LOCUS       sequence-488492-        5644 bp    DNA     circular SYN 06-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 5644)
  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 5644)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 6, 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(dsCsgA)-dsCsgA-Mfp3-A
FEATURES             Location/Qualifiers
     source          1..5644
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     236..255
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(322..424)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(541..559)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     rep_origin      1880..2651
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             2652..3314
                     /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(3881..3928)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     3924..3942
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(4001..4021)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     RBS             5074..5096
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    5111..5135
                     /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(5135..5154)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5136..5154)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5198..5219
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 actccagagc gatgaaaacg tttcagtttg ctcatggaaa acggtgtaac aagggtgaac
       61 actatcccat atcaccagct caccgtcttt cattgccata cggaattccg gatgagcatt
      121 catcaggcgg gcaagaatgt gaataaaggc cggataaaac ttgtgcttat ttttctttac
      181 ggtctttaaa aaggccgtaa tatccagctg aacggtctgg ttataggtac attgagcaac
      241 tgactgaaat gcctcaaaat gttctttacg atgccattgg gatatatcaa cggtggtata
      301 tccagtgatt tttttctcca ttttagcttc cttagctcct gaaaatctcg ataactcaaa
      361 aaatacgccc ggtagtgatc ttatttcatt atggtgaaag ttggaacctc ttacgtgccg
      421 atcaacgtct cattttcgcc aaaagttggc ccagggcttc ccggtatcaa cagggacacc
      481 aggatttatt tattctgcga agtgatcttc cgtcacaggt atttattcga agacgaaagg
      541 gcctcgtgat acgcctattt ttataggtta atgatcgcca tgtaagccca ctgcaagcta
      601 cctgctttct ctttgcgctt gcgttttccc ttgtccagat agcccagtag ctgacattca
      661 tcccaggtgg cacttttcgg ggaaatgtgc gcgcccgcgt tcctgctggc gctgggcctg
      721 tttctggcgc tggacttccc gctgttccgt cagcagcttt tcgcccacgg ccttgatgat
      781 cgcggcggcc ttggcctgca tatcccgatt caacggcccc agggcgtcca gaacgggctt
      841 caggcgctcc cgaaggtctc gggccgtctc ttgggcttga tcggccttct tgcgcatctc
      901 acgcgctcct gcggcggcct gtagggcagg ctcatacccc tgccgaaccg cttttgtcag
      961 ccggtcggcc acggcttccg gcgtctcaac gcgctttgag attcccagct tttcggccaa
     1021 tccctgcggt gcataggcgc gtggctcgac cgcttgcggg ctgatggtga cgtggcccac
     1081 tggtggccgc tccagggcct cgtagaacgc ctgaatgcgc gtgtgacgtg ccttgctgcc
     1141 ctcgatgccc cgttgcagcc ctagatcggc cacagcggcc gcaaacgtgg tctggtcgcg
     1201 ggtcatctgc gctttgttgc cgatgaactc cttggccgac agcctgccgt cctgcgtcag
     1261 cggcaccacg aacgcggtca tgtgcgggct ggtttcgtca cggtggatgc tggccgtcac
     1321 gatgcgatcc gccccgtact tgtccgccag ccacttgtgc gccttctcga agaacgccgc
     1381 ctgctgttct tggctggccg acttccacca ttccgggctg gccgtcatga cgtactcgac
     1441 cgccaacaca gcgtccttgc gccgcttctc tggcagcaac tcgcgcagtc ggcccatcgc
     1501 ttcatcggtg ctgctggccg cccagtgctc gttctctggc gtcctgctgg cgtcagcgtt
     1561 gggcgtctcg cgctcgcggt aggcgtgctt gagactggcc gccacgttgc ccattttcgc
     1621 cagcttcttg catcgcatga tcgcgtatgc cgccatgcct gcccctccct tttggtgtcc
     1681 aaccggctcg acgggggcag cgcaaggcgg tgcctccggc gggccactca atgcttgagt
     1741 atactcacta gactttgctt cgcaaagtcg tgaccgccta cggcggctgc ggcgccctac
     1801 gggcttgctc tccgggcttc gccctgcgcg gtcgctgcgc tcccttgcca gcccgtggat
     1861 atgtggacga tggccgcgag cggccaccgg ctggctcgct tcgctcggcc cgtggacaac
     1921 cctgctggac aagctgatgg acaggctgcg cctgcccacg agcttgacca cagggattgc
     1981 ccaccggcta cccagccttc gaccacatac ccaccggctc caactgcgcg gcctgcggcc
     2041 ttgccccatc aattttttta attttctctg gggaaaagcc tccggcctgc ggcctgcgcg
     2101 cttcgcttgc cggttggaca ccaagtggaa ggcgggtcaa ggctcgcgca gcgaccgcgc
     2161 agcggcttgg ccttgacgcg cctggaacga cccaagccta tgcgagtggg ggcagtcgaa
     2221 gggcgaagcc cgcccgcctg ccccccgagc ctcacggcgg cgagtgcggg ggttccaagg
