LOCUS       Exported                6058 bp DNA     circular SYN 30-MAY-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6058)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6058
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          join(4930..6058,1..4929)
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          join(4930..6058,1..4929)
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          join(5175..6058,1..5174)
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          join(5175..6058,1..5174)
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    151..172
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        187..217
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    225..241
                     /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)."
     primer_bind     249..265
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        286..304
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     334..350
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(445..1320)
                     /codon_start=1
                     /label=araC
                     /note="L-arabinose regulatory protein"
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1347..1631
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     complement(1671..1687)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(1720..1738)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1748..1764)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(2476..3135)
                     /codon_start=1
                     /label=pBBR1 Rep
                     /note="replication protein for the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica"
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     rep_origin      complement(3136..3907)
                     /direction=LEFT
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     promoter        4012..4063
                     /label=promoter for mob
     misc_feature    4030..4052
                     /label=RSA
                     /note="transfer origins (also called recombination site A 
                     [RSA]), is known as the specific site necessary for 
                     mobilization and recombination mediated by a Mob/Pre 
                     protein. "
     CDS             4131..5177
                     /codon_start=1
                     /product="MobV family relaxase from Pseudomonadota"
                     /label=mobV
                     /translation="MAAYAIMRCKKLAKMGNVAASLKHAYRERETPNADASRTPENEHW
                     AASSTDEAMGRLRELLPEKRRKDAVLAVEYVMTASPEWWKSASQEQQAAFFEKAHKWLA
                     DKYGADRIVTASIHRDETSPHMTAFVVPLTQDGRLSAKEFIGNKAQMTRDQTTFAAAVA
                     DLGLQRGIEGSKARHTRIQAFYEALERPPVGHVTISPQAVEPRAYAPQGLAEKLGISKR
                     VETPEAVADRLTKAVRQGYEPALQAAAGAREMRKKADQAQETARDLRERLKPVLDALGP
                     LNRDMQAKAAAIIKAVGEKLLTEQREVQRQKQAQRQQERGRAHFPEKCHLRRCDNLPNN
                     SAAGKPISA"
     CDS             complement(5192..5722)
                     /codon_start=1
                     /label=GmR
                     /note="gentamycin acetyltransferase"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(5911..5939)
                     /label=Pc promoter
                     /note="class 1 integron promoter"
ORIGIN
        1 ccatgtcggc agaatgctta atgaattaca acagttttta tgcatgcgcc caatacgcaa
       61 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga
      121 ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc
      181 ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca
      241 atttcacaca ggaaacagct atgaccatga ttacgccaag cgcgcaatta accctcacta
