LOCUS Exported 5545 bp ds-DNA circular SYN 18-DEC-2018
DEFINITION S. pombe expression vector pLYS1K DNA, complete sequence.
ACCESSION AB364152
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5545)
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 5545)
AUTHORS Matsuyama A, Shirai A, Yoshida M.
TITLE Fission yeast integration vector pLYS1K
JOURNAL Published Only in Database (2007)
REFERENCE 3 (bases 1 to 5545)
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 5545)
AUTHORS .
TITLE Direct Submission
COMMENT Fission yeast expression vector.
FEATURES Location/Qualifiers
source 1..5545
/organism="Schizosaccharomyces pombe expression vector
pLYS1K"
/mol_type="other DNA"
/db_xref="taxon:477430"
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..956
/pseudo
/gene="lys1"
/label=derived from Schizosaccharomyces pombe
/note="derived from Schizosaccharomyces pombe"
CDS 242..956
/pseudo
/gene="lys1"
/label=lys1
/note="NotI site is created at the middle of this fragment
(613-620)."
gene 242..956
/pseudo
/gene="lys1"
/label=lys1
regulatory 959..1341
/regulatory_class="promoter"
/gene="TEF1"
gene 959..1341
/gene="TEF1"
/label=TEF1
gene 998..2354
/label=kanMX
/note="yeast selectable marker conferring kanamycin
resistance (Wach et al., 1994)"
promoter 998..1341
/label=TEF promoter
/note="Ashbya gossypii TEF promoter"
CDS 1342..2151
/codon_start=1
/gene="kanR"
/product="G418-resistance gene"
/label=kanR
/protein_id="BAF90991