LOCUS       sequence-310443-        5013 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pBAV1k-lacI-Trc-mCherry base, lacI repression vector for ADP1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5013)
  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 5013)
  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: pBWB162
FEATURES             Location/Qualifiers
     source          1..5013
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             34..45
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     terminator      complement(247..341)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(357..1151)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEEYRCVKDTEGMSPAKVYKLVGENEILYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEGYED
                     EQSPEKTIELYAECIRLFHSIDISDCPYTNSLDGRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     misc_feature    1357..1376
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that start with ""ATG"""
     promoter        1403..1480
                     /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             1481..2563
                     /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(1500..1519)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(2795..2800,2801..2817,2818..2824)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    2832..2848
                     /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)."
     primer_bind     2837..2859
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             2869..3579
                     /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(3014..3032)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3282..3301
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     terminator      3630..3716
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ccctcatact cccttgagcc tcctccaacc gaaatagaag ggcgctgcgc ttattatttc
       61 attcagtcat cggctttcat aatctaacag acaacatctt cgctgcaaag ccacgctacg
      121 ctcaagggct tttacgctac gataacgcct gttttaacga ttatgccgat aactaaacga
      181 aataaacgct aaaacgtctc agaaacgatt ttgagacgtt ttaataaaaa atcgcctagt
      241 gcttggattc tcaccaataa aaaacgcccg gcggcaaccg agcgttctga acaaatccag
      301 atggagttct gaggtcatta ctggatctat caacgggagt ccaagcgagc tcggtactaa
      361 aacaattcat ccagtaaaat ataatatttt attttctccc aatcaggctt gatccccagt
      421 aagtcaaaaa atagctcgac atactgttct tccccgatat cctccctgat cgaccggacg
      481 cagaaggcaa tgtcatacca cttgtccgcc ctgccgcttc tcccaagatc aataaagcca
      541 cttactttgc catctttcac aaagatgttg ctgtctccca ggtcgccgtg ggaaaagaca
      601 agttcctctt cgggcttttc cgtctttaaa aaatcataca gctcgcgcgg atctttaaat
      661 ggagtgtctt cttcccagtt ttcgcaatcc acatcggcca gatcgttatt cagtaagtaa
      721 tccaattcgg ctaagcggcc gtctaagcta ttcgtatagg gacaatccga tatgtcgatg
      781 gagtgaaaga gcctgatgca ctccgcatac agctcgatag tcttttcagg gctttgttca
      841 tcttcatacc cttccgagca aaggacgcca tcggcctcac tcatgagcag attgctccag
      901 ccatcatgcc gttcaaagtg caggaccttt ggaacaggca gctttccttc cagccatagc
      961 atcatgtcct tttcccgttc cacatcatag gtggtccctt tataccggct gtccgtcatt
     1021 tttaaatata ggatttcatt ttctcccacc agcttatata ccttagcagg agacattcct
     1081 tccgtatctt ttacgcagcg gtattcttcg atcagttttt tcaattccgg tgatattctc
     1141 attttagcca tttattattt ccttcctctt ttctacagta tttaaagata ccccaagaag
     1201 ctaattataa caagacgaac tccaattcac tgttccttgc attctaaaac cttaaataca
     1261 gaaaacagcc ttttcaaagt tgttttcaaa gttggcgtat aacatagtat cgacggagcc
     1321 gattttgaaa ccacaattat gatagaattt gacgtcgaat tcgcggccgc ttctagaact
     1381 atagctagca tgcatttacg ttgacaccat cgaatggtgc aaaacctttc gcggtatggc
     1441 atgatagcgc ccggaagaga gtcaattcag ggtggtgaat gtgaaaccag taacgttata
     1501 cgatgtcgca gagtatgccg gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc
     1561 cagccacgtt tctgcgaaaa cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta
     1621 cattcccaac cgcgtggcac aacaactggc gggcaaacag tcgttgctga ttggcgttgc
     1681 cacctccagt ctggccctgc acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc
     1741 cgatcaactg ggtgccagcg tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg
