LOCUS       40924_42659        7682 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pTAOR4-Rev-boxAB DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7682)
  AUTHORS   Fujii Y, Norihisa K, Fujii T, Aritoku Y, Kagawa Y, Sallam KI, Johdo 
            O, Arisawa A, Tamura T.
  TITLE     Construction of a novel expression vector in Pseudonocardia 
            autotrophica and its application to efficient biotransformation of 
            compactin to pravastatin, a specific HMG-CoA reductase inhibitor
  JOURNAL   Biochem. Biophys. Res. Commun. 404 (1), 511-516 (2011)
  PUBMED    21144838
REFERENCE   2  (bases 1 to 7682)
  AUTHORS   Arisawa A, Fujii Y, Tamura T.
  TITLE     Direct Submission
  JOURNAL   Submitted (10-NOV-2010) Contact:Akira Arisawa Mercian Bioresource 
            Laboratories; Nakaizumi, Iwata, Shizuoka 438-0078, Japan
REFERENCE   3  (bases 1 to 7682)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7682)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Biochem. 
            Biophys. Res. Commun."; date: "2011"; volume: "404"; issue: "1"; 
            pages: "511-516"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (10-NOV-2010) Contact:Akira Arisawa Mercian Bioresource 
            Laboratories; Nakaizumi, Iwata, Shizuoka 438-0078, Japan"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7682
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     oriT            complement(224..333)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     promoter        complement(568..658)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     promoter        complement(852..942)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     CDS             1308..2108
                     /label=ApmR
                     /note="aminoglycoside 3-N-acetyltransferase type IV"
     promoter        complement(2390..2408)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2426..2442)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2450..2466)
                     /label=lac operator
                     /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)."
     promoter        complement(2474..2504)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2519..2540)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(2828..3416)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     regulatory      3493..3687
                     /label=downstream of cloning site
                     /note="downstream of cloning site"
                     /regulatory_class="terminator"
     CDS             complement(3688..3879)
                     /codon_start=1
                     /gene="boxB"
                     /product="ferredoxin"
                     /label=boxB
                     /protein_id="BAJ53866.1"
                     /translation="MRVTADREVCVGAGLCALTAPEVFDQDDDGVVTVLAAEPGEAGRA
                     AALEAGALCPSGAVRVVE"
     gene            complement(3688..3879)
                     /gene="boxB"
                     /label=boxB
     CDS             complement(3900..5117)
                     /codon_start=1
                     /gene="boxA"
                     /product="cytochrome P450"
                     /label=boxA
                     /note="compactin 6beta-hydroxylase"
                     /protein_id="BAJ53867.1"
                     /translation="MTETVTTPTSGAPAFPSDRTCPYHLPDRYNDLRDREGSLQRVTLY
                     DGRQAWLVTGYDTARKLLADPRLSSDRTHADFPATSGRVESFRDRRPAFISLDPPEHGP
                     KRRMTISEFTVRRIKGMRADVEQIVHGFLDEMIAGGPPADLVSQFALPVPSLVICRLLG
