LOCUS       Exported                9273 bp DNA     circular SYN 03-SEP-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9273)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9273
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          2442..2498
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     oriT            618..727
                     /note="incP origin of transfer"
     CDS             760..1131
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     rep_origin      1258..1969
                     /label=oriV
                     /note="incP origin of replication"
     misc_feature    2442..2498
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     promoter        complement(2505..2523)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2541..2557)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2565..2581
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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(2589..2619)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2634..2655
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2902..2930
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             2978..4168
                     /codon_start=1
                     /gene="tet"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSIIGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     CDS             4665..5813
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     CDS             complement(6942..7913)
                     /codon_start=1
                     /gene="repA"
                     /product="replication protein of plasmid pSa"
                     /label=RepA
                     /translation="MPKNNKAPGHRINEIIKTSLALEMEDAREAGLVGYMARCLVQATM
                     PHTDPKTSYFERTNGIVTLSIMGKPSIGLPYGSMPRTLLAWICTEAVRTKDPVLNLGRS
                     QSEFLQRLGMHTDGRYTATLRNQAQRLFSSMISLAGEQGNDFGIENVVIAKRAFLFWNP
                     KRPEDRALWDSTLTLTGDFFEEVTRSPVPIRIDYLHALRQSPLAMDIYTWLTYRVFLLR
                     AKGRPFVQIPWVALQAQFGSSYGSRARNSPELDDKARERAERAALASFKYNFKKRLREV
                     LIVYPEASDCIEDDGECLRIKSTRLHVTRAPGKGARIGPPPT"
     CDS             8560..9210
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline resistance regulatory protein"
                     /label=TetR
                     /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
                     RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
                     PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
                     QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKGDD"
ORIGIN
        1 cccctttcca ccgcgtcaga cgcccgtagc agcccgctac gggctttttc atgccctgcc
       61 ctagcgtcca agcctcacgg ccgcgctcgg cctctctggc ggccttctgg cgctcttccg
      121 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
      181 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
      241 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
      301 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
      361 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
      421 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
      481 cgcttttccg ctgcataacc ctgcttcggg gtcattatag cgattttttc ggtatatcca
      541 tcctttttcg cacgatatac aggattttgc caaagggttc gtgtagactt tccttggtgt
      601 atccaacggc gtcagccggg caggataggt gaagtaggcc cacccgcgag cgggtgttcc
      661 ttcttcactg tcccttattc gcacctggcg gtgctcaacg ggaatcctgc tctgcgaggc
      721 tggccggcta ccgccggcgt aacagatgag ggcaagcgga tggctgatga aaccaagcca
