LOCUS       sequence-488486-        5782 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 5782)
  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 5782)
  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(aphII)-dsCsgA-Mfp3-S
FEATURES             Location/Qualifiers
     source          1..5782
                     /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             2993..3015
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    3030..3054
                     /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(3054..3073)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3055..3073)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3255..3276
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     3937..3956
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4023..4125)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(4242..4260)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     rep_origin      join(5581..5782,1..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 gtagccgccc caggggccgt tgttccaccc cttgttccag
     1981 ccataccggc catggtagcc gttatagccg ttgtagccgt agtagccatt attgtacggc
     2041 cacggcgcgt tatagcccag gtcgtatccg tagcccccac tgccaccgcc accgctaccg
     2101 ccaccgcccc agggcacctc cttgccggcc cagtcgaaaa atccgccggt ctgcgcggcg
     2161 gtgacgccgt ccatgacccg cacgaggttg gccgcggcct ccgcgggtgc cacggcgggg
     2221 tgtcgtccga ggtatttcgc ggtgaacggc gtggcgaccg tgccgggatg cagagcaatg
     2281 acggccgcct gcttgtgggt gcgggaaatc tcgatcgccg ccgtatggac gatctggttc
     2341 accgcagcct tggcggcccg gtaactgtac caaccgccga gccggttgtc gccgatcgaa
     2401 ccgacccgcg ccgacagcac ggcgaatacg cagggggtat ccttgggcaa cagccgctgc
     2461 gcgtgccgaa gcaccagtgc cggccccatc gcattcaggg cgaactgatc caggagaccc
     2521 tgcggcgaga ggtcgcgcag ggacttttcc ggtgccgccc cgccgatctc gagcgcgccg
     2581 gtggcgacca cgacccgcgc gaaaggaccc tccagacggc tcagatgggc gttgaccgag
     2641 tcctcgtccg tgatgtcgaa cccgtcgacg ctccgcgaca ggcccttgat ttcgtgtgtt
     2701 tttgttagct ccgcaaccag cgcggcacca atgccgccgg aactgccgag cacgagggcg
     2761 cgaggggcca tggcttctct ccttcacagg agggtgacac ttaggtcgca gcggcaatct
     2821 gccgaggctt tttattaacc ttttcgcggt agcgtttttg caaggctgag ggaggcgaca
     2881 tagagtgagt ttgacctatg accagactgc tattggccgc gcttttgacc gcgctgctcg
     2941 ggggcccggc ctccgcgatt gaactgcacc tgctgcggag ctcatatgta tatctccttc
     3001 ttaaagttaa acaaaattat ttctagaggg gaattgttat ccgctcacaa ttcccctata
     3061 gtgagtcgta ttaatttcgc gggatcgaga tcaattccgc gagcgaaacg atcctcatcc
     3121 tgtctcttga tcagatcttg atcccctgcg ccatcagatc cttggcggca agaaagccat
     3181 ccagtttact ttgcagggct tcccaacctt accagagggc gccccagctg gcaattccgg
     3241 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     3301 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     3361 ggcggtttgc gtattgggcg catgcataaa aactgttgta attcattaag cattctgccg
     3421 acatggaagc catcacaaac ggcatgatga acctgaatcg ccagcggcat cagcaccttg
