LOCUS       sequence-408629-        3614 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 2 destination vector containing pBBR1 origin of replication 
            and CIDAR AG 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_AG
FEATURES             Location/Qualifiers
     source          1..3614
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      26..797
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             798..1460
                     /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            1471..1579
                     /note="incP origin of transfer"
     misc_feature    1612..1633
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     CDS             complement(1628..1951)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCRRHLLRP
                     TSL"
     primer_bind     1846..1868
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1860..1877
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1861..1877
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1878..1934
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(1947..1963)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1947..1963)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1960..1982)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1971..1987
                     /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(1995..2001,2002..2019,2020..2025))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2040..2061
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_feature    2161..2181
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     promoter        2228..2332
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2554..3345
                     /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"
ORIGIN
        1 gtggatatgt ggacgatggc cgcgagcggc caccggctgg ctcgcttcgc tcggcccgtg
       61 gacaaccctg ctggacaagc tgatggacag gctgcgcctg cccacgagct tgaccacagg
      121 gattgcccac cggctaccca gccttcgacc acatacccac cggctccaac tgcgcggcct
      181 gcggccttgc cccatcaatt tttttaattt tctctgggga aaagcctccg gcctgcggcc
      241 tgcgcgcttc gcttgccggt tggacaccaa gtggaaggcg ggtcaaggct cgcgcagcga
      301 ccgcgcagcg gcttggcctt gacgcgcctg gaacgaccca agcctatgcg agtgggggca
      361 gtcgaagggc gaagcccgcc cgcctgcccc ccgagcctca cggcggcgag tgcgggggtt
      421 ccaagggggc agcgccacct tgggcaaggc cgaaggccgc gcagtcgatc aacaagcccc
      481 ggaggggcca ctttttgccg gagggggagc cgcgccgaag gcgtggggga accccgcagg
      541 ggtgcccttc tttgggcacc aaagaactag atatagggcg aaatgcgaaa gacttaaaaa
      601 tcaacaactt aaaaaagggg ggtacgcaac agctcattgc ggcacccccc gcaatagctc
      661 attgcgtagg ttaaagaaaa tctgtaattg actgccactt ttacgcaacg cataattgtt
      721 gtcgcgctgc cgaaaagttg cagctgattg cgcatggtgc cgcaaccgtg cggcacccct
      781 accgcatgga gataagcatg gccacgcagt ccagagaaat cggcattcaa gccaagaaca
      841 agcccggtca ctgggtgcaa acggaacgca aagcgcatga ggcgtgggcc gggcttattg
      901 cgaggaaacc cacggcggca atgctgctgc atcacctcgt ggcgcagatg ggccaccaga
      961 acgccgtggt ggtcagccag aaaacacttt ccaagctcat cggacgttct ttgcggacgg
     1021 tccaatacgc agtcaaggac ttggtggccg agcgctggat ctccgtcgtg aagctcaacg
     1081 gccccggcac cgtgtcggcc tacgtggtca atgaccgcgt ggcgtggggc cagccccgcg
     1141 accagttgcg cctgtcggtg ttcagtgccg ccgtggtggt tgatcacgac gaccaggacg
     1201 aatcgctgtt ggggcatggc gacctgcgcc gcatcccgac cctgtatccg ggcgagcagc
     1261 aactaccgac cggccccggc gaggagccgc ccagccagcc cggcattccg ggcatggaac
     1321 cagacctgcc agccttgacc gaaacggagg aatgggaacg gcgcgggcag cagcgcctgc
     1381 cgatgcccga tgagccgtgt tttctggacg atggcgagcc gttggagccg ccgacacggg
     1441 taacgctgcc gcgccggtag ggccggccta cggccagcct cgcagagcag gattcccgtt
     1501 gagcaccgcc aggtgcgaat aagggacagt gaagaaggaa cacccgctcg cgggtgggcc
     1561 tacttcacct atcctgccct tgtgtctcaa aatctctgag gatgatttct ggaattcgcg
     1621 gccgcttcta gagactagtg ggtctcagga gatgtcttct gcaccatatg cggtgtgaaa
     1681 taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc
     1741 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag
     1801 ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt
     1861 gtaaaacgac ggccagtgaa ttcgagctcg gtacccgggg atcctctaga gtcgacctgc
     1921 aggcatgcaa gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     1981 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     2041 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     2101 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggaagacgtt gccagagacc
     2161 tactagtagc ggccgctgca ggcttcctcg ctcactgact ccccagaact ctgtcagacc
     2221 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     2281 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgcgctc
     2341 acgcaactgg tccagaacct tgaccgaacg cagcggtggt aacggcgcag tggcggtttt
     2401 catggcttgt tatgactgtt tttttttggg gtacagtcta tgcctcgggc atccaagcag
     2461 caagcgcgtt acgccgtggg tcgatgtttg atgttatgga gcagcaacga tgttacgcag
     2521 cagggcagtc gccctaaaac aaagttaaac atcatgaggg aagcggtgat cgccgaagta
     2581 tcgactcaac tatcagaggt agttggcgtc atcgagcgcc atctcgaacc gacgttgctg
     2641 gccgtacatt tgtacggctc cgcagtggat ggcggcctga agccacacag cgatattgat
     2701 ttgctggtta cggtgaccgt aaggcttgat gaaacaacgc ggcgagcttt gatcaacgac
     2761 cttttggaaa cttcggcttc ccctggagag agcgagattc tccgcgctgt agaagtcacc
     2821 attgttgtgc acgacgacat cattccgtgg cgttatccag ctaagcgcga actgcaattt
     2881 ggagaatggc agcgcaatga cattcttgca ggtatcttcg agccagccac gatcgacatt
     2941 gatctggcta tcttgctgac aaaagcaaga gaacatagcg ttgccttggt aggtccagcg
     3001 gcggaggaac tctttgatcc ggttcctgaa caggatctat ttgaggcgct aaatgaaacc
     3061 ttaacgctat ggaactcgcc gcccgactgg gctggcgatg agcgaaatgt agtgcttacg
     3121 ttgtcccgca tttggtacag cgcagtaacc ggcaaaatcg cgccgaagga tgtcgctgcc
     3181 gactgggcaa tggagcgcct gccggcccag tatcagcccg tcatacttga agctagacag
     3241 gcttatcttg gacaagaaga agatcgcttg gcctcgcgcg cagatcagtt ggaagaattt
     3301 gtccattacg taaaaggcga gatcaccaag gtagtcggca aataactgtc agaccaagtt
     3361 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     3421 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     3481 gcgtcagacc ccgtagaaaa gatcaaagga tcttctagga caaatccgcc gccctagaca
     3541 gctgggcgcg ccccccctac gggcttgctc tccgggcttc gccctgcgcg gtcgctgcgc
     3601 tcccttgcca gccc
//
