LOCUS       sequence-488487-        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-S
FEATURES             Location/Qualifiers
     source          1..5644
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             915..937
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    952..976
                     /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(976..995)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(977..995)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1039..1060
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     1721..1740
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1807..1909)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(2026..2044)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     rep_origin      3365..4136
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             4137..4799
                     /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(5366..5413)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     5409..5427
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(5486..5506)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
ORIGIN
        1 actgccaccg ccaccgctac cgccaccgcc ccagggcacc tccttgccgg cccagtcgaa
       61 aaatccgccg gtctgcgcgg cggtgacgcc gtccatgacc cgcacgaggt tggccgcggc
      121 ctccgcgggt gccacggcgg ggtgtcgtcc gaggtatttc gcggtgaacg gcgtggcgac
      181 cgtgccggga tgcagagcaa tgacggccgc ctgcttgtgg gtgcgggaaa tctcgatcgc
      241 cgccgtatgg acgatctggt tcaccgcagc cttggcggcc cggtaactgt accaaccgcc
      301 gagccggttg tcgccgatcg aaccgacccg cgccgacagc acggcgaaga cgcagggggt
      361 atccttgggc aacagccgct gcgcgtgccg aagcaccagt gccggcccca tcgcattcag
      421 ggcgaactga tccaggagac cctgcggcga gaggtcgcgc agggactttt ccggtgccgc
      481 cccgccgatc tcgagcgcgc cggtggcgac cacgacccgc gcgaaaggac cctccagacg
      541 gctcagatgg gcgttgaccg agtcctcgtc cgtgatgtcg aacccgtcga cgctccgcga
      601 caggcccttg atttcgtgtg tttttgttag ctccgcaacc agcgcggcac caatgccgcc
      661 ggaactgccg agcacgaggg cgcgaggggc catggcttct ctccttcaca ggagggtgac
      721 acttaggtcg cagcggcaat ctgccgaggc tttttattaa ccttttcgcg gtagcgtttt
      781 tgcaaggctg agggaggcga catagagtga gtttgaccta tgaccagact gctattggcc
      841 gcgcttttga ccgcgctgct cgggggcccg gcctccgcga ttgaactgca cctgctgcgg
      901 agctcatatg tatatctcct tcttaaagtt aaacaaaatt atttctagag gggaattgtt
      961 atccgctcac aattccccta tagtgagtcg tattaatttc gcgggatcga gatcaattcc
     1021 gcgagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     1081 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     1141 gagaggcggt ttgcgtattg ggcgcatgca taaaaactgt tgtaattcat taagcattct
     1201 gccgacatgg aagccatcac aaacggcatg atgaacctga atcgccagcg gcatcagcac
     1261 cttgtcgcct tgcgtataat atttgcccat ggtgaaaacg ggggcgaaga agttgtccat
     1321 attggccacg tttaaatcaa aactggtgaa actcacccag ggattggctg agacgaaaaa
     1381 catattctca ataaaccctt tagggaaata ggccaggttt tcaccgtaac acgccacatc
     1441 ttgcgaatat atgtgtagaa actgccggaa atcgtcgtgg tattcactcc agagcgatga
     1501 aaacgtttca gtttgctcat ggaaaacggt gtaacaaggg tgaacactat cccatatcac
     1561 cagctcaccg tctttcattg ccatacggaa ttccggatga gcattcatca ggcgggcaag
     1621 aatgtgaata aaggccggat aaaacttgtg cttatttttc tttacggtct ttaaaaaggc
     1681 cgtaatatcc agctgaacgg tctggttata ggtacattga gcaactgact gaaatgcctc
     1741 aaaatgttct ttacgatgcc attgggatat atcaacggtg gtatatccag tgattttttt
     1801 ctccatttta gcttccttag ctcctgaaaa tctcgataac tcaaaaaata cgcccggtag
     1861 tgatcttatt tcattatggt gaaagttgga acctcttacg tgccgatcaa cgtctcattt
     1921 tcgccaaaag ttggcccagg gcttcccggt atcaacaggg acaccaggat ttatttattc
     1981 tgcgaagtga tcttccgtca caggtattta ttcgaagacg aaagggcctc gtgatacgcc
     2041 tatttttata ggttaatgat cgccatgtaa gcccactgca agctacctgc tttctctttg
     2101 cgcttgcgtt ttcccttgtc cagatagccc agtagctgac attcatccca ggtggcactt
     2161 ttcggggaaa tgtgcgcgcc cgcgttcctg ctggcgctgg gcctgtttct ggcgctggac
     2221 ttcccgctgt tccgtcagca gcttttcgcc cacggccttg atgatcgcgg cggccttggc
     2281 ctgcatatcc cgattcaacg gccccagggc gtccagaacg ggcttcaggc gctcccgaag
     2341 gtctcgggcc gtctcttggg cttgatcggc cttcttgcgc atctcacgcg ctcctgcggc
