LOCUS       Exported                6204 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector lacI-ptac-gusA-pBAV1k, complete sequence.
ACCESSION   JF828584
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6204)
  AUTHORS   Murin CD, Segal K, Bryksin A, Matsumura I.
  TITLE     Expression Vectors for Acinetobacter baylyi ADP1
  JOURNAL   Appl. Environ. Microbiol. 78 (1), 280-283 (2012)
  PUBMED    22020504
REFERENCE   2  (bases 1 to 6204)
  AUTHORS   Murin CD, Segal K, Bryksin A, Matsumura I.
  TITLE     Direct Submission
  JOURNAL   Submitted (22-APR-2011) Biochemistry, Emory University, 1510 Clifton
            Rd NE, Atlanta, GA 30322, USA
REFERENCE   3  (bases 1 to 6204)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6204
                     /organism="Expression vector lacI-ptac-gusA-pBAV1k"
                     /mol_type="other DNA"
                     /db_xref="taxon:1009850"
     regulatory      187..266
                     /regulatory_class="promoter"
                     /note="from Ptac"
     regulatory      192..198
                     /regulatory_class="minus_35_signal"
     promoter        193..222
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     regulatory      216..222
                     /regulatory_class="minus_10_signal"
     misc_feature    228..248
                     /label=lac operator/trp repressor binding site
                     /note="lac operator/trp repressor binding site"
     misc_feature    228
                     /label=+1 (Ptac)
                     /note="+1 (Ptac)"
     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)."
     CDS             267..2135
                     /codon_start=1
                     /gene="gusA"
                     /product="GusA"
                     /label=gusA
                     /note="6-His beta-D-glucuronidase"
                     /protein_id="AEI53610.1"
                     /translation="MGSSHHHHHHSQDPALRAVETPTREIKKLDGLWAFSLDRENCGID
                     QRWWESALQESRAIAVPGSFNDQFADADIRNYAGNVWYQREVFIPKGWAGQRIVLRFDA
                     VTHYGKVWVNNQEVMEHQGGYTPFEADVTPYVIAGKSVRITVCVNNELNWQTIPPGMVI
                     TDENGKKKQSYFHDFFNYAGIHRSVMLYTTPNTWVDDITVVTHVAQDCNHASVDWQVVA
                     NGDVSVELRDADQQVVATGQGTSGTLQVVNPHLWQPGEGYLYELCVTAKSQTECDIYPL
                     RVGIRSVAVKGEQFLINHKPFYFTGFGRHEDADLRGKGFDNVLMVHDHALMDWIGANSY
                     RTSHYPYAEEMLDWADEHGIVVIDETAAVGFNLSLGIGFEAGNKPKELYSEEAVNGETQ
                     QAHLQAIKELIARDKNHPSVVMWSIANEPDTRPQGAREYFAPLAEATRKLDPTRPITCV
                     NVMFCDAHTDTISDLFDVLCLNRYYGWYVQSGDLETAEKVLEKELLAWQEKLHQPIIIT
                     EYGVDTLAGLHSMYTDMWSEEYQCAWLDMYHRVFDRVSAVVGEQVWNFADFATSQGILR
                     VGGNKKGIFTRDRKPKSAAFLLQKRWTGMNFGEKPQQGGKQGLCGR"
     gene            267..2135
                     /gene="gusA"
                     /label=gusA
     CDS             279..296
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    2148..2168
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     regulatory      2213..2291
                     /regulatory_class="terminator"
                     /note="t1"
     terminator      2219..2305
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2326..3024)
                     /codon_start=1
                     /gene="repA"
                     /product="RepA"
                     /label=repA
                     /note="replication initiator protein"
                     /protein_id="AEI53606.1"
                     /translation="MAIKNTKARNFGFLLYPDSIPNDWKEKLESLGVSMAVSPLHDMDE
                     KKDKDTWNSSDVIRNGKHYKKPHYHVIYIARNPVTIESVRNKIKRKLGNSSVAHVEILD
