LOCUS       Exported                7456 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pJKPN, complete sequence.
ACCESSION   JN795551
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7456)
  AUTHORS   Gong Z, Lv H.
  TITLE     Vector pJKPN for targeted integration at the Leu1-32 of 
            Schizosaccharomyces pombe
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7456)
  AUTHORS   Gong Z, Lv H.
  TITLE     Direct Submission
  JOURNAL   Submitted (28-SEP-2011) School of Life Science, Fudan University, 
            Room 618, Science Building, Fudan University, Handan Road 220, 
            Shanghai, Shanghai 200433, China
REFERENCE   3  (bases 1 to 7456)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7456
                     /organism="Cloning vector pJKPN"
                     /mol_type="other DNA"
                     /db_xref="taxon:1147118"
     gene            187..2258
                     /gene="Leu1"
                     /label=Leu1
                     /note="derived from Schizosaccharomyces pombe Leu1"
     5UTR           187..306
                     /gene="Leu1"
     CDS             307..1422
                     /codon_start=1
                     /gene="Leu1"
                     /product="3-isopropylmalate dehydrogenase"
                     /label=Leu1
                     /protein_id="AFA55176.1"
                     /translation="MCAKKIVVLPGDHIGPEIVASALEVLKVVEKKRPELKLEFEEHKI
                     GGASIDAYGTPLTDETVKACLEADGVLLGAVGGPEWTNPNCRPEQGLLKLRKSMGVWAN
                     LRPCNFASKSLVKYSPLKPEIVEGVDFCVVRELTGGCYFGERTEDNGSGYAMDTWPYSL
                     EEVSRIARLAAWLAETSNPPAPVTLLDKANVLATSRLWRKTVAKIFKEEYPHLTLKNQL
                     IDSAAMLLVKSPRTLNGVVLTDNLFGDIISDEASVIPGSLGLLPSASLSGVVGKSEEKV
                     HCLVEPIHGSAPDIAGKGIVNPVGTILSASLLLRYGLNAPKEAEAIEAAVRKVLDDTSI
                     GGRGLYTRDLGGEASTADITKAVVEELEKIL"
     3UTR           1423..2258
                     /gene="Leu1"
     rep_origin      complement(2308..2763)
                     /direction=LEFT
                     /label=f1 origin
                     /note="f1 origin"
     primer_bind     2910..2926
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2936..2954
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2965..4157
                     /regulatory_class="promoter"
                     /note="NMT1 promoter P1"
     promoter        2995..4159
                     /label=nmt1 promoter
                     /note="wild-type nmt1 promoter from Schizosaccharomyces 
                     pombe, conferring strong thiamine-repressible expression"
     regulatory      4181..5177
                     /regulatory_class="terminator"
                     /note="NMT1 terminator"
     promoter        complement(5196..5214)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5235..5251)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5259..5275
                     /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(5283..5313)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     rep_origin      5622..6241
                     /label=pBR322 origin
                     /note="pBR322 origin"
     rep_origin      complement(5637..6225)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6396..7256)
                     /codon_start=1
                     /gene="ampR"
                     /product="beta-lactamase"
                     /label=ampR
                     /protein_id="AFA55177.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFFHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(6396..7256)
                     /gene="ampR"
                     /label=ampR
     promoter        complement(7257..7361)
                     /gene="bla"
                     /label=AmpR promoter