LOCUS       Exported                6366 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector lacI-T5-gusA-pBAV1k, complete sequence.
ACCESSION   JF828582
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6366)
  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 6366)
  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 6366)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6366
                     /organism="Expression vector lacI-T5-gusA-pBAV1k"
                     /mol_type="other DNA"
                     /db_xref="taxon:1009848"
     regulatory      64..141
                     /regulatory_class="terminator"
                     /note="t1"
     terminator      70..156
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(177..875)
                     /codon_start=1
                     /gene="repA"
                     /product="RepA"
                     /label=repA
                     /note="replication initiator protein"
                     /protein_id="AEI53594.1"
                     /translation="MAIKNTKARNFGFLLYPDSIPNDWKEKLESLGVSMAVSPLHDMDE
                     KKDKDTWNSSDVIRNGKHYKKPHYHVIYIARNPVTIESVRNKIKRKLGNSSVAHVEILD
                     YIKGSYEYLTHESKDAIAKNKHIYDKKDILNINDFDIDRYITLDESQKRELKNLLLDIV
                     DDYNLVNTKDLMAFIRLRGAEFGILNTNDVKDIVSTNSSAFRLWFEGNYQCGYRASYAK
                     VLDAETGEIK"
     gene            complement(177..875)
                     /gene="repA"
                     /label=repA
     CDS             complement(942..1103)
                     /codon_start=1
                     /product="hypothetical protein"
                     /label=hypothetical protein
                     /note="ORFC"
                     /protein_id="AEI53595.1"
                     /translation="MVISESKKRVMISLTKEQDKKLTDMAKQKGFSKSAVAALAIEEYA
                     RKESEQKK"
     CDS             complement(1144..1353)
                     /codon_start=1
                     /product="hypothetical protein"
                     /label=hypothetical protein
                     /note="ORFB"
                     /protein_id="AEI53596.1"
                     /translation="MGGKEANFASVLRPPIKCRVPIFVPKTLYPNWLKGLRGFSIANES
                     PTFSPTFFINLYLSSFIVVFMITK"
     repeat_region   1306..1354
                     /label=IRIII
                     /note="IRIII"
     repeat_region   1438..1460
                     /label=IRII
                     /note="IRII"
     CDS             1487..1498
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     repeat_region   1507..1638
                     /label=IRI
                     /note="IRI"
     regulatory      1691..1804
                     /regulatory_class="terminator"
                     /note="t0"
     terminator      1700..1794
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1810..2604)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEEYRCVKDTEGMSPAKVYKLVGENEILYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEGYED
                     EQSPEKTIELYAECIRLFHSIDISDCPYTNSLDGRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     CDS             complement(1810..2472)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase APH (3')-IIIA"
                     /label=aminoglycoside phosphotransferase APH
                     /note="APH(3)(5)-IIIa"
                     /protein_id="AEI53597.1"
                     /translation="MTDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLL
                     MSEADGVLCSEGYEDEQSPEKTIELYAECIRLFHSIDISDCPYTNSLDGRLAELDYLLN
                     NDLADVDCENWEEDTPFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFI
                     DLGRSGRADKWYDIAFCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     misc_feature    2810..2829
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that start with ""ATG"""
     promoter        2856..2933
                     /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             2934..4016
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /protein_id="AEI53599.1"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     gene            2934..4016
                     /gene="lacI"
                     /label=lacI
     protein_bind    4029..4050
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4065..4095
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4103..4119
                     /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     4127..4143
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4159..4175)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        4375..4419
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     regulatory      4388..4395
                     /regulatory_class="minus_35_signal"
     misc_feature    4395..4413
                     /label=lac operator
                     /note="lac operator"
     protein_bind    4395..4411
                     /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      4413..4453
                     /regulatory_class="ribosome_binding_site"
     regulatory      4413..4418
                     /regulatory_class="minus_10_signal"
     misc_feature    4427..4445
                     /label=lac operator
                     /note="lac operator"
     protein_bind    4427..4443
                     /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)."