     2281 gggcagcgcc accttgggca aggccgaagg ccgcgcagtc gatcaacaag ccccggaggg
     2341 gccacttttt gccggagggg gagccgcgcc gaaggcgtgg gggaaccccg caggggtgcc
     2401 cttctttggg caccaaagaa ctagatatag ggcgaaatgc gaaagactta aaaatcaaca
     2461 acttaaaaaa ggggggtacg caacagctca ttgcggcacc ccccgcaata gctcattgcg
     2521 taggttaaag aaaatctgta attgactgcc acttttacgc aacgcataat tgttgtcgcg
     2581 ctgccgaaaa gttgcagctg attgcgcatg gtgccgcaac cgtgcggcac ccctaccgca
     2641 tggagataag catggccacg cagtccagag aaatcggcat tcaagccaag aacaagcccg
     2701 gtcactgggt gcaaacggaa cgcaaagcgc atgaggcgtg ggccgggctt attgcgagga
     2761 aacccacggc ggcaatgctg ctgcatcacc tcgtggcgca gatgggccac cagaacgccg
     2821 tggtggtcag ccagaagaca ctttccaagc tcatcggacg ttctttgcgg acggtccaat
     2881 acgcagtcaa ggacttggtg gccgagcgct ggatctccgt cgtgaagctc aacggccccg
     2941 gcaccgtgtc ggcctacgtg gtcaatgacc gcgtggcgtg gggccagccc cgcgaccagt
     3001 tgcgcctgtc ggtgttcagt gccgccgtgg tggttgatca cgacgaccag gacgaatcgc
     3061 tgttggggca tggcgacctg cgccgcatcc cgaccctgta tccgggcgag cagcaactac
     3121 cgaccggccc cggcgaggag ccgcccagcc agcccggcat tccgggcatg gaaccagacc
     3181 tgccagcctt gatcgaaacg gaggaatggg aacggcgcgg gcagcagcgc ctgccgatgc
     3241 ccgatgagcc gtgttttctg gacgatggcg agccgttgga gccgccgaca cgggtcacgc
     3301 tgccgcgccg gtagcacttg ggttgcgcag caacccgtaa gtgcgctgtt ccagactatc
     3361 ggctgtagcc gcctcgccgc cctatacctt gtctgcctcc ccgcgttgcg tcgcggtgca
     3421 tggagccggg ccacctcgac ctgaatggaa gccggcggca cctcgctaac ggattcaccg
     3481 tttttatcag gctctgggag gcagaataaa tgatcatatc gtcaattatt acctccacgg
     3541 ggagagcctg agcaaactgg cctcaggcat ttgagaagca cacggtcaca ctgcttccgg
     3601 tagtcaataa accggtaaac cagcaataga cataagcggc tatttaacga ccctgccctg
     3661 aaccgacgac cgggtcgaat ttgctttcga atttctgcca ttcatccgct tattatcact
     3721 tattcaggcg tagcaaccag gcgtttaagg gcaccaataa ctgccttaaa aaaattacgc
     3781 cccgccctgc cactcatcgc agtacggcct attggttaaa aaatgagctg atttaacaaa
     3841 aatttaacgc gaattttacg gatatagttc ctcctttcag caaaaaaccc ctcaagaccc
     3901 gtttagaggc cccaaggggt tatgctagtt attgctcagc ggtggcagca gccaactcag
     3961 cttcctttcg ggctttgtta gcagccggat ctcactatta atggtgatgg tgatggtgat
     4021 gataatactt tcgtccccat cgacctcttt tccaaccatt gttccagcct ttatacccgc
     4081 cataacgtct gccatatcta ttgtagttgc caccaccgta gcgtcttgga ggaccatact
     4141 ttggaccata ataatcagca ctgccaccgc caccgctacc gccaccgccc cagggcacct
     4201 ccttgccggc ccagtcgaaa aatccgccgg tctgcgcggc ggtgacgccg tccatgaccc
     4261 gcacgaggtt ggccgcggcc tccgcgggtg ccacggcggg gtgtcgtccg aggtatttcg
     4321 cggtgaacgg cgtggcgacc gtgccgggat gcagagcaat gacggccgcc tgcttgtggg
     4381 tgcgggaaat ctcgatcgcc gccgtatgga cgatctggtt caccgcagcc ttggcggccc
     4441 ggtaactgta ccaaccgccg agccggttgt cgccgatcga accgacccgc gccgacagca
     4501 cggcgaagac gcagggggta tccttgggca acagccgctg cgcgtgccga agcaccagtg
     4561 ccggccccat cgcattcagg gcgaactgat ccaggagacc ctgcggcgag aggtcgcgca
     4621 gggacttttc cggtgccgcc ccgccgatct cgagcgcgcc ggtggcgacc acgacccgcg
     4681 cgaaaggacc ctccagacgg ctcagatggg cgttgaccga gtcctcgtcc gtgatgtcga
     4741 acccgtcgac gctccgcgac aggcccttga tttcgtgtgt ttttgttagc tccgcaacca
     4801 gcgcggcacc aatgccgccg gaactgccga gcacgagggc gcgaggggcc atggcttctc
     4861 tccttcacag gagggtgaca cttaggtcgc agcggcaatc tgccgaggct ttttattaac
     4921 cttttcgcgg tagcgttttt gcaaggctga gggaggcgac atagagtgag tttgacctat
     4981 gaccagactg ctattggccg cgcttttgac cgcgctgctc gggggcccgg cctccgcgat
     5041 tgaactgcac ctgctgcgga gctcatatgt atatctcctt cttaaagtta aacaaaatta
     5101 tttctagagg ggaattgtta tccgctcaca attcccctat agtgagtcgt attaatttcg
     5161 cgggatcgag atcaattccg cgagcctggg gtgcctaatg agtgagctaa ctcacattaa
     5221 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     5281 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgcatgcat aaaaactgtt
     5341 gtaattcatt aagcattctg ccgacatgga agccatcaca aacggcatga tgaacctgaa
     5401 tcgccagcgg catcagcacc ttgtcgcctt gcgtataata tttgcccatg gtgaaaacgg
     5461 gggcgaagaa gttgtccata ttggccacgt ttaaatcaaa actggtgaaa ctcacccagg
     5521 gattggctga gacgaaaaac atattctcaa taaacccttt agggaaatag gccaggtttt
     5581 caccgtaaca cgccacatct tgcgaatata tgtgtagaaa ctgccggaaa tcgtcgtggt
     5641 attc
//
