LOCUS       sequence-39772-e        6213 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6213)
  AUTHORS   Murin CD, Segal K, Bryksin A, Matsumura I
  TITLE     Expression vectors for the engineering of genes and genomes in 
            Acinetobacter baylyi ADP1.
  JOURNAL   Appl Environ Microbiol. 2011 Oct 21.
  PUBMED    22020504
REFERENCE   2  (bases 1 to 6213)
  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: "Appl 
            Environ Microbiol. 2011 Oct 21."
COMMENT     Sequence Label: lacI-Ptac-gusA-pBAV1k
FEATURES             Location/Qualifiers
     source          1..6213
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(193..198,199..215,216..222)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    230..246
                     /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     235..257
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             279..296
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             321..2126
                     /codon_start=1
                     /gene="uidA (or gusA) from E. coli"
                     /product="beta-glucuronidase"
                     /label=GUS
                     /translation="LRPVETPTREIKKLDGLWAFSLDRENCGIDQRWWESALQESRAIA
                     VPGSFNDQFADADIRNYAGNVWYQREVFIPKGWAGQRIVLRFDAVTHYGKVWVNNQEVM
                     EHQGGYTPFEADVTPYVIAGKSVRITVCVNNELNWQTIPPGMVITDENGKKKQSYFHDF
                     FNYAGIHRSVMLYTTPNTWVDDITVVTHVAQDCNHASVDWQVVANGDVSVELRDADQQV
                     VATGQGTSGTLQVVNPHLWQPGEGYLYELCVTAKSQTECDIYPLRVGIRSVAVKGEQFL
                     INHKPFYFTGFGRHEDADLRGKGFDNVLMVHDHALMDWIGANSYRTSHYPYAEEMLDWA
                     DEHGIVVIDETAAVGFNLSLGIGFEAGNKPKELYSEEAVNGETQQAHLQAIKELIARDK
                     NHPSVVMWSIANEPDTRPQGAREYFAPLAEATRKLDPTRPITCVNVMFCDAHTDTISDL
                     FDVLCLNRYYGWYVQSGDLETAEKVLEKELLAWQEKLHQPIIITEYGVDTLAGLHSMYT
                     DMWSEEYQCAWLDMYHRVFDRVSAVVGEQVWNFADFATSQGILRVGGNKKGIFTRDRKP
                     KSAAFLLQKRWTGMNFGEKPQQGGKQ"
     misc_feature    2157..2177
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      2228..2314
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             3645..3656
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     terminator      complement(3858..3952)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3968..4762)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEEYRCVKDTEGMSPAKVYKLVGENEILYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEGYED
                     EQSPEKTIELYAECIRLFHSIDISDCPYTNSLDGRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     misc_feature    4968..4987
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that start with ""ATG"""
     promoter        5014..5091
                     /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             5092..6174
                     /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(5111..5130)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 gtttgacagc ttatcatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc
       61 ggaagctgtg gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc
      121 gcactcccgt tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc
      181 tgaaatgagc tgttgacaat taatcatccg gctcgtataa tgtgtggaat tgtgagcgga
      241 taacaatttc acacaggaaa cagaccatgg gcagcagcca tcaccatcat caccacagcc
      301 aggatccggc cctgagggcc ttacgtcctg tagaaacccc aacccgtgaa atcaaaaaac
      361 tcgacggcct gtgggcattc agtctggatc gcgaaaactg tggaattgat cagcgttggt
      421 gggaaagcgc gttacaagaa agccgggcaa ttgctgtgcc aggcagtttt aacgatcagt
