LOCUS       62056_16715        9347 bp DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9347)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9347
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             860..1486
                     /codon_start=1
                     /label=pVS1 StaA
                     /note="stability protein from the Pseudomonas plasmid pVS1
                     (Heeb et al., 2000)"
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             1918..2988
                     /codon_start=1
                     /label=pVS1 RepA
                     /note="replication protein from the Pseudomonas plasmid
                     pVS1 (Heeb et al., 2000)"
                     /translation="VSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     rep_origin      3057..3251
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid
                     pVS1 (Heeb et al., 2000)"
     rep_origin      3961..4506
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin."
     protein_bind    complement(5170..5186)
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(5194..5222)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     promoter        5456..5533
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             complement(7504..8151)
                     /codon_start=1
                     /label=TetR
                     /note="tetracycline resistance regulatory protein"
                     /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
                     RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
                     PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
                     QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKGDD"
     promoter        8257..8285
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             8474..9004
                     /codon_start=1
                     /label=GmR
                     /note="gentamycin acetyltransferase"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
ORIGIN
        1 accggcacca ggcgcgaccg cccggagctg gccaggatgc ttgaccacct acgccctggc
       61 gacgttgtga cagtgaccag gctagaccgc ctggcccgca gcacccgcga cctactggac
      121 attgccgagc gcatccagga ggccggcgcg ggcctgcgta gcctggcaga gccgtgggcc
      181 gacaccacca cgccggccgg ccgcatggtg ttgaccgtgt tcgccggcat tgccgagttc
      241 gagcgttccc taatcatcga ccgcacccgg agcgggcgcg aggccgccaa ggcccgaggc
      301 gtgaagtttg gcccccgccc taccctcacc ccggcacaga tcgcgcacgc ccgcgagctg
      361 atcgaccagg aaggccgcac cgtgaaagag gcggctgcac tgcttggcgt gcatcgctcg
      421 accctgtacc gcgcacttga gcgcagcgag gaagtgacgc ccaccgaggc caggcggcgc
      481 ggtgccttcc gtgaggacgc attgaccgag gccgacgccc tggcggccgc cgagaatgaa
      541 cgccaagagg aacaagcatg aaaccgcacc aggacggcca ggacgaaccg tttttcatta
      601 ccgaagagat cgaggcggag atgatcgcgg ccgggtacgt gttcgagccg cccgcgcacg
      661 tctcaaccgt gcggctgcat gaaatcctgg ccggtttgtc tgatgccaag ctggcggcct
      721 ggccggccag cttggccgct gaagaaaccg agcgccgccg tctaaaaagg tgatgtgtat
      781 ttgagtaaaa cagcttgcgt catgcggtcg ctgcgtatat gatgcgatga gtaaataaac
      841 aaatacgcaa ggggaacgca tgaaggttat cgctgtactt aaccagaaag gcgggtcagg
      901 caagacgacc atcgcaaccc atctagcccg cgccctgcaa ctcgccgggg ccgatgttct
      961 gttagtcgat tccgatcccc agggcagtgc ccgcgattgg gcggccgtgc gggaagatca
     1021 accgctaacc gttgtcggca tcgaccgccc gacgattgac cgcgacgtga aggccatcgg