                     VPYADHDFFQDASARLIQSPDAAGARAARDDLESYLGALVDSLRGESRPGLLSTLVREQ
                     LEKGAIDREELVSTAILLLVAGHETTASMTSLSVITLLEHPDQHAALRADPSLVPGAVE
                     ELLRYLAIADIAGGRIATADIEIDGQRIRAGEGVIVTNSIANRDGSVFADPDAFDVRRE
                     ARHHLAFGYGVHQCLGQNLARLELEVILTALFERLPGLRLAVPVDRLTLRPGTTIQGVN
                     ELPVTW"
     gene            complement(3900..5117)
                     /gene="boxA"
                     /label=boxA
     regulatory      complement(5121..5568)
                     /gene="aceA"
                     /note="acetone inducible promoter from Pseudonocardia 
                     autotrophica"
                     /experiment="SDS-PAGE of gene product in acetone induction"
                     /regulatory_class="promoter"
     gene            complement(5121..5568)
                     /gene="aceA"
                     /label=aceA
     rep_origin      5995..7293
                     /label=replication origin of Pseudonocardia autotrophica
                     /note="replication origin of Pseudonocardia autotrophica"
ORIGIN
        1 tgtacatcgc ggacgtgctc atagtccacg acgcccgtga ttttgtagcc ctggccgacg
       61 gccagcaggt aggccgacag gctcatgccg gccgccgccg ccttttcctc aatcgctctt
      121 cgttcgtctg gaaggcagta caccttgata ggtgggctgc ccttcctggt tggcttggtt
      181 tcatcagcca tccgcttgcc ctcatctgtt acgccggcgg tagccggcca gcctcgcaga
      241 gcaggattcc cgttgagcac cgccaggtgc gaataaggga cagtgaagaa ggaacacccg
      301 ctcgcgggtg ggcctacttc acctatcctg ccccgctgac gccgttggat acaccaagga
      361 aagtctacac gaaccctttg gcaaaatcct gtatatcgtg cgaaaaagga tggatatacc
      421 gaaaaaatcg ctataatgac cccgaagcag ggttatgcag cggaaaagcg ctgcttccct
      481 gctgttttgt ggaatatcta ccgactggaa acaggcaaat gcaggaaatt actgaactga
      541 ggggacaggc gagagacgat gccaaagagc tcctgaaaat ctcgataact caaaaaatac
      601 gcccggtagt gatcttattt cattatggtg aaagttggaa cctcttacgt gccgatcaac
      661 gtctcatttt cgccaaaagt tggcccaggg cttcccggta tcaacaggga caccaggatt
      721 tatttattct gcgaagtgat cttccgtcac aggtatttat tcggcgcaaa gtgcgtcggg
      781 tgatgctgcc aacttactga tttagtgtat gatggtgttt ttgaggtgct ccagtggctt
      841 ctgtttctat cagctcctga aaatctcgat aactcaaaaa atacgcccgg tagtgatctt
      901 atttcattat ggtgaaagtt ggaacctctt acgtgccgat caacgtctca ttttcgccaa
      961 aagttggccc agggcttccc ggtatcaaca gggacaccag gatttattta ttctgcgaag
     1021 tgatcttccg tcacaggtat ttattcggcg caaagtgcgt cgggtgatgc tgccaactta
     1081 ctgatttagt gtatgatggt gtttttgagg tgctccagtg gcttctgttt ctatcagggg
     1141 atccatgccg tatttgcagt accagcgtac ggcccacaga atgatgtcac gctgaaaatg
     1201 ccggcctttg aatgggttca tgtgcagctc catcagcaaa aggggatgat aagtttatca
     1261 ccaccgacta tttgcaacag tgccgttgat cgtgctatga tcgactgatg tcatcagcgg
     1321 tggagtgcaa tgtcgtgcaa tacgaatggc gaaaagccga gctcatcggt cagcttctca
     1381 accttggggt tacccccggc ggtgtgctgc tggtccacag ctccttccgt agcgtccggc
     1441 ccctcgaaga tgggccactt ggactgatcg aggccctgcg tgctgcgctg ggtccgggag
     1501 ggacgctcgt catgccctcg tggtcaggtc tggacgacga gccgttcgat cctgccacgt
     1561 cgcccgttac accggacctt ggagttgtct ctgacacatt ctggcgcctg ccaaatgtaa
     1621 agcgcagcgc ccatccattt gcctttgcgg cagcggggcc acaggcagag cagatcatct
     1681 ctgatccatt gcccctgcca cctcactcgc ctgcaagccc ggtcgcccgt gtccatgaac
     1741 tcgatgggca ggtacttctc ctcggcgtgg gacacgatgc caacacgacg ctgcatcttg
     1801 ccgagttgat ggcaaaggtt ccctatgggg tgccgagaca ctgcaccatt cttcaggatg
     1861 gcaagttggt acgcgtcgat tatctcgaga atgaccactg ctgtgagcgc tttgccttgg