      781 accaggaagg gcagcccacc tatcaaggtg tactgccttc cagacgaacg aagagcgatt
      841 gaggaaaagg cggcggcggc cggcatgagc ctgtcggcct acctgctggc cgtcggccag
      901 ggctacaaaa tcacgggcgt cgtggactat gagcacgtcc gcgagctggc ccgcatcaat
      961 ggcgacctgg gccgcctggg cggcctgctg aaactctggc tcaccgacga cccgcgcacg
     1021 gcgcggttcg gtgatgccac gatcctcgcc ctgctggcga agatcgaaga gaagcaggac
     1081 gagcttggca aggtcatgat gggcgtggtc cgcccgaggg cagagccatg acttttttag
     1141 ccgctaaaac ggccgggggg tgcgcgtgat tgccaagcac gtccccatgc gctccatcaa
     1201 gaagagcgac ttcgcggagc tggtgaagta catcaccgac gagcaaggca agaccgagcg
     1261 cctttccgac gctcaccggg ctggttgccc tcgccgctgg gctggcggcc gtctatggcc
     1321 ctgcaaacgc gccagaaacg ccgtcgaagc cgtgtgcgag acaccgcggc cgccggcgtt
     1381 gtggatacct cgcggaaaac ttggccctca ctgacagatg aggggcggac gttgacactt
     1441 gaggggccga ctcacccggc gcggcgttga cagatgaggg gcaggctcga tttcggccgg
     1501 cgacgtggag ctggccagcc tcgcaaatcg gcgaaaacgc ctgattttac gcgagtttcc
     1561 cacagatgat gtggacaagc ctggggataa gtgccctgcg gtattgacac ttgaggggcg
     1621 cgactactga cagatgaggg gcgcgatcct tgacacttga ggggcagagt gctgacagat
     1681 gaggggcgca cctattgaca tttgaggggc tgtccacagg cagaaaatcc agcatttgca
     1741 agggtttccg cccgtttttc ggccaccgct aacctgtctt ttaacctgct tttaaaccaa
     1801 tatttataaa ccttgttttt aaccagggct gcgccctgtg cgcgtgaccg cgcacgccga
     1861 aggggggtgc ccccccttct cgaaccctcc cggcccgcta acgcgggcct cccatccccc
     1921 caggggctgc gcccctcggc cgcgaacggc ctcaccccaa aaatggcagc gctggcagtc
     1981 cttgccattg ccgggatcgg ggcagtaacg ggatgggcga tcagcccgag cgcgacgccc
     2041 ggaagcattg acgtgccgca ggtgctggca tcgacattca gcgaccaggt gccgggcagt
     2101 gagggcggcg gcctgggtgg cggcctgccc ttcacttcgg ccgtcggggc attcacggac
     2161 ttcatggcgg ggccggcaat ttttaccttg ggcattcttg gcatagtggt cgcgggtgcc
     2221 gtgctcgtgt tcgggggtgc gataaaccca gcgaaccatt tgaggtgata ggtaagatta
     2281 taccgaggta tgaaaacgag aattggacct ttacagaatt actctatgaa gcgccatatt
     2341 taaaaagcta ccaagacgaa gaggatgaag aggatgagga ggcagattgc cttgaatata
     2401 ttgacaatac tgataagata atatatcttt tatatagaag ataattcgag ctcggtaccc
     2461 ggggatcctc tagagtcgac ctgcaggcat gcaagcttga gtattctata gtgtcaccta
     2521 aatagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     2581 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     2641 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     2701 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     2761 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     2821 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     2881 aacatgaatt aattctcatg tttgacagct tatcatcgat tagctttaat gcggtagttt
     2941 atcacagtta aattgctaac gcagtcaggc accgtgtatg aaatctaaca atgcgctcat
     3001 cgtcatcctc ggcaccgtca ccctagatgc tgtaggcata ggcttggtta tgccggtact
     3061 gccgggcctc ttgcgggata tcgtccattc cgacagcatc gccagtcact atggcgtgct
     3121 gctagcgcta tatgcgttga tgcaatttct atgcgcaccc gttctcggag cactgtccga
     3181 ccgctttggc cgccgcccag tcctgctcgc ttcgctactt ggagccacta tcgactacgc
     3241 gatcatggcg accacacccg tcctgtggat tctctacgcc ggacgcatcg tggccggcat
     3301 caccggcgcc acaggtgcgg ttgctggcgc ctatatcgcc gacatcaccg atggggaaga
     3361 tcgggctcgc cacttcgggc tcatgagcgc ttgtttcggc gtgggtatgg tggcaggccc
     3421 cgtggccggg ggactgttgg gcgccatctc cttacatgca ccattccttg cggcggcggt
     3481 gctcaacggc ctcaacctac tactgggctg cttcctaatg caggagtcgc ataagggaga
     3541 gcgccgaccc atgcccttga gagccttcaa cccagtcagc tccttccggt gggcgcgggg
     3601 catgactatc gtcgccgcac ttatgactgt cttctttatc atgcaactcg taggacaggt
