From: GenBank-Updates@genbank.bio.net
Newsgroups: bionet.molbio.genbank.updates
Subject: Cloning vector (C.perfringens,E.coli) replication enzyme (rep)
Date: 15 Sep 92 10:06:14 GMT
Distribution: bionet
Checksum: 35818    15

LOCUS       SYNCLOSHP    7358 bp ds-DNA   Circular  SYN       15-SEP-1992
DEFINITION  Cloning vector (C.perfringens,E.coli) replication enzyme (rep)
            gene, complete cds; chloramphenicol acetyltransferase (catP) gene,
            complete cds; RNA methylase (ermBP) gene, complete cds; and
            beta-glactosidase alpha-peptide (lacZ') gene, complete cds.
ACCESSION   M77169
KEYWORDS    RNA methylase; beta-galactosidase alpha-peptide; catP gene;
            chloramphenicol acetyltransferase; ermBP gene; lacZ' gene;
            rep gene; replication enzyme; shuttle plasmid.
SOURCE      Cloning vector DNA.
  ORGANISM  Cloning vector
            Artificial sequences; Cloning vehicles.
REFERENCE   1  (bases 1 to 7358)
  AUTHORS   Sloan,J., Warner,T.A., Scott,P.T., Bannam,T.L., Berryman,D.I. and
            Rood,J.I.
  TITLE     Construction of a sequenced Clostridium perfringens - Escherichia
            coli shuttle plasmid
  JOURNAL   Plasmid 27, 207-219 (1992)
  STANDARD  simple staff_entry
FEATURES             Location/Qualifiers
     CDS             join(7345..7358,1..310)
                     /standard_name="lacZ'"
                     /note="derived from pUC18"
                     /product="beta-galactosidase alpha-peptide"
                     /gene="lacZ'"
                     /evidence=EXPERIMENTAL
                     /codon_start=1
                     /translation="MTMITNSSSVPGDPLESTCRHASLALAVVLQRRDWENPGVTQLN
                     RLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTI
                     CSDAA"
     misc_feature    1..57
                     /standard_name="multiple cloning site"
                     /evidence=EXPERIMENTAL
     misc_feature    1190..1419
                     /function="C. perfringens pIP404 replication origin"
                     /standard_name="ori"
                     /note="derived from complete sequence of pIP404 (Garnier &
                     Cole, Plasmid 19, 134-150, 1988; GenBank Accession number
                     M32882)"
                     /evidence=EXPERIMENTAL
     CDS             complement(1482..2702)
                     /function="pIP404 replication"
                     /standard_name="rep"
                     /note="derived from complete sequence of pIP404 (Garnier &
                     Cole, Plasmid 19, 134-150, 1988; GenBank Accession number
                     M32882)"
                     /product="replication enzyme"
                     /gene="rep"
                     /evidence=EXPERIMENTAL
                     /codon_start=1
                     /translation="VRNALKLNKKNYIDNIHSRSKGWITRSVIDKKGYSQWHYKYAEL
                     KDLDMSDENIYITLNTFYKPCRRLENIKELNTLFIDLDYYKTGKTKDQVLMDLEKNYF
                     NQSIPIPNYVIDSGRGMYLIWIINAVPSKALPLWKAVQEYLYNQLKYFGADRQALDAT
                     RILRVPGSINSKSKTVVNILDEYEYIYDLREIQNGFLPELKPYERKKGRPSKINYIYR
                     ERSLYYGRIQDIIKLCELREYDLKGHRELILFLYRYYLCSFTEDIEKALNDVLELNSM
                     FRQHLSEREVIRATRSAERCYLDKNKQYKYKNETLIELLEITEEEQKYMTIIISKKEY
                     KRRENIRGKKNYQEQLKAKGKATKKEELNVLRKKIKALKEKGFKNKEITLMLEVPIKT
                     LERHITYMKKNGLL"
                     /partial
     CDS             complement(4207..4830)
                     /standard_name="CAT"
                     /note="derived from C. perfringens catP (T. Bannam & J.
