LOCUS       sequence-388775-        3218 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 3218)
  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 3218)
  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_DVC_AF
FEATURES             Location/Qualifiers
     source          1..3218
                     /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"
     CDS             670..1329
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(736..755)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      1597..2368
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             2369..3031
                     /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            3042..3150
                     /note="incP origin of transfer"
     misc_feature    3183..3204
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     CDS             complement(join(3199..3218,1..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 acccacgtaa gaggttccaa ctttcaccat
      601 aatgaaataa gatcactacc gggcgtattt tttgagttat cgagattttc aggagctaag
      661 gaagctaaaa tggagaaaaa aatcactgga tataccaccg ttgatatatc ccaatggcat
      721 cgtaaagaac attttgaggc atttcagtca gttgctcaat gtacctataa ccagaccgtt
      781 cagctggata ttacggcctt tttaaagacc gtaaagaaaa ataagcacaa gttttatccg
      841 gcctttattc acattcttgc ccgcctgatg aatgctcatc cggaatttcg tatggcaatg
      901 aaagacggtg agctggtgat atgggatagt gttcaccctt gttacaccgt tttccatgag
      961 caaactgaaa cgttttcatc gctctggagt gaataccacg acgatttccg gcagtttcta
     1021 cacatatatt cgcaagatgt ggcgtgttac ggtgaaaacc tggcctattt ccctaaaggg
     1081 tttattgaga atatgttttt cgtctcagcc aatccctggg tgagtttcac cagttttgat
     1141 ttaaacgtgg ccaatatgga caacttcttc gcccccgttt tcaccatggg caaatattat
     1201 acgcaaggcg acaaggtgct gatgccgctg gcgattcagg ttcatcatgc cgtttgtgat
     1261 ggcttccatg tcggcagaat gcttaatgaa ttacaacagt actgcgatga gtggcagggc
     1321 ggggcgtaat ttgatatcga gctcgcttgg actgctgttg atagatccag taatgacctc
     1381 agaactctgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     1441 taatttaaaa ggatctaggt gcagcttgcc tttaggacaa atccgccgcc ctagacagct
     1501 gggcgcgccc cccctacggg cttgctctcc gggcttcgcc ctgcgcggtc gctgcgctcc
     1561 cttgccagcc cgtggatatg tggacgatgg ccgcgagcgg ccaccggctg gctcgcttcg
     1621 ctcggcccgt ggacaaccct gctggacaag ctgatggaca ggctgcgcct gcccacgagc
     1681 ttgaccacag ggattgccca ccggctaccc agccttcgac cacataccca ccggctccaa
     1741 ctgcgcggcc tgcggccttg ccccatcaat ttttttaatt ttctctgggg aaaagcctcc
     1801 ggcctgcggc ctgcgcgctt cgcttgccgg ttggacacca agtggaaggc gggtcaaggc
     1861 tcgcgcagcg accgcgcagc ggcttggcct tgacgcgcct ggaacgaccc aagcctatgc
     1921 gagtgggggc agtcgaaggg cgaagcccgc ccgcctgccc cccgagcctc acggcggcga
     1981 gtgcgggggt tccaaggggg cagcgccacc ttgggcaagg ccgaaggccg cgcagtcgat
     2041 caacaagccc cggaggggcc actttttgcc ggagggggag ccgcgccgaa ggcgtggggg
     2101 aaccccgcag gggtgccctt ctttgggcac caaagaacta gatatagggc gaaatgcgaa
     2161 agacttaaaa atcaacaact taaaaaaggg gggtacgcaa cagctcattg cggcaccccc
     2221 cgcaatagct cattgcgtag gttaaagaaa atctgtaatt gactgccact tttacgcaac
     2281 gcataattgt tgtcgcgctg ccgaaaagtt gcagctgatt gcgcatggtg ccgcaaccgt
     2341 gcggcacccc taccgcatgg agataagcat ggccacgcag tccagagaaa tcggcattca
     2401 agccaagaac aagcccggtc actgggtgca aacggaacgc aaagcgcatg aggcgtgggc
     2461 cgggcttatt gcgaggaaac ccacggcggc aatgctgctg catcacctcg tggcgcagat
     2521 gggccaccag aacgccgtgg tggtcagcca gaaaacactt tccaagctca tcggacgttc
     2581 tttgcggacg gtccaatacg cagtcaagga cttggtggcc gagcgctgga tctccgtcgt
     2641 gaagctcaac ggccccggca ccgtgtcggc ctacgtggtc aatgaccgcg tggcgtgggg
     2701 ccagccccgc gaccagttgc gcctgtcggt gttcagtgcc gccgtggtgg ttgatcacga
     2761 cgaccaggac gaatcgctgt tggggcatgg cgacctgcgc cgcatcccga ccctgtatcc
     2821 gggcgagcag caactaccga ccggccccgg cgaggagccg cccagccagc ccggcattcc
     2881 gggcatggaa ccagacctgc cagccttgac cgaaacggag gaatgggaac ggcgcgggca
     2941 gcagcgcctg ccgatgcccg atgagccgtg ttttctggac gatggcgagc cgttggagcc
     3001 gccgacacgg gtaacgctgc cgcgccggta gggccggcct acggccagcc tcgcagagca
     3061 ggattcccgt tgagcaccgc caggtgcgaa taagggacag tgaagaagga acacccgctc
     3121 gcgggtgggc ctacttcacc tatcctgccc ttgtgtctca aaatctctga ggatgatttc
     3181 tggaattcgc ggccgcttct agagactagt gggtctca
//
