LOCUS       sequence-39783-e        6202 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 6202)
  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 6202)
  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-PT7-gusA-pBAV1k
FEATURES             Location/Qualifiers
     source          1..6202
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    3..27
                     /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             83..100
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             125..1930
                     /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    1961..1981
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      2032..2118
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             3449..3460
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     terminator      complement(3662..3756)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3772..4566)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEEYRCVKDTEGMSPAKVYKLVGENEILYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEGYED
                     EQSPEKTIELYAECIRLFHSIDISDCPYTNSLDGRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     misc_feature    4772..4791
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that start with ""ATG"""
     CDS             complement(4980..6062)
                     /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     6024..6043
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6063..6140)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     join(6186..6202,1..3)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        join(6186..6202,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaataag
       61 gagatatacc atgggcagca gccatcacca tcatcaccac agccaggatc cggccctgag
      121 ggccttacgt cctgtagaaa ccccaacccg tgaaatcaaa aaactcgacg gcctgtgggc
      181 attcagtctg gatcgcgaaa actgtggaat tgatcagcgt tggtgggaaa gcgcgttaca
      241 agaaagccgg gcaattgctg tgccaggcag ttttaacgat cagttcgccg atgcagatat
      301 tcgtaattat gcgggcaacg tctggtatca gcgcgaagtc tttataccga aaggttgggc
      361 aggccagcgt atcgtgctgc gtttcgatgc ggtcactcat tacggcaaag tgtgggtcaa
      421 taatcaggaa gtgatggagc atcagggcgg ctatacgcca tttgaagccg atgtcacgcc
      481 gtatgttatt gccgggaaaa gtgtacgtat caccgtttgt gtgaacaacg aactgaactg
      541 gcagactatc ccgccgggaa tggtgattac cgacgaaaac ggcaagaaaa agcagtctta
      601 cttccatgat ttctttaact atgccggaat ccatcgcagc gtaatgctct acaccacgcc
      661 gaacacctgg gtggacgata tcaccgtggt gacgcatgtc gcgcaagact gtaaccacgc
      721 gtctgttgac tggcaggtgg tggccaatgg tgatgtcagc gttgaactgc gtgatgcgga
      781 tcaacaggtg gttgcaactg gacaaggcac tagcgggact ttgcaagtgg tgaatccgca
      841 cctctggcaa ccgggtgaag gttatctcta tgaactgtgc gtcacagcca aaagccagac
      901 agagtgtgat atctacccgc ttcgcgtcgg catccggtca gtggcagtga agggcgaaca
      961 gttcctgatt aaccacaaac cgttctactt tactggcttt ggtcgtcatg aagatgcgga
     1021 cttgcgtggc aaaggattcg ataacgtgct gatggtgcac gaccacgcat taatggactg
     1081 gattggggcc aactcctacc gtacctcgca ttacccttac gctgaagaga tgctcgactg
     1141 ggcagatgaa catggcatcg tggtgattga tgaaactgct gctgtcggct ttaacctctc