                     Rood, unpublished results; GenBank Accession number
                     M74769)"
                     /product="chloramphenicol acetyltransferase"
                     /gene="catP"
                     /evidence=EXPERIMENTAL
                     /codon_start=1
                     /translation="MVFEKIDKNSWNRKEYFDHYFASVPCTYSMTVKVDITQIKEKGM
                     KLYPAMLYYIAMIVNRHSEFRTAINQDGELGIYDEMIPSYTIFHNDTETFSSLWTECK
                     SDFKSFLADYESDTQRYGNNHRMEGKPNAPENIFNVSMIPWSTFDGFNLNLQKGYDYL
                     IPIFTMGKYYKEDNKIILPLAIQVHHAVCDGFHICRFVNELQELINS"
     CDS             complement(5165..5902)
                     /function="methylation of 23S rRNA"
                     /standard_name="RNA methylase"
                     /note="derived from sequence of C. perfringens ermBP gene
                     region (D. Berryman & J. Rood, unpublished results; EMBL
                     Accession number X58285)"
                     /product="RNA methylase"
                     /gene="ermBP"
                     /evidence=EXPERIMENTAL
                     /codon_start=1
                     /translation="MNKNIKYSQNFLTSEKVLNQIIKQLNLKETDTVYEIGTGKGHLT
                     TKLAKISKQVTSIELDSHLFNLSSEKLKLNTRVTLIHQDILQFQFPNKQRYKIVGSIP
                     YHLSTQIIKKVVFESHASDIYLIVEEGFYKRTLDIHRTLGLLLHTQVSIQQLLKLPAE
                     CFHPKPKVNSVLIKLTRHTTDVPDKYWKLYTYFVSKWVNREYRQLFTKNQFHQAMKHA
                     KVNNLSTVTYEQVLSIFNSYLLFNGRK"
     misc_feature    6780..7000
                     /function="replication region from pUC18"
                     /standard_name="oriEC"
                     /evidence=EXPERIMENTAL
BASE COUNT     2236 a   1404 c   1156 g   2562 t
ORIGIN
        1 gaattcgagc tcggtacccg gggatcctct agagtcgacc tgcaggcatg caagcttggc
       61 actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg
      121 ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg
      181 cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt attttctcct
      241 tacgcatctg tgcggtattt cacaccgcat atggtgcact ctcagtacaa tctgctctga
      301 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
      361 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
      421 tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct
      481 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
      541 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
      601 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
      661 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
      721 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
      781 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
      841 acgttttcca atgatgagca cttttaaatt aaaaatgaag ttttaaaact tcatttttaa
      901 tttaaattaa aaatgaagtt ttatcaaaaa aatttccaat aatcccactc taagccacaa
      961 acacgcccta taaaatcccg ctttaatccc actttgagac acatgtaata ttactttacg
     1021 ccctagtata gtgataattt tttacattca atgccacgca aaaaaataaa ggggcactat
     1081 aataaaagtt ccttcggaac taactaaagt aaaaaattat ctttacaacc tccccaaaaa
     1141 aaagaacagg tacaaagtac cctataatac aagcgtaaaa aaatgagggt aaaaataaaa
     1201 aaataaaaaa ataaaaaaat aaaaaaataa aaaaaataaa aaaataaaaa aataaaaaaa
     1261 taaaaaaata aaaaaataaa aaaataaaaa aataaaaaaa tataaaaata aaaaaatata
     1321 aaaataaaaa aatataaaaa taaaaaaata taaaaataaa aaaataaaaa aatataaaaa
     1381 taaaaaaata aaaaaatata aaaatatttt ttatttaaag tttgaaaaaa atttttttat
     1441 attatataat ctttgaagaa aagaatataa aaaatgagcc tttataaaag cccatttttt
     1501 ttcatatacg taatatgacg ttctaatgtt tttattggta cttctaacat tagagtaatt
