LOCUS       Exported                4664 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Conjugative vector pHW001, complete sequence.
ACCESSION   DQ457004
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4664)
  AUTHORS   Yang H, Yang Z.
  TITLE     A novel way overcoming blunt-ligation in the construction of 
            Streptomyces conjugative vector pHW001
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4664)
  AUTHORS   Yang H, Yang Z.
  TITLE     Direct Submission
  JOURNAL   Submitted (21-MAR-2006) Life Science and Technology Department, 
            Biochemistry, Huazhong Agricultural University, 1 Shizishan Street, 
            Wuhan, Hubei Province 430070, China
REFERENCE   3  (bases 1 to 4664)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4664
                     /organism="Conjugative vector pHW001"
                     /focus
                     /lab_host="Streptomyces"
                     /mol_type="other DNA"
                     /db_xref="taxon:380046"
     source          1..455
                     /organism="Cloning vector pUC18"
                     /mol_type="other DNA"
                     /db_xref="taxon:72227"
     source          2334..4664
                     /organism="Cloning vector pUC18"
                     /mol_type="other DNA"
                     /db_xref="taxon:72227"
     misc_feature    complement(149..454)
                     /label=lacZa
                     /note="lacZa"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    399..455
                     /label=multiple cloning site
                     /note="multiple cloning site"
     oriT            complement(608..719)
                     /direction=LEFT
                     /label=RP4 from pHZ132
                     /note="RP4 from pHZ132"
     oriT            610..719
                     /note="incP origin of transfer"
     CDS             752..1123
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     gene            complement(1407..2400)
                     /gene="tsr"
                     /label=tsr
                     /note="derived from pHZ132"
     CDS             complement(1407..2216)
                     /codon_start=1
                     /gene="tsr"
                     /product="thiostrepton resistance"
                     /label=tsr
                     /protein_id="ABE60866.1"
                     /translation="MTELDTIANPSDPAVQRIIDVTKPSRSNIKTTLIEDVEPLMHSIA
                     AGVEFIEVYGSDSSPFPSELLDLCGRQNIPVRLIDSSIVNQLFKGERKAKTFGIARVPR
                     PARFGDIASRRGDVVVLDGVKIVGNIGAIVRTSLALGASGIILVDSDITSIADRRLQRA
                     SRGYVFSLPVVLSGREEAIAFIRDSGMQLMTLKADGDISVKELGDNPDRLALLFGSEKG
                     GPSDLFEEASSASVSIPMMSQTESLNVSVSLGIALHERIDRNLAANR"
     regulatory      complement(2267..2272)
                     /regulatory_class="minus_10_signal"
                     /gene="tsr"
                     /note="tandem promoter tsrp1"
     regulatory      complement(2395..2400)
                     /regulatory_class="minus_10_signal"
                     /gene="tsr"
                     /note="tandem promoter tsrp2"
     primer_bind     complement(2443..2459)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2467..2483
                     /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      complement(2486..2573)
                     /regulatory_class="promoter"
                     /note="lac promoter"
     promoter        complement(2491..2521)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2536..2557
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(2845..3433)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      2845
                     /note="pUC18 Ori; RNaseH cleavage point"
     gene            complement(3604..4505)
                     /gene="bla"
                     /label=bla
     CDS             complement(3604..4464)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactmase"
                     /label=bla
                     /note="ampicillin resistance gene"
                     /protein_id="ABE60867.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4465..4569)
                     /gene="bla"
                     /label=AmpR promoter
     regulatory      complement(4499..4505)
                     /regulatory_class="promoter"
                     /gene="bla"