     1081 ccggcgcgac ttcgtagtga tcgacggagc gccccaggcg gcggacttgg ctgtgtccgc
     1141 gatcaaggca gccgacttcg tgctgattcc ggtgcagcca agcccttacg acatatgggc
     1201 caccgccgac ctggtggagc tggttaagca gcgcattgag gtcacggatg gaaggctaca
     1261 agcggccttt gtcgtgtcgc gggcgatcaa aggcacgcgc atcggcggtg aggttgccga
     1321 ggcgctggcc gggtacgagc tgcccattct tgagtcccgt atcacgcagc gcgtgagcta
     1381 cccaggcact gccgccgccg gcacaaccgt tcttgaatca gaacccgagg gcgacgctgc
     1441 ccgcgaggtc caggcgctgg ccgctgaaat taaatcaaaa ctcatttgag ttaatgaggt
     1501 aaagagaaaa tgagcaaaag cacaaacacg ctaagtgccg gccgtccgag cgcacgcagc
     1561 agcaaggctg caacgttggc cagcctggca gacacgccag ccatgaagcg ggtcaacttt
     1621 cagttgccgg cggaggatca caccaagctg aagatgtacg cggtacgcca aggcaagacc
     1681 attaccgagc tgctatctga atacatcgcg cagctaccag agtaaatgag caaatgaata
     1741 aatgagtaga tgaattttag cggctaaagg aggcggcatg gaaaatcaag aacaaccagg
     1801 caccgacgcc gtggaatgcc ccatgtgtgg aggaacgggc ggttggccag gcgtaagcgg
     1861 ctgggttgtc tgccggccct gcaatggcac tggaaccccc aagcccgagg aatcggcgtg
     1921 agcggtcgca aaccatccgg cccggtacaa atcggcgcgg cgctgggtga tgacctggtg
     1981 gagaagttga aggccgcgca ggccgcccag cggcaacgca tcgaggcaga agcacgcccc
     2041 ggtgaatcgt ggcaagcggc cgctgatcga atccgcaaag aatcccggca accgccggca
     2101 gccggtgcgc cgtcgattag gaagccgccc aagggcgacg agcaaccaga ttttttcgtt
     2161 ccgatgctct atgacgtggg cacccgcgat agtcgcagca tcatggacgt ggccgttttc
     2221 cgtctgtcga agcgtgaccg acgagctggc gaggtgatcc gctacgagct tccagacggg
     2281 cacgtagagg tttccgcagg gccggccggc atggccagtg tgtgggatta cgacctggta
     2341 ctgatggcgg tttcccatct aaccgaatcc atgaaccgat accgggaagg gaagggagac
     2401 aagcccggcc gcgtgttccg tccacacgtt gcggacgtac tcaagttctg ccggcgagcc
     2461 gatggcggaa agcagaaaga cgacctggta gaaacctgca ttcggttaaa caccacgcac
     2521 gttgccatgc agcgtacgaa gaaggccaag aacggccgcc tggtgacggt atccgagggt
     2581 gaagccttga ttagccgcta caagatcgta aagagcgaaa ccgggcggcc ggagtacatc
     2641 gagatcgagc tagctgattg gatgtaccgc gagatcacag aaggcaagaa cccggacgtg
     2701 ctgacggttc accccgatta ctttttgatc gatcccggca tcggccgttt tctctaccgc
     2761 ctggcacgcc gcgccgcagg caaggcagaa gccagatggt tgttcaagac gatctacgaa
     2821 cgcagtggca gcgccggaga gttcaagaag ttctgtttca ccgtgcgcaa gctgatcggg
     2881 tcaaatgacc tgccggagta cgatttgaag gaggaggcgg ggcaggctgg cccgatccta
     2941 gtcatgcgct accgcaacct gatcgagggc gaagcatccg ccggttccta atgtacggag
     3001 cagatgctag ggcaaattgc cctagcaggg gaaaaaggtc gaaaaggtct ctttcctgtg
     3061 gatagcacgt acattgggaa cccaaagccg tacattggga accggaaccc gtacattggg
     3121 aacccaaagc cgtacattgg gaaccggtca cacatgtaag tgactgatat aaaagagaaa
     3181 aaaggcgatt tttccgccta aaactcttta aaacttatta aaactcttaa aacccgcctg
     3241 gcctgtgcat aactgtctgg ccagcgcaca gccgaagagc tgcaaaaagc gcctaccctt
     3301 cggtcgctgc gctccctacg ccccgccgct tcgcgtcggc ctatcgcggc cgctggccgc
     3361 tcaaaaatgg ctggcctacg gccaggcaat ctaccagggc gcggacaagc cgcgccgtcg
     3421 ccactcgacc gccggcgctg aggtctgcct cgtgaagaag gtgttgctga ctcataccag
     3481 ggcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     3541 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     3601 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     3661 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     3721 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     3781 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     3841 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     3901 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccttaataa gatgatcttc
