LOCUS       Exported                6193 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector lacI-T7-gusA-pBAV1k, complete sequence.
ACCESSION   JF828583
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6193)
  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 6193)
  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 6193)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6193
                     /organism="Expression vector lacI-T7-gusA-pBAV1k"
                     /mol_type="other DNA"
                     /db_xref="taxon:1009849"
     misc_feature    1
                     /label=T7 +1
                     /note="T7 +1"
     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)."
     regulatory      58..64
                     /regulatory_class="ribosome_binding_site"
     CDS             71..1939
                     /codon_start=1
                     /gene="gusA"
                     /product="GusA"
                     /label=gusA
                     /note="6-His beta-D-glucuronidase"
                     /db_xref="ASAP:ABE-0005410"
                     /db_xref="EcoGene:EG11055"
                     /db_xref="UniProtKB/Swiss-Prot:P05804"
                     /protein_id="AEI53604.1"
                     /translation="MGSSHHHHHHSQDPALRAVETPTREIKKLDGLWAFSLDRENCGID
                     QRWWESALQESRAIAVPGSFNDQFADADIRNYAGNVWYQREVFIPKGWAGQRIVLRFDA
                     VTHYGKVWVNNQEVMEHQGGYTPFEADVTPYVIAGKSVRITVCVNNELNWQTIPPGMVI
                     TDENGKKKQSYFHDFFNYAGIHRSVMLYTTPNTWVDDITVVTHVAQDCNHASVDWQVVA
                     NGDVSVELRDADQQVVATGQGTSGTLQVVNPHLWQPGEGYLYELCVTAKSQTECDIYPL
                     RVGIRSVAVKGEQFLINHKPFYFTGFGRHEDADLRGKGFDNVLMVHDHALMDWIGANSY
                     RTSHYPYAEEMLDWADEHGIVVIDETAAVGFNLSLGIGFEAGNKPKELYSEEAVNGETQ
                     QAHLQAIKELIARDKNHPSVVMWSIANEPDTRPQGAREYFAPLAEATRKLDPTRPITCV
                     NVMFCDAHTDTISDLFDVLCLNRYYGWYVQSGDLETAEKVLEKELLAWQEKLHQPIIIT
                     EYGVDTLAGLHSMYTDMWSEEYQCAWLDMYHRVFDRVSAVVGEQVWNFADFATSQGILR
                     VGGNKKGIFTRDRKPKSAAFLLQKRWTGMNFGEKPQQGGKQGLCGR"
     gene            71..1939
                     /gene="gusA"
                     /label=gusA
     CDS             83..100
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    1952..1972
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     regulatory      2017..2094
                     /regulatory_class="terminator"
                     /note="t1"
     terminator      2023..2109
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2130..2828)
                     /codon_start=1
                     /gene="repA"
                     /product="RepA"
                     /label=repA
                     /note="replication initiator protein"
                     /protein_id="AEI53600.1"
                     /translation="MAIKNTKARNFGFLLYPDSIPNDWKEKLESLGVSMAVSPLHDMDE
                     KKDKDTWNSSDVIRNGKHYKKPHYHVIYIARNPVTIESVRNKIKRKLGNSSVAHVEILD
                     YIKGSYEYLTHESKDAIAKNKHIYDKKDILNINDFDIDRYITLDESQKRELKNLLLDIV
                     DDYNLVNTKDLMAFIRLRGAEFGILNTNDVKDIVSTNSSAFRLWFEGNYQCGYRASYAK
                     VLDAETGEIK"
     gene            complement(2130..2828)
                     /gene="repA"
                     /label=repA
     CDS             complement(2895..3056)
                     /codon_start=1
                     /product="hypothetical protein"
                     /label=hypothetical protein
                     /note="ORFC"
                     /protein_id="AEI53601.1"
