LOCUS       sequence-322865-        5073 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pBAV1k-lacI-Trc-BCD9-mCherry. BCD RBS..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5073)
  AUTHORS   Biggs BW, Bedore SR, Arvay E, Huang S, Subramanian H, McIntyre EA, 
            Duscent-Maitland CV, Neidle EL, Tyo KEJ
  TITLE     Development of a genetic toolset for the highly engineerable and 
            metabolically versatile Acinetobacter baylyi ADP1.
  JOURNAL   Nucleic Acids Res. 2020 Apr 4. pii: 5815822. doi: 
            10.1093/nar/gkaa167.
  PUBMED    32246719
REFERENCE   2  (bases 1 to 5073)
  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: "Nucleic 
            Acids Res. 2020 Apr 4. pii: 5815822. doi: 10.1093/nar/gkaa167."
COMMENT     Sequence Label: pBWB294
FEATURES             Location/Qualifiers
     source          1..5073
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(692..697,698..714,715..721)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    729..745
                     /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)."
     CDS             838..1548
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(983..1001)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1251..1270
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     terminator      1599..1685
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             3004..3015
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     terminator      complement(3217..3311)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3327..4121)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEEYRCVKDTEGMSPAKVYKLVGENEILYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEGYED
                     EQSPEKTIELYAECIRLFHSIDISDCPYTNSLDGRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     misc_feature    4327..4346
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that start with ""ATG"""
     promoter        4373..4450
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             join(4451..5073,1..460)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(4470..4489)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 tcaaattcag ccgatagcgg aacgggaagg cgactggagt gccatgtccg gttttcaaca
       61 aaccatgcaa atgctgaatg agggcatcgt tcccactgcg atgctggttg ccaacgatca
      121 gatggcgctg ggcgcaatgc gcgccattac cgagtccggg ctgcgcgttg gtgcggatat
      181 ctcggtagtg ggatacgacg ataccgaaga cagctcatgt tatatcccgc cgttaaccac
      241 catcaaacag gattttcgcc tgctggggca aaccagcgtg gaccgcttgc tgcaactctc
      301 tcagggccag gcggtgaagg gcaatcagct gttgcccgtc tcactggtga aaagaaaaac
      361 caccctggcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
      421 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtaa
      481 gttagcgcga attgatctgg tttgacagct tatcatcgac tgcacggtgc accaatgctt
      541 ctggcgtcag gcagccatcg gaagctgtgg tatggctgtg caggtcgtaa atcactgcat
      601 aattcgtgtc gctcaaggcg cactcccgtt ctggataatg ttttttgcgc cgacatcata
      661 acggttctgg caaatattct gaaatgagct gttgacaatt aatcatccgg ctcgtataat
      721 gtgtggaatt gtgagcggat aacaatttca cagggcccaa gttcacttaa aaaggagatc
      781 aacaatgaaa gcaattttcg tactgaaaca tcttaatcat gcagaggagt ctttctaatg
      841 gtgagcaagg gcgaggagga taacatggcc atcatcaagg agttcatgcg cttcaaggtt
      901 cacatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc
      961 ccctacgagg gcacccagac cgccaagctg aaggtgacca agggtggccc cctgcccttc
     1021 gcctgggaca tcctgtcccc tcagttcatg tacggctcca aggcctacgt gaagcacccc
     1081 gccgacatcc ccgactactt gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg
     1141 atgaacttcg aggacggcgg cgtggtgacc gtgacccagg actcctccct gcaagacggc
     1201 gagttcatct acaaggtgaa gctgcgcggc accaacttcc cctccgacgg ccccgtaatg
     1261 cagaagaaga ctatgggctg ggaggcctcc tccgagcgga tgtaccccga ggacggcgcg
     1321 ctgaagggcg agatcaagca gaggctgaag ctgaaggacg gcggccacta cgacgctgag
     1381 gtcaagacca cctacaaggc caagaagccc gtgcaactgc ccggcgcgta caacgtcaac
     1441 atcaagttgg acatcacctc ccacaacgag gactacacca tcgtggaaca gtacgaacgc
     1501 gccgagggcc gccactccac cggcggcatg gacgagctgt ataagtaagc ctcagggccc
     1561 gatcgatgcc gccgcttaat taattaatcc agaggcatca aataaaacga aaggctcagt
     1621 cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc ctgagtagga