     3481 tcgccttgcg tataatattt gcccatggtg aaaacggggg cgaagaagtt gtccatattg
     3541 gccacgttta aatcaaaact ggtgaaactc acccagggat tggctgagac gaaaaacata
     3601 ttctcaataa accctttagg gaaataggcc aggttttcac cgtaacacgc cacatcttgc
     3661 gaatatatgt gtagaaactg ccggaaatcg tcgtggtatt cactccagag cgatgaaaac
     3721 gtttcagttt gctcatggaa aacggtgtaa caagggtgaa cactatccca tatcaccagc
     3781 tcaccgtctt tcattgccat acggaattcc ggatgagcat tcatcaggcg ggcaagaatg
     3841 tgaataaagg ccggataaaa cttgtgctta tttttcttta cggtctttaa aaaggccgta
     3901 atatccagct gaacggtctg gttataggta cattgagcaa ctgactgaaa tgcctcaaaa
     3961 tgttctttac gatgccattg ggatatatca acggtggtat atccagtgat ttttttctcc
     4021 attttagctt ccttagctcc tgaaaatctc gataactcaa aaaatacgcc cggtagtgat
     4081 cttatttcat tatggtgaaa gttggaacct cttacgtgcc gatcaacgtc tcattttcgc
     4141 caaaagttgg ctcagggctt cccggtatca acagggacac caggatttat ttattctgcg
     4201 aagtgatctt ccgtcacagg tatttattcg aagacgaaag ggcctcgtga tacgcctatt
     4261 tttataggtt aatgatcgcc atgtaagccc actgcaagct acctgctttc tctttgcgct
     4321 tgcgttttcc cttgtccaga tagcccagta gctgacattc atcccaggtg gcacttttcg
     4381 gggaaatgtg cgcgcccgcg ttcctgctgg cgctgggcct gtttctggcg ctggacttcc
     4441 cgctgttccg tcagcagctt ttcgcccacg gccttgatga tcgcggcggc cttggcctgc
     4501 atatcccgat tcaacggccc cagggcgtcc agaacgggct tcaggcgctc ccgaaggtct
     4561 cgggccgtct cttgggcttg atcggccttc ttgcgcatct cacgcgctcc tgcggcggcc
     4621 tgtagggcag gctcataccc ctgccgaacc gcttttgtca gccggtcggc cacggcttcc
     4681 ggcgtctcaa cgcgctttga gattcccagc ttttcggcca atccctgcgg tgcataggcg
     4741 cgtggctcga ccgcttgcgg gctgatggtg acgtggccca ctggtggccg ctccagggcc
     4801 tcgtagaacg cctgaatgcg cgtgtgacgt gccttgctgc cctcgatgcc ccgttgcagc
     4861 cctagatcgg ccacagcggc cgcaaacgtg gtctggtcgc gggtcatctg cgctttgttg
     4921 ccgatgaact ccttggccga cagcctgccg tcctgcgtca gcggcaccac gaacgcggtc
     4981 atgtgcgggc tggtttcgtc acggtggatg ctggccgtca cgatgcgatc cgccccgtac
     5041 ttgtccgcca gccacttgtg cgccttctcg aagaacgccg cctgctgttc ttggctggcc
     5101 gacttccacc attccgggct ggccgtcatg acgtactcga ccgccaacac agcgtccttg
     5161 cgccgcttct ctggcagcaa ctcgcgcagt cggcccatcg cttcatcggt gctgctggcc
     5221 gcccagtgct cgttctctgg cgtcctgctg gcgtcagcgt tgggcgtctc gcgctcgcgg
     5281 taggcgtgct tgagactggc cgccacgttg cccattttcg ccagcttctt gcatcgcatg
     5341 atcgcgtatg ccgccatgcc tgcccctccc ttttggtgtc caaccggctc gacgggggca
     5401 gcgcaaggcg gtgcctccgg cgggccactc aatgcttgag tatactcact agactttgct
     5461 tcgcaaagtc gtgaccgcct acggcggctg cggcgcccta cgggcttgct ctccgggctt
     5521 cgccctgcgc ggtcgctgcg ctcccttgcc agcccgtgga tatgtggacg atggccgcga
     5581 gcggccaccg gctggctcgc ttcgctcggc ccgtggacaa ccctgctgga caagctgatg
     5641 gacaggctgc gcctgcccac gagcttgacc acagggattg cccaccggct acccagcctt
     5701 cgaccacata cccaccggct ccaactgcgc ggcctgcggc cttgccccat caattttttt
     5761 aattttctct ggggaaaagc ct
//