     2401 ggcctgtagg gcaggctcat acccctgccg aaccgctttt gtcagccggt cggccacggc
     2461 ttccggcgtc tcaacgcgct ttgagattcc cagcttttcg gccaatccct gcggtgcata
     2521 ggcgcgtggc tcgaccgctt gcgggctgat ggtgacgtgg cccactggtg gccgctccag
     2581 ggcctcgtag aacgcctgaa tgcgcgtgtg acgtgccttg ctgccctcga tgccccgttg
     2641 cagccctaga tcggccacag cggccgcaaa cgtggtctgg tcgcgggtca tctgcgcttt
     2701 gttgccgatg aactccttgg ccgacagcct gccgtcctgc gtcagcggca ccacgaacgc
     2761 ggtcatgtgc gggctggttt cgtcacggtg gatgctggcc gtcacgatgc gatccgcccc
     2821 gtacttgtcc gccagccact tgtgcgcctt ctcgaagaac gccgcctgct gttcttggct
     2881 ggccgacttc caccattccg ggctggccgt catgacgtac tcgaccgcca acacagcgtc
     2941 cttgcgccgc ttctctggca gcaactcgcg cagtcggccc atcgcttcat cggtgctgct
     3001 ggccgcccag tgctcgttct ctggcgtcct gctggcgtca gcgttgggcg tctcgcgctc
     3061 gcggtaggcg tgcttgagac tggccgccac gttgcccatt ttcgccagct tcttgcatcg
     3121 catgatcgcg tatgccgcca tgcctgcccc tcccttttgg tgtccaaccg gctcgacggg
     3181 ggcagcgcaa ggcggtgcct ccggcgggcc actcaatgct tgagtatact cactagactt
     3241 tgcttcgcaa agtcgtgacc gcctacggcg gctgcggcgc cctacgggct tgctctccgg
     3301 gcttcgccct gcgcggtcgc tgcgctccct tgccagcccg tggatatgtg gacgatggcc
     3361 gcgagcggcc accggctggc tcgcttcgct cggcccgtgg acaaccctgc tggacaagct
     3421 gatggacagg ctgcgcctgc ccacgagctt gaccacaggg attgcccacc ggctacccag
     3481 ccttcgacca catacccacc ggctccaact gcgcggcctg cggccttgcc ccatcaattt
     3541 ttttaatttt ctctggggaa aagcctccgg cctgcggcct gcgcgcttcg cttgccggtt
     3601 ggacaccaag tggaaggcgg gtcaaggctc gcgcagcgac cgcgcagcgg cttggccttg
     3661 acgcgcctgg aacgacccaa gcctatgcga gtgggggcag tcgaagggcg aagcccgccc
     3721 gcctgccccc cgagcctcac ggcggcgagt gcgggggttc caagggggca gcgccacctt
     3781 gggcaaggcc gaaggccgcg cagtcgatca acaagccccg gaggggccac tttttgccgg
     3841 agggggagcc gcgccgaagg cgtgggggaa ccccgcaggg gtgcccttct ttgggcacca
     3901 aagaactaga tatagggcga aatgcgaaag acttaaaaat caacaactta aaaaaggggg
     3961 gtacgcaaca gctcattgcg gcaccccccg caatagctca ttgcgtaggt taaagaaaat
     4021 ctgtaattga ctgccacttt tacgcaacgc ataattgttg tcgcgctgcc gaaaagttgc
     4081 agctgattgc gcatggtgcc gcaaccgtgc ggcaccccta ccgcatggag ataagcatgg
     4141 ccacgcagtc cagagaaatc ggcattcaag ccaagaacaa gcccggtcac tgggtgcaaa
     4201 cggaacgcaa agcgcatgag gcgtgggccg ggcttattgc gaggaaaccc acggcggcaa
     4261 tgctgctgca tcacctcgtg gcgcagatgg gccaccagaa cgccgtggtg gtcagccaga
     4321 agacactttc caagctcatc ggacgttctt tgcggacggt ccaatacgca gtcaaggact
     4381 tggtggccga gcgctggatc tccgtcgtga agctcaacgg ccccggcacc gtgtcggcct
     4441 acgtggtcaa tgaccgcgtg gcgtggggcc agccccgcga ccagttgcgc ctgtcggtgt
     4501 tcagtgccgc cgtggtggtt gatcacgacg accaggacga atcgctgttg gggcatggcg
     4561 acctgcgccg catcccgacc ctgtatccgg gcgagcagca actaccgacc ggccccggcg
     4621 aggagccgcc cagccagccc ggcattccgg gcatggaacc agacctgcca gccttgatcg
     4681 aaacggagga atgggaacgg cgcgggcagc agcgcctgcc gatgcccgat gagccgtgtt
     4741 ttctggacga tggcgagccg ttggagccgc cgacacgggt cacgctgccg cgccggtagc
     4801 acttgggttg cgcagcaacc cgtaagtgcg ctgttccaga ctatcggctg tagccgcctc
     4861 gccgccctat accttgtctg cctccccgcg ttgcgtcgcg gtgcatggag ccgggccacc
     4921 tcgacctgaa tggaagccgg cggcacctcg ctaacggatt caccgttttt atcaggctct
     4981 gggaggcaga ataaatgatc atatcgtcaa ttattacctc cacggggaga gcctgagcaa
     5041 actggcctca ggcatttgag aagcacacgg tcacactgct tccggtagtc aataaaccgg
     5101 taaaccagca atagacataa gcggctattt aacgaccctg ccctgaaccg acgaccgggt
     5161 cgaatttgct ttcgaatttc tgccattcat ccgcttatta tcacttattc aggcgtagca
     5221 accaggcgtt taagggcacc aataactgcc ttaaaaaaat tacgccccgc cctgccactc
     5281 atcgcagtac ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     5341 ttacggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
     5401 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
     5461 tgttagcagc cggatctcac tattaatggt gatggtgatg gtgatggtag ccgccccagg
     5521 ggccgttgtt ccaccccttg ttccagccat accggccatg gtagccgtta tagccgttgt
     5581 agccgtagta gccattattg tacggccacg gcgcgttata gcccaggtcg tatccgtagc
     5641 cccc
//
