LOCUS       sequence-272915-        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             1200..1211
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     terminator      complement(1413..1507)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1523..2317)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEEYRCVKDTEGMSPAKVYKLVGENEILYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEGYED
                     EQSPEKTIELYAECIRLFHSIDISDCPYTNSLDGRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     misc_feature    2523..2542
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that start with ""ATG"""
     promoter        2569..2646
                     /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             2647..3729
                     /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
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(2666..2685)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(3961..3966,3967..3983,3984..3990)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    3998..4014
                     /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     4003..4025
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             4035..4745
                     /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(4180..4198)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4448..4467
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     terminator      4796..4882
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 aatatctttt acatcattcg tatttaaaat tccaaactcc gctcccctaa ggcgaataaa
       61 agccattaaa tcttttgtat ttaccaaatt atagtcatcc actatatcta agagtaaatt
      121 cttcaattct cttttttggc tttcatcaag tgttatatag cggtcaatat caaaatcatt
      181 aatgttcaaa atatcttttt tgtcgtatat atgtttattc ttagcaatag cgtcctttga
      241 ttcatgagtc aaatattcat atgaaccttt gatataatca agtatctcaa catgagcaac
      301 tgaactattc cccaattttc gcttaatctt gttcctaacg ctttctattg ttacaggatt
      361 tcgtgcaata tatataacgt gatagtgtgg ttttttatag tgctttccat ttcgtataac
      421 atcactacta ttccatgtat ctttatcttt tttttcgtcc atatcgtgta aaggactgac
      481 agccatagat acgcccaaac tctctaattt ttccttccaa tcattaggaa ttgagtcagg
      541 atataataaa aatccaaaat ttctagcttt agtattttta atagccatga tataattacc
      601 ttatcaaaaa caagtagcga aaactcgtat ccttctaaaa acgcgagctt tcgcttattt
      661 tttttgttct gattcctttc ttgcatattc ttctatagct aacgccgcaa ccgcagattt
      721 tgaaaaacct ttttgtttcg ccatatctgt taatttttta tcttgctctt ttgtcagaga
      781 aatcataact ctttttttcg attctgaaat caccatttaa aaaactccaa tcaaataatt
      841 ttataaagtt agtgtatcac tttgtaatca taaaaacaac aataaagcta cttaaatata
      901 gatttataaa aaacgttggc gaaaacgttg gcgattcgtt ggcgattgaa aaacccctta
      961 aacccttgag ccagttggga tagagcgttt ttggcacaaa aattggcact cggcacttaa
     1021 tggggggtcg tagtacggaa gcaaaattcg cttcctttcc ccccattttt ttccaaattc
     1081 caaatttttt tcaaaaattt tccagcgcta ccgctcggca aaattgcaag caatttttaa
     1141 aatcaaaccc atgagggaat ttcattccct catactccct tgagcctcct ccaaccgaaa
     1201 tagaagggcg ctgcgcttat tatttcattc agtcatcggc tttcataatc taacagacaa
     1261 catcttcgct gcaaagccac gctacgctca agggctttta cgctacgata acgcctgttt
     1321 taacgattat gccgataact aaacgaaata aacgctaaaa cgtctcagaa acgattttga
     1381 gacgttttaa taaaaaatcg cctagtgctt ggattctcac caataaaaaa cgcccggcgg
     1441 caaccgagcg ttctgaacaa atccagatgg agttctgagg tcattactgg atctatcaac
     1501 aggagtccaa gcgagctcgg tactaaaaca attcatccag taaaatataa tattttattt
     1561 tctcccaatc aggcttgatc cccagtaagt caaaaaatag ctcgacatac tgttcttccc
     1621 cgatatcctc cctgatcgac cggacgcaga aggcaatgtc ataccacttg tccgccctgc
     1681 cgcttctccc aagatcaata aagccactta ctttgccatc tttcacaaag atgttgctgt
     1741 ctcccaggtc gccgtgggaa aagacaagtt cctcttcggg cttttccgtc tttaaaaaat
     1801 catacagctc gcgcggatct ttaaatggag tgtcttcttc ccagttttcg caatccacat
     1861 cggccagatc gttattcagt aagtaatcca attcggctaa gcggccgtct aagctattcg
