LOCUS       Exported                3417 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pGEX-PUC-3T DNA, complete sequence.
ACCESSION   AB014641
VERSION     .
KEYWORDS    cloning vector; GST-fusion protein; GST-stuffer fusion protein.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3417)
  AUTHORS   Fukunaga R, Hunter T.
  TITLE     MNK1, a new MAP kinase-activated protein kinase, isolated by a novel
            expression screening method for identifying protein kinase 
            substrates
  JOURNAL   EMBO J. 16 (8), 1921-1933 (1997)
  PUBMED    9155018
REFERENCE   2  (bases 1 to 3417)
  AUTHORS   Fukunaga R.
  TITLE     Direct Submission
  JOURNAL   Submitted (22-MAY-1998) Rikiro Fukunaga, Osaka University Medical 
            School, Department of Genetics B-3; 2-2 Yamadaoka, Suita, Osaka 
            565-0871, Japan (E-mail:fukunaga@genetic.med.osaka-u.ac.jp, 
            Tel:81-6-879-3318, Fax:81-6-879-3319)
REFERENCE   3  (bases 1 to 3417)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..3417
                     /organism="Cloning vector pGEX-PUC-3T"
                     /mol_type="other DNA"
                     /db_xref="taxon:77040"
     misc_feature    9..1997
                     /label=pUC119
                     /note="pUC119"
     promoter        36..140
                     /gene="bla"
                     /label=AmpR promoter
     CDS             141..1001
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      1172..1760
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    2005..2953
                     /label=pGEX-KG
                     /note="pGEX-KG"
     promoter        2171..2199
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    2207..2223
                     /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)."
     CDS             2246..3142
                     /codon_start=1
                     /gene="GST-Stuffer fusion"
                     /product="GST-stuffer fusion protein"
                     /label=GST-Stuffer fusion
                     /protein_id="BAA28713.1"
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPKSDLV
                     PRGSPGISGGGGGIGPAGLEGILLHDCGHPGAQAAVSWVWDTWPEGILPVGAATEPGSS
                     TEAGEQQQEAEQAQDDV"
     gene            2246..3142
                     /gene="GST-Stuffer fusion"
                     /label=GST-Stuffer fusion
     CDS             2246..2899
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     CDS             2906..2923
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     misc_feature    2966..3385
                     /note="Stuffer:that the stuffer DNA is derived from the 
                     420bp PvuII fragment(1802-2221) of the human G-CSF receptor
                     cDNA (Acc# No.M38025)"