     3661 gccggcagcg ctctgggtca ttttcggcga ggaccgcttt cgctggagcg cgacgatgat
     3721 cggcctgtcg cttgcggtat tcggaatctt gcacgccctc gctcaagcct tcgtcactgg
     3781 tcccgccacc aaacgtttcg gcgagaagca ggccattatc gccggcatgg cggccgacgc
     3841 gctgggctac gtcttgctgg cgttcgcgac gcgaggctgg atggccttcc ccattatgat
     3901 tcttctcgct tccggcggca tcgggatgcc cgcgttgcag gccatgctgt ccaggcaggt
     3961 agatgacgac catcagggac agcttcaagg atcgctcgcg gctcttacca gcctaacttc
     4021 gatcattgga ccgctgatcg tcacggcgat ttatgccgcc tcggcgagca catggaacgg
     4081 gttggcatgg attgtaggcg ccgccctata ccttgtctgc ctccccgcgt tgcgtcgcgg
     4141 tgcatggagc cgggccacct cgacctgaat ggaagccggc ggcacctcgc taacggattc
     4201 accactccaa gaattggagc caatcaattc ttgcggagaa ctgtgaatgc gcaaaccaac
     4261 ccttgatcgg ggaagaacag tatgtcgagc tattttttga cttactgggg atcaagcctg
     4321 attgggagaa aataaaatat tatattttac tggatgaatt gttttagtac ctagatgtgg
     4381 cgcaacgatg ccggcgacaa gcaggagcgc accgacttct tccgatcaag tgttttggct
     4441 ctcaggccga ggcccacggc aagtatttgg gcaaggggtc gctggtattc gtgcagggca
     4501 agattcggaa taccaagtac gagaaggacg gccagacggt ctacgggacc gacttcattg
     4561 ccgataaggt ggattatctg gacaccaagg caccaggcgg gtcaaatcag gaataagggc
     4621 acattgcccc ggcgtgagtc ggggcaatcc cgcaaggagg gtgaatgaat cggacgtttg
     4681 accggaaggc atacaggcaa gaactgatcg acgcggggtt ttccgccgag gatgccgaaa
     4741 ccatcgcaag ccgcaccgtc atgcgtgcgc cccgcgaaac cttccagtcc gtcggctcga
     4801 tggtccagca agctacggcc aagatcgagc gcgacagcgt gcaactggct ccccctgccc
     4861 tgcccgcgcc atcggccgcc gtggagcgtt cgcgtcgtct cgaacaggag gcggcaggtt
     4921 tggcgaagtc gatgaccatc gacacgcgag gaactatgac gaccaagaag cgaaaaaccg
     4981 ccggcgagga cctggcaaaa caggtcagcg aggccaagca ggccgcgttg ctgaaacaca
     5041 cgaagcagca gatcaaggaa atgcagcttt ccttgttcga tattgcgccg tggccggaca
     5101 cgatgcgagc gatgccaaac gacacggccc gctctgccct gttcaccacg cgcaacaaga
     5161 aaatcccgcg cgaggcgctg caaaacaagg tcattttcca cgtcaacaag gacgtgaaga
     5221 tcacctacac cggcgtcgag ctgcgggccg acgatgacga actggtgtgg cagcaggtgt
     5281 tggagtacgc gaagcgcacc cctatcggcg agccgatcac cttcacgttc tacgagcttt
     5341 gccaggacct gggctggtcg atcaatggcc ggtattacac gaaggccgag gaatgcctgt
     5401 cgcgcctaca ggcgacggcg atgggcttca cgtccgaccg cgttgggcac ctggaatcgg
     5461 tgtcgctgct gcaccgcttc cgcgtcctgg accgtggcaa gaaaacgtcc cgttgccagg
     5521 tcctgatcga cgaggaaatc gtcgtgctgt ttgctggcga ccactacacg aaattcatat
     5581 gggagaagta ccgcaagctg tcgccgacgg cccgacggat gttcgactat ttcagctcgc
     5641 accgggagcc gtacccgctc aagctggaaa ccttccgcct catgtgcgga tcggattcca
     5701 cccgcgtgaa gaagtggcgc gagcaggtcg gcgaagcctg cgaagagttg cgaggcagcg
     5761 gcctggtgga acacgcctgg gtcaatgatg acctggtgca ttgcaaacgc tagggccttg
     5821 tggggtcagt tccggctggg ggttcagcag ccagcgcttt actggcattt caggaacaag
     5881 cgggcactgc tcgacgcact tgcttcgctc agtatcgctc gggacgcacg gcgcgctcta
     5941 cgaactgccg ataaacagag gattaaaatt gacaattgtg attaaggctc agattcgacg
     6001 gcttggagcg gccgacgtgc aggatttccg cgagatccga ttgtcggccc tgaagaaagc
     6061 tccagagatg ttcgggtccg tttacgagca cgaggagaaa aagcccatgg aggcgttcgc
     6121 tgaacggttg cgagatgccg tggcattcgg cgcctacatc gacggcgaga tcattgggct
     6181 gtcggtcttc aaacaggagg acggccccaa ggacgctcac aaggcgcatc tgtccggcgt
     6241 tttcgtggag cccgaacagc gaggccgagg ggtcgccggt atgctgctgc gggcgttgcc
     6301 ggcgggttta ttgctcgtga tgatcgtccg acagattcca acgggaatct ggtggatgcg
     6361 catcttcatc ctcggcgcac ttaatatttc gctattctgg agcttgttgt ttatttcggt
     6421 ctaccgcctg ccgggcgggg tcgcggcgac ggtaggcgct gtgcagccgc tgatggtcgt
