LOCUS       sequence-488566-        5719 bp    DNA     circular SYN 07-OCT-2025
DEFINITION  pBBR1MCS replicative vector for increasing biofilm attachment.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5719)
  AUTHORS   Nevot G, Pol Cros M, Toloza L, Campama-Sanz N, Artigues-Lleixa M, 
            Aguilera L, Guell M
  TITLE     Engineered Marine Biofilms for Ocean Environment Monitoring.
  JOURNAL   ACS Synth Biol. 2025 Jun 22. doi: 10.1021/acssynbio.5c00192.
  PUBMED    40545707
REFERENCE   2  (bases 1 to 5719)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 7, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "ACS Synth 
            Biol. 2025 Jun 22. doi: 10.1021/acssynbio.5c00192."
COMMENT     Sequence Label: pDS_P(aphII)-dsCsgA-Mfp3-SP
FEATURES             Location/Qualifiers
     source          1..5719
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             571..1233
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALIETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     terminator      complement(1800..1847)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     1843..1861
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(1920..1940)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     RBS             2930..2952
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    2967..2991
                     /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)."
     primer_bind     complement(2991..3010)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2992..3010)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3192..3213
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     3874..3893
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(3960..4062)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(4179..4197)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     rep_origin      join(5518..5719,1..570)
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
ORIGIN
        1 ccggcctgcg gcctgcgcgc ttcgcttgcc ggttggacac caagtggaag gcgggtcaag
       61 gctcgcgcag cgaccgcgca gcggcttggc cttgacgcgc ctggaacgac ccaagcctat
      121 gcgagtgggg gcagtcgaag ggcgaagccc gcccgcctgc cccccgagcc tcacggcggc
      181 gagtgcgggg gttccaaggg ggcagcgcca ccttgggcaa ggccgaaggc cgcgcagtcg
      241 atcaacaagc cccggagggg ccactttttg ccggaggggg agccgcgccg aaggcgtggg
      301 ggaaccccgc aggggtgccc ttctttgggc accaaagaac tagatatagg gcgaaatgcg
      361 aaagacttaa aaatcaacaa cttaaaaaag gggggtacgc aacagctcat tgcggcaccc
      421 cccgcaatag ctcattgcgt aggttaaaga aaatctgtaa ttgactgcca cttttacgca
      481 acgcataatt gttgtcgcgc tgccgaaaag ttgcagctga ttgcgcatgg tgccgcaacc
      541 gtgcggcacc cctaccgcat ggagataagc atggccacgc agtccagaga aatcggcatt
      601 caagccaaga acaagcccgg tcactgggtg caaacggaac gcaaagcgca tgaggcgtgg
      661 gccgggctta ttgcgaggaa acccacggcg gcaatgctgc tgcatcacct cgtggcgcag
      721 atgggccacc agaacgccgt ggtggtcagc cagaagacac tttccaagct catcggacgt
      781 tctttgcgga cggtccaata cgcagtcaag gacttggtgg ccgagcgctg gatctccgtc
      841 gtgaagctca acggccccgg caccgtgtcg gcctacgtgg tcaatgaccg cgtggcgtgg
