LOCUS       sequence-388778-        3614 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 2 destination vector containing pBBR1 origin of replication 
            and CIDAR AF scars.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3614)
  AUTHORS   Keating KW, Young EM
  TITLE     Systematic Part Transfer by Extending a Modular Toolkit to Diverse 
            Bacteria.
  JOURNAL   ACS Synth Biol. 2023 Jun 9. doi: 10.1021/acssynbio.3c00104.
  PUBMED    37294017
REFERENCE   2  (bases 1 to 3614)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "ACS Synth 
            Biol. 2023 Jun 9. doi: 10.1021/acssynbio.3c00104."
COMMENT     Sequence Label: pBBR1_DVS_AF
FEATURES             Location/Qualifiers
     source          1..3614
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     199..221
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     213..230
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     214..230
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    231..287
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(300..316)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(300..316)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(313..335)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    324..340
                     /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)."
     promoter        complement(join(348..354,355..372,373..378))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    393..414
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_feature    514..534
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     promoter        581..685
                     /gene="bla"
                     /label=AmpR promoter
     CDS             907..1698
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      1993..2764
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             2765..3427
                     /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            3438..3546
                     /note="incP origin of transfer"
     misc_feature    3579..3600
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     CDS             complement(join(3595..3614,1..304))
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCRRHLLRP
                     TSL"
ORIGIN
        1 ggagatgtct tctgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat
       61 accgcatcag gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
      121 gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt
      181 gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt gaattcgagc
      241 tcggtacccg gggatcctct agagtcgacc tgcaggcatg caagcttggc gtaatcatgg
      301 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
      361 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
      421 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
      481 ggccaacgcg cggggaagac gtcgctagag acctactagt agcggccgct gcaggcttcc
      541 tcgctcactg actccccaga actctgtcag accaaatgtg cgcggaaccc ctatttgttt
      601 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
      661 tcaataatat tgaaaaagga agagtatgcg ctcacgcaac tggtccagaa ccttgaccga
      721 acgcagcggt ggtaacggcg cagtggcggt tttcatggct tgttatgact gttttttttt
      781 ggggtacagt ctatgcctcg ggcatccaag cagcaagcgc gttacgccgt gggtcgatgt
      841 ttgatgttat ggagcagcaa cgatgttacg cagcagggca gtcgccctaa aacaaagtta
      901 aacatcatga gggaagcggt gatcgccgaa gtatcgactc aactatcaga ggtagttggc
      961 gtcatcgagc gccatctcga accgacgttg ctggccgtac atttgtacgg ctccgcagtg
     1021 gatggcggcc tgaagccaca cagcgatatt gatttgctgg ttacggtgac cgtaaggctt
     1081 gatgaaacaa cgcggcgagc tttgatcaac gaccttttgg aaacttcggc ttcccctgga
     1141 gagagcgaga ttctccgcgc tgtagaagtc accattgttg tgcacgacga catcattccg
     1201 tggcgttatc cagctaagcg cgaactgcaa tttggagaat ggcagcgcaa tgacattctt
     1261 gcaggtatct tcgagccagc cacgatcgac attgatctgg ctatcttgct gacaaaagca
     1321 agagaacata gcgttgcctt ggtaggtcca gcggcggagg aactctttga tccggttcct
     1381 gaacaggatc tatttgaggc gctaaatgaa accttaacgc tatggaactc gccgcccgac
     1441 tgggctggcg atgagcgaaa tgtagtgctt acgttgtccc gcatttggta cagcgcagta
     1501 accggcaaaa tcgcgccgaa ggatgtcgct gccgactggg caatggagcg cctgccggcc
     1561 cagtatcagc ccgtcatact tgaagctaga caggcttatc ttggacaaga agaagatcgc
     1621 ttggcctcgc gcgcagatca gttggaagaa tttgtccatt acgtaaaagg cgagatcacc
     1681 aaggtagtcg gcaaataact gtcagaccaa gtttactcat atatacttta gattgattta
     1741 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     1801 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     1861 ggatcttcta ggacaaatcc gccgccctag acagctgggc gcgccccccc tacgggcttg
     1921 ctctccgggc ttcgccctgc gcggtcgctg cgctcccttg ccagcccgtg gatatgtgga
     1981 cgatggccgc gagcggccac cggctggctc gcttcgctcg gcccgtggac aaccctgctg
     2041 gacaagctga tggacaggct gcgcctgccc acgagcttga ccacagggat tgcccaccgg
     2101 ctacccagcc ttcgaccaca tacccaccgg ctccaactgc gcggcctgcg gccttgcccc
     2161 atcaattttt ttaattttct ctggggaaaa gcctccggcc tgcggcctgc gcgcttcgct
     2221 tgccggttgg acaccaagtg gaaggcgggt caaggctcgc gcagcgaccg cgcagcggct
     2281 tggccttgac gcgcctggaa cgacccaagc ctatgcgagt gggggcagtc gaagggcgaa
     2341 gcccgcccgc ctgccccccg agcctcacgg cggcgagtgc gggggttcca agggggcagc
     2401 gccaccttgg gcaaggccga aggccgcgca gtcgatcaac aagccccgga ggggccactt
     2461 tttgccggag ggggagccgc gccgaaggcg tgggggaacc ccgcaggggt gcccttcttt
     2521 gggcaccaaa gaactagata tagggcgaaa tgcgaaagac ttaaaaatca acaacttaaa
     2581 aaaggggggt acgcaacagc tcattgcggc accccccgca atagctcatt gcgtaggtta
     2641 aagaaaatct gtaattgact gccactttta cgcaacgcat aattgttgtc gcgctgccga
     2701 aaagttgcag ctgattgcgc atggtgccgc aaccgtgcgg cacccctacc gcatggagat
     2761 aagcatggcc acgcagtcca gagaaatcgg cattcaagcc aagaacaagc ccggtcactg
     2821 ggtgcaaacg gaacgcaaag cgcatgaggc gtgggccggg cttattgcga ggaaacccac
     2881 ggcggcaatg ctgctgcatc acctcgtggc gcagatgggc caccagaacg ccgtggtggt
     2941 cagccagaaa acactttcca agctcatcgg acgttctttg cggacggtcc aatacgcagt
     3001 caaggacttg gtggccgagc gctggatctc cgtcgtgaag ctcaacggcc ccggcaccgt
     3061 gtcggcctac gtggtcaatg accgcgtggc gtggggccag ccccgcgacc agttgcgcct
     3121 gtcggtgttc agtgccgccg tggtggttga tcacgacgac caggacgaat cgctgttggg
     3181 gcatggcgac ctgcgccgca tcccgaccct gtatccgggc gagcagcaac taccgaccgg
     3241 ccccggcgag gagccgccca gccagcccgg cattccgggc atggaaccag acctgccagc
     3301 cttgaccgaa acggaggaat gggaacggcg cgggcagcag cgcctgccga tgcccgatga
     3361 gccgtgtttt ctggacgatg gcgagccgtt ggagccgccg acacgggtaa cgctgccgcg
     3421 ccggtagggc cggcctacgg ccagcctcgc agagcaggat tcccgttgag caccgccagg
     3481 tgcgaataag ggacagtgaa gaaggaacac ccgctcgcgg gtgggcctac ttcacctatc
     3541 ctgcccttgt gtctcaaaat ctctgaggat gatttctgga attcgcggcc gcttctagag
     3601 actagtgggt ctca
//