     6481 gttcatctct gccgctctgc taggtagccc gatacgattg atggcggtcc tgggggctat
     6541 ttgcggaact gcgggcgtgg cgctgttggt gttgacacca aacgcagcgc tagatcctgt
     6601 cggcgtcgca gcgggcctgg cgggggcggt ttccatggcg ttcggaaccg tgctgacccg
     6661 caagtggcaa cctcccgtgc ctctgctcac ctttaccgcc tggcaactgg cggccggagg
     6721 acttctgctc gttccagtag ctttagtgtt tgatccgcca atcccgatgc ctacaggaac
     6781 caatgttctc ggcctggcgt ggctcggcct gatcggagcg ggtttaacct acttcctttg
     6841 gttccggggg atctcgcgac tcgaacctac agttgtttcc ttactgggct ttctcagccc
     6901 cagatcgatc cttgaccaag ccaagcgtag cgttggcctg gtcaagtcgg aggggggccg
     6961 atgcgagcgc ccttgccggg tgcgcgggtg acatgcaggc gtgtggattt gatgcgcagg
     7021 cattcgccgt catcttcgat gcagtcgctt gcctcgggat agacaatcaa cacttcgcgt
     7081 aggcgctttt tgaagttgta tttgaagctg gcgagtgctg cccgctctgc ccgctctcgg
     7141 gccttatcgt ccagttcggg cgagttgcgt gcgcggctgc cataggatga gccgaattgc
     7201 gcttgcaggg cgacccaagg gatttgcacg aaggggcggc ccttggcccg caacaggaac
     7261 acgcgatagg tcagccacgt gtaaatgtcc atcgcaagcg gagactgccg caaggcgtgc
     7321 aggtagtcga ttcggatagg aaccggtgag cgggtgactt cctcgaagaa atcgcctgtg
     7381 agggtgaggg tgctatccca tagcgcccga tcttctggcc gcttgggatt ccagaataga
     7441 aaagcgcgct tggcaatgac gacgttctca atgccgaagt cattgccttg ctcgccggca
     7501 agcgaaatca tggatgaaaa caggcgttgc gcctgattgc gaagggtggc cgtgtaacgg
     7561 ccatcggtgt gcattccgag cctttgtaga aattccgatt gcgaccggcc aaggttcaac
     7621 acggggtctt tcgttcgcac ggcctcggtg catatccaag caagcaaggt gcgcggcata
     7681 gaaccgtagg gcaggccgat gctcggcttg cccatgatcg acaaggtgac gatgccattg
     7741 gtgcgctcaa agtagctggt cttggggtcg gtgtggggca tggtcgcttg cacaaggcaa
     7801 cgggccatgt agccgactaa gccagcttcg cgggcatcct ccatttcgag cgcgaggctc
     7861 gtcttgatga tctcgttgat acgatggccg ggggctttgt tgttcttagg catgttgttc
     7921 cctccccggc atggtgatgg ttggtctagt gtttgtgggt ttgatgttcc ggcgtttgat
     7981 gaacaggcgc aaggtgtgag ggctgacgcc taacaactcg gctgcgcgac tttgcggcaa
     8041 gccaaggttc acgtatgcct gtacttcatc aatacggctg tccagcttca aggcgctcga
     8101 tttgctgccc ttgggtcgcc cgagcgtctt gccgcgctct ctggcgactt gtagcgcctc
     8161 ggtggtacgt gcctgaatga aatgccgctc gatcctctag gatctggcgc cggccagcga
     8221 gacgagcaag attggccgcc gcccgaaacg atccgacagc gcgcccagca caggtgcgca
     8281 ggcaaattgc accaacgcat acagcgccag cagaatgcca tagtgggcgg tgacgtcgtt
     8341 cgagtgaacc agatcgcgca ggaggcccgg cagcaccggc ataatcaggc cgatgccgac
     8401 agcgtcgagc gcgacagtgc tcagaattac gatcaggggt atgttgggtt tcacgtctgg
     8461 cctccggacc agcctccgct ggtccgattg aacgcgcgga ttctttatca ctgataagtt
     8521 ggtggacata ttatgtttat cagtgataaa gtgtcaagca tgacaaagtt gcagccgaat
     8581 acagtgatcc gtgccgccct ggacctgttg aacgaggtcg gcgtagacgg tctgacgaca
     8641 cgcaaactgg cggaacggtt gggggttcag cagccggcgc tttactggca cttcaggaac
     8701 aagcgggcgc tgctcgacgc actggccgaa gccatgctgg cggagaatca tacgcattcg
     8761 gtgccgagag ccgacgacga ctggcgctca tttctgatcg ggaatgcccg cagcttcagg
     8821 caggcgctgc tcgcctaccg cgatggcgcg cgcatccatg ccggcacgcg accgggcgca
     8881 ccgcagatgg aaacggccga cgcgcagctt cgcttcctct gcgaggcggg tttttcggcc
     8941 ggggacgccg tcaatgcgct gatgacaatc agctacttca ctgttggggc cgtgcttgag
     9001 gagcaggccg gcgacagcga tgccggcgag cgcggcggca ccgttgaaca ggctccgctc
     9061 tcgccgctgt tgcgggccgc gatagacgcc ttcgacgaag ccggtccgga cgcagcgttc
     9121 gagcagggac tcgcggtgat tgtcgatgga ttggcgaaaa ggaggctcgt tgtcaggaac
     9181 gttgaaggac cgagaaaggg tgacgattga tcaggaccgc tgccggagcg caacccactc
     9241 actacagcag agccatgtag acaacatccc ctc
//