     1921 cggacaggtg gctcaaggag aagagccttc agaaggaagg tccagtcggt catgcctttg
     1981 ctcggttgat ccgctcccgc gacattgtgg cgacagccct gggtcaactg ggccgagatc
     2041 cgttgatctt cctgcatccg ccagaggcgg gatgcgaaga atgcgatgcc gctcgccagt
     2101 cgattggctg agctcatgag cggagaacga gatgacgttg gaggggcaag gtcgcgctga
     2161 ttgctggggc aacacgtgga gcggatcggg gattgtcttt cttcagctcg ctgatgatat
     2221 gctgacgctc aatgccgttt ggcctccgac taacgaaaat cccgcatttg gacggctgat
     2281 ccgattggca cggcggacgg cgaatggcgg agcagacgct cgtccggggg caatgagata
     2341 tgaaaaagcc tgaactcacc gcgacgtaag ctttaagcag cttgagtatt ctatagtgtc
     2401 acctaaatag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     2461 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     2521 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     2581 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     2641 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     2701 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     2761 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     2821 tggcgttttt cgataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     2881 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     2941 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     3001 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     3061 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     3121 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     3181 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     3241 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
     3301 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     3361 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     3421 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     3481 ttttggggta cctagagtcc cgctgaggcg gcgtagcagg tcagccgccc cagcggtggt
     3541 caccaaccgg ggtggaacgg cgccggtatc gggtgtgtcc gtggcgctca ttccaacctc
     3601 cgtgtgtttg tgcaggtttc gcgtgttgca gtccctcgca ccggcacccg cagcgagggg
     3661 ctcacgggtg ccggtgggtc gactagtcta ctcgacgacg cgtaccgcgc cggacgggca
     3721 cagcgcgccg gcttcgagtg ccgccgcacg gccggcctcg ccgggttccg cggccagcac
     3781 cgtcaccaca ccgtcgtcgt cctggtcgaa gacctccggc gccgtcaagg cgcacaggcc
     3841 cgctcccacg cagacctccc ggtcggccgt cacacgcatg gctgctcctt tcgccgcggt
     3901 caccaggtga ccgggagttc gttcacgccc tggatcgtcg tgcccgggcg cagggtcagc
     3961 cggtccaccg gcaccgccag ccgcagaccg ggcagccgct cgaacagcgc cgtgaggatg
     4021 acctccagtt cgaggcgggc caggttctgg ccgaggcact gatgcacccc gtagccgaac
     4081 gccaggtggt ggcgggcctc gcgccgcacg tcgaaggcgt ccgggtcggc gaagacggag
     4141 ccgtcgcggt tggcgatcga gttggtgacg atgaccccct cccccgcccg gatgcgctgc
     4201 ccgtcgatct cgatgtccgc cgtcgcgatc cgcccgccgg cgatgtcggc gatggccaga
     4261 tagcgcagca gctcctccac cgcgccgggc accagcgacg gatcggcgcg caacgcggcg
     4321 tgctggtcgg gatgttcgag gagggtgatg acgctgagcg acgtcatcga cgccgtcgtc
     4381 tcgtgtccgg cgaccagcag caggatcgcc gtcgacacca gctcctcccg gtcgatcgcg
     4441 cccttctcca gctgctccct gacgagcgtg ctcagcaggc ccggccggga ctcgcctcgc
     4501 aggctgtcca ccagagcgcc cagatagctc tccaggtcgt cccgggcggc acgcgcaccc
     4561 gccgcgtccg gggactggat cagccgtgcg ctggcgtcct ggaagaagtc gtggtccgcg
     4621 tagggcacac cgagcagacg gcagatcacc agggacggga cgggcagcgc gaactggctg
     4681 accaggtcgg cgggcgggcc gcctgcgatc atctcgtcca ggaagccgtg cacgatctgc
     4741 tcgacgtcgg cccgcatgcc cttgatgcgc cggacggtga actcgctgat ggtcatgcgc
     4801 cgtttcggcc cgtgctcggg cgggtccagg ctgatgaacg ccggccggcg gtcccggaag
     4861 ctctccaccc gcccggaggt ggcggggaag tcggcgtgtg tccggtcgga cgagagccgg