                     /translation="MVISESKKRVMISLTKEQDKKLTDMAKQKGFSKSAVAALAIEEYA
                     RKESEQKK"
     CDS             complement(3097..3306)
                     /codon_start=1
                     /product="hypothetical protein"
                     /label=hypothetical protein
                     /note="ORFB"
                     /protein_id="AEI53602.1"
                     /translation="MGGKEANFASVLRPPIKCRVPIFVPKTLYPNWLKGLRGFSIANES
                     PTFSPTFFINLYLSSFIVVFMITK"
     repeat_region   3259..3307
                     /label=IRIII
                     /note="IRIII"
     repeat_region   3391..3413
                     /label=IRII
                     /note="IRII"
     CDS             3440..3451
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     repeat_region   3460..3591
                     /label=IRI
                     /note="IRI"
     regulatory      3644..3757
                     /regulatory_class="terminator"
                     /note="t0"
     terminator      3653..3747
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3763..4557)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEEYRCVKDTEGMSPAKVYKLVGENEILYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEGYED
                     EQSPEKTIELYAECIRLFHSIDISDCPYTNSLDGRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     CDS             complement(3763..4425)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase APH (3')-IIIA"
                     /label=aminoglycoside phosphotransferase APH
                     /note="APH(3)(5)-IIIa"
                     /protein_id="AEI53603.1"
                     /translation="MTDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLL
                     MSEADGVLCSEGYEDEQSPEKTIELYAECIRLFHSIDISDCPYTNSLDGRLAELDYLLN
                     NDLADVDCENWEEDTPFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFI
                     DLGRSGRADKWYDIAFCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     misc_feature    4763..4782
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that start with ""ATG"""
     CDS             complement(4971..6053)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /protein_id="AEI53605.1"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     gene            complement(4971..6053)
                     /gene="lacI"
                     /label=lacI
     promoter        complement(6054..6131)
                     /gene="lacI"
                     /label=lacI promoter
     regulatory      6156..6162
                     /regulatory_class="minus_35_signal"
                     /note="from T7"
     promoter        6177..2
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      6177..6193
                     /regulatory_class="promoter"
                     /note="from T7"
     regulatory      6181..6187
                     /regulatory_class="minus_10_signal"
                     /note="from T7"
ORIGIN
        1 ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaataag
       61 gagatatacc atgggcagca gccatcacca tcatcaccac agccaggatc cggccctgag
      121 ggccgtagaa accccaaccc gtgaaatcaa aaaactcgac ggcctgtggg cattcagtct
      181 ggatcgcgaa aactgtggaa ttgatcagcg ttggtgggaa agcgcgttac aagaaagccg
      241 ggcaattgct gtgccaggca gttttaacga tcagttcgcc gatgcagata ttcgtaatta
      301 tgcgggcaac gtctggtatc agcgcgaagt ctttataccg aaaggttggg caggccagcg
      361 tatcgtgctg cgtttcgatg cggtcactca ttacggcaaa gtgtgggtca ataatcagga
      421 agtgatggag catcagggcg gctatacgcc atttgaagcc gatgtcacgc cgtatgttat
      481 tgccgggaaa agtgtacgta tcaccgtttg tgtgaacaac gaactgaact ggcagactat