     1561 tctttatttt taaagccttt ttctttaagg gcttttattt tttttcttaa tacatttaat
     1621 tcctcttttt ttgttgcttt tcctttagct tttaattgct cttgataatt ttttttacct
     1681 ctaatatttt ctcttctctt atattccttt ttagaaatta ttattgtcat atatttttgt
     1741 tcttcttctg taatttctaa taactctata agagtttcat tcttatactt atattgctta
     1801 tttttatcta aataacatct ttcagcactt ctagttgctc ttataacttc tctttcactt
     1861 aaatgttgtc taaacatact attaagttct aaaacatcat ttaatgcctt ctcaatgtct
     1921 tctgtaaagc tacaaagata atatctatat aaaaataata taagctctct gtgtcctttt
     1981 aaatcatatt ctcttagttc acaaagtttt attatgtctt gtattcttcc ataatataaa
     2041 cttctttctc tataaatata atttattttg cttggtctac cctttttcct ttcatatggt
     2101 tttaattcag gtaaaaatcc attttgtatt tctcttaagt cataaatata ttcgtactca
     2161 tctaatatat tgactactgt ttttgattta gagtttatac ttcctggaac tcttaatatt
     2221 ctggttgcat ctaaggcttg tctatctgct ccaaagtatt ttaattgatt atataaatat
     2281 tcttgaaccg ctttccataa tggtaatgct ttactaggta ctgcatttat tatccatatt
     2341 aaatacattc ctcttccact atctattaca tagtttggta taggaatact ttgattaaaa
     2401 taattctttt ctaagtccat taatacctgg tctttagttt tgccagtttt ataataatcc
     2461 aagtctataa acagtgtatt taactctttt atattttcta atcgcctaca cggcttataa
     2521 aaggtattta gagttatata gatattttca tcactcatat ctaaatcttt taattcagcg
     2581 tatttatagt gccattggct atatcctttt ttatctataa cgctcctggt tatccaccct
     2641 ttacttctac tatgaatatt atctatatag ttctttttat tcagctttaa tgcgtttctc
     2701 acttattcac ctccccttct gtaaaactaa gaaaattata tcatattttc aataattatt
     2761 aactattctt aaactcttaa taaaaaatag agtaagtccc caattgaaac ttaatctatt
     2821 ttttatgttt taatttatta tttttattaa aatattttaa actaaattaa atgattcttt
     2881 ttaatttttt actatttcat tccataatat attactataa ttatttacaa ataatatttc
     2941 ttcatttgta atatttagat gatttactaa ttttagtttt tatatattaa ataattaatg
     3001 tataatttat ataaaaaatc aaaggagctt ataaattatg attatttcca aagatactaa
     3061 agatttaatt ttttcaattt taacaatact ttttgtaata ttatgtttaa atttaattgt
     3121 atttttttca tataataaag ccgttgaagt aaaccaatcc attttcctta tgatgttatt
     3181 attaaattta agttttataa taatatcttt attatattta ttgtttttaa aaaaactagt
     3241 gaaatttccg gctttattaa acttattttt aggaatttta ttttcatttt catctttaca
     3301 ggatttgatt atatctttaa atatgtttta tcaaatatta tctttttcta aatttatata
     3361 tatttttatt atatttatta ttatatatat tttattttta agtttctttc taacagctat
     3421 taaaaagaaa cttaaaaata aaaacacgta ctctaaacca ataaataaaa ctatttttat
     3481 tattgctgcc ttgattggaa tagtttttag taaaattaat ttcaatattc cacaatatta
     3541 tattataagc tagctttgca ttgtactttt caatcgcttc acgaatgcgg ttatctccga
     3601 aagataaagt cttttcatct tccttgatga agataagatt ttctccgtct ccgccggcag
     3661 aattgaagcg gggtactacg gtatcgtctg cgtcatcttc cgttgtctga tagatgatag
     3721 tcataggctc attttcttcc gtttcggtaa aggggatagg ttcgcccttt gagagcaggg
     3781 cggcgatgga aagcattaac ttgcttttcc catcgcccgg atctccctgc aatagcgtaa
     3841 ctttgccaaa cggaatatac ggataccaca gccactttac ttctttcggc tcgatttcac
     3901 ttgccttgat gatttcaaga ggtacgctga aattcatttc gttttcattt agtttcattt
     3961 tttcttgttc tccttttctc tgaaaatata aaaaccacag attgatacta aaaccttggt
     4021 tgtgttgctt ttcggggctt aaatcaagga aaaatccttg ttttaagcct ttcaaaaaga
     4081 aacacaaggt ctttgtacta acctgtggtt atgtataaaa ttgtagattt tagggtaaca
     4141 aaaaacaccg tatttctacg atgtttttgc ttaaatactt gtttttagtt acagacaaac
     4201 ctgaagttaa ctatttatca attcctgcaa ttcgtttaca aaacggcaaa tgtgaaatcc
     4261 gtcacatact gcgtgatgaa cttgaattgc caaaggaagt ataattttgt tatcttcttt
     4321 ataatatttc cccatagtaa aaataggaat caaataatca tatcctttct gcaaattcag
