LOCUS       sequence-440374-        6366 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pBBR1 Origin, KanR, with maACR gene under Vhb promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6366)
  AUTHORS   Mehrer CR, Incha MR, Politz MC, Pfleger BF
  TITLE     Anaerobic production of medium-chain fatty alcohols via a 
            beta-reduction pathway.
  JOURNAL   Metab Eng. 2018 Jul;48:63-71. doi: 10.1016/j.ymben.2018.05.011. Epub
            2018 May 25.
  PUBMED    29807110
REFERENCE   2  (bases 1 to 6366)
  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: "Metab 
            Eng."; date: "2018-07"; volume: "48:63-71. doi"; pages: " 
            10.1016/j.ymben.2018.05.011. Epub 2018 May 25"
COMMENT     Sequence Label: pBTRCk-pVhb-maACR
FEATURES             Location/Qualifiers
     source          1..6366
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             55..720
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRRT"
     CDS             810..1625
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNGDRVFRLAQAQSRMNNGLVGA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(883..902)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      complement(1642..1673)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             1895..2995
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELVKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQT"
     primer_bind     2969..2992
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             3044..3055
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             5057..5080
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     primer_bind     complement(5118..5135)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      5288..5374
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5466..5493
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(5607..5643)
                     /label=soxR terminator
                     /note="bidirectional E. coli soxR transcription terminator"
     rep_origin      join(5649..6366,1..54)
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
ORIGIN
        1 ctgattgcgc atggtgccgc aaccgtgcgg cacccctacc gcatggagat aagcatggcc
       61 acgcagtcca gagaaatcgg cattcaagcc aagaacaagc ccggtcactg ggtgcaaacg
      121 gaacgcaaag cgcatgaggc gtgggccggg cttattgcga ggaaacccac ggcggcaatg
      181 ctgctgcatc acctcgtggc gcagatgggc caccagaacg ccgtggtggt cagccagaag
      241 acactttcca agctcatcgg acgttctttg cggacggtcc aatacgcagt caaggacttg
      301 gtggccgagc gctggatctc cgtcgtgaag ctcaacggcc ccggcaccgt gtcggcctac
      361 gtggtcaatg accgcgtggc gtggggccag ccccgcgacc agttgcgcct gtcggtgttc
      421 agtgccgccg tggtggttga tcacgacgac caggacgaat cgctgttggg gcatggcgac
      481 ctgcgccgca tcccgaccct gtatccgggc gagcagcaac taccgaccgg ccccggcgag
      541 gagccgccca gccagcccgg cattccgggc atggaaccag acctgccagc cttgaccgaa
      601 acggaggaat gggaacgacg cgggcagcag cgcctgccga tgcccgatga gccgtgtttt
      661 ctggacgatg gcgagccgtt ggagccgccg acacgggtca cgctgccgcg ccgtacgtaa
      721 gaggttccaa ctttcaccat aatgaaataa gatcactacc gggcgtattt tttgagttat
      781 cgagattttc aggagctaag gaagctaaaa tgagccatat tcaacgggaa acgtcttgct
      841 cgaggccgcg attaaattcc aacatggatg ctgatttata tgggtataaa tgggctcgcg
      901 ataatgtcgg gcaatcaggt gcgacaatct atcgattgta tgggaagccc gatgcgccag
      961 agttgtttct gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtca
