaagtaca   3180 

tgtttcgcat gggaacccca gaccctgagt acccagatga ctacagccaa ggtgggacca   3240 

ggctggacgg gaagaatctg gtgcaggaat ggctggcgaa gcgccagggt gcccggtatg   3300 

tgtggaaccg cactgagctc atgcaggctt ccctggaccc gtctgtgacc catctcatgg   3360 

gtctctttga gcctggagac atgaaatacg agatccaccg agactccaca ctggacccct   3420 

ccctgatgga gatgacagag gctgccctgc gcctgctgag caggaacccc cgcggcttct   3480 

tcctcttcgt ggagggtggt cgcatcgacc atggtcatca tgaaagcagg gcttaccggg   3540 

cactgactga gacgatcatg ttcgacgacg ccattgagag ggcgggccag ctcaccagcg   3600 

aggaggacac gctgagcctc gtcactgccg accactccca cgtcttctcc ttcggaggct   3660 

accccctgcg agggagctcc atcttcgggc tggcccctgg caaggcccgg gacaggaagg   3720 

cctacacggt cctcctatac ggaaacggtc caggctatgt gctcaaggac ggcgcccggc   3780 

cggatgttac cgagagcgag agcgggagcc ccgagtatcg gcagcagtca gcagtgcccc   3840 

tggacgaaga gacccacgca ggcgaggacg tggcggtgtt cgcgcgcggc ccgcaggcgc   3900 

acctggttca cggcgtgcag gagcagacct tcatagcgca cgtcatggcc ttcgccgcct   3960 

gcctggagcc ctacaccgcc tgcgacctgg cgccccccgc cggcaccacc gacgccgcgc   4020 

acccgggtta ctctagagtc ggggcggccg gccgcttcga gcagacatct cccgggaatc   4080 

cgcggctgca ggtcgacgaa caaaaactca tctcagaaga ggatctgaat gctgtgggcc   4140 

aggacacgca ggaggtcatc gtggtgccac actccttgcc ctttaaggtg gtggtgatct   4200 

cagccatcct ggccctggtg gtgctcacca tcatctccct tatcatcctc atcatgcttt   4260 

ggcagaagaa gccacgttag gcggccgctc gagatcagct agagggcccg tttaaacccg   4320 

ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt   4380 

gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat   4440 

tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag   4500 

caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc   4560 

ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg   4620 

cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc   4680 

cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc   4740 

ccgtcaagct ctaaatcggg gcatcccttt agggttccga tttagtgctt tacggcacct   4800 

cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac   4860 

ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac   4920 

tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttggggat   4980 

ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg   5040 

tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccaggcag gcagaagtat   5100 

gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc   5160 

aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac   5220 

tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact   5280 

aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta   5340 

gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc   5400 

cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg   5460 

attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca   5520 

acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt   5580 

tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg   5640 

gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga   5700 

agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca   5760 

ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct   5820 

tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac   5880 

tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc   5940 

gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt   6000 

gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt   6060 

catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg   6120 

tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat   6180 

cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc   6240 

gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc   6300 

gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc   6360 

tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt   6420 

attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca   6480 

tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc   6540 

tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg   6600 

tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa   6660 

gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct   6720 

ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga   6780 

ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc   6840 

gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa   6900 

tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt   6960 

aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa   7020 

aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt   7080 

ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg   7140 

tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc   7200 

agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc   7260 

gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta   7320 

tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct   7380 

acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc   7440 

tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa   7500 

caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa   7560 

aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa   7620 

aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt   7680 

ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac   7740 

agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc   7800 

atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc   7860 

cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata   7920 

aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc   7980 

cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc   8040 

aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca   8100 

ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa   8160 

gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca   8220 

ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt   8280 

tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt   8340 

tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg   8400 

ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga   8460 

tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc   8520 

agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg   8580 

acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag   8640 

ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg   8700 

gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg              8750