     misc_feature    4427
                     /label=+1 (PT5)
                     /note="+1 (PT5)"
     RBS             4466..4477
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             4484..6352
                     /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="AEI53598.1"
                     /translation="MGSSHHHHHHSQDPALRAVETPTREIKKLDGLWAFSLDRENCGID
                     QRWWESALQESRAIAVPGSFNDQFADADIRNYAGNVWYQREVFIPKGWAGQRIVLRFDA
                     VTHYGKVWVNNQEVMEHQGGYTPFEADVTPYVIAGKSVRITVCVNNELNWQTIPPGMVI
                     TDENGKKKQSYFHDFFNYAGIHRSVMLYTTPNTWVDDITVVTHVAQDCNHASVDWQVVA
                     NGDVSVELRDADQQVVATGQGTSGTLQVVNPHLWQPGEGYLYELCVTAKSQTECDIYPL
                     RVGIRSVAVKGEQFLINHKPFYFTGFGRHEDADLRGKGFDNVLMVHDHALMDWIGANSY
                     RTSHYPYAEEMLDWADEHGIVVIDETAAVGFNLSLGIGFEAGNKPKELYSEEAVNGETQ
                     QAHLQAIKELIARDKNHPSVVMWSIANEPDTRPQGAREYFAPLAEATRKLDPTRPITCV
                     NVMFCDAHTDTISDLFDVLCLNRYYGWYVQSGDLETAEKVLEKELLAWQEKLHQPIIIT
                     EYGVDTLAGLHSMYTDMWSEEYQCAWLDMYHRVFDRVSAVVGEQVWNFADFATSQGILR
                     VGGNKKGIFTRDRKPKSAAFLLQKRWTGMNFGEKPQQGGKQGLCGR"
     gene            4484..6352
                     /gene="gusA"
                     /label=gusA
     CDS             4496..4513
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    6365..19
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
ORIGIN
        1 ctagtagcgg ccgctgcagg cctcagggcc cgatcgatgc cgccgcttaa ttaattaatc
       61 cagaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt
      121 tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc cgccctagac ctagtgtcat
      181 tttatttccc ccgtttcagc atcaagaacc tttgcataac ttgctctata tccacactga
      241 taattgccct caaaccataa tctaaaggcg ctagagtttg ttgaaacaat atcttttaca
      301 tcattcgtat ttaaaattcc aaactccgct cccctaaggc gaataaaagc cattaaatct
      361 tttgtattta ccaaattata gtcatccact atatctaaga gtaaattctt caattctctt
      421 ttttggcttt catcaagtgt tatatagcgg tcaatatcaa aatcattaat gttcaaaata
      481 tcttttttgt cgtatatatg tttattctta gcaatagcgt cctttgattc atgagtcaaa
      541 tattcatatg aacctttgat ataatcaagt atctcaacat gagcaactga actattcccc
      601 aattttcgct taatcttgtt cctaacgctt tctattgtta caggatttcg tgcaatatat
      661 ataacgtgat agtgtggttt tttatagtgc tttccatttc gtataacatc actactattc
      721 catgtatctt tatctttttt ttcgtccata tcgtgtaaag gactgacagc catagatacg
      781 cccaaactct ctaatttttc cttccaatca ttaggaattg agtcaggata taataaaaat
      841 ccaaaatttc tagctttagt atttttaata gccatgatat aattacctta tcaaaaacaa
      901 gtagcgaaaa ctcgtatcct tctaaaaacg cgagctttcg cttatttttt ttgttctgat
      961 tcctttcttg catattcttc tatagctaac gccgcaaccg cagattttga aaaacctttt
     1021 tgtttcgcca tatctgttaa ttttttatct tgctcttttg tcagagaaat cataactctt
     1081 tttttcgatt ctgaaatcac catttaaaaa actccaatca aataatttta taaagttagt
     1141 gtatcacttt gtaatcataa aaacaacaat aaagctactt aaatatagat ttataaaaaa
     1201 cgttggcgaa aacgttggcg attcgttggc gattgaaaaa ccccttaaac ccttgagcca
     1261 gttgggatag agcgtttttg gcacaaaaat tggcactcgg cacttaatgg ggggtcgtag
     1321 tacggaagca aaattcgctt cctttccccc catttttttc caaattccaa atttttttca
