LOCUS       Exported                9283 bp DNA     circular SYN 
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9283)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 9283)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 9283)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 9283)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     SGRef: number: 2; type: "Journal Article"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..9283
                     /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      3960..4505
                     /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."
     misc_feature    5106..5151
                     /label=MCS
     protein_bind    complement(5168..5184)
                     /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(5192..5220)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     promoter        5454..5531
                     /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             5655..6614
                     /codon_start=1
                     /label=LacI (partial)
                     /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSR
                     ADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPA
                     LFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKY
                     LTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLR
                     VGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVS
                     LVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ"
     CDS             complement(7441..8088)
                     /codon_start=1
                     /label=TetR
                     /note="tetracycline resistance regulatory protein"
                     /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
                     RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
                     PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
                     QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKGDD"
     promoter        8194..8222
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             8411..8941
                     /codon_start=1
                     /label=GmR
                     /note="gentamycin acetyltransferase"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     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 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     3541 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     3601 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     3661 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     3721 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     3781 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     3841 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     3901 atcccttaac gtgagttttc gttccactga gcgtcagacc ccttaataag atgatcttct
     3961 tgagatcgtt ttggtctgcg cgtaatctct tgctctgaaa acgaaaaaac cgccttgcag
     4021 ggcggttttt cgaaggttct ctgagctacc aactctttga accgaggtaa ctggcttgga
     4081 ggagcgcagt caccaaaact tgtcctttca gtttagcctt aaccggcgca tgacttcaag
     4141 actaactcct ctaaatcaat taccagtggc tgctgccagt ggtgcttttg catgtctttc
     4201 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcggact gaacgggggg
     4261 ttcgtgcata cagtccagct tggagcgaac tgcctacccg gaactgagtg tcaggcgtgg
     4321 aatgagacaa acgcggccat aacagcggaa tgacaccggt aaaccgaaag gcaggaacag
     4381 gagagcgcac gagggagccg ccagggggaa acgcctggta tctttatagt cctgtcgggt
     4441 ttcgccacca ctgatttgag cgtcagattt cgtgatgctt gtcagggggg cggagcctat
     4501 ggaaaaacgg ctttgccgcg gccctctcac ttccctgtta agtatcttcc tggcatcttc
     4561 caggaaatct ccgccccgtt cgtaagccat ttccgctcgc cgcagtcgaa cgaccgagcg
     4621 tagcgagtca gtgagcgagg aagcggaata tatcctgtat cacatattct gctgacgcac
     4681 cggtgcagcc ttttttctcc tgccacatga agcacttcac tgacaccctc atcagtgcca
     4741 acatagtaag ccagtataca ctccgctagc gctgaggtct gcctcgtgaa gaaggtgttg
     4801 ctgactcata ccaggtagtc cgaggagctt tatgcttgta aaccgttttg tgaaaaaatt
     4861 tttaaaataa aaaaggggac ctctagggtc cccaattaat tagtaatata atctattaaa
     4921 ggtcattcaa aaggtcatcc accggatcaa ttcccctgct cgcgcaggct gggtgccaag
     4981 ctctcgggta acatcaaggc ccgatccttg gagcccttgc cctcccgcac gatgatcgtg
     5041 ccgtgatcga aatccagatc cttgacccgc agttgcaaac cctcactgat ccgctagtcc
     5101 gaggcctcga gatctatcga tgcatgccat ggtacccggg agctcgaatt ctgtttcctg
     5161 tgtgaaattg ttatccgctc acaattccac acattatacg agccgatgat taattgtcaa
     5221 cagctcattt cagaatattt gccagaaccg ttatgatgtc ggcgcaaaaa acattatcca
     5281 gaacgggagt gcgccttgag cgacacgaat tatgcagtga tttacgacct gcacagccat
     5341 accacagctt ccgatggctg cctgacgcca gaagcattgg tgcaccgtgc agtcgataag
     5401 cccggatcag cttgcaattc gcgcgcgaag gcgaagcggc atgcatttac gttgacacca