      541 cccgccggga atggtgatta ccgacgaaaa cggcaagaaa aagcagtctt acttccatga
      601 tttctttaac tatgccggaa tccatcgcag cgtaatgctc tacaccacgc cgaacacctg
      661 ggtggacgat atcaccgtgg tgacgcatgt cgcgcaagac tgtaaccacg cgtctgttga
      721 ctggcaggtg gtggccaatg gtgatgtcag cgttgaactg cgtgatgcgg atcaacaggt
      781 ggttgcaact ggacaaggca ctagcgggac tttgcaagtg gtgaatccgc acctctggca
      841 accgggtgaa ggttatctct atgaactgtg cgtcacagcc aaaagccaga cagagtgtga
      901 tatctacccg cttcgcgtcg gcatccggtc agtggcagtg aagggcgaac agttcctgat
      961 taaccacaaa ccgttctact ttactggctt tggtcgtcat gaagatgcgg acttgcgtgg
     1021 caaaggattc gataacgtgc tgatggtgca cgaccacgca ttaatggact ggattggggc
     1081 caactcctac cgtacctcgc attaccctta cgctgaagag atgctcgact gggcagatga
     1141 acatggcatc gtggtgattg atgaaactgc tgctgtcggc tttaacctct ctttaggcat
     1201 tggtttcgaa gcgggcaaca agccgaaaga actgtacagc gaagaggcag tcaacgggga
     1261 aactcagcaa gcgcacttac aggcgattaa agagctgata gcgcgtgaca aaaaccaccc
     1321 aagcgtggtg atgtggagta ttgccaacga accggatacc cgtccgcaag gtgcacggga
     1381 atatttcgcg ccactggcgg aagcaacgcg taaactcgac ccgacgcgtc cgatcacctg
     1441 cgtcaatgta atgttctgcg acgctcacac cgataccatc agcgatctct ttgatgtgct
     1501 gtgcctgaac cgttattacg gatggtatgt ccaaagcggc gatttggaaa cggcagagaa
     1561 ggtactggaa aaagaacttc tggcctggca ggagaaactg catcagccga ttatcatcac
     1621 cgaatacggc gtggatacgt tagccgggct gcactcaatg tacaccgaca tgtggagtga
     1681 agagtatcag tgtgcatggc tggatatgta tcaccgcgtc tttgatcgcg tcagcgccgt
     1741 cgtcggtgaa caggtatgga atttcgccga ttttgcgacc tcgcaaggca tattgcgcgt
     1801 tggcggtaac aagaaaggga tcttcactcg cgaccgcaaa ccgaagtcgg cggcttttct
     1861 gctgcaaaaa cgctggactg gcatgaactt cggtgaaaaa ccgcagcagg gaggcaaaca
     1921 aggcctatgc ggccggtaga aaggagaagc ttactagtag cggccgctgc aggcctcagg
     1981 gcccgatcga tgccgccgct taattaatta atccagaggc atcaaataaa acgaaaggct
     2041 cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt
     2101 aggacaaatc cgccgcccta gacctagtgt cattttattt cccccgtttc agcatcaaga
     2161 acctttgcat aacttgctct atatccacac tgataattgc cctcaaacca taatctaaag
     2221 gcgctagagt ttgttgaaac aatatctttt acatcattcg tatttaaaat tccaaactcc
     2281 gctcccctaa ggcgaataaa agccattaaa tcttttgtat ttaccaaatt atagtcatcc
     2341 actatatcta agagtaaatt cttcaattct cttttttggc tttcatcaag tgttatatag
     2401 cggtcaatat caaaatcatt aatgttcaaa atatcttttt tgtcgtatat atgtttattc
     2461 ttagcaatag cgtcctttga ttcatgagtc aaatattcat atgaaccttt gatataatca
     2521 agtatctcaa catgagcaac tgaactattc cccaattttc gcttaatctt gttcctaacg
     2581 ctttctattg ttacaggatt tcgtgcaata tatataacgt gatagtgtgg ttttttatag
     2641 tgctttccat ttcgtataac atcactacta ttccatgtat ctttatcttt tttttcgtcc
     2701 atatcgtgta aaggactgac agccatagat acgcccaaac tctctaattt ttccttccaa
     2761 tcattaggaa ttgagtcagg atataataaa aatccaaaat ttctagcttt agtattttta
     2821 atagccatga tataattacc ttatcaaaaa caagtagcga aaactcgtat ccttctaaaa
     2881 acgcgagctt tcgcttattt tttttgttct gattcctttc ttgcatattc ttctatagct
     2941 aacgccgcaa ccgcagattt tgaaaaacct ttttgtttcg ccatatctgt taatttttta
     3001 tcttgctctt ttgtcagaga aatcataact ctttttttcg attctgaaat caccatttaa
     3061 aaaactccaa tcaaataatt ttataaagtt agtgtatcac tttgtaatca taaaaacaac
     3121 aataaagcta cttaaatata gatttataaa aaacgttggc gaaaacgttg gcgattcgtt
     3181 ggcgattgaa aaacccctta aacccttgag ccagttggga tagagcgttt ttggcacaaa
     3241 aattggcact cggcacttaa tggggggtcg tagtacggaa gcaaaattcg cttcctttcc
     3301 ccccattttt ttccaaattc caaatttttt tcaaaaattt tccagcgcta ccgctcggca
     3361 aaattgcaag caatttttaa aatcaaaccc atgagggaat ttcattccct catactccct