     1381 aaaattttcc agcgctaccg ctcggcaaaa ttgcaagcaa tttttaaaat caaacccatg
     1441 agggaatttc attccctcat actcccttga gcctcctcca accgaaatag aagggcgctg
     1501 cgcttattat ttcattcagt catcggcttt cataatctaa cagacaacat cttcgctgca
     1561 aagccacgct acgctcaagg gcttttacgc tacgataacg cctgttttaa cgattatgcc
     1621 gataactaaa cgaaataaac gctaaaacgt ctcagaaacg attttgagac gttttaataa
     1681 aaaatcgcct agtgcttgga ttctcaccaa taaaaaacgc ccggcggcaa ccgagcgttc
     1741 tgaacaaatc cagatggagt tctgaggtca ttactggatc tatcaacagg agtccaagcg
     1801 agctcggtac taaaacaatt catccagtaa aatataatat tttattttct cccaatcagg
     1861 cttgatcccc agtaagtcaa aaaatagctc gacatactgt tcttccccga tatcctccct
     1921 gatcgaccgg acgcagaagg caatgtcata ccacttgtcc gccctgccgc ttctcccaag
     1981 atcaataaag ccacttactt tgccatcttt cacaaagatg ttgctgtctc ccaggtcgcc
     2041 gtgggaaaag acaagttcct cttcgggctt ttccgtcttt aaaaaatcat acagctcgcg
     2101 cggatcttta aatggagtgt cttcttccca gttttcgcaa tccacatcgg ccagatcgtt
     2161 attcagtaag taatccaatt cggctaagcg gccgtctaag ctattcgtat agggacaatc
     2221 cgatatgtcg atggagtgaa agagcctgat gcactccgca tacagctcga tagtcttttc
     2281 agggctttgt tcatcttcat acccttccga gcaaaggacg ccatcggcct cactcatgag
     2341 cagattgctc cagccatcat gccgttcaaa gtgcaggacc tttggaacag gcagctttcc
     2401 ttccagccat agcatcatgt ccttttcccg ttccacatca taggtggtcc ctttataccg
     2461 gctgtccgtc atttttaaat ataggatttc attttctccc accagcttat ataccttagc
     2521 aggagacatt ccttccgtat cttttacgca gcggtattct tcgatcagtt ttttcaattc
     2581 cggtgatatt ctcattttag ccatttatta tttccttcct cttttctaca gtatttaaag
     2641 ataccccaag aagctaatta taacaagacg aactccaatt cactgttcct tgcattctaa
     2701 aaccttaaat acagaaaaca gccttttcaa agttgttttc aaagttggcg tataacatag
     2761 tatcgacgga gccgattttg aaaccacaat tatgatagaa tttgacgtcg aattcgcggc
     2821 cgcttctaga actatagcta gcatgcattt acgttgacac catcgaatgg tgcaaaacct
     2881 ttcgcggtat ggcatgatag cgcccggaag agagtcaatt cagggtggtg aatgtgaaac
     2941 cagtaacgtt atacgatgtc gcagagtatg ccggtgtctc ttatcagacc gtttcccgcg
     3001 tggtgaacca ggccagccac gtttctgcga aaacgcggga aaaagtggaa gcggcgatgg
     3061 cggagctgaa ttacattccc aaccgcgtgg cacaacaact ggcgggcaaa cagtcgttgc
     3121 tgattggcgt tgccacctcc agtctggccc tgcacgcgcc gtcgcaaatt gtcgcggcga
     3181 ttaaatctcg cgccgatcaa ctgggtgcca gcgtggtggt gtcgatggta gaacgaagcg
     3241 gcgtcgaagc ctgtaaagcg gcggtgcaca atcttctcgc gcaacgcgtc agtgggctga
     3301 tcattaacta tccgctggat gaccaggatg ccattgctgt ggaagctgcc tgcactaatg
     3361 ttccggcgtt atttcttgat gtctctgacc agacacccat caacagtatt attttctccc
     3421 atgaagacgg tacgcgactg ggcgtggagc atctggtcgc attgggtcac cagcaaatcg
     3481 cgctgttagc gggcccatta agttctgtct cggcgcgtct gcgtctggct ggctggcata
     3541 aatatctcac tcgcaatcaa attcagccga tagcggaacg ggaaggcgac tggagtgcca
     3601 tgtccggttt tcaacaaacc atgcaaatgc tgaatgaggg catcgttccc actgcgatgc
     3661 tggttgccaa cgatcagatg gcgctgggcg caatgcgcgc cattaccgag tccgggctgc
     3721 gcgttggtgc ggatatctcg gtagtgggat acgacgatac cgaagacagc tcatgttata
     3781 tcccgccgtt aaccaccatc aaacaggatt ttcgcctgct ggggcaaacc agcgtggacc