      901 ggccagcccc gcgaccagtt gcgcctgtcg gtgttcagtg ccgccgtggt ggttgatcac
      961 gacgaccagg acgaatcgct gttggggcat ggcgacctgc gccgcatccc gaccctgtat
     1021 ccgggcgagc agcaactacc gaccggcccc ggcgaggagc cgcccagcca gcccggcatt
     1081 ccgggcatgg aaccagacct gccagccttg atcgaaacgg aggaatggga acggcgcggg
     1141 cagcagcgcc tgccgatgcc cgatgagccg tgttttctgg acgatggcga gccgttggag
     1201 ccgccgacac gggtcacgct gccgcgccgg tagcacttgg gttgcgcagc aacccgtaag
     1261 tgcgctgttc cagactatcg gctgtagccg cctcgccgcc ctataccttg tctgcctccc
     1321 cgcgttgcgt cgcggtgcat ggagccgggc cacctcgacc tgaatggaag ccggcggcac
     1381 ctcgctaacg gattcaccgt ttttatcagg ctctgggagg cagaataaat gatcatatcg
     1441 tcaattatta cctccacggg gagagcctga gcaaactggc ctcaggcatt tgagaagcac
     1501 acggtcacac tgcttccggt agtcaataaa ccggtaaacc agcaatagac ataagcggct
     1561 atttaacgac cctgccctga accgacgacc gggtcgaatt tgctttcgaa tttctgccat
     1621 tcatccgctt attatcactt attcaggcgt agcaaccagg cgtttaaggg caccaataac
     1681 tgccttaaaa aaattacgcc ccgccctgcc actcatcgca gtacggccta ttggttaaaa
     1741 aatgagctga tttaacaaaa atttaacgcg aattttacgg atatagttcc tcctttcagc
     1801 aaaaaacccc tcaagacccg tttagaggcc ccaaggggtt atgctagtta ttgctcagcg
     1861 gtggcagcag ccaactcagc ttcctttcgg gctttgttag cagccggatc tcactattaa
     1921 tggtgatggt gatggtgatg atacccgttc cagcccttgt tccagccata ccggtagccg
     1981 ttatagccat acggccagtt atacccgtcg tagccactgc caccgccacc gctaccgcca
     2041 ccgccccagg gcacctcctt gccggcccag tcgaaaaatc cgccggtctg cgcggcggtg
     2101 acgccgtcca tgacccgcac gaggttggcc gcggcctccg cgggtgccac ggcggggtgt
     2161 cgtccgaggt atttcgcggt gaacggcgtg gcgaccgtgc cgggatgcag agcaatgacg
     2221 gccgcctgct tgtgggtgcg ggaaatctcg atcgccgccg tatggacgat ctggttcacc
     2281 gcagccttgg cggcccggta actgtaccaa ccgccgagcc ggttgtcgcc gatcgaaccg
     2341 acccgcgccg acagcacggc gaatacgcag ggggtatcct tgggcaacag ccgctgcgcg
     2401 tgccgaagca ccagtgccgg ccccatcgca ttcagggcga actgatccag gagaccctgc
     2461 ggcgagaggt cgcgcaggga cttttccggt gccgccccgc cgatctcgag cgcgccggtg
     2521 gcgaccacga cccgcgcgaa aggaccctcc agacggctca gatgggcgtt gaccgagtcc
     2581 tcgtccgtga tgtcgaaccc gtcgacgctc cgcgacaggc ccttgatttc gtgtgttttt
     2641 gttagctccg caaccagcgc ggcaccaatg ccgccggaac tgccgagcac gagggcgcga
     2701 ggggccatgg cttctctcct tcacaggagg gtgacactta ggtcgcagcg gcaatctgcc
     2761 gaggcttttt attaaccttt tcgcggtagc gtttttgcaa ggctgaggga ggcgacatag
     2821 agtgagtttg acctatgacc agactgctat tggccgcgct tttgaccgcg ctgctcgggg
     2881 gcccggcctc cgcgattgaa ctgcacctgc tgcggagctc atatgtatat ctccttctta
     2941 aagttaaaca aaattatttc tagaggggaa ttgttatccg ctcacaattc ccctatagtg
     3001 agtcgtatta atttcgcggg atcgagatca attccgcgag cgaaacgatc ctcatcctgt
     3061 ctcttgatca gatcttgatc ccctgcgcca tcagatcctt ggcggcaaga aagccatcca
     3121 gtttactttg cagggcttcc caaccttacc agagggcgcc ccagctggca attccgggcc
     3181 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     3241 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     3301 ggtttgcgta ttgggcgcat gcataaaaac tgttgtaatt cattaagcat tctgccgaca
