LOCUS       Exported                8703 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression shuttle vector pIMB4, complete sequence.
ACCESSION   HM756283
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8703)
  AUTHORS   Zhu Y, Wang L, Du Y, Wang S, Yu T, Hong B.
  TITLE     Heterologous expression of human interleukin-6 in Streptomyces 
            lividans TK24 using novel secretory expression vectors
  JOURNAL   Biotechnol. Lett. 33 (2), 253-261 (2011)
  PUBMED    20931350
REFERENCE   2  (bases 1 to 8703)
  AUTHORS   Zhu Y, Wang L, Du Y, Wang S, Yu T, Hong B.
  TITLE     Direct Submission
  JOURNAL   Submitted (13-JUL-2010) Key Laboratory of Biotechnology of 
            Antibiotics of Ministry of Health, Institute of Medicinal 
            Biotechnology, Chinese Academy of Medical Sciences and Peking Union 
            Medical College, No. 1 Tiantan Xili, Beijing, Beijing 100050, China
REFERENCE   3  (bases 1 to 8703)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8703
                     /organism="Expression shuttle vector pIMB4"
                     /mol_type="other DNA"
                     /label=used for expression in Streptomyces lividans
                     /note="used for expression in Streptomyces lividans"
                     /db_xref="taxon:876193"
     CDS             177..977
                     /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             204..980
                     /codon_start=1
                     /gene="aac(3)IV"
                     /product="AprR"
                     /label=aac(3)IV
                     /note="apramycin resistance protein"
                     /protein_id="ADM21210.1"
                     /translation="MQYEWRKAELIGQLLNLGVTPGGVLLVHSSFRSVRPLEDGPLGLI
                     EALRAALGPGGTLVMPSWSGLDDEPFDPATSPVTPDLGVVSDTFWRLPNVKRSAHPFAF
                     AAAGPQAEQIISDPLPLPPHSPASPVARVHELDGQVLLLGVGHDANTTLHLAELMAKVP
                     YGVPRHCTILQDGKLVRVDYLENDHCCERFALADRWLKEKSLQKEGPVGHAFARLIRSR
                     DIVATALGQLGRDPLIFLHPPEAGCEECDAARQSIG"
     gene            204..980
                     /gene="aac(3)IV"
                     /label=aac(3)IV
     misc_feature    1452..1481
                     /label=multiple cloning site
                     /note="multiple cloning site"
     misc_feature    complement(1482..1580)
                     /label=signal peptide of CagA
                     /note="signal peptide of CagA"
     regulatory      complement(1581..1839)
                     /regulatory_class="promoter"
                     /note="cagA promoter"
     regulatory      complement(1846..2057)
                     /regulatory_class="promoter"
                     /note="ermE* promoter"
     primer_bind     complement(2066..2082)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(2112..2130)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2151..2167)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2175..2191
                     /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(2199..2229)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2244..2265
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      2497..3164
                     /label=pUC origin of replication for use in E. coli
                     /note="pUC origin of replication for use in E. coli"
     rep_origin      complement(2553..3141)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3312..4172)
                     /codon_start=1
                     /gene="bla"
                     /product="beta lactamase"
                     /label=bla
                     /note="ampicillin resistance protein"
                     /protein_id="ADM21211.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(3312..4172)
                     /gene="bla"
                     /label=bla
     promoter        complement(4173..4277)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4303..4758)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     4900..4916
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        4926..4944
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(5303..6739)
                     /codon_start=1
                     /gene="rep"
                     /product="Rep"
                     /label=rep
                     /note="replication protein of pSGL1 in Streptomyces"
                     /protein_id="ADM21212.1"
                     /translation="MRTRDARWTLREGLQRVTEDKGLKGCGRSAIAGGVGVVLNGKTAH
                     LSGVATCGKIHLCPVCSAKIRAARSEEIDTVTGAWQAAEHGLAMMTLTLRHCRRMPLGT
                     IRRAERGGLIGVQPGHLHIHTIVLWIRARWVRSVEGQGATGPTTVDSGSTSPGRSAQQF
                     SRYLFKSQDGKARFERWAPGAELTSADKAGRKASRMPFEVAAGAAAGLAEDVPLWQSTS
                     TAPMGSGQSTGPTACGHCSASSCKLDERTDEEIASEDVGGTGVALIPAETWYGVILRHR
                     GRSLDLIRGRRDRRRRRGPVAHHRMGAGVGPGRSPGPGDPDRLTLFWPGGSSRPPGRIG
                     LHRICAGRTGSASRNPLRAPGRARAPGARRRRPIRRISWHGRSRPADERDDDGREPVRL
                     VRRAGPTVRDRAQARVLRADASGAGLPGTAAAAGGGRGGCPGARGRSVGIVSCQLSKRG
                     PRNGVPIRQLSKRLRGRSAP"
     gene            complement(5303..6739)
                     /gene="rep"
                     /label=rep
     rep_origin      complement(6740..7139)
                     /direction=LEFT
                     /label=origin of pSGL1
                     /note="origin of pSGL1"
     CDS             complement(8039..8410)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            8443..8552
                     /note="incP origin of transfer"
     oriT            8444..8554
                     /note="oriT; RK2 origin of transfer"