     1801 taaagcggcg gtgcacaatc ttctcgcgca acgcgtcagt gggctgatca ttaactatcc
     1861 gctggatgac caggatgcca ttgctgtgga agctgcctgc actaatgttc cggcgttatt
     1921 tcttgatgtc tctgaccaga cacccatcaa cagtattatt ttctcccatg aagacggtac
     1981 gcgactgggc gtggagcatc tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg
     2041 cccattaagt tctgtctcgg cgcgtctgcg tctggctggc tggcataaat atctcactcg
     2101 caatcaaatt cagccgatag cggaacggga aggcgactgg agtgccatgt ccggttttca
     2161 acaaaccatg caaatgctga atgagggcat cgttcccact gcgatgctgg ttgccaacga
     2221 tcagatggcg ctgggcgcaa tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga
     2281 tatctcggta gtgggatacg acgataccga agacagctca tgttatatcc cgccgttaac
     2341 caccatcaaa caggattttc gcctgctggg gcaaaccagc gtggaccgct tgctgcaact
     2401 ctctcagggc caggcggtga agggcaatca gctgttgccc gtctcactgg tgaaaagaaa
     2461 aaccaccctg gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     2521 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
     2581 taagttagcg cgaattgatc tggtttgaca gcttatcatc gactgcacgg tgcaccaatg
     2641 cttctggcgt caggcagcca tcggaagctg tggtatggct gtgcaggtcg taaatcactg
     2701 cataattcgt gtcgctcaag gcgcactccc gttctggata atgttttttg cgccgacatc
     2761 ataacggttc tggcaaatat tctgaaatga gctgttgaca attaatcatc cggctcgtat
     2821 aatgtgtgga attgtgagcg gataacaatt tcacacagga aacagaccat ggtgagcaag
     2881 ggcgaggagg ataacatggc catcatcaag gagttcatgc gcttcaaggt tcacatggag
     2941 ggctccgtga acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag
     3001 ggcacccaga ccgccaagct gaaggtgacc aagggtggcc ccctgccctt cgcctgggac
     3061 atcctgtccc ctcagttcat gtacggctcc aaggcctacg tgaagcaccc cgccgacatc
     3121 cccgactact tgaagctgtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc
     3181 gaggacggcg gcgtggtgac cgtgacccag gactcctccc tgcaagacgg cgagttcatc
     3241 tacaaggtga agctgcgcgg caccaacttc ccctccgacg gccccgtaat gcagaagaag
     3301 actatgggct gggaggcctc ctccgagcgg atgtaccccg aggacggcgc gctgaagggc
     3361 gagatcaagc agaggctgaa gctgaaggac ggcggccact acgacgctga ggtcaagacc
     3421 acctacaagg ccaagaagcc cgtgcaactg cccggcgcgt acaacgtcaa catcaagttg
     3481 gacatcacct cccacaacga ggactacacc atcgtggaac agtacgaacg cgccgagggc
     3541 cgccactcca ccggcggcat ggacgagctg tataagtaag cctcagggcc cgatcgatgc
     3601 cgccgcttaa ttaattaatc cagaggcatc aaataaaacg aaaggctcag tcgaaagact
     3661 gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc
     3721 cgccctagac ctagtgtcat tttatttccc ccgtttcagc atcaagaacc tttgcataac
     3781 ttgctctata tccacactga taattgccct caaaccataa tctaaaggcg ctagagtttg
     3841 ttgaaacaat atcttttaca tcattcgtat ttaaaattcc aaactccgct cccctaaggc
     3901 gaataaaagc cattaaatct tttgtattta ccaaattata gtcatccact atatctaaga
     3961 gtaaattctt caattctctt ttttggcttt catcaagtgt tatatagcgg tcaatatcaa
     4021 aatcattaat gttcaaaata tcttttttgt cgtatatatg tttattctta gcaatagcgt
     4081 cctttgattc atgagtcaaa tattcatatg aacctttgat ataatcaagt atctcaacat
     4141 gagcaactga actattcccc aattttcgct taatcttgtt cctaacgctt tctattgtta
     4201 caggatttcg tgcaatatat ataacgtgat agtgtggttt tttatagtgc tttccatttc
     4261 gtataacatc actactattc catgtatctt tatctttttt ttcgtccata tcgtgtaaag
     4321 gactgacagc catagatacg cccaaactct ctaatttttc cttccaatca ttaggaattg
     4381 agtcaggata taataaaaat ccaaaatttc tagctttagt atttttaata gccatgatat
     4441 aattacctta tcaaaaacaa gtagcgaaaa ctcgtatcct tctaaaaacg cgagctttcg
     4501 cttanttttt ttgttctgat tcctttcttg catattcttc tatagctaac gccgcaaccg
     4561 cagattttga aaaacctttt tgtttcgcca tatctgttaa ttttttatct tgctcttttg
     4621 tcagagaaat cataactctt tttttcgatt ctgaaatcac catttaaaaa actccaatca
     4681 aataatttta taaagttagt gtatcacttt gtaatcataa aaacaacaat aaagctactt
     4741 aaatatagat ttataaaaaa cgttggcgaa aacgttggcg attcgttggc gattgaaaaa
     4801 ccccttaaac ccttgagcca gttgggatag agcgtttttg gcacaaaaat tggcactcgg
     4861 cacttaatgg ggggtcgtag tacggaagca aaattcgctt cctttccccc catttttttc
     4921 caaattccag atttttttca aaaattttcc agcgctaccg ctcggcaaaa ttgcaagcaa
     4981 tttttaaaat caaacccatg agggaatttc att
//