     4381 attaaagcca tcgaaggttg accacggtat catagataca ttaaaaatgt tttccggagc
     4441 atttggcttt ccttccattc tatgattgtt tccataccgt tgcgtatcac tttcataatc
     4501 tgctaaaaat gatttaaagt cagacttaca ctcagtccaa aggctggaaa atgtttcagt
     4561 atcattgtga aatattgtat agcttggtat catctcatca tatatcccca attcaccatc
     4621 ttgattgatt gccgtcctaa actctgaatg gcggtttaca atcattgcaa tataataaag
     4681 cattgcagga tatagtttca ttcccttttc ctttatttgt gtgatatcca ctttaacggt
     4741 catgctgtat gtacaaggta cacttgcaaa gtagtggtca aaatactctt ttctgttcca
     4801 actattttta tcaatttttt caaataccat ctaagttccc tctcaaattc aagtttatcg
     4861 ctctaatgaa caaagatatt ataccacatt tttgtgaatt tttcaacttg cccacttcga
     4921 ctgcactccc gacttaataa cttcttgaac acttgccgaa aaagaaaaac tgccgggtac
     4981 gtacccggga tcgatccccg ccgagcgctt agtgggaatt tgtacccctt atcgatacaa
     5041 attccccgta ggcgctaggg acctctttag ctccttggaa gctgtcagta gtatacctaa
     5101 taatttatct acattccctt tagtaacgtg taactttcca aatttacaaa agcgactcat
     5161 agaattattt cctcccgtta aataatagat aactattaaa aatagacaat acttgctcat
     5221 aagtaacggt acttaaattg tttactttgg cgtgtttcat tgcttgatga aactgatttt
     5281 tagtaaacag ttgacgatat tctcgattga cccattttga aacaaagtac gtatatagct
     5341 tccaatattt atctggaaca tctgtggtat ggcgggtaag ttttattaag acactgttta
     5401 cttttggttt aggatgaaag cattccgctg gcagcttaag caattgctga atcgagactt
     5461 gagtgtgcaa gagcaaccct agtgttcggt gaatatccaa ggtacgcttg tagaatcctt
     5521 cttcaacaat cagatagatg tcagacgcat ggctttcaaa aaccactttt ttaataattt
     5581 gtgtgcttaa atggtaagga atactcccaa caattttata cctctgtttg ttagggaatt
     5641 gaaactgtag aatatcttgg tgaattaaag tgacacgagt attcagtttt aatttttctg
     5701 acgataagtt gaatagatga ctgtctaatt caatagacgt tacctgttta cttattttag
     5761 ccagtttcgt cgttaaatgc cctttacctg ttccaatttc gtaaacggta tcggtttctt
     5821 ttaaattcaa ttgttttatt atttggttga gtactttttc actcgttaaa aagttttgag
     5881 aatattttat atttttgttc atgtaatcac tccttcttaa ttacaaattt ttagcatcta
     5941 atttaacttc aattcctatt atacaaaatt ttaagatact gcactatcaa cacactctta
     6001 agtttgcttc taagtcttat ttccataact tcttttacgt ttccgggtac aattcgtaat
     6061 catgtcatag ctgtttcctg tgtgaaattc ttatccgctc acaattccac acaacatacg
     6121 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     6181 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccaga aaacttcatt
     6241 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     6301 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     6361 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     6421 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     6481 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
     6541 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     6601 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     6661 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     6721 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     6781 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     6841 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     6901 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     6961 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     7021 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     7081 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
     7141 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
     7201 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg
     7261 caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat
     7321 aacaatttca cacaggaaac agctatgacc atgattac
//