     3961 ttgagatcgt tttggtctgc gcgtaatctc ttgctctgaa aacgaaaaaa ccgccttgca
     4021 gggcggtttt tcgaaggttc tctgagctac caactctttg aaccgaggta actggcttgg
     4081 aggagcgcag tcaccaaaac ttgtcctttc agtttagcct taaccggcgc atgacttcaa
     4141 gactaactcc tctaaatcaa ttaccagtgg ctgctgccag tggtgctttt gcatgtcttt
     4201 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcggac tgaacggggg
     4261 gttcgtgcat acagtccagc ttggagcgaa ctgcctaccc ggaactgagt gtcaggcgtg
     4321 gaatgagaca aacgcggcca taacagcgga atgacaccgg taaaccgaaa ggcaggaaca
     4381 ggagagcgca cgagggagcc gccaggggga aacgcctggt atctttatag tcctgtcggg
     4441 tttcgccacc actgatttga gcgtcagatt tcgtgatgct tgtcaggggg gcggagccta
     4501 tggaaaaacg gctttgccgc ggccctctca cttccctgtt aagtatcttc ctggcatctt
     4561 ccaggaaatc tccgccccgt tcgtaagcca tttccgctcg ccgcagtcga acgaccgagc
     4621 gtagcgagtc agtgagcgag gaagcggaat atatcctgta tcacatattc tgctgacgca
     4681 ccggtgcagc cttttttctc ctgccacatg aagcacttca ctgacaccct catcagtgcc
     4741 aacatagtaa gccagtatac actccgctag cgctgaggtc tgcctcgtga agaaggtgtt
     4801 gctgactcat accaggctag tccgaggagc tttatgcttg taaaccgttt tgtgaaaaaa
     4861 tttttaaaat aaaaaagggg acctctaggg tccccaatta attagtaata taatctatta
     4921 aaggtcattc aaaaggtcat ccaccggatc aattcccctg ctcgcgcagg ctgggtgcca
     4981 agctctcggg taacatcaag gcccgatcct tggagccctt gccctcccgc acgatgatcg
     5041 tgccgtgatc gaaatccaga tccttgaccc gcagttgcaa accctcactg atccgctagt
     5101 ccgaggcctc gagatctatc gatgcatgcc atggtacccg ggagctcgaa ttctgtttcc
     5161 tgtgtgaaat tgttatccgc tcacaattcc acacattata cgagccgatg attaattgtc
     5221 aacagctcat ttcagaatat ttgccagaac cgttatgatg tcggcgcaaa aaacattatc
     5281 cagaacggga gtgcgccttg agcgacacga attatgcagt gatttacgac ctgcacagcc
     5341 ataccacagc ttccgatggc tgcctgacgc cagaagcatt ggtgcaccgt gcagtcgata
     5401 agcccggatc agcttgcaat tcgcgcgcga aggcgaagcg gcatgcattt acgttgacac
     5461 catcgaatgg tgcaaaacct ttcgcggtat ggcatgatag cgcccggaag agagtcaatt
     5521 cagggtggtg aatgtgaaac cagtaacgtt atacgatgtc gcagagtatg ccggtgtctc
     5581 ttatcagacc gtttcccgcg tggtgaacca ggccagccac gtttctgcga aaacgcggga
     5641 aaaagtggaa gcggcgatgg cggagctgaa ttacattccc aaccgcgtgg cacaacaact
     5701 ggcgggcaaa cagtcgttgc tgattggcgt tgccacctcc agtctggccc tgcacgcgcc
     5761 gtcgcaaatt gtcgcggcga ttaaatctcg cgccgatcaa ctgggtgcca gcgtggtggt
     5821 gtcgatggta gaacgaagcg gcgtcgaagc ctgtaaagcg gcggtgcaca atcttctcgc
     5881 gcaacgcgtc agtgggctga tcattaacta tccgctggat gaccaggatg ccattgctgt
     5941 ggaagctgcc tgcactaatg ttccggcgtt atttcttgat gtctctgacc agacacccat
     6001 caacagtatt attttctccc atgaagacgg tacgcgactg ggcgtggagc atctggtcgc
     6061 attgggtcac cagcaaatcg cgctgttagc gggcccatta agttctgtct cggcgcgtct
     6121 gcgtctggct ggctggcata aatatctcac tcgcaatcaa attcagccga tagcggaacg
     6181 ggaaggcgac tggagtgcca tgtccggttt tcaacaaacc atgcaaatgc tgaatgaggg
     6241 catcgttccc actgcgatgc tggttgccaa cgatcagatg gcgctgggcg caatgcgcgc
     6301 cattaccgag tccgggctgc gcgttggtgc ggatatctcg gtagtgggat acgacgatac
     6361 cgaagacagc tcatgttata tcccgccgtt aaccaccatc aaacaggatt ttcgcctgct
     6421 ggggcaaacc acgtggaccg cttgctgcaa ctctctcagg gccaggcggt gaagggcaat
     6481 cagctgttgc ccgtctcact ggtgaaaaga aaaaccaccc tggcgcccaa tacgcaaacc
     6541 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