      481 tcgccgatgc agatattcgt aattatgcgg gcaacgtctg gtatcagcgc gaagtcttta
      541 taccgaaagg ttgggcaggc cagcgtatcg tgctgcgttt cgatgcggtc actcattacg
      601 gcaaagtgtg ggtcaataat caggaagtga tggagcatca gggcggctat acgccatttg
      661 aagccgatgt cacgccgtat gttattgccg ggaaaagtgt acgtatcacc gtttgtgtga
      721 acaacgaact gaactggcag actatcccgc cgggaatggt gattaccgac gaaaacggca
      781 agaaaaagca gtcttacttc catgatttct ttaactatgc cgggatccat cgcagcgtaa
      841 tgctctacac cacgccgaac acctgggtgg acgatatcac cgtggtgacg catgtcgcgc
      901 aagactgtaa ccacgcgtct gttgactggc aggtggtggc caatggtgat gtcagcgttg
      961 aactgcgtga tgcggatcaa caggtggttg caactggaca aggcactagc gggactttgc
     1021 aagtggtgaa tccgcacctc tggcaaccgg gtgaaggtta tctctatgaa ctgtgcgtca
     1081 cagccaaaag ccagacagag tgtgatatct acccgcttcg cgtcggcatc cggtcagtgg
     1141 cagtgaaggg cgaacagttc ctgattaacc acaaaccgtt ctactttact ggctttggtc
     1201 gtcatgaaga tgcggacttg cgtggcaaag gattcgataa cgtgctgatg gtgcacgacc
     1261 acgcattaat ggactggatt ggggccaact cctaccgtac ctcgcattac ccttacgctg
     1321 aagagatgct cgactgggca gatgaacatg gcatcgtggt gattgatgaa actgctgctg
     1381 tcggctttaa cctctcttta ggcattggtt tcgaagcggg caacaagccg aaagaactgt
     1441 acagcgaaga ggcagtcaac ggggaaactc agcaagcgca cttacaggcg attaaagagc
     1501 tgatagcgcg tgacaaaaac cacccaagcg tggtgatgtg gagtattgcc aacgaaccgg
     1561 atacccgtcc gcaaggtgca cgggaatatt tcgcgccact ggcggaagca acgcgtaaac
     1621 tcgacccgac gcgtccgatc acctgcgtca atgtaatgtt ctgcgacgct cacaccgata
     1681 ccatcagcga tctctttgat gtgctgtgcc tgaaccgtta ttacggatgg tatgtccaaa
     1741 gcggcgattt ggaaacggca gagaaggtac tggaaaaaga acttctggcc tggcaggaga
     1801 aactgcatca gccgattatc atcaccgaat acggcgtgga tacgttagcc gggctgcact
     1861 caatgtacac cgacatgtgg agtgaagagt atcagtgtgc atggctggat atgtatcacc
     1921 gcgtctttga tcgcgtcagc gccgtcgtcg gtgaacaggt atggaatttc gccgattttg
     1981 cgacctcgca aggcatattg cgcgttggcg gtaacaagaa agggatcttc actcgcgacc
     2041 gcaaaccgaa gtcggcggct tttctgctgc aaaaacgctg gactggcatg aacttcggtg
     2101 aaaaaccgca gcagggaggc aaacaaggcc tatgcggccg gtagaaagga gaagcttact
     2161 agtagcggcc gctgcaggcc tcagggcccg atcgatgccg ccgcttaatt aattaatcca
     2221 gaggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg
     2281 tttgtcggtg aacgctctcc tgagtaggac aaatccgccg ccctagacct agtgtcattt
     2341 tatttccccc gtttcagcat caagaacctt tgcataactt gctctatatc cacactgata
     2401 attgccctca aaccataatc taaaggcgct agagtttgtt gaaacaatat cttttacatc
     2461 attcgtattt aaaattccaa actccgctcc cctaaggcga ataaaagcca ttaaatcttt
     2521 tgtatttacc aaattatagt catccactat atctaagagt aaattcttca attctctttt
     2581 ttggctttca tcaagtgtta tatagcggtc aatatcaaaa tcattaatgt tcaaaatatc
     2641 ttttttgtcg tatatatgtt tattcttagc aatagcgtcc tttgattcat gagtcaaata
     2701 ttcatatgaa cctttgatat aatcaagtat ctcaacatga gcaactgaac tattccccaa
     2761 ttttcgctta atcttgttcc taacgctttc tattgttaca ggatttcgtg caatatatat
     2821 aacgtgatag tgtggttttt tatagtgctt tccatttcgt ataacatcac tactattcca
     2881 tgtatcttta tctttttttt cgtccatatc gtgtaaagga ctgacagcca tagatacgcc
     2941 caaactctct aatttttcct tccaatcatt aggaattgag tcaggatata ataaaaatcc
     3001 aaaatttcta gctttagtat ttttaatagc catgatataa ttaccttatc aaaaacaagt
     3061 agcgaaaact cgtatccttc taaaaacgcg agctttcgct tatttttttt gttctgattc
     3121 ctttcttgca tattcttcta tagctaacgc cgcaaccgca gattttgaaa aacctttttg
     3181 tttcgccata tctgttaatt ttttatcttg ctcttttgtc agagaaatca taactctttt
     3241 tttcgattct gaaatcacca tttaaaaaac tccaatcaaa taattttata aagttagtgt
     3301 atcactttgt aatcataaaa acaacaataa agctacttaa atatagattt ataaaaaacg