     4921 gggtcggcca gcagcttgcg tgccgtgtcg tacccggtca ccagccatgc ctgccggccg
     4981 tcgtagaggg tgacccgctg cagcgaaccc tcccggtccc ggaggtcgtt gtaccggtcg
     5041 gggaggtggt aggggcaggt gcggtcactg gggaaggcgg gggcgcctga tgtgggcgtc
     5101 gtaacggtct cggtcatatg cgatggtcct tcccgtgggg gatgtccgtg ggggatgtcc
     5161 gtggagatat cactgcattg cgcgtgatgc cggacactag gaaggtgggg gttgcacagg
     5221 ggttgcacgc agatgcggaa cctgaacctc cggagagtgc cgatcccgcg gattcgattc
     5281 aggtaaggct gagctaacca tagggtgagt gtcgtgacgc gtgaacgagg cgggctggag
     5341 cgggcgcgtg gcgccgcgct gaccggttcg gccgcaccgg aggtgcgccc ggagatccgg
     5401 gcgtcctggg agcgttcgct ggccgccgcc gtcgatccgg agcggtcccg gccctcgcac
     5461 acctaccagc cgggcgaggt cgccgagctg cgcgccggac atccgctggc gcaggtcctg
     5521 ccgctgctgc gggagaccct cgtcgccgcg gccgagcagg ccaggcacgg taccctcccg
     5581 ccgccccgca cggcacgccg tcatgacggt caaccgtcgg cgtcgtccta tcgacgacag
     5641 caccggatgc cggacgggga acaggagcac cgcccgtctc gaccgctgcc cttcctcgat
     5701 acaaggccgc acgagtgcgg ggcaggttca gtcaagggtc ggcgcagccg atcgcgtagc
     5761 gacgccgcag gcgcccttgg cgggttctga cctgcattcg acacttagcc gccatcgagg
     5821 tagggcccac cgcagcacgc gctacagcac cggcaccgag aacaccctca gctctcgcgc
     5881 cgcaggcgcg cgcccggtcc ggacgggccc ggcccgcggg ccggaggcag gagcgggccg
     5941 gagcccggcc cggccgggcg ccggccacag cggcccgatc gctggcggtg ctcgatgacc
     6001 gccgcgctga cgcgcgtcga cgcgggcgtg cccgcgcttg gtactgacgc gaaaagtgcg
     6061 gccaccgcag gtcaggtccc cgtgggggac tgggcaggga gcttctggga gcggcaggac
     6121 cgcgccttgc gggagaagta ccgggcccgc cgtgagctgg ccaggatcac gacgctgcgt
     6181 cgcgttgcac gctgcggacg ctcctcgatg aacgacggcg gagacgtcgt cctgcgctac
     6241 tcgcccggca ccggggagga cggctcggcg tcggcgggtt tcggcggact ggtgacctgc
     6301 ggcagcctgt gggcctgccc ggtctgctcg gccaagatca gcgcccgccg ggcccgggag
     6361 ctggaacacc tgatcacctg gaacgccgcc cgcggcggca ccgtcgcgct gctcagcctg
     6421 accatgcgcc accacagcgg ccaccgcctg cgcgacctgc gccgagggct gagcgccgcg
     6481 tggcgccacg tcaccagctc ccgcgggtgg aagcgctgga agagcgtctt gggaatggac
     6541 tacgtccgcg ggattgaggc cacccacgga gcgaacggct ggcatctgca catccacgcc
     6601 ctgctgatct tccccggcga cgtcacggag gagatgcacg ccctcaccgc cgagatctgg
     6661 acccgctggt cgaccggcct gcggcgcaag ggcttcgacg ccacgatcgc ccacggcgtc
     6721 gacgtccggg tcggcaccgg cgccctcgaa cagctcggcc gctacatctc caaactggcc
     6781 ttcgagacct ccggcggccg gtggaagctg ggcaagaacg gcagccgtac cccgttccag
     6841 atcctcgccg acgccctgga ccgagcccgc gaccgagacc tcgcgctctg ggcggagtgg
     6901 gagcaggcca gccacggcat gcagcagctc gtgtggtcca acggactcaa ggcggcctgc
     6961 cagctcgacg agatcgacga cgagacgatc gcggaggagg acgacggtgg tgagttcgtc
     7021 gcccagctcc cccgccgcac ctgggagaag gtctaccccg tcgccgaaga cctgatcatc
     7081 gccacccgca ctggtggccc cgaagcaggc cgcgcctggc ttgacgcccg cggcctggcc
     7141 tatgaccacg agcgcgacac gagcgaacga gcagtcctgc tcgacgagcc ggacccgccg
     7201 ttcgcgtggc tgagggccgc tctcgcggcc gaagaccccg agcagcgccg ggagcgacgc
     7261 cgccgctact accgcaccgc acagaccaac tgagctcgat gaggagcaga accacgatgg
     7321 ccgacacctg gacgatcagc ccgctgatcc gccccctgct gcgcaagctc gatgagcgcg
     7381 gaggctgcgc ctacccgcag gacttcccgc actgccacgg cgccgagatc cagcacaccg
     7441 acggcaccgc cgaatgcttc aaccccggcc agccctgccc ctacccacga cccggcgcgc
     7501 acgccttcgt ccacacctgc gctgacgtca cccaccggct cacccaccgc tgcacccgct
     7561 gccgctgatt caccacaggc catgcaccta tgcattgccc aatgatgatc ttcggctgct
     7621 gacgtcgtcg atacgacgac attactcaat tctgttgtcc cataaagcat ctggcctcta
     7681 at
//