     3421 tgagcctcct ccaaccgaaa tagaagggcg ctgcgcttat tatttcattc agtcatcggc
     3481 tttcataatc taacagacaa catcttcgct gcaaagccac gctacgctca agggctttta
     3541 cgctacgata acgcctgttt taacgattat gccgataact aaacgaaata aacgctaaaa
     3601 cgtctcagaa acgattttga gacgttttaa taaaaaatcg cctagtgctt ggattctcac
     3661 caataaaaaa cgcccggcgg caaccgagcg ttctgaacaa atccagatgg agttctgagg
     3721 tcattactgg atctatcaac aggagtccaa gcgagctcgg tactaaaaca attcatccag
     3781 taaaatataa tattttattt tctcccaatc aggcttgatc cccagtaagt caaaaaatag
     3841 ctcgacatac tgttcttccc cgatatcctc cctgatcgac cggacgcaga aggcaatgtc
     3901 ataccacttg tccgccctgc cgcttctccc aagatcaata aagccactta ctttgccatc
     3961 tttcacaaag atgttgctgt ctcccaggtc gccgtgggaa aagacaagtt cctcttcggg
     4021 cttttccgtc tttaaaaaat catacagctc gcgcggatct ttaaatggag tgtcttcttc
     4081 ccagttttcg caatccacat cggccagatc gttattcagt aagtaatcca attcggctaa
     4141 gcggccgtct aagctattcg tatagggaca atccgatatg tcgatggagt gaaagagcct
     4201 gatgcactcc gcatacagct cgatagtctt ttcagggctt tgttcatctt catacccttc
     4261 cgagcaaagg acgccatcgg cctcactcat gagcagattg ctccagccat catgccgttc
     4321 aaagtgcagg acctttggaa caggcagctt tccttccagc catagcatca tgtccttttc
     4381 ccgttccaca tcataggtgg tccctttata ccggctgtcc gtcattttta aatataggat
     4441 ttcattttct cccaccagct tatatacctt agcaggagac attccttccg tatcttttac
     4501 gcagcggtat tcttcgatca gttttttcaa ttccggtgat attctcattt tagccattta
     4561 ttatttcctt cctcttttct acagtattta aagatacccc aagaagctaa ttataacaag
     4621 acgaactcca attcactgtt ccttgcattc taaaacctta aatacagaaa acagcctttt
     4681 caaagttgtt ttcaaagttg gcgtataaca tagtatcgac ggagccgatt ttgaaaccac
     4741 aattatgata gaatttgacg tcgaattcgc ggccgcttct agatttcagt gcaatttatc
     4801 tcttcaaatg tagcacctga agtcagcccc atacgatata agttgtaatt ctcatgttag
     4861 tcatgccccg cgcccaccgg aaggagctga ctgggttgaa ggctctcaag ggcatcggtc
     4921 gagatcccgg tgcctaatga gtgagctaac ttacattaat tgcgttgcgc tcactgcccg
     4981 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     5041 gaggcggttt gcgtattggg cgccagggtg gtttttcttt tcaccagtga gacgggcaac
     5101 agctgattgc ccttcaccgc ctggccctga gagagttgca gcaagcggtc cacgctggtt
     5161 tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata acatgagctg
     5221 tcttcggtat cgtcgtatcc cactaccgag atgtccgcac caacgcgcag cccggactcg
     5281 gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg caaccagcat cgcagtggga
     5341 acgatgccct cattcagcat ttgcatggtt tgttgaaaac cggacatggc actccagtcg
     5401 ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga gatatttatg ccagccagcc
     5461 agacgcagac gcgccgagac agaacttaat gggcccgcta acagcgcgat ttgctggtga
     5521 cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt cttcatggga gaaaataata
     5581 ctgttgatgg gtgtctggtc agagacatca agaaataacg ccggaacatt agtgcaggca
     5641 gcttccacag caatggcatc ctggtcatcc agcggatagt taatgatcag cccactgacg
     5701 cgttgcgcga gaagattgtg caccgccgct ttacaggctt cgacgccgct tcgttctacc
     5761 atcgacacca ccacgctggc acccagttga tcggcgcgag atttaatcgc cgcgacaatt
     5821 tgcgacggcg cgtgcagggc cagactggag gtggcaacgc caatcagcaa cgactgtttg
     5881 cccgccagtt gttgtgccac gcggttggga atgtaattca gctccgccat cgccgcttcc
     5941 actttttccc gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc
     6001 tgataagaga caccggcata ctctgcgaca tcgtataacg ttactggttt cacattcacc
     6061 accctgaatt gactctcttc cgggcgctat catgccatac cgcgaaaggt tttgcgccat
     6121 tcgatggtgt ccgggatctc gacgctctcc cttatgcgac tcctgcatta ggaaattaat
     6181 acgactcact ata
//