     1201 tttaggcatt ggtttcgaag cgggcaacaa gccgaaagaa ctgtacagcg aagaggcagt
     1261 caacggggaa actcagcaag cgcacttaca ggcgattaaa gagctgatag cgcgtgacaa
     1321 aaaccaccca agcgtggtga tgtggagtat tgccaacgaa ccggataccc gtccgcaagg
     1381 tgcacgggaa tatttcgcgc cactggcgga agcaacgcgt aaactcgacc cgacgcgtcc
     1441 gatcacctgc gtcaatgtaa tgttctgcga cgctcacacc gataccatca gcgatctctt
     1501 tgatgtgctg tgcctgaacc gttattacgg atggtatgtc caaagcggcg atttggaaac
     1561 ggcagagaag gtactggaaa aagaacttct ggcctggcag gagaaactgc atcagccgat
     1621 tatcatcacc gaatacggcg tggatacgtt agccgggctg cactcaatgt acaccgacat
     1681 gtggagtgaa gagtatcagt gtgcatggct ggatatgtat caccgcgtct ttgatcgcgt
     1741 cagcgccgtc gtcggtgaac aggtatggaa tttcgccgat tttgcgacct cgcaaggcat
     1801 attgcgcgtt ggcggtaaca agaaagggat cttcactcgc gaccgcaaac cgaagtcggc
     1861 ggcttttctg ctgcaaaaac gctggactgg catgaacttc ggtgaaaaac cgcagcaggg
     1921 aggcaaacaa ggcctatgcg gccggtagaa aggagaagct tactagtagc ggccgctgca
     1981 ggcctcaggg cccgatcgat gccgccgctt aattaattaa tccagaggca tcaaataaaa
     2041 cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct
     2101 ctcctgagta ggacaaatcc gccgccctag acctagtgtc attttatttc ccccgtttca
     2161 gcatcaagaa cctttgcata acttgctcta tatccacact gataattgcc ctcaaaccat
     2221 aatctaaagg cgctagagtt tgttgaaaca atatctttta catcattcgt atttaaaatt
     2281 ccaaactccg ctcccctaag gcgaataaaa gccattaaat cttttgtatt taccaaatta
     2341 tagtcatcca ctatatctaa gagtaaattc ttcaattctc ttttttggct ttcatcaagt
     2401 gttatatagc ggtcaatatc aaaatcatta atgttcaaaa tatctttttt gtcgtatata
     2461 tgtttattct tagcaatagc gtcctttgat tcatgagtca aatattcata tgaacctttg
     2521 atataatcaa gtatctcaac atgagcaact gaactattcc ccaattttcg cttaatcttg
     2581 ttcctaacgc tttctattgt tacaggattt cgtgcaatat atataacgtg atagtgtggt
     2641 tttttatagt gctttccatt tcgtataaca tcactactat tccatgtatc tttatctttt
     2701 ttttcgtcca tatcgtgtaa aggactgaca gccatagata cgcccaaact ctctaatttt
     2761 tccttccaat cattaggaat tgagtcagga tataataaaa atccaaaatt tctagcttta
     2821 gtatttttaa tagccatgat ataattacct tatcaaaaac aagtagcgaa aactcgtatc
     2881 cttctaaaaa cgcgagcttt cgcttatttt ttttgttctg attcctttct tgcatattct
     2941 tctatagcta acgccgcaac cgcagatttt gaaaaacctt tttgtttcgc catatctgtt
     3001 aattttttat cttgctcttt tgtcagagaa atcataactc tttttttcga ttctgaaatc
     3061 accatttaaa aaactccaat caaataattt tataaagtta gtgtatcact ttgtaatcat
     3121 aaaaacaaca ataaagctac ttaaatatag atttataaaa aacgttggcg aaaacgttgg
     3181 cgattcgttg gcgattgaaa aaccccttaa acccttgagc cagttgggat agagcgtttt
     3241 tggcacaaaa attggcactc ggcacttaat ggggggtcgt agtacggaag caaaattcgc
     3301 ttcctttccc cccatttttt tccaaattcc aaattttttt caaaaatttt ccagcgctac
     3361 cgctcggcaa aattgcaagc aatttttaaa atcaaaccca tgagggaatt tcattccctc
     3421 atactccctt gagcctcctc caaccgaaat agaagggcgc tgcgcttatt atttcattca
     3481 gtcatcggct ttcataatct aacagacaac atcttcgctg caaagccacg ctacgctcaa
     3541 gggcttttac gctacgataa cgcctgtttt aacgattatg ccgataacta aacgaaataa
     3601 acgctaaaac gtctcagaaa cgattttgag acgttttaat aaaaaatcgc ctagtgcttg
     3661 gattctcacc aataaaaaac gcccggcggc aaccgagcgt tctgaacaaa tccagatgga