     5461 tcgaatggtg caaaaccttt cgcggtatgg catgatagcg cccggaagag agtcaattca
     5521 gggtggtgaa tgtgaaacca gtaacgttat acgatgtcgc agagtatgcc ggtgtctctt
     5581 atcagaccgt ttcccgcgtg gtgaaccagg ccagccacgt ttctgcgaaa acgcgggaaa
     5641 aagtggaagc ggcgatggcg gagctgaatt acattcccaa ccgcgtggca caacaactgg
     5701 cgggcaaaca gtcgttgctg attggcgttg ccacctccag tctggccctg cacgcgccgt
     5761 cgcaaattgt cgcggcgatt aaatctcgcg ccgatcaact gggtgccagc gtggtggtgt
     5821 cgatggtaga acgaagcggc gtcgaagcct gtaaagcggc ggtgcacaat cttctcgcgc
     5881 aacgcgtcag tgggctgatc attaactatc cgctggatga ccaggatgcc attgctgtgg
     5941 aagctgcctg cactaatgtt ccggcgttat ttcttgatgt ctctgaccag acacccatca
     6001 acagtattat tttctcccat gaagacggta cgcgactggg cgtggagcat ctggtcgcat
     6061 tgggtcacca gcaaatcgcg ctgttagcgg gcccattaag ttctgtctcg gcgcgtctgc
     6121 gtctggctgg ctggcataaa tatctcactc gcaatcaaat tcagccgata gcggaacggg
     6181 aaggcgactg gagtgccatg tccggttttc aacaaaccat gcaaatgctg aatgagggca
     6241 tcgttcccac tgcgatgctg gttgccaacg atcagatggc gctgggcgca atgcgcgcca
     6301 ttaccgagtc cgggctgcgc gttggtgcgg atatctcggt agtgggatac gacgataccg
     6361 aagacagctc atgttatatc ccgccgttaa ccaccatcaa acaggatttt cgcctgctgg
     6421 ggcaaaccag cgtggaccgc ttgctgcaac tctctcaggg ccaggcggtg aagggcaatc
     6481 agctgttgcc cgtctcactg gtgaaaagaa aaaccaccct ggcgcccaat acgcaaaccg
     6541 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     6601 aaagcgggca gtgagcgcaa cgcaattaat gtaagttagc gcgaattgca agctggccga
     6661 cgcgctgggc tacgtcttgc tggcgttcgg atccgatatc gccgtggcgg ccagcttttg
     6721 ccattctcac cggattcagt cgtcactcat ggtgatttct cacttgataa ccttattttt
     6781 gacgagggga aattaatagg ttgtattgat gttggacgag tcggaatcgc agaccgatac
     6841 caggatcttg ccatcctatg gaactgcctc ggtgagtttt ctccttcatt acagaaacgg
     6901 ctttttcaaa aatatggtat tgataatcct gatatgaata aattgcagtt tcatttgatg
     6961 ctcgatgagt ttttctaatc agaattggtt aattggttgt aacactggca gagcattacg
     7021 ctgacttgac gggacggcgg ctttgttgaa taaatcgaac ttttgctgag ttgaaggatc
     7081 agatcacgca tcttcccgac aacgcagacc gttccgtggc aaagcaaaag ttcaaaatca
     7141 ccaactggtc cacctacaac aaagctctca tcaaccgtgg ctccctcact ttctggctgg
     7201 atgatggggc gattcaggcc tggtatgagt cagcaacacc ttcttcacga ggcagacctc
     7261 agcgccagaa ggccgccaga gaggccgagc gcggccgtga ggcttggacg ctagggcagg
     7321 gcatgaaaaa gcccgtagcg ggctgctacg ggcgtctgac gcggtggaaa gggggagggg
     7381 atgttgtcta catggctctg ctgtagtgag tgggttgcgc tccggcagcg gtcctgatca
     7441 atcgtcaccc tttctcggtc cttcaacgtt cctgacaacg agcctccttt tcgccaatcc
     7501 atcgacaatc accgcgagtc cctgctcgaa cgctgcgtcc ggaccggctt cgtcgaaggc
     7561 gtctatcgcg gcccgcaaca gcggcgagag cggagcctgt tcaacggtgc cgccgcgctc
     7621 gccggcatcg ctgtcgccgg cctgctcctc aagcacggcc ccaacagtga agtagctgat
     7681 tgtcatcagc gcattgacgg cgtccccggc cgaaaaaccc gcctcgcaga ggaagcgaag
     7741 ctgcgcgtcg gccgtttcca tctgcggtgc gcccggtcgc gtgccggcat ggatgcgcgc
     7801 gccatcgcgg taggcgagca gcgcctgcct gaagctgcgg gcattcccga tcagaaatga
     7861 gcgccagtcg tcgtcggctc tcggcaccga atgcgtatga ttctccgcca gcatggcttc
     7921 ggccagtgcg tcgagcagcg cccgcttgtt cctgaagtgc cagtaaagcg ccggctgctg
     7981 aacccccaac cgttccgcca gtttgcgtgt cgtcagaccg tctacgccga cctcgttcaa
     8041 caggtccagg gcggcacgga tcactgtatt cggctgcaac tttgtcatgc ttgacacttt
     8101 atcactgata aacataatat gtccaccaac ttatcagtga taaagaatcc gcgcgttcaa
     8161 tcggaccagc ggaggctggt ccggaggcca gacttgacat aagcctgttc ggttcgtaaa
     8221 ctgtaatgca agtagcgtat gcgctcacgc aactggtcca gaaccttgac cgaacgcagc
     8281 ggtggtaacg gcgcagtggc ggttttcatg gcttgttatg actgtttttt tgtacagtct
     8341 atgcctcggg catccaagca gcaagcgcgt tacgccgtgg gtcgatgttt gatgttatgg
     8401 agcagcaacg atgttacgca gcagcaacga tgttacgcag cagggcagtc gccctaaaac
     8461 aaagttaggt ggctcaagta tgggcatcat tcgcacatgt aggctcggcc ctgaccaagt
     8521 caaatccatg cgggctgctc ttgatctttt cggtcgtgag ttcggagacg tagccaccta
     8581 ctcccaacat cagccggact ccgattacct cgggaacttg ctccgtagta agacattcat
     8641 cgcgcttgct gccttcgacc aagaagcggt tgttggcgct ctcgcggctt acgttctgcc
     8701 caagtttgag cagccgcgta gtgagatcta tatctatgat ctcgcagtct ccggagagca
     8761 ccggaggcag ggcattgcca ccgcgctcat caatctcctc aagcatgagg ccaacgcgct
     8821 tggtgcttat gtgatctacg tgcaagcaga ttacggtgac gatcccgcag tggctctcta
     8881 tacaaagttg ggcatacggg aagaagtgat gcactttgat atcgacccaa gtaccgccac
     8941 ctaatctaga actagctaga actagatcca acccctccgc tgctatagtg cagtcggctt
     9001 ctgacgttca gtgcagccgt cttctgaaaa cgacatgtcg cacaagtcct aagttacgcg
     9061 acaggctgcc gccctgccct tttcctggcg ttttcttgtc gcgtgtttta gtcgcataaa
     9121 gtagaatact tgcgactaga accggagaca ttacgccatg aacaagagcg ccgccgctgg
     9181 cctgctgggc tatgcccgcg tcagcaccga cgaccaggac ttgaccaacc aacgggccga
     9241 actgcacgcg gccggctgca ccaagctgtt ttccgagaag atc
//