      301 aagggaacaa aagctgggta ccgggccccc cctcgaggtc gacggtatcg atgcataatg
      361 tgcctgtcaa atggacgaag cagggattct gcaaacccta tgctactccg tcaagccgtc
      421 aattgtctga ttcgttacca attatgacaa cttgacggct acatcattca ctttttcttc
      481 acaaccggca cggaactcgc tcgggctggc cccggtgcat tttttaaata cccgcgagaa
      541 atagagttga tcgtcaaaac caacattgcg accgacggtg gcgataggca tccgggtggt
      601 gctcaaaagc agcttcgcct ggctgatacg ttggtcctcg cgccagctta agacgctaat
      661 ccctaactgc tggcggaaaa gatgtgacag acgcgacggc gacaagcaaa catgctgtgc
      721 gacgctggcg atatcaaaat tgctgtctgc caggtgatcg ctgatgtact gacaagcctc
      781 gcgtacccga ttatccatcg gtggatggag cgactcgtta atcgcttcca tgcgccgcag
      841 taacaattgc tcaagcagat ttatcgccag cagctccgaa tagcgccctt ccccttgccc
      901 ggcgttaatg atttgcccaa acaggtcgct gaaatgcggc tggtgcgctt catccgggcg
      961 aaagaacccc gtattggcaa atattgacgg ccagttaagc cattcatgcc agtaggcgcg
     1021 cggacgaaag taaacccact ggtgatacca ttcgcgagcc tccggatgac gaccgtagtg
     1081 atgaatctct cctggcggga acagcaaaat atcacccggt cggcaaacaa attctcgtcc
     1141 ctgatttttc accaccccct gaccgcgaat ggtgagattg agaatataac ctttcattcc
     1201 cagcggtcgg tcgataaaaa aatcgagata accgttggcc tcaatcggcg ttaaacccgc
     1261 caccagatgg gcattaaacg agtatcccgg cagcagggga tcattttgcg cttcagccat
     1321 acttttcata ctcccgccat tcagagaaga aaccaattgt ccatattgca tcagacattg
     1381 ccgtcactgc gtcttttact ggctcttctc gctaaccaaa ccggtaaccc cgcttattaa
     1441 aagcattctg taacaaagcg ggaccaaagc catgacaaaa acgcgtaaca aaagtgtcta
     1501 taatcacggc agaaaagtcc acattgatta tttgcacggc gtcacacttt gctatgccat
     1561 agcattttta tccataagat tagcggatcc tacctgacgc tttttatcgc aactctctac
     1621 tgtttctcca tacccgtttt tttgggctag cgaattcctg cagcccgggg gatccactag
     1681 ttctagagcg gccgccaccg cggtggagct ccaattcgcc ctatagtgag tcgtattacg
     1741 cgcgctcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac
     1801 ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca
     1861 ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggaaattg taagcgttaa
     1921 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     1981 cgtactgcga tgagtggcag ggcggggcgt aattttttta aggcagttat tggtgccctt
     2041 aaacgcctgg ttgctacgcc tgaataagtg ataataagcg gatgaatggc agaaattcga
     2101 aagcaaattc gacccggtcg tcggttcagg gcagggtcgt taaatagccg cttatgtcta
     2161 ttgctggttt accggtttat tgactaccgg aagcagtgtg accgtgtgct tctcaaatgc
     2221 ctgaggccag tttgctcagg ctctccccgt ggaggtaata attgacgata tgatcattta
     2281 ttctgcctcc cagagcctga taaaaacggt gaatccgtta gcgaggtgcc gccggcttcc
     2341 attcaggtcg aggtggcccg gctccatgca ccgcgacgca acgcggggag gcagacaagg
     2401 tatagggcgg cgaggcggct acagccgata gtctggaaca gcgcacttac gggttgctgc
     2461 gcaacccaag tgctaccggc gcggcagcgt gacccgtgtc ggcggctcca acggctcgcc
     2521 atcgtccaga aaacacggct catcgggcat cggcaggcgc tgctgcccgc gccgttccca
     2581 ttcctccgtt tcggtcaagg ctggcaggtc tggttccatg cccggaatgc cgggctggct
     2641 gggcggctcc tcgccggggc cggtcggtag ttgctgctcg cccggataca gggtcgggat
     2701 gcggcgcagg tcgccatgcc ccaacagcga ttcgtcctgg tcgtcgtgat caaccaccac
     2761 ggcggcactg aacaccgaca ggcgcaactg gtcgcggggc tggccccacg ccacgcggtc
     2821 attgaccacg taggccgaca cggtgccggg gccgttgagc ttcacgacgg agatccagcg
     2881 ctcggccacc aagtccttga ctgcgtattg gaccgtccgc aaagaacgtc cgatgagctt
     2941 ggaaagtgtc ttctggctga ccaccacggc gttctggtgg cccatctgcg ccacgaggtg
     3001 atgcagcagc attgccgccg tgggtttcct cgcaataagc ccggcccacg cctcatgcgc
     3061 tttgcgttcc gtttgcaccc agtgaccggg cttgttcttg gcttgaatgc cgatttctct
     3121 ggactgcgtg gccatgctta tctccatgcg gtaggggtgc cgcacggttg cggcaccatg