     3721 gttctgaggt cattactgga tctatcaaca ggagtccaag cgagctcggt actaaaacaa
     3781 ttcatccagt aaaatataat attttatttt ctcccaatca ggcttgatcc ccagtaagtc
     3841 aaaaaatagc tcgacatact gttcttcccc gatatcctcc ctgatcgacc ggacgcagaa
     3901 ggcaatgtca taccacttgt ccgccctgcc gcttctccca agatcaataa agccacttac
     3961 tttgccatct ttcacaaaga tgttgctgtc tcccaggtcg ccgtgggaaa agacaagttc
     4021 ctcttcgggc ttttccgtct ttaaaaaatc atacagctcg cgcggatctt taaatggagt
     4081 gtcttcttcc cagttttcgc aatccacatc ggccagatcg ttattcagta agtaatccaa
     4141 ttcggctaag cggccgtcta agctattcgt atagggacaa tccgatatgt cgatggagtg
     4201 aaagagcctg atgcactccg catacagctc gatagtcttt tcagggcttt gttcatcttc
     4261 atacccttcc gagcaaagga cgccatcggc ctcactcatg agcagattgc tccagccatc
     4321 atgccgttca aagtgcagga cctttggaac aggcagcttt ccttccagcc atagcatcat
     4381 gtccttttcc cgttccacat cataggtggt ccctttatac cggctgtccg tcatttttaa
     4441 atataggatt tcattttctc ccaccagctt atatacctta gcaggagaca ttccttccgt
     4501 atcttttacg cagcggtatt cttcgatcag ttttttcaat tccggtgata ttctcatttt
     4561 agccatttat tatttccttc ctcttttcta cagtatttaa agatacccca agaagctaat
     4621 tataacaaga cgaactccaa ttcactgttc cttgcattct aaaaccttaa atacagaaaa
     4681 cagccttttc aaagttgttt tcaaagttgg cgtataacat agtatcgacg gagccgattt
     4741 tgaaaccaca attatgatag aatttgacgt cgaattcgcg gccgcttcta gatttcagtg
     4801 caatttatct cttcaaatgt agcacctgaa gtcagcccca tacgatataa gttgtaattc
     4861 tcatgttagt catgccccgc gcccaccgga aggagctgac tgggttgaag gctctcaagg
     4921 gcatcggtcg agatcccggt gcctaatgag tgagctaact tacattaatt gcgttgcgct
     4981 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     5041 gcgcggggag aggcggtttg cgtattgggc gccagggtgg tttttctttt caccagtgag
     5101 acgggcaaca gctgattgcc cttcaccgcc tggccctgag agagttgcag caagcggtcc
     5161 acgctggttt gccccagcag gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa
     5221 catgagctgt cttcggtatc gtcgtatccc actaccgaga tgtccgcacc aacgcgcagc
     5281 ccggactcgg taatggcgcg cattgcgccc agcgccatct gatcgttggc aaccagcatc
     5341 gcagtgggaa cgatgccctc attcagcatt tgcatggttt gttgaaaacc ggacatggca
     5401 ctccagtcgc cttcccgttc cgctatcggc tgaatttgat tgcgagtgag atatttatgc
     5461 cagccagcca gacgcagacg cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt
     5521 tgctggtgac ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag
     5581 aaaataatac tgttgatggg tgtctggtca gagacatcaa gaaataacgc cggaacatta
     5641 gtgcaggcag cttccacagc aatggcatcc tggtcatcca gcggatagtt aatgatcagc
     5701 ccactgacgc gttgcgcgag aagattgtgc accgccgctt tacaggcttc gacgccgctt
     5761 cgttctacca tcgacaccac cacgctggca cccagttgat cggcgcgaga tttaatcgcc
     5821 gcgacaattt gcgacggcgc gtgcagggcc agactggagg tggcaacgcc aatcagcaac
     5881 gactgtttgc ccgccagttg ttgtgccacg cggttgggaa tgtaattcag ctccgccatc
     5941 gccgcttcca ctttttcccg cgttttcgca gaaacgtggc tggcctggtt caccacgcgg
     6001 gaaacggtct gataagagac accggcatac tctgcgacat cgtataacgt tactggtttc
     6061 acattcacca ccctgaattg actctcttcc gggcgctatc atgccatacc gcgaaaggtt
     6121 ttgcgccatt cgatggtgtc cgggatctcg acgctctccc ttatgcgact cctgcattag
     6181 gaaattaata cgactcacta ta
//