     1681 caaatccgcc gccctagacc tagtgtcatt ttatttcccc cgtttcagca tcaagaacct
     1741 ttgcataact tgctctatat ccacactgat aattgccctc aaaccataat ctaaaggcgc
     1801 tagagtttgt tgaaacaata tcttttacat cattcgtatt taaaattcca aactccgctc
     1861 ccctaaggcg aataaaagcc attaaatctt ttgtatttac caaattatag tcatccacta
     1921 tatctaagag taaattcttc aattctcttt tttggctttc atcaagtgtt atatagcggt
     1981 caatatcaaa atcattaatg ttcaaaatat cttttttgtc gtatatatgt ttattcttag
     2041 caatagcgtc ctttgattca tgagtcaaat attcatatga acctttgata taatcaagta
     2101 tctcaacatg agcaactgaa ctattcccca attttcgctt aatcttgttc ctaacgcttt
     2161 ctattgttac aggatttcgt gcaatatata taacgtgata gtgtggtttt ttatagtgct
     2221 ttccatttcg tataacatca ctactattcc atgtatcttt atcttttttt tcgtccatat
     2281 cgtgtaaagg actgacagcc atagatacgc ccaaactctc taatttttcc ttccaatcat
     2341 taggaattga gtcaggatat aataaaaatc caaaatttct agctttagta tttttaatag
     2401 ccatgatata attaccttat caaaaacaag tagcgaaaac tcgtatcctt ctaaaaacgc
     2461 gagctttcgc ttattttttt tgttctgatt cctttcttgc atattcttct atagctaacg
     2521 ccgcaaccgc agattttgaa aaaccttttt gtttcgccat atctgttaat tttttatctt
     2581 gctcttttgt cagagaaatc ataactcttt ttttcgattc tgaaatcacc atttaaaaaa
     2641 ctccaatcaa ataattttat aaagttagtg tatcactttg taatcataaa aacaacaata
     2701 aagctactta aatatagatt tataaaaaac gttggcgatt cgttggcgat tgaaaaaccc
     2761 cttaaaccct tgagccagtt gggatagagc gtttttggca caaaaattgg cactcggcac
     2821 ttaatggggg gtcgtagtac ggaagcaaaa ttcgcttcct ttccccccat ttttttccaa
     2881 attccagatt tttttcaaaa attttccagc gctaccgctc ggcaaaattg caagcaattt
     2941 ttaaaatcaa acccatgagg gaatttcatt ccctcatact cccttgagcc tcctccaacc
     3001 gaaatagaag ggcgctgcgc ttattatttc attcagtcat cggctttcat aatctaacag
     3061 acaacatctt cgctgcaaag ccacgctacg ctcaagggct tttacgctac gataacgcct
     3121 gttttaacga ttatgccgat aactaaacga aataaacgct aaaacgtctc agaaacgatt
     3181 ttgagacgtt ttaataaaaa atcgcctagt gcttggattc tcaccaataa aaaacgcccg
     3241 gcggcaaccg agcgttctga acaaatccag atggagttct gaggtcatta ctggatctat
     3301 caacgggagt ccaagcgagc tcggtactaa aacaattcat ccagtaaaat ataatatttt
     3361 attttctccc aatcaggctt gatccccagt aagtcaaaaa atagctcgac atactgttct
     3421 tccccgatat cctccctgat cgaccggacg cagaaggcaa tgtcatacca cttgtccgcc
     3481 ctgccgcttc tcccaagatc aataaagcca cttactttgc catctttcac aaagatgttg
     3541 ctgtctccca ggtcgccgtg ggaaaagaca agttcctctt cgggcttttc cgtctttaaa
     3601 aaatcataca gctcgcgcgg atctttaaat ggagtgtctt cttcccagtt ttcgcaatcc
     3661 acatcggcca gatcgttatt cagtaagtaa tccaattcgg ctaagcggcc gtctaagcta
     3721 ttcgtatagg gacaatccga tatgtcgatg gagtgaaaga gcctgatgca ctccgcatac
     3781 agctcgatag tcttttcagg gctttgttca tcttcatacc cttccgagca aaggacgcca
     3841 tcggcctcac tcatgagcag attgctccag ccatcatgcc gttcaaagtg caggaccttt
     3901 ggaacaggca gctttccttc cagccatagc atcatgtcct tttcccgttc cacatcatag
     3961 gtggtccctt tataccggct gtccgtcatt tttaaatata ggatttcatt ttctcccacc
     4021 agcttatata ccttagcagg agacattcct tccgtatctt ttacgcagcg gtattcttcg
     4081 atcagttttt tcaattccgg tgatattctc attttagcca tttattattt ccttcctctt
     4141 ttctacagta tttaaagata ccccaagaag ctaattataa caagacgaac tccaattcac
     4201 tgttccttgc attctaaaac cttaaataca gaaaacagcc ttttcaaagt tgttttcaaa
     4261 gttggcgtat aacatagtat cgacggagcc gattttgaaa ccacaattat gatagaattt
     4321 gacgtcgaat tcgcggccgc ttctagaact atagctagca tgcatttacg ttgacaccat
     4381 cgaatggtgc aaaacctttc gcggtatggc atgatagcgc ccggaagaga gtcaattcag
     4441 ggtggtgaat gtgaaaccag taacgttata cgatgtcgca gagtatgccg gtgtctctta
     4501 tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa
     4561 agtggaagcg gcgatggcgg agctgaatta cattcccaac cgcgtggcac aacaactggc
     4621 gggcaaacag tcgttgctga ttggcgttgc cacctccagt ctggccctgc acgcgccgtc
     4681 gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc
     4741 gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc ttctcgcgca
     4801 acgcgtcagt gggctgatca ttaactatcc gctggatgac caggatgcca ttgctgtgga
     4861 agctgcctgc actaatgttc cggcgttatt tcttgatgtc tctgaccaga cacccatcaa
     4921 cagtattatt ttctcccatg aagacggtac gcgactgggc gtggagcatc tggtcgcatt
     4981 gggtcaccag caaatcgcgc tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg
     5041 tctggctggc tggcataaat atctcactcg caa
//