     3181 cgcaatcagc tgcaactttt cggcagcgcg acaacaatta tgcgttgcgt aaaagtggca
     3241 gtcaattaca gattttcttt aacctacgca atgagctatt gcggggggtg ccgcaatgag
     3301 ctgttgcgta cccccctttt ttaagttgtt gatttttaag tctttcgcat ttcgccctat
     3361 atctagttct ttggtgccca aagaagggca cccctgcggg gttcccccac gccttcggcg
     3421 cggctccccc tccggcaaaa agtggcccct ccggggcttg ttgatcgact gcgcggcctt
     3481 cggccttgcc caaggtggcg ctgccccctt ggaacccccg cactcgccgc cgtgaggctc
     3541 ggggggcagg cgggcgggct tcgcccttcg actgccccca ctcgcatagg cttgggtcgt
     3601 tccaggcgcg tcaaggccaa gccgctgcgc ggtcgctgcg cgagccttga cccgccttcc
     3661 acttggtgtc caaccggcaa gcgaagcgcg caggccgcag gccggaggct tttccccaga
     3721 gaaaattaaa aaaattgatg gggcaaggcc gcaggccgcg cagttggagc cggtgggtat
     3781 gtggtcgaag gctgggtagc cggtgggcaa tccctgtggt caagctcgtg ggcaggcgca
     3841 gcctgtccat cagcttgtcc agcagggttg tccacgggcc gagcgaagcg agccagccgg
     3901 tggccgctcg cggccatcgt ccacatatcc acgggctggc aagggagcgc agcgaccgcg
     3961 cagggcgaag cccggagagc aagcccgtag ggcgccgcag ccgccgtagg cggtcacgac
     4021 tttgcgaagc aaagtctagt gagtatactc aagcattgag tggcccgccg gaggcaccgc
     4081 cttgcgctgc ccccgtcgag ccggttggac accaaaaggg aggggcaggc atggcggcat
     4141 acgcgatcat gcgatgcaag aagctggcga aaatgggcaa cgtggcggcc agtctcaagc
     4201 acgcctaccg cgagcgcgag acgcccaacg ctgacgccag caggacgcca gagaacgagc
     4261 actgggcggc cagcagcacc gatgaagcga tgggccgact gcgcgagttg ctgccagaga
     4321 agcggcgcaa ggacgctgtg ttggcggtcg agtacgtcat gacggccagc ccggaatggt
     4381 ggaagtcggc cagccaagaa cagcaggcgg cgttcttcga gaaggcgcac aagtggctgg
     4441 cggacaagta cggggcggat cgcatcgtga cggccagcat ccaccgtgac gaaaccagcc
     4501 cgcacatgac cgcgttcgtg gtgccgctga cgcaggacgg caggctgtcg gccaaggagt
     4561 tcatcggcaa caaagcgcag atgacccgcg accagaccac gtttgcggcc gctgtggccg
     4621 atctagggct gcaacggggc atcgagggca gcaaggcacg tcacacgcgc attcaggcgt
     4681 tctacgaggc cctggagcgg ccaccagtgg gccacgtcac catcagcccg caagcggtcg
     4741 agccacgcgc ctatgcaccg cagggattgg ccgaaaagct gggaatctca aagcgcgttg
     4801 agacgccgga agccgtggcc gaccggctga caaaagcggt tcggcagggg tatgagcctg
     4861 ccctacaggc cgccgcagga gcgcgtgaga tgcgcaagaa ggccgatcaa gcccaagaga
     4921 cggcccgaga ccttcgggag cgcctgaagc ccgttctgga cgccctgggg ccgttgaatc
     4981 gggatatgca ggccaaggcc gccgcgatca tcaaggccgt gggcgaaaag ctgctgacgg
     5041 aacagcggga agtccagcgc cagaaacagg cccagcgcca gcaggaacgc gggcgcgcac
     5101 atttccccga aaagtgccac ctgcggcgtt gtgacaattt accgaacaac tccgcggccg
     5161 ggaagccgat ctcggcttga acgaattgtt aggtggcggt acttgggtcg atatcaaagt
     5221 gcatcacttc ttcccgtatg cccaactttg tatagagagc cactgcggga tcgtcaccgt
     5281 aatctgcttg cacgtagatc acataagcac caagcgcgtt ggcctcatgc ttgaggagat
     5341 tgatgagcgc ggtggcaatg ccctgcctcc ggtgctctcc ggagactgcg agatcataga
     5401 tatagatctc actacgcggc tgctcaaact tgggcagaac gtaagccgcg agagcgccaa
     5461 caaccgcttc ttggtcgaag gcagcaagcg cgatgaatgt cttactacgg agcaagttcc
     5521 cgaggtaatc ggagtccggc tgatgttggg agtaggtggc tacgtctccg aactcacgac
     5581 cgaaaagatc aagagcagcc cgcatggatt tgacttggtc agggccgagc ctacatgtgc
     5641 gaatgatgcc catacttgag ccacctaact ttgttttagg gcgactgccc tgctgcgtaa
     5701 catcgttgct gctgcgtaac atcgttgctg ctccataaca tcaaacatcg acccacggcg
     5761 taacgcgctt gctgcttgga tgcccgaggc atagactgta caaaaaaaca gtcataacaa
     5821 gccatgaaaa ccgccactgc gccgttacca ccgctgcgtt cggtcaaggt tctggaccag
     5881 ttgcgtgagc gcatacgcta cttgcattac agtttacgaa ccgaacaggc ttatgtcaac
     5941 tgggttcgtg ccttcatccg tttccacggt gtgcgtccat gggcaaatat tatacgcaag
     6001 gcgacaaggt gctgatgccg ctggcgattc aggttcatca tgccgtttgt gatggctt
//