                     YIKGSYEYLTHESKDAIAKNKHIYDKKDILNINDFDIDRYITLDESQKRELKNLLLDIV
                     DDYNLVNTKDLMAFIRLRGAEFGILNTNDVKDIVSTNSSAFRLWFEGNYQCGYRASYAK
                     VLDAETGEIK"
     gene            complement(2326..3024)
                     /gene="repA"
                     /label=repA
     CDS             complement(3091..3252)
                     /codon_start=1
                     /product="hypothetical protein"
                     /label=hypothetical protein
                     /note="ORFC"
                     /protein_id="AEI53607.1"
                     /translation="MVISESKKRVMISLTKEQDKKLTDMAKQKGFSKSAVAALAIEEYA
                     RKESEQKK"
     CDS             complement(3293..3502)
                     /codon_start=1
                     /product="hypothetical protein"
                     /label=hypothetical protein
                     /note="ORFB"
                     /protein_id="AEI53608.1"
                     /translation="MGGKEANFASVLRPPIKCRVPIFVPKTLYPNWLKGLRGFSIANES
                     PTFSPTFFINLYLSSFIVVFMITK"
     repeat_region   3455..3503
                     /label=IRIII
                     /note="IRIII"
     repeat_region   3587..3609
                     /label=IRII
                     /note="IRII"
     CDS             3636..3647
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     repeat_region   3656..3787
                     /label=IRI
                     /note="IRI"
     regulatory      3840..3953
                     /regulatory_class="terminator"
                     /note="t0"
     terminator      3849..3943
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3959..4753)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEEYRCVKDTEGMSPAKVYKLVGENEILYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEGYED
                     EQSPEKTIELYAECIRLFHSIDISDCPYTNSLDGRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     CDS             complement(3959..4621)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase APH (3')-IIIA"
                     /label=aminoglycoside phosphotransferase APH
                     /note="APH(3)(5)-IIIa"
                     /protein_id="AEI53609.1"
                     /translation="MTDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLL
                     MSEADGVLCSEGYEDEQSPEKTIELYAECIRLFHSIDISDCPYTNSLDGRLAELDYLLN
                     NDLADVDCENWEEDTPFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFI
                     DLGRSGRADKWYDIAFCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     misc_feature    4959..4978
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that start with ""ATG"""
     promoter        5005..5082
                     /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             5083..6165
                     /codon_start=1
                     /gene="lacI"
                     /product="lacI transcriptional repressor"
                     /label=lacI
                     /db_xref="GOA:C5W1L3"
                     /db_xref="InterPro:IPR000843"
                     /db_xref="InterPro:IPR001761"
                     /db_xref="InterPro:IPR010982"
                     /protein_id="AEI53611.1"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     gene            5083..6165
                     /gene="lacI"
                     /label=lacI
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 gtagaaaccc caacccgtga aatcaaaaaa ctcgacggcc
      361 tgtgggcatt cagtctggat cgcgaaaact gtggaattga tcagcgttgg tgggaaagcg
      421 cgttacaaga aagccgggca attgctgtgc caggcagttt taacgatcag ttcgccgatg