     3361 ttggcgaaaa cgttggcgat tcgttggcga ttgaaaaacc ccttaaaccc ttgagccagt
     3421 tgggatagag cgtttttggc acaaaaattg gcactcggca cttaatgggg ggtcgtagta
     3481 cggaagcaaa attcgcttcc tttcccccca tttttttcca aattccaaat ttttttcaaa
     3541 aattttccag cgctaccgct cggcaaaatt gcaagcaatt tttaaaatca aacccatgag
     3601 ggaatttcat tccctcatac tcccttgagc ctcctccaac cgaaatagaa gggcgctgcg
     3661 cttattattt cattcagtca tcggctttca taatctaaca gacaacatct tcgctgcaaa
     3721 gccacgctac gctcaagggc ttttacgcta cgataacgcc tgttttaacg attatgccga
     3781 taactaaacg aaataaacgc taaaacgtct cagaaacgat tttgagacgt tttaataaaa
     3841 aatcgcctag tgcttggatt ctcaccaata aaaaacgccc ggcggcaacc gagcgttctg
     3901 aacaaatcca gatggagttc tgaggtcatt actggatcta tcaacaggag tccaagcgag
     3961 ctcggtacta aaacaattca tccagtaaaa tataatattt tattttctcc caatcaggct
     4021 tgatccccag taagtcaaaa aatagctcga catactgttc ttccccgata tcctccctga
     4081 tcgaccggac gcagaaggca atgtcatacc acttgtccgc cctgccgctt ctcccaagat
     4141 caataaagcc acttactttg ccatctttca caaagatgtt gctgtctccc aggtcgccgt
     4201 gggaaaagac aagttcctct tcgggctttt ccgtctttaa aaaatcatac agctcgcgcg
     4261 gatctttaaa tggagtgtct tcttcccagt tttcgcaatc cacatcggcc agatcgttat
     4321 tcagtaagta atccaattcg gctaagcggc cgtctaagct attcgtatag ggacaatccg
     4381 atatgtcgat ggagtgaaag agcctgatgc actccgcata cagctcgata gtcttttcag
     4441 ggctttgttc atcttcatac ccttccgagc aaaggacgcc atcggcctca ctcatgagca
     4501 gattgctcca gccatcatgc cgttcaaagt gcaggacctt tggaacaggc agctttcctt
     4561 ccagccatag catcatgtcc ttttcccgtt ccacatcata ggtggtccct ttataccggc
     4621 tgtccgtcat ttttaaatat aggatttcat tttctcccac cagcttatat accttagcag
     4681 gagacattcc ttccgtatct tttacgcagc ggtattcttc gatcagtttt ttcaattccg
     4741 gtgatattct cattttagcc atttattatt tccttcctct tttctacagt atttaaagat
     4801 accccaagaa gctaattata acaagacgaa ctccaattca ctgttccttg cattctaaaa
     4861 ccttaaatac agaaaacagc cttttcaaag ttgttttcaa agttggcgta taacatagta
     4921 tcgacggagc cgattttgaa accacaatta tgatagaatt tgacgtcgaa ttcgcggccg
     4981 cttctagaac tatagctagc atgcatttac gttgacacca tcgaatggtg caaaaccttt
     5041 cgcggtatgg catgatagcg cccggaagag agtcaattca gggtggtgaa tgtgaaacca
     5101 gtaacgttat acgatgtcgc agagtatgcc ggtgtctctt atcagaccgt ttcccgcgtg
     5161 gtgaaccagg ccagccacgt ttctgcgaaa acgcgggaaa aagtggaagc ggcgatggcg
     5221 gagctgaatt acattcccaa ccgcgtggca caacaactgg cgggcaaaca gtcgttgctg
     5281 attggcgttg ccacctccag tctggccctg cacgcgccgt cgcaaattgt cgcggcgatt
     5341 aaatctcgcg ccgatcaact gggtgccagc gtggtggtgt cgatggtaga acgaagcggc
     5401 gtcgaagcct gtaaagcggc ggtgcacaat cttctcgcgc aacgcgtcag tgggctgatc
     5461 attaactatc cgctggatga ccaggatgcc attgctgtgg aagctgcctg cactaatgtt
     5521 ccggcgttat ttcttgatgt ctctgaccag acacccatca acagtattat tttctcccat
     5581 gaagacggta cgcgactggg cgtggagcat ctggtcgcat tgggtcacca gcaaatcgcg
     5641 ctgttagcgg gcccattaag ttctgtctcg gcgcgtctgc gtctggctgg ctggcataaa
     5701 tatctcactc gcaatcaaat tcagccgata gcggaacggg aaggcgactg gagtgccatg
     5761 tccggttttc aacaaaccat gcaaatgctg aatgagggca tcgttcccac tgcgatgctg
     5821 gttgccaacg atcagatggc gctgggcgca atgcgcgcca ttaccgagtc cgggctgcgc
     5881 gttggtgcgg atatctcggt agtgggatac gacgataccg aagacagctc atgttatatc
     5941 ccgccgtcaa ccaccatcaa acaggatttt cgcctgctgg ggcaaaccag cgtggaccgc
     6001 ttgctgcaac tctctcaggg ccaggcggtg aagggcaatc agctgttgcc cgtctcactg
     6061 gtgaaaagaa aaaccaccct ggcgcccaat acgcaaaccg cctctccccg cgcgttggcc
     6121 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
     6181 cgcaattaat gtgagttagc gcgaattgat ctg
//