     6601 gaaagcgggc agtgagcgca acgcaattaa tgtaagttag cgcgaattgc aagctggccg
     6661 acgcgctggg ctacgtcttg ctggcgttcg gatccgatat cgccgtggcg gcttttgcca
     6721 ttctcaccgg attcagtcgt cactcatggt gatttctcac ttgataacct tatttttgac
     6781 gaggggaaat taataggttg tattgatgtt ggacgagtcg gaatcgcaga ccgataccag
     6841 gatcttgcca tcctatggaa ctgcctcggt gagttttctc cttcattaca gaaacggctt
     6901 tttcaaaaat atggtattga taatcctgat atgaataaat tgcagtttca tttgatgctc
     6961 gatgagtttt tctaatcaga attggttaat tggttgtaac actggcagag cattacgctg
     7021 acttgacggg acggcggctt tgttgaataa atcgaacttt tgctgagttg aaggatcaga
     7081 tcacgcatct tcccgacaac gcagaccgtt ccgtggcaaa gcaaaagttc aaaatcacca
     7141 actggtccac ctacaacaaa gctctcatca accgtggctc cctcactttc tggctggatg
     7201 atggggcgat tcaggcctgg tatgagtcag caacaccttc ttcacgaggc agacctcagc
     7261 gccaggcgca cgaggccggc gcgcggaccc gcgccacggc ccacgagcgg acgccgcagc
     7321 aggagcgcca gaaggccgcc agagaggccg agcgcggccg tgaggcttgg acgctagggc
     7381 agggcatgaa aaagcccgta gcgggctgct acgggcgtct gacgcggtgg aaagggggag
     7441 gggatgttgt ctacatggct ctgctgtagt gagtgggttg cgctccggca gcggtcctga
     7501 tcaatcgtca ccctttctcg gtccttcaac gttcctgaca acgagcctcc ttttcgccaa
     7561 tccatcgaca atcaccgcga gtccctgctc gaacgctgcg tccggaccgg cttcgtcgaa
     7621 ggcgtctatc gcggcccgca acagcggcga gagcggagcc tgttcaacgg tgccgccgcg
     7681 ctcgccggca tcgctgtcgc cggcctgctc ctcaagcacg gccccaacag tgaagtagct
     7741 gattgtcatc agcgcattga cggcgtcccc ggccgaaaaa cccgcctcgc agaggaagcg
     7801 aagctgcgcg tcggccgttt ccatctgcgg tgcgcccggt cgcgtgccgg catggatgcg
     7861 cgcgccatcg cggtaggcga gcagcgcctg cctgaagctg cgggcattcc cgatcagaaa
     7921 tgagcgccag tcgtcgtcgg ctctcggcac cgaatgcgta tgattctccg ccagcatggc
     7981 ttcggccagt gcgtcgagca gcgcccgctt gttcctgaag tgccagtaaa gcgccggctg
     8041 ctgaaccccc aaccgttccg ccagtttgcg tgtcgtcaga ccgtctacgc cgacctcgtt
     8101 caacaggtcc agggcggcac ggatcactgt attcggctgc aactttgtca tgcttgacac
     8161 tttatcactg ataaacataa tatgtccacc aacttatcag tgataaagaa tccgcgcgtt
     8221 caatcggacc agcggaggct ggtccggagg ccagacttga cataagcctg ttcggttcgt
     8281 aaactgtaat gcaagtagcg tatgcgctca cgcaactggt ccagaacctt gaccgaacgc
     8341 agcggtggta acggcgcagt ggcggttttc atggcttgtt atgactgttt ttttgtacag
     8401 tctatgcctc gggcatccaa gcagcaagcg cgttacgccg tgggtcgatg tttgatgtta
     8461 tggagcagca acgatgttac gcagcagcaa cgatgttacg cagcagggca gtcgccctaa
     8521 aacaaagtta ggtggctcaa gtatgggcat cattcgcaca tgtaggctcg gccctgacca
     8581 agtcaaatcc atgcgggctg ctcttgatct tttcggtcgt gagttcggag acgtagccac
     8641 ctactcccaa catcagccgg actccgatta cctcgggaac ttgctccgta gtaagacatt
     8701 catcgcgctt gctgccttcg accaagaagc ggttgttggc gctctcgcgg cttacgttct
     8761 gcccaggttt gagcagccgc gtagtgagat ctatatctat gatctcgcag tctccggcga
     8821 gcaccggagg cagggcattg ccaccgcgct catcaatctc ctcaagcatg aggccaacgc
     8881 gcttggtgct tatgtgatct acgtgcaagc agattacggt gacgatcccg cagtggctct
     8941 ctatacaaag ttgggcatac gggaagaagt gatgcacttt gatatcgacc caagtaccgc
     9001 cacctaatct agaactagct agaactatga tccaacccct ccgctgctat agtgcagtcg
     9061 gcttctgacg ttcagtgcag ccgtcttctg aaaacgacat gtcgcacaag tcctaagtta
     9121 cgcgacaggc tgccgccctg cccttttcct ggcgttttct tgtcgcgtgt tttagtcgca
     9181 taaagtagaa tacttgcgac tagaaccgga gacattacgc catgaacaag agcgccgccg
     9241 ctggcctgct gggctatgcc cgcgtcagca ccgacgacca ggacttgacc aaccaacggg
     9301 ccgaactgca cgcggccggc tgcaccaagc tgttttccga gaagatc
//