      481 cagatattcg taattatgcg ggcaacgtct ggtatcagcg cgaagtcttt ataccgaaag
      541 gttgggcagg ccagcgtatc gtgctgcgtt tcgatgcggt cactcattac ggcaaagtgt
      601 gggtcaataa tcaggaagtg atggagcatc agggcggcta tacgccattt gaagccgatg
      661 tcacgccgta tgttattgcc gggaaaagtg tacgtatcac cgtttgtgtg aacaacgaac
      721 tgaactggca gactatcccg ccgggaatgg tgattaccga cgaaaacggc aagaaaaagc
      781 agtcttactt ccatgatttc tttaactatg ccggaatcca tcgcagcgta atgctctaca
      841 ccacgccgaa cacctgggtg gacgatatca ccgtggtgac gcatgtcgcg caagactgta
      901 accacgcgtc tgttgactgg caggtggtgg ccaatggtga tgtcagcgtt gaactgcgtg
      961 atgcggatca acaggtggtt gcaactggac aaggcactag cgggactttg caagtggtga
     1021 atccgcacct ctggcaaccg ggtgaaggtt atctctatga actgtgcgtc acagccaaaa
     1081 gccagacaga gtgtgatatc tacccgcttc gcgtcggcat ccggtcagtg gcagtgaagg
     1141 gcgaacagtt cctgattaac cacaaaccgt tctactttac tggctttggt cgtcatgaag
     1201 atgcggactt gcgtggcaaa ggattcgata acgtgctgat ggtgcacgac cacgcattaa
     1261 tggactggat tggggccaac tcctaccgta cctcgcatta cccttacgct gaagagatgc
     1321 tcgactgggc agatgaacat ggcatcgtgg tgattgatga aactgctgct gtcggcttta
     1381 acctctcttt aggcattggt ttcgaagcgg gcaacaagcc gaaagaactg tacagcgaag
     1441 aggcagtcaa cggggaaact cagcaagcgc acttacaggc gattaaagag ctgatagcgc
     1501 gtgacaaaaa ccacccaagc gtggtgatgt ggagtattgc caacgaaccg gatacccgtc
     1561 cgcaaggtgc acgggaatat ttcgcgccac tggcggaagc aacgcgtaaa ctcgacccga
     1621 cgcgtccgat cacctgcgtc aatgtaatgt tctgcgacgc tcacaccgat accatcagcg
     1681 atctctttga tgtgctgtgc ctgaaccgtt attacggatg gtatgtccaa agcggcgatt
     1741 tggaaacggc agagaaggta ctggaaaaag aacttctggc ctggcaggag aaactgcatc
     1801 agccgattat catcaccgaa tacggcgtgg atacgttagc cgggctgcac tcaatgtaca
     1861 ccgacatgtg gagtgaagag tatcagtgtg catggctgga tatgtatcac cgcgtctttg
     1921 atcgcgtcag cgccgtcgtc ggtgaacagg tatggaattt cgccgatttt gcgacctcgc
     1981 aaggcatatt gcgcgttggc ggtaacaaga aagggatctt cactcgcgac cgcaaaccga
     2041 agtcggcggc ttttctgctg caaaaacgct ggactggcat gaacttcggt gaaaaaccgc
     2101 agcagggagg caaacaaggc ctatgcggcc ggtagaaagg agaagcttac tagtagcggc
     2161 cgctgcaggc ctcagggccc gatcgatgcc gccgcttaat taattaatcc agaggcatca
     2221 aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt
     2281 gaacgctctc ctgagtagga caaatccgcc gccctagacc tagtgtcatt ttatttcccc
     2341 cgtttcagca tcaagaacct ttgcataact tgctctatat ccacactgat aattgccctc
     2401 aaaccataat ctaaaggcgc tagagtttgt tgaaacaata tcttttacat cattcgtatt
     2461 taaaattcca aactccgctc ccctaaggcg aataaaagcc attaaatctt ttgtatttac
     2521 caaattatag tcatccacta tatctaagag taaattcttc aattctcttt tttggctttc
     2581 atcaagtgtt atatagcggt caatatcaaa atcattaatg ttcaaaatat cttttttgtc
     2641 gtatatatgt ttattcttag caatagcgtc ctttgattca tgagtcaaat attcatatga
     2701 acctttgata taatcaagta tctcaacatg agcaactgaa ctattcccca attttcgctt
     2761 aatcttgttc ctaacgcttt ctattgttac aggatttcgt gcaatatata taacgtgata
     2821 gtgtggtttt ttatagtgct ttccatttcg tataacatca ctactattcc atgtatcttt
     2881 atcttttttt tcgtccatat cgtgtaaagg actgacagcc atagatacgc ccaaactctc
     2941 taatttttcc ttccaatcat taggaattga gtcaggatat aataaaaatc caaaatttct
     3001 agctttagta tttttaatag ccatgatata attaccttat caaaaacaag tagcgaaaac
     3061 tcgtatcctt ctaaaaacgc gagctttcgc ttattttttt tgttctgatt cctttcttgc
     3121 atattcttct atagctaacg ccgcaaccgc agattttgaa aaaccttttt gtttcgccat
     3181 atctgttaat tttttatctt gctcttttgt cagagaaatc ataactcttt ttttcgattc
     3241 tgaaatcacc atttaaaaaa ctccaatcaa ataattttat aaagttagtg tatcactttg
     3301 taatcataaa aacaacaata aagctactta aatatagatt tataaaaaac gttggcgaaa
