LOCUS       sequence-500861-        4857 bp    DNA     circular SYN 23-FEB-2026
DEFINITION  pBBR1 plasmid with constituitive lambda-CI (J23101-Bujard-Lambda-CI)
            and PirWT (PTet-RJ64-Pir4150) expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4857)
  AUTHORS   Hina-Nilesh Joshi S, Jenkins C, Ulaeto D, Gorochowski TE
  TITLE     SHARK: A Specialized Host for Assembling R6K Plasmids.
  JOURNAL   ACS Synth Biol. 2026 Jan 13. doi: 10.1021/acssynbio.5c00749.
  PUBMED    41529903
REFERENCE   2  (bases 1 to 4857)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Feb 23, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "ACS Synth 
            Biol. 2026 Jan 13. doi: 10.1021/acssynbio.5c00749."
COMMENT     Sequence Label: pSHARK6
FEATURES             Location/Qualifiers
     source          1..4857
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      88..859
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             860..1522
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     oriT            1533..1641
                     /note="incP origin of transfer"
     CDS             complement(1902..2693)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRTDQLEEFVHYVKGEITKVVGK"
     promoter        2835..2869
                     /label=BBa_J23101
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.7"
     RBS             2883..2894
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             2902..3615
                     /codon_start=1
                     /gene="cI"
                     /product="phage lambda repressor"
                     /label=lambda repressor
                     /translation="MSTKKKPLTQEQLEDARRLKAIYEKKKNELGLSQESVADKMGMGQ
                     SGVGALFNGINALNAYNAALLAKILKVSVEEFSPSIAREIYEMYEAVSMQPSLRSEYEY
                     PVFSHVQAGMFSPELRTFTKGDAERWVSTTKKASDSAFWLEVEGNSMTAPTGSKPSFPD
                     GMLILVDPEQAVEPGDFCIARLGGDEFTFKKLIRDSGQVFLQPLNPQYPMIPCNESCSV
                     VGKVIASQWPEETFG"
     protein_bind    3714..3732
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3739..3757
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_RNA        3774..3823
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
ORIGIN
        1 ccccctacgg gcttgctctc cgggcttcgc cctgcgcggt cgctgcgctc ccttgccagc
       61 ccgtggatat gtggacgatg gccgcgagcg gccaccggct ggctcgcttc gctcggcccg
      121 tggacaaccc tgctggacaa gctgatggac aggctgcgcc tgcccacgag cttgaccaca
      181 gggattgccc accggctacc cagccttcga ccacataccc accggctcca actgcgcggc
      241 ctgcggcctt gccccatcaa tttttttaat tttctctggg gaaaagcctc cggcctgcgg
      301 cctgcgcgct tcgcttgccg gttggacacc aagtggaagg cgggtcaagg ctcgcgcagc
      361 gaccgcgcag cggcttggcc ttgacgcgcc tggaacgacc caagcctatg cgagtggggg
      421 cagtcgaagg gcgaagcccg cccgcctgcc ccccgagcct cacggcggcg agtgcggggg
      481 ttccaagggg gcagcgccac cttgggcaag gccgaaggcc gcgcagtcga tcaacaagcc
      541 ccggaggggc cactttttgc cggaggggga gccgcgccga aggcgtgggg gaaccccgca
      601 ggggtgccct tctttgggca ccaaagaact agatataggg cgaaatgcga aagacttaaa
      661 aatcaacaac ttaaaaaagg ggggtacgca acagctcatt gcggcacccc ccgcaatagc
      721 tcattgcgta ggttaaagaa aatctgtaat tgactgccac ttttacgcaa cgcataattg
      781 ttgtcgcgct gccgaaaagt tgcagctgat tgcgcatggt gccgcaaccg tgcggcaccc
      841 ctaccgcatg gagataagca tggccacgca gtccagagaa atcggcattc aagccaagaa
      901 caagcccggt cactgggtgc aaacggaacg caaagcgcat gaggcgtggg ccgggcttat
      961 tgcgaggaaa cccacggcgg caatgctgct gcatcacctc gtggcgcaga tgggccacca
     1021 gaacgccgtg gtggtcagcc agaagacact ttccaagctc atcggacgtt ctttgcggac
     1081 ggtccaatac gcagtcaagg acttggtggc cgagcgctgg atctccgtcg tgaagctcaa
     1141 cggccccggc accgtgtcgg cctacgtggt caatgaccgc gtggcgtggg gccagccccg
     1201 cgaccagttg cgcctgtcgg tgttcagtgc cgccgtggtg gttgatcacg acgaccagga
     1261 cgaatcgctg ttggggcatg gcgacctgcg ccgcatcccg accctgtatc cgggcgagca
     1321 gcaactaccg accggccccg gcgaggagcc gcccagccag cccggcattc cgggcatgga
     1381 accagacctg ccagccttga ccgaaacgga ggaatgggaa cggcgcgggc agcagcgcct
     1441 gccgatgccc gatgagccgt gttttctgga cgatggcgag ccgttggagc cgccgacacg
     1501 ggtaacgctg ccgcgccggt agggccggcc tacggccagc ctcgcagagc aggattcccg