     1021 ggctaaactg gctgacggaa tttatgcctc ttccgaccat caagcatttt atccgtactc
     1081 ctgatgatgc atggttactc accactgcga tcccagggaa aacagcattc caggtattag
     1141 aagaatatcc tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt
     1201 tgcattcgat tcctgtttgt aattgtcctt ttaacggcga tcgcgtattt cgtctcgctc
     1261 aggcgcaatc acgaatgaat aacggtttgg ttggtgcgag tgattttgat gacgagcgta
     1321 atggctggcc tgttgaacaa gtctggaaag aaatgcataa gcttttgcca ttctcaccgg
     1381 attcagtcgt cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat
     1441 taataggttg tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca
     1501 tcctatggaa ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat
     1561 atggtattga taatcctgat atgaataaat tgcagtttca cttgatgctc gatgagtttt
     1621 tctgagggcg gatccccctc aagtcaaaag cctccggtcg gaggtttttg actttctgct
     1681 atggaggtca ggtatgattt aaatggtcag tattgagcga tgcatttacg ttaagcttac
     1741 aggacgctgg ggttaaaagt atttgagttt tgatgtggat taagttttaa gaggcaataa
     1801 agattataat aagtgctgct acaccatact gatgtatggc aaaaccatat aatgaactta
     1861 aggctcaact cacaataaac aagcggcaaa agaaatgaaa atcgaagaag gtaaactggt
     1921 aatctggatt aacggcgata aaggctataa cggtctcgct gaagtcggta agaaattcga
     1981 gaaagatacc ggaattaaag tcaccgttga gcatccggat aaactggaag agaaattccc
     2041 acaggttgcg gcaactggcg atggccctga cattatcttc tgggcacacg accgctttgg
     2101 tggctacgct caatctggcc tgttggctga aatcaccccg gacaaagcgt tccaggacaa
     2161 gctgtatccg tttacctggg atgccgtacg ttacaacggc aagctgattg cttacccgat
     2221 cgctgttgaa gcgttatcgc tgatttataa caaagatctg ctgccgaacc cgccaaaaac
     2281 ctgggaagag atcccggcgc tggataaaga actgaaagcg aaaggtaaga gcgcgctgat
     2341 gttcaacctg caagaaccgt acttcacctg gccgctgatt gctgctgacg ggggttatgc
     2401 gttcaagtat gaaaacggca agtacgacat taaagacgtg ggcgtggata acgctggcgc
     2461 gaaagcgggt ctgaccttcc tggttgacct gattaaaaac aaacacatga atgcagacac
     2521 cgattactcc atcgcagaag ctgcctttaa taaaggcgaa acagcgatga ccatcaacgg
     2581 cccgtgggca tggtccaaca tcgacaccag caaagtgaat tatggtgtaa cggtactgcc
     2641 gaccttcaag ggtcaaccat ccaaaccgtt cgttggcgtg ctgagcgcag gtattaacgc
     2701 cgccagtccg aacaaagagc tggcaaaaga gttcctcgaa aactatctgc tgactgatga
     2761 aggtctggaa gcggttaata aagacaaacc gctgggtgcc gtagcgctga agtcttacga
     2821 ggaagagttg gtgaaagatc cgcggattgc cgccactatg gaaaacgccc agaaaggtga
     2881 aatcatgccg aacatcccgc agatgtccgc tttctggtat gccgtgcgta ctgcggtgat
     2941 caacgccgcc agcggtcgtc agactgtcga tgaagccctg aaagacgcgc agactaattc
     3001 gagctcgaac aacaacaaca ataacaataa caacaacctc gggatcgagg gaaggatttc
     3061 agaattcaat tatttcctga caggcggcac cggttttatc ggtcgttttc tggttgagaa
     3121 actcttggcg cgcggcggca ccgtgtatgt tctggttcgc gagcagtccc aggacaagct
     3181 ggagcggctc cgggagcgct ggggtgcaga cgacaagcaa gtgaaggctg tgatcggcga
     3241 cctcaccagc aaaaaccttg gtattgacgc gaaaacgctg aaatcactga aaggaaatat
     3301 cgaccacgta ttccatcttg ccgcggtcta cgacatgggc gcagacgaag aagcccaggc
     3361 cgccaccaat atcgaaggca ccagggcggc tgttcaggcc gccgaagcca tgggcgccaa
     3421 gcatttccat catgtgtcat ccatcgcggc agcgggtctg ttcaagggta tcttccggga
     3481 ggatatgttc gaagaagccg agaagcttga tcatccttac ctgcgcacca agcacgaatc
     3541 cgaaaaagtt gtgcgtgaag aatgcaaggt tccgttccgc atctaccgcc ctggtatggt
     3601 cattggccat tcggaaaccg gcgaaatgga caaggttgac gggccctatt acttcttcaa