     3361 acgttggcga ttcgttggcg attgaaaaac cccttaaacc cttgagccag ttgggataga
     3421 gcgtttttgg cacaaaaatt ggcactcggc acttaatggg gggtcgtagt acggaagcaa
     3481 aattcgcttc ctttcccccc atttttttcc aaattccaaa tttttttcaa aaattttcca
     3541 gcgctaccgc tcggcaaaat tgcaagcaat ttttaaaatc aaacccatga gggaatttca
     3601 ttccctcata ctcccttgag cctcctccaa ccgaaataga agggcgctgc gcttattatt
     3661 tcattcagtc atcggctttc ataatctaac agacaacatc ttcgctgcaa agccacgcta
     3721 cgctcaaggg cttttacgct acgataacgc ctgttttaac gattatgccg ataactaaac
     3781 gaaataaacg ctaaaacgtc tcagaaacga ttttgagacg ttttaataaa aaatcgccta
     3841 gtgcttggat tctcaccaat aaaaaacgcc cggcggcaac cgagcgttct gaacaaatcc
     3901 agatggagtt ctgaggtcat tactggatct atcaacagga gtccaagcga gctcggtact
     3961 aaaacaattc atccagtaaa atataatatt ttattttctc ccaatcaggc ttgatcccca
     4021 gtaagtcaaa aaatagctcg acatactgtt cttccccgat atcctccctg atcgaccgga
     4081 cgcagaaggc aatgtcatac cacttgtccg ccctgccgct tctcccaaga tcaataaagc
     4141 cacttacttt gccatctttc acaaagatgt tgctgtctcc caggtcgccg tgggaaaaga
     4201 caagttcctc ttcgggcttt tccgtcttta aaaaatcata cagctcgcgc ggatctttaa
     4261 atggagtgtc ttcttcccag ttttcgcaat ccacatcggc cagatcgtta ttcagtaagt
     4321 aatccaattc ggctaagcgg ccgtctaagc tattcgtata gggacaatcc gatatgtcga
     4381 tggagtgaaa gagcctgatg cactccgcat acagctcgat agtcttttca gggctttgtt
     4441 catcttcata cccttccgag caaaggacgc catcggcctc actcatgagc agattgctcc
     4501 agccatcatg ccgttcaaag tgcaggacct ttggaacagg cagctttcct tccagccata
     4561 gcatcatgtc cttttcccgt tccacatcat aggtggtccc tttataccgg ctgtccgtca
     4621 tttttaaata taggatttca ttttctccca ccagcttata taccttagca ggagacattc
     4681 cttccgtatc ttttacgcag cggtattctt cgatcagttt tttcaattcc ggtgatattc
     4741 tcattttagc catttattat ttccttcctc ttttctacag tatttaaaga taccccaaga
     4801 agctaattat aacaagacga actccaattc actgttcctt gcattctaaa accttaaata
     4861 cagaaaacag ccttttcaaa gttgttttca aagttggcgt ataacatagt atcgacggag
     4921 ccgattttga aaccacaatt atgatagaat ttgacgtcga attcgcggcc gcttctagaa
     4981 ctatagctag catgcattta cgttgacacc atcgaatggt gcaaaacctt tcgcggtatg
     5041 gcatgatagc gcccggaaga gagtcaattc agggtggtga atgtgaaacc agtaacgtta
     5101 tacgatgtcg cagagtatgc cggtgtctct tatcagaccg tttcccgcgt ggtgaaccag
     5161 gccagccacg tttctgcgaa aacgcgggaa aaagtggaag cggcgatggc ggagctgaat
     5221 tacattccca accgcgtggc acaacaactg gcgggcaaac agtcgttgct gattggcgtt
     5281 gccacctcca gtctggccct gcacgcgccg tcgcaaattg tcgcggcgat taaatctcgc
     5341 gccgatcaac tgggtgccag cgtggtggtg tcgatggtag aacgaagcgg cgtcgaagcc
     5401 tgtaaagcgg cggtgcacaa tcttctcgcg caacgcgtca gtgggctgat cattaactat
     5461 ccgctggatg accaggatgc cattgctgtg gaagctgcct gcactaatgt tccggcgtta
     5521 tttcttgatg tctctgacca gacacccatc aacagtatta ttttctccca tgaagacggt
     5581 acgcgactgg gcgtggagca tctggtcgca ttgggtcacc agcaaatcgc gctgttagcg
     5641 ggcccattaa gttctgtctc ggcgcgtctg cgtctggctg gctggcataa atatctcact
     5701 cgcaatcaaa ttcagccgat agcggaacgg gaaggcgact ggagtgccat gtccggtttt
     5761 caacaaacca tgcaaatgct gaatgagggc atcgttccca ctgcgatgct ggttgccaac
     5821 gatcagatgg cgctgggcgc aatgcgcgcc attaccgagt ccgggctgcg cgttggtgcg
     5881 gatatctcgg tagtgggata cgacgatacc gaagacagct catgttatat cccgccgtca
     5941 accaccatca aacaggattt tcgcctgctg gggcaaacca gcgtggaccg cttgctgcaa
     6001 ctctctcagg gccaggcggt gaagggcaat cagctgttgc ccgtctcact ggtgaaaaga
     6061 aaaaccaccc tggcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     6121 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
     6181 tgtgagttag cgcgaattga tctg
//
