LOCUS       Exported                7682 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pTAOR4-Rev-boxAB DNA, complete sequence.
ACCESSION   AB600173
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)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7682
                     /organism="Expression vector pTAOR4-Rev-boxAB"
                     /mol_type="other DNA"
                     /db_xref="taxon:927590"
     oriT            224..333
                     /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
                     /codon_start=1
                     /gene="aac(3)IV"
                     /product="aminoglycoside 3-N-acetyltransferase type IV     
                               "
                     /label=ApmR
                     /note="confers resistance to apramycin (Paget and Davies, 
                     1996)"
                     /translation="MSSAVECNVVQYEWRKAELIGQLLNLGVTPGGVLLVHSSFRSVRP
                     LEDGPLGLIEALRAALGPGGTLVMPSWSGLDDEPFDPATSPVTPDLGVVSDTFWRLPNV
                     KRSAHPFAFAAAGPQAEQIISDPLPLPPHSPASPVARVHELDGQVLLLGVGHDANTTLH
                     LAELMAKVPYGVPRHCTILQDGKLVRVDYLENDHCCERFALADRWLKEKSLQKEGPVGH
                     AFARLIRSRDIVATALGQLGRDPLIFLHPPEAGCEECDAARQSIG"
     CDS             1335..2111
                     /codon_start=1
                     /gene="aprr"
                     /product="apramycin resistance protein"
                     /label=aprr
                     /protein_id="BAJ53865.1"
                     /translation="MQYEWRKAELIGQLLNLGVTPGGVLLVHSSFRSVRPLEDGPLGLI
                     EALRAALGPGGTLVMPSWSGLDDEPFDPATSPVTPDLGVVSDTFWRLPNVKRSAHPFAF
                     AAAGPQAEQIISDPLPLPPHSPASPVARVHELDGQVLLLGVGHDANTTLHLAELMAKVP
                     YGVPRHCTILQDGKLVRVDYLENDHCCERFALADRWLKEKSLQKEGPVGHAFARLIRSR
                     DIVATALGQLGRDPLIFLHPPEAGCEECDAARQSIG"
     gene            1335..2111
                     /gene="aprr"
                     /label=aprr
     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    2450..2466
                     /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)."
     promoter        complement(2474..2504)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2519..2540
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      2769..3442
                     /label=replication origin of Escherichia coli
                     /note="replication origin of Escherichia coli"
     rep_origin      complement(2828..3416)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     regulatory      3493..3687
                     /regulatory_class="terminator"
                     /note="downstream of cloning site"
     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)
                     /regulatory_class="promoter"
                     /gene="aceA"
                     /note="acetone inducible promoter from Pseudonocardia 
                     autotrophica"
                     /experiment="SDS-PAGE of gene product in acetone induction"
     gene            complement(5121..5568)
                     /gene="aceA"
                     /label=aceA
     rep_origin      5995..7293
                     /label=replication origin of Pseudonocardia autotrophica
                     /note="replication origin of Pseudonocardia autotrophica"