LOCUS       pBBR1MCS        4711 bp DNA     circular SYN 18-JUL-2025
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4711)
  AUTHORS   caoheibi
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 4711)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 4711)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4711
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1023..1794
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             1795..2454
                     /codon_start=1
                     /label=pBBR1 Rep
                     /note="replication protein for the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica"
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     primer_bind     3166..3182
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3192..3210
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    3219..3326
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     promoter        complement(3339..3357)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3378..3394)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3402..3418)
                     /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(3426..3456)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3471..3492)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(3606..4238)
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQF"
     promoter        complement(4239..4341)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
ORIGIN
        1 ctcgggccgt ctcttgggct tgatcggcct tcttgcgcat ctcacgcgct cctgcggcgg
       61 cctgtagggc aggctcatac ccctgccgaa ccgcttttgt cagccggtcg gccacggctt
      121 ccggcgtctc aacgcgcttt gagattccca gcttttcggc caatccctgc ggtgcatagg
      181 cgcgtggctc gaccgcttgc gggctgatgg tgacgtggcc cactggtggc cgctccaggg
      241 cctcgtagaa cgcctgaatg cgcgtgtgac gtgccttgct gccctcgatg ccccgttgca
      301 gccctagatc ggccacagcg gccgcaaacg tggtctggtc gcgggtcatc tgcgctttgt
      361 tgccgatgaa ctccttggcc gacagcctgc cgtcctgcgt cagcggcacc acgaacgcgg
      421 tcatgtgcgg gctggtttcg tcacggtgga tgctggccgt cacgatgcga tccgccccgt
      481 acttgtccgc cagccacttg tgcgccttct cgaagaacgc cgcctgctgt tcttggctgg
      541 ccgacttcca ccattccggg ctggccgtca tgacgtactc gaccgccaac acagcgtcct
      601 tgcgccgctt ctctggcagc aactcgcgca gtcggcccat cgcttcatcg gtgctgctgg
      661 ccgcccagtg ctcgttctct ggcgtcctgc tggcgtcagc gttgggcgtc tcgcgctcgc
      721 ggtaggcgtg cttgagactg gccgccacgt tgcccatttt cgccagcttc ttgcatcgca
      781 tgatcgcgta tgccgccatg cctgcccctc ccttttggtg tccaaccggc tcgacggggg
      841 cagcgcaagg cggtgcctcc ggcgggccac tcaatgcttg agtatactca ctagactttg
      901 cttcgcaaag tcgtgaccgc ctacggcggc tgcggcgccc tacgggcttg ctctccgggc
      961 ttcgccctgc gcggtcgctg cgctcccttg ccagcccgtg gatatgtgga cgatggccgc
     1021 gagcggccac cggctggctc gcttcgctcg gcccgtggac aaccctgctg gacaagctga
     1081 tggacaggct gcgcctgccc acgagcttga ccacagggat tgcccaccgg ctacccagcc
     1141 ttcgaccaca tacccaccgg ctccaactgc gcggcctgcg gccttgcccc atcaattttt
     1201 ttaattttct ctggggaaaa gcctccggcc tgcggcctgc gcgcttcgct tgccggttgg
     1261 acaccaagtg gaaggcgggt caaggctcgc gcagcgaccg cgcagcggct tggccttgac
     1321 gcgcctggaa cgacccaagc ctatgcgagt gggggcagtc gaagggcgaa gcccgcccgc
     1381 ctgccccccg agcctcacgg cggcgagtgc gggggttcca agggggcagc gccaccttgg
     1441 gcaaggccga aggccgcgca gtcgatcaac aagccccgga ggggccactt tttgccggag
     1501 ggggagccgc gccgaaggcg tgggggaacc ccgcaggggt gcccttcttt gggcaccaaa
     1561 gaactagata tagggcgaaa tgcgaaagac ttaaaaatca acaacttaaa aaaggggggt
     1621 acgcaacagc tcattgcggc accccccgca atagctcatt gcgtaggtta aagaaaatct
     1681 gtaattgact gccactttta cgcaacgcat aattgttgtc gcgctgccga aaagttgcag
     1741 ctgattgcgc atggtgccgc aaccgtgcgg cacccctacc gcatggagat aagcatggcc
     1801 acgcagtcca gagaaatcgg cattcaagcc aagaacaagc ccggtcactg ggtgcaaacg
     1861 gaacgcaaag cgcatgaggc gtgggccggg cttattgcga ggaaacccac ggcggcaatg
     1921 ctgctgcatc acctcgtggc gcagatgggc caccagaacg ccgtggtggt cagccagaag
     1981 acactttcca agctcatcgg acgttctttg cggacggtcc aatacgcagt caaggacttg
     2041 gtggccgagc gctggatctc cgtcgtgaag ctcaacggcc ccggcaccgt gtcggcctac
     2101 gtggtcaatg accgcgtggc gtggggccag ccccgcgacc agttgcgcct gtcggtgttc
     2161 agtgccgccg tggtggttga tcacgacgac caggacgaat cgctgttggg gcatggcgac
     2221 ctgcgccgca tcccgaccct gtatccgggc gagcagcaac taccgaccgg ccccggcgag
     2281 gagccgccca gccagcccgg cattccgggc atggaaccag acctgccagc cttgaccgaa
     2341 acggaggaat gggaacggcg cgggcagcag cgcctgccga tgcccgatga gccgtgtttt
     2401 ctggacgatg gcgagccgtt ggagccgccg acacgggtca cgctgccgcg ccggtagcac
     2461 ttgggttgcg cagcaacccg taagtgcgct gttccagact atcggctgta gccgcctcgc
     2521 cgccctatac cttgtctgcc tccccgcgtt gcgtcgcggt gcatggagcc gggccacctc
     2581 gacctgaatg gaagccggcg gcacctcgct aacggattca ccgtttttat caggctctgg
     2641 gaggcagaat aaatgatcat atcgtcaatt attacctcca cggggagagc ctgagcaaac
     2701 tggcctcagg catttgagaa gcacacggtc acactgcttc cggtagtcaa taaaccggta
     2761 aaccagcaat agacataagc ggctatttaa cgaccctgcc ctgaaccgac gaccgggtcg
     2821 aatttgcttt cgaatttctg ccattcatcc gcttattatc acttattcag gcgtagcaac
     2881 caggcgttta agggcaccaa taactgcctt aaaaaaatta cgccccgccc tgccactcat
     2941 cgcagtacgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     3001 aacaaaatat taacgcttac aatttccatt cgccattcag gctgcgcaac tgttgggaag
     3061 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
     3121 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
     3181 gtgagcgcgc gtaatacgac tcactatagg gcgaattgga gctccaccgc ggtggcggcc
     3241 gctctagaac tagtggatcc cccgggctgc aggaattcga tatcaagctt atcgataccg
     3301 tcgacctcga gggggggccc ggtacccagc ttttgttccc tttagtgagg gttaattgcg
     3361 cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     3421 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     3481 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     3541 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgcatg
     3601 cataaaaact gttgtaattc attaagcatt ctgccgacat ggaagccatc acaaacggca
     3661 tgatgaacct gaatcgccag cggcatcagc accttgtcgc cttgcgtata atatttgccc
     3721 atggtgaaaa cgggggcgaa gaagttgtcc atattggcca cgtttaaatc aaaactggtg
     3781 aaactcaccc agggattggc tgagacgaaa aacatattct caataaaccc tttagggaaa
     3841 taggccaggt tttcaccgta acacgccaca tcttgcgaat atatgtgtag aaactgccgg
     3901 aaatcgtcgt ggtattcact ccagagcgat gaaaacgttt cagtttgctc atggaaaacg
     3961 gtgtaacaag ggtgaacact atcccatatc accagctcac cgtctttcat tgccatacgg
     4021 aattccggat gagcattcat caggcgggca agaatgtgaa taaaggccgg ataaaacttg
     4081 tgcttatttt tctttacggt ctttaaaaag gccgtaatat ccagctgaac ggtctggtta
     4141 taggtacatt gagcaactga ctgaaatgcc tcaaaatgtt ctttacgatg ccattgggat
     4201 atatcaacgg tggtatatcc agtgattttt ttctccattt tagcttcctt agctcctgaa
     4261 aatctcgata actcaaaaaa tacgcccggt agtgatctta tttcattatg gtgaaagttg
     4321 gaacctctta cgtgccgatc aacgtctcat tttcgccaaa agttggccca gggcttcccg
     4381 gtatcaacag ggacaccagg atttatttat tctgcgaagt gatcttccgt cacaggtatt
     4441 tattcgaaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
     4501 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcgccc gcgttcctgc
     4561 tggcgctggg cctgtttctg gcgctggact tcccgctgtt ccgtcagcag cttttcgccc
     4621 acggccttga tgatcgcggc ggccttggcc tgcatatccc gattcaacgg ccccagggcg
     4681 tccagaacgg gcttcaggcg ctcccgaagg t
//