     1561 ttgagcaccg ccaggtgcga ataagggaca gtgaagaagg aacacccgct cgcgggtggg
     1621 cctacttcac ctatcctgcc cggctgacgc cgttggatac accaaggaaa gtctacacga
     1681 accctttggc aaaatcctgt atatcgtgcg aaaaaggatg gatataccga aaaaatcgct
     1741 ataatgaccc cgaagcaggg ttatgcagcg gaaaaggcaa gaagttccta ttctcaagaa
     1801 agaataggaa cttcaggtgg caaacgcgaa gtaatctttt cggttttaaa gaaaaagggc
     1861 agggtggtga caccttgccc gtttttttgc cggagctagt attatttgcc gactaccttg
     1921 gtgatctcgc ctttcacgta gtggacaaat tcttccaact gatctgtgcg cgaggccaag
     1981 cgatcttctt cttgtccaag ataagcctgt ctagcttcaa gtatgacggg ctgatactgg
     2041 gccggcaggc gctccattgc ccagtcggca gcgacatcct tcggcgcgat tttgccggtt
     2101 actgcgctgt accaaatgcg ggacaacgta agcactacat ttcgctcatc gccagcccag
     2161 tcgggcggcg agttccatag cgttaaggtt tcatttagcg cctcaaatag atcctgttca
     2221 ggaaccggat caaagagttc ctccgccgct ggacctacca aggcaacgct atgttctctt
     2281 gcttttgtca gcaagatagc cagatcaatg tcgatcgtgg ctggctcgaa gatacctgca
     2341 agaatgtcat tgcgctgcca ttctccaaat tgcagttcgc gcttagctgg ataacgccac
     2401 ggaatgatgt cgtcgtgcac aacaatggtg acttctacag cgcggagaat ctcgctctct
     2461 ccaggggaag ccgaagtttc caaaaggtcg ttgatcaaag ctcgccgcgt tgtttcatca
     2521 agccttacgg tcaccgtaac cagcaaatca atatcactgt gtggcttcag gccgccatcc
     2581 actgcggagc cgtacaaatg tacggccagc aacgtcggtt cgagatggcg ctcgatgacg
     2641 ccaactacct ctgatagttg agtcgatact tcggcgatca ccgcttccct cattttagct
     2701 tccttagctc ctgaaaatct cgataactca aaaaatacgc ccggtagtga tcttatttca
     2761 ttatggtgaa agttggaacc tcttacgtgc ccgatcgaag ttcctattct caagaaagaa
     2821 taggaacttc ggagtttaca gctagctcag tcctaggtat tatgctagct actgaattca
     2881 ttaaagagga gaaaggtacc aatgagcaca aaaaagaaac cattaacaca agagcagctt
     2941 gaggacgcac gtcgccttaa agcaatttat gaaaaaaaga aaaatgaact tggcttatcc
     3001 caggaatctg tcgcagacaa gatggggatg gggcagtcag gcgttggtgc tttatttaat
     3061 ggcatcaatg cattaaatgc ttataacgcc gcattgcttg caaaaattct caaagttagc
     3121 gttgaagaat ttagcccttc aatcgccaga gaaatctacg agatgtatga agcggttagt
     3181 atgcagccgt cacttagaag tgagtatgag taccctgttt tttctcatgt tcaggcaggg
     3241 atgttctcac ctgagcttag aacctttacc aaaggtgatg cggagagatg ggtaagcaca
     3301 accaaaaaag ccagtgattc tgcattctgg cttgaggttg aaggtaattc catgaccgca
     3361 ccaacaggct ccaagccaag ctttcctgac ggaatgttaa ttctcgttga ccctgagcag
     3421 gctgttgagc caggtgattt ctgcatagcc agacttgggg gtgatgagtt taccttcaag
     3481 aaactgatca gggatagcgg tcaggtgttt ttacaaccac taaacccaca gtacccaatg
     3541 atcccatgca atgagagttg ttccgttgtg gggaaagtta tcgctagtca gtggcctgaa
     3601 gagacgtttg gctgaaggtt tcagccaaaa aacttaagac cgccggtctt gtccactacc
     3661 ttgcagtaat gcggtggaca ggatcggcgg ttttcttttc tcttctcaat acttccctat
     3721 cagtgataga gattgacatc cctatcagtg atagagatac tgagcactac tagctgtcac
     3781 cggatgtgct ttccggtctg atgagtccgt gaggacgaaa cagcctctac aaataatttt
     3841 gtttaatact agagaaagag gggaaatact agaatgagac tcaaggtcat gatggacgtg
     3901 aacaaaaaaa cgaaaattcg ccaccgaaac gagctaaatc acaccctggc tcaacttcct
     3961 ttgcccgcaa agcgagtgat gtatatggcg cttgctccca ttgatagcaa agaacctctt
     4021 gaacgagggc gagttttcaa aattagggct gaagatcttg cagcgctcgc caaaatcacc
     4081 ccatcgcttg cttatcgaca attaaaagag ggtggtaaat tacttggtgc cagcaaaatt
     4141 tcgctaagag gggatgatat cattgcttta gctaaagagc ttaacctgcc ctttattgct
     4201 aaaaactcct ctgaagagtt agatcttaac attattgagt ggatagctta ttcaaatgat
     4261 gaaggatact tgtctttaaa attcaccaga accatagaac catatatctc tagccttatt
     4321 gggaaaaaaa ataaattcac aacgcaattg ttaacggcaa gcttacgctt aagtagccag
     4381 tattcatctt ctctttatca acttatcagg aagcattact ctaattttaa gaagaaaaat
     4441 tattttatta tttccgttga tgagttaaag gaagagttaa tagcttatac ttttgataaa
     4501 gatggaaata ttgagtacaa ataccctgac tttcctattt ttaaaaggga tgtgttaaat
     4561 aaagccattg ctgaaattaa aaagaaaaca gaaatatcgt ttgttggctt cactgttcat
     4621 gaaaaagaag gaagaaaaat tagtaagctg aagttcgaat ttgtcgttga tgaagatgaa
     4681 ttttctggcg ataaagatga tgaagctttt tttatgaatt tatctgaagc tgatgcagct
     4741 tttctcaagg tatttgatga aaccgtacct cccaaaaaag ctaaggggtg aaggtggcca
     4801 attattgaag gcctccctaa cggggggcct ttttttgttt ctggtctgcc ctcgctt
//