     1921 tatagggaca atccgatatg tcgatggagt gaaagagcct gatgcactcc gcatacagct
     1981 cgatagtctt ttcagggctt tgttcatctt catacccttc cgagcaaagg acgccatcgg
     2041 cctcactcat gagcagattg ctccagccat catgccgttc aaagtgcagg acctttggaa
     2101 caggcagctt tccttccagc catagcatca tgtccttttc ccgttccaca tcataggtgg
     2161 tccctttata ccggctgtcc gtcattttta aatataggat ttcattttct cccaccagct
     2221 tatatacctt agcaggagac attccttccg tatcttttac gcagcggtat tcttcgatca
     2281 gttttttcaa ttccggtgat attctcattt tagccattta ttatttcctt cctcttttct
     2341 acagtattta aagatacccc aagaagctaa ttataacaag acgaactcca attcactgtt
     2401 ccttgcattc taaaacctta aatacagaaa acagcctttt caaagttgtt ttcaaagttg
     2461 gcgtataaca tagtatcgac ggagccgatt ttgaaaccac aattatgata gaatttgacg
     2521 tcgaattcgc ggccgcttct agaactatag ctagcatgca tttacgttga caccatcgaa
     2581 tggtgcaaaa cctttcgcgg tatggcatga tagcgcccgg aagagagtca attcagggtg
     2641 gtgaatgtga aaccagtaac gttatacgat gtcgcagagt atgccggtgt ctcttatcag
     2701 accgtttccc gcgtggtgaa ccaggccagc cacgtttctg cgaaaacgcg ggaaaaagtg
     2761 gaagcggcga tggcggagct gaattacatt cccaaccgcg tggcacaaca actggcgggc
     2821 aaacagtcgt tgctgattgg cgttgccacc tccagtctgg ccctgcacgc gccgtcgcaa
     2881 attgtcgcgg cgattaaatc tcgcgccgat caactgggtg ccagcgtggt ggtgtcgatg
     2941 gtagaacgaa gcggcgtcga agcctgtaaa gcggcggtgc acaatcttct cgcgcaacgc
     3001 gtcagtgggc tgatcattaa ctatccgctg gatgaccagg atgccattgc tgtggaagct
     3061 gcctgcacta atgttccggc gttatttctt gatgtctctg accagacacc catcaacagt
     3121 attattttct cccatgaaga cggtacgcga ctgggcgtgg agcatctggt cgcattgggt
     3181 caccagcaaa tcgcgctgtt agcgggccca ttaagttctg tctcggcgcg tctgcgtctg
     3241 gctggctggc ataaatatct cactcgcaat caaattcagc cgatagcgga acgggaaggc
     3301 gactggagtg ccatgtccgg ttttcaacaa accatgcaaa tgctgaatga gggcatcgtt
     3361 cccactgcga tgctggttgc caacgatcag atggcgctgg gcgcaatgcg cgccattacc
     3421 gagtccgggc tgcgcgttgg tgcggatatc tcggtagtgg gatacgacga taccgaagac
     3481 agctcatgtt atatcccgcc gtcaaccacc atcaaacagg attttcgcct gctggggcaa
     3541 accagcgtgg accgcttgct gcaactctct cagggccagg cggtgaaggg caatcagctg
     3601 ttgcccgtct cactggtgaa aagaaaaacc accctggcgc ccaatacgca aaccgcctct
     3661 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
     3721 gggcagtgag cgcaacgcaa ttaatgtgag ttagcgcgaa ttgatctggt ttgacagctt
     3781 atcatcgact gcacggtgca ccaatgcttc tggcgtcagg cagccatcgg aagctgtggt
     3841 atggctgtgc aggtcgtaaa tcactgcata attcgtgtcg ctcaaggcgc actcccgttc
     3901 tggataatgt tttttgcgcc gacatcataa cggttctggc aaatattctg aaatgagctg
     3961 ttgacaatta atcatccggc tcgtataatg tgtggaattg tgagcggata acaatttcac
     4021 acaggaaaca gaccatggtg agcaagggcg aggaggataa catggccatc atcaaggagt
     4081 tcatgcgctt caaggttcac atggagggct ccgtgaacgg ccacgagttc gagatcgagg
     4141 gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag gtgaccaagg
     4201 gtggccccct gcccttcgcc tgggacatcc tgtcccctca gttcatgtac ggctccaagg
     4261 cctacgtgaa gcaccccgcc gacatccccg actacttgaa gctgtccttc cccgagggct
     4321 tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg acccaggact
     4381 cctccctgca agacggcgag ttcatctaca aggtgaagct gcgcggcacc aacttcccct
     4441 ccgacggccc cgtaatgcag aagaagacta tgggctggga ggcctcctcc gagcggatgt
     4501 accccgagga cggcgcgctg aagggcgaga tcaagcagag gctgaagctg aaggacggcg
     4561 gccactacga cgctgaggtc aagaccacct acaaggccaa gaagcccgtg caactgcccg
     4621 gcgcgtacaa cgtcaacatc aagttggaca tcacctccca caacgaggac tacaccatcg
     4681 tggaacagta cgaacgcgcc gagggccgcc actccaccgg cggcatggac gagctgtata
     4741 agtaagcctc agggcccgat cgatgccgcc gcttaattaa ttaatccaga ggcatcaaat
     4801 aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa
     4861 cgctctcctg agtaggacaa atccgccgcc ctagacctag tgtcatttta tttcccccgt
     4921 ttcagcatca agaacctttg cataacttgc tctatatcca cactgataat tgccctcaaa
     4981 ccataatcta aaggcgctag agtttgttga aac
//