     3361 tggaagccat cacaaacggc atgatgaacc tgaatcgcca gcggcatcag caccttgtcg
     3421 ccttgcgtat aatatttgcc catggtgaaa acgggggcga agaagttgtc catattggcc
     3481 acgtttaaat caaaactggt gaaactcacc cagggattgg ctgagacgaa aaacatattc
     3541 tcaataaacc ctttagggaa ataggccagg ttttcaccgt aacacgccac atcttgcgaa
     3601 tatatgtgta gaaactgccg gaaatcgtcg tggtattcac tccagagcga tgaaaacgtt
     3661 tcagtttgct catggaaaac ggtgtaacaa gggtgaacac tatcccatat caccagctca
     3721 ccgtctttca ttgccatacg gaattccgga tgagcattca tcaggcgggc aagaatgtga
     3781 ataaaggccg gataaaactt gtgcttattt ttctttacgg tctttaaaaa ggccgtaata
     3841 tccagctgaa cggtctggtt ataggtacat tgagcaactg actgaaatgc ctcaaaatgt
     3901 tctttacgat gccattggga tatatcaacg gtggtatatc cagtgatttt tttctccatt
     3961 ttagcttcct tagctcctga aaatctcgat aactcaaaaa atacgcccgg tagtgatctt
     4021 atttcattat ggtgaaagtt ggaacctctt acgtgccgat caacgtctca ttttcgccaa
     4081 aagttggctc agggcttccc ggtatcaaca gggacaccag gatttattta ttctgcgaag
     4141 tgatcttccg tcacaggtat ttattcgaag acgaaagggc ctcgtgatac gcctattttt
     4201 ataggttaat gatcgccatg taagcccact gcaagctacc tgctttctct ttgcgcttgc
     4261 gttttccctt gtccagatag cccagtagct gacattcatc ccaggtggca cttttcgggg
     4321 aaatgtgcgc gcccgcgttc ctgctggcgc tgggcctgtt tctggcgctg gacttcccgc
     4381 tgttccgtca gcagcttttc gcccacggcc ttgatgatcg cggcggcctt ggcctgcata
     4441 tcccgattca acggccccag ggcgtccaga acgggcttca ggcgctcccg aaggtctcgg
     4501 gccgtctctt gggcttgatc ggccttcttg cgcatctcac gcgctcctgc ggcggcctgt
     4561 agggcaggct catacccctg ccgaaccgct tttgtcagcc ggtcggccac ggcttccggc
     4621 gtctcaacgc gctttgagat tcccagcttt tcggccaatc cctgcggtgc ataggcgcgt
     4681 ggctcgaccg cttgcgggct gatggtgacg tggcccactg gtggccgctc cagggcctcg
     4741 tagaacgcct gaatgcgcgt gtgacgtgcc ttgctgccct cgatgccccg ttgcagccct
     4801 agatcggcca cagcggccgc aaacgtggtc tggtcgcggg tcatctgcgc tttgttgccg
     4861 atgaactcct tggccgacag cctgccgtcc tgcgtcagcg gcaccacgaa cgcggtcatg
     4921 tgcgggctgg tttcgtcacg gtggatgctg gccgtcacga tgcgatccgc cccgtacttg
     4981 tccgccagcc acttgtgcgc cttctcgaag aacgccgcct gctgttcttg gctggccgac
     5041 ttccaccatt ccgggctggc cgtcatgacg tactcgaccg ccaacacagc gtccttgcgc
     5101 cgcttctctg gcagcaactc gcgcagtcgg cccatcgctt catcggtgct gctggccgcc
     5161 cagtgctcgt tctctggcgt cctgctggcg tcagcgttgg gcgtctcgcg ctcgcggtag
     5221 gcgtgcttga gactggccgc cacgttgccc attttcgcca gcttcttgca tcgcatgatc
     5281 gcgtatgccg ccatgcctgc ccctcccttt tggtgtccaa ccggctcgac gggggcagcg
     5341 caaggcggtg cctccggcgg gccactcaat gcttgagtat actcactaga ctttgcttcg
     5401 caaagtcgtg accgcctacg gcggctgcgg cgccctacgg gcttgctctc cgggcttcgc
     5461 cctgcgcggt cgctgcgctc ccttgccagc ccgtggatat gtggacgatg gccgcgagcg
     5521 gccaccggct ggctcgcttc gctcggcccg tggacaaccc tgctggacaa gctgatggac
     5581 aggctgcgcc tgcccacgag cttgaccaca gggattgccc accggctacc cagccttcga
     5641 ccacataccc accggctcca actgcgcggc ctgcggcctt gccccatcaa tttttttaat
     5701 tttctctggg gaaaagcct
//
