LOCUS       Exported                5399 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  S. pombe expression vector pHIS3K DNA, complete sequence.
ACCESSION   AB364151
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5399)
  AUTHORS   Matsuyama A, Shirai A, Yoshida M.
  TITLE     A novel series of vectors for chromosomal integration in fission 
            yeast
  JOURNAL   Biochem. Biophys. Res. Commun. 374 (2), 315-319 (2008)
  PUBMED    18634753
REFERENCE   2  (bases 1 to 5399)
  AUTHORS   Matsuyama A, Shirai A, Yoshida M.
  TITLE     Fission yeast integration vector pHIS3K
  JOURNAL   Published Only in Database (2007)
REFERENCE   3  (bases 1 to 5399)
  AUTHORS   Matsuyama A, Shirai A, Yoshida M.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-OCT-2007) Contact:Akihisa Matsuyama RIKEN, Chemical 
            Genetics Laboratory; Hirosawa 2-1, Wako, Saitama 351-0198, Japan URL
            :http://cgl.riken.go.jp/
REFERENCE   4  (bases 1 to 5399)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     Fission yeast expression vector.
FEATURES             Location/Qualifiers
     source          1..5399
                     /organism="Schizosaccharomyces pombe expression vector
                     pHIS3K"
                     /mol_type="other DNA"
                     /db_xref="taxon:477429"
     regulatory      1..237
                     /regulatory_class="terminator"
                     /gene="ADH1"
     misc_feature    1..237
                     /gene="ADH1"
                     /label=derived from Saccharomyces cerevisiae
                     /note="derived from Saccharomyces cerevisiae"
     gene            1..237
                     /gene="ADH1"
                     /label=ADH1
     terminator      21..208
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     misc_feature    242..808
                     /pseudo
                     /gene="his3"
                     /label=derived from Schizosaccharomyces pombe
                     /note="derived from Schizosaccharomyces pombe"
     CDS             242..808
                     /pseudo
                     /gene="his3"
                     /label=his3
                     /note="NotI site is created at the middle of this fragment 
                     (487-494)."
     gene            242..808
                     /pseudo
                     /gene="his3"
                     /label=his3
     regulatory      813..1195
                     /regulatory_class="promoter"
                     /gene="TEF1"
     gene            813..1195
                     /gene="TEF1"
                     /label=TEF1
     gene            852..2208
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        852..1195
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1196..2005
                     /codon_start=1
                     /gene="kanR"
                     /product="G418-resistance gene"
                     /label=kanR
                     /protein_id="BAF90990.1"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     gene            1196..2005
                     /gene="kanR"
                     /label=kanR
     regulatory      2006..2246
                     /regulatory_class="terminator"
                     /gene="TEF1"
     gene            2006..2246
                     /gene="TEF1"
                     /label=TEF1
     terminator      2011..2208
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     misc_feature    2247..5399
                     /label=derived from Cloning vector pUC119
                     /note="derived from Cloning vector pUC119"
     primer_bind     complement(2259..2275)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      2488..2943
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        3225..3329
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3330..4190
                     /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      4361..4949
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    5237..5258
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5273..5303
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5311..5327
                     /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)."
     primer_bind     5335..5351
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"