     3661 gatgattcag aagatccgtc atgcgttgcc ccagtgggta cccaccatcg gtattgaagg
     3721 tggccggctg aacattgtgc cggtggattt cgtggtcgat gcactggatc acattgccca
     3781 tctggaaggc gaagatggca actgtttcca tctggtggac tccgatccgt ataaggtggg
     3841 tgagatcctc aatattttct gcgaggccgg ccatgccccc cgcatgggta tgcgcatcga
     3901 ttcccggatg ttcggtttta ttccgccgtt tattcgccag agcatcaaga atctgcctcc
     3961 ggtcaagcgc attactggtg cgcttctgga tgacatgggc attccgccct cggtgatgtc
     4021 cttcattaat tacccgaccc gttttgatac ccgggagctg gagcgggttc tgaagggcac
     4081 agacattgag gtgccgcgtc tgccgtccta tgccccggtt atctgggact actgggagcg
     4141 caatctggac ccggacctgt tcaaggaccg caccctcaag ggcacggttg aaggtaaggt
     4201 ttgcgtggtc accggcgcga cctcgggtat tggcctggca acggcagaga agctggcaga
     4261 ggccggtgcc attctggtca ttggtgcgcg caccaaggaa actctggatg aagtggcggc
     4321 cagtctggag gccaagggtg gcaacgtgca tgcgtaccag tgcgactttt cggacatgga
     4381 cgactgcgac cgctttgtga agacggtgct ggataatcac ggccacgtgg atgtactggt
     4441 gaataacgcg ggtcgctcca tccgccgctc gctggcgttg tcttttgacc ggttccacga
     4501 ttttgagcgg accatgcagc tgaactactt tggctccgtt cggctgatca tgggctttgc
     4561 gccagccatg ctggagcgtc gccgcgggca cgtggtgaat atttcttcca tcggggtact
     4621 taccaacgct ccgcgtttct cggcctatgt ctcctcgaaa tccgcactgg acgcgttcag
     4681 ccgctgtgcc gctgcagaat ggtcggatcg caacgtgacc ttcaccacca tcaacatgcc
     4741 gttggtgaaa acgccgatga tcgcgcccac caagatctac gattccgtgc cgacgctgac
     4801 gccggatgaa gccgcccaga tggtggcgga tgcgattgtg taccggccca agcgcattgc
     4861 cacccgtctt ggcgtgttcg cgcaggttct gcatgcgctg gcaccgaaga tgggtgagat
     4921 cattatgaac actggctacc ggatgttccc ggattctcca gcagccgctg gcagcaagtc
     4981 cggcgaaaag ccgaaagtct ctaccgagca ggtggccttt gcggcgatta tgcgggggat
     5041 atactggagt ctagaccatc atcaccatca tcaccatcac tgataagctg ttttggcgga
     5101 tgagagaaga ttttcagcct gatacagatt aaatcagaac gcagaagcgg tctgataaaa
     5161 cagaatttgc ctggcggcag tagcgcggtg gtcccacctg accccatgcc gaactcagaa
     5221 gtgaaacgcc gtagcgccga tggtagtgtg gggtctcccc atgcgagagt agggaactgc
     5281 caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg
     5341 tttgtcggtg aacgctctcc tgagtaggac aaatccgccg ggagcggatt tgaacgttgc
     5401 gaagcaacgg cccggagggt ggcgggcagg acgcccgcca taaactgcca ggcatcaaat
     5461 taagcagaag gccatcctga cggatggcct ttttgcgttt ctacaaactc ttttgtttat
     5521 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataacatcat tcaggacgag
     5581 cctcagactc cagcgtaact ggactgaaaa caaactaaag cgcccttgtg gcgctttagt
     5641 tttgttccgc ggccaccggc tggctcgctt cgctcggccc gtggacaacc ctgctggaca
     5701 agctgatgga caggctgcgc ctgcccacga gcttgaccac agggattgcc caccggctac
     5761 ccagccttcg accacatacc caccggctcc aactgcgcgg cctgcggcct tgccccatca
     5821 atttttttaa ttttctctgg ggaaaagcct ccggcctgcg gcctgcgcgc ttcgcttgcc
     5881 ggttggacac caagtggaag gcgggtcaag gctcgcgcag cgaccgcgca gcggcttggc
     5941 cttgacgcgc ctggaacgac ccaagcctat gcgagtgggg gcagtcgaag ggcgaagccc
     6001 gcccgcctgc cccccgagcc tcacggcggc gagtgcgggg gttccaaggg ggcagcgcca
     6061 ccttgggcaa ggccgaaggc cgcgcagtcg atcaacaagc cccggagggg ccactttttg
     6121 ccggaggggg agccgcgccg aaggcgtggg ggaaccccgc aggggtgccc ttctttgggc
     6181 accaaagaac tagatatagg gcgaaatgcg aaagacttaa aaatcaacaa cttaaaaaag
     6241 gggggtacgc aacagctcat tgcggcaccc cccgcaatag ctcattgcgt aggttaaaga
     6301 aaatctgtaa ttgactgcca cttttacgca acgcataatt gttgtcgcgc tgccgaaaag
     6361 ttgcag
//