     3841 gcttgctgca actctctcag ggccaggcgg tgaagggcaa tcagctgttg cccgtctcac
     3901 tggtgaaaag aaaaaccacc ctggcgccca atacgcaaac cgcctctccc cgcgcgttgg
     3961 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     4021 aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt
     4081 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat
     4141 gaccatgatt acggattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg
     4201 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
     4261 agaggcccgc accgatcgcc cttcccaaca gttgcctaag aaaccattat tatcatgaca
     4321 ttaacctata aaaataggcg tatcacgagg ccctttcgtc ttcacctcga gaaatcataa
     4381 aaaatttatt tgctttgtga gcggataaca attataatag attcaattgt gagcggataa
     4441 caatttcaca cagaattaat tcattaaaga ggagaaatta accatgggca gcagccatca
     4501 ccatcatcac cacagccagg atccggccct gagggccgta gaaaccccaa cccgtgaaat
     4561 caaaaaactc gacggcctgt gggcattcag tctggatcgc gaaaactgtg gaattgatca
     4621 gcgttggtgg gaaagcgcgt tacaagaaag ccgggcaatt gctgtgccag gcagttttaa
     4681 cgatcagttc gccgatgcag atattcgtaa ttatgcgggc aacgtctggt atcagcgcga
     4741 agtctttata ccgaaaggtt gggcaggcca gcgtatcgtg ctgcgtttcg atgcggtcac
     4801 tcattacggc aaagtgtggg tcaataatca ggaagtgatg gagcatcagg gcggctatac
     4861 gccatttgaa gccgatgtca cgccgtatgt tattgccggg aaaagtgtac gtatcaccgt
     4921 ttgtgtgaac aacgaactga actggcagac tatcccgccg ggaatggtga ttaccgacga
     4981 aaacggcaag aaaaagcagt cttacttcca tgatttcttt aactatgccg gaatccatcg
     5041 cagcgtaatg ctctacacca cgccgaacac ctgggtggac gatatcaccg tggtgacgca
     5101 tgtcgcgcaa gactgtaacc acgcgtctgt tgactggcag gtggtggcca atggtgatgt
     5161 cagcgttgaa ctgcgtgatg cggatcaaca ggtggttgca actggacaag gcactagcgg
     5221 gactttgcaa gtggtgaatc cgcacctctg gcaaccgggt gaaggttatc tctatgaact
     5281 gtgcgtcaca gccaaaagcc agacagagtg tgatatctac ccgcttcgcg tcggcatccg
     5341 gtcagtggca gtgaagggcg aacagttcct gattaaccac aaaccgttct actttactgg
     5401 ctttggtcgt catgaagatg cggacttgcg tggcaaagga ttcgataacg tgctgatggt
     5461 gcacgaccac gcattaatgg actggattgg ggccaactcc taccgtacct cgcattaccc
     5521 ttacgctgaa gagatgctcg actgggcaga tgaacatggc atcgtggtga ttgatgaaac
     5581 tgctgctgtc ggctttaacc tctctttagg cattggtttc gaagcgggca acaagccgaa
     5641 agaactgtac agcgaagagg cagtcaacgg ggaaactcag caagcgcact tacaggcgat
     5701 taaagagctg atagcgcgtg acaaaaacca cccaagcgtg gtgatgtgga gtattgccaa
     5761 cgaaccggat acccgtccgc aaggtgcacg ggaatatttc gcgccactgg cggaagcaac
     5821 gcgtaaactc gacccgacgc gtccgatcac ctgcgtcaat gtaatgttct gcgacgctca
     5881 caccgatacc atcagcgatc tctttgatgt gctgtgcctg aaccgttatt acggatggta
     5941 tgtccaaagc ggcgatttgg aaacggcaga gaaggtactg gaaaaagaac ttctggcctg
     6001 gcaggagaaa ctgcatcagc cgattatcat caccgaatac ggcgtggata cgttagccgg
     6061 gctgcactca atgtacaccg acatgtggag tgaagagtat cagtgtgcat ggctggatat
     6121 gtatcaccgc gtctttgatc gcgtcagcgc cgtcgtcggt gaacaggtat ggaatttcgc
     6181 cgattttgcg acctcgcaag gcatattgcg cgttggcggt aacaagaaag ggatcttcac
     6241 tcgcgaccgc aaaccgaagt cggcggcttt tctgctgcaa aaacgctgga ctggcatgaa
     6301 cttcggtgaa aaaccgcagc agggaggcaa acaaggccta tgcggccggt agaaaggaga
     6361 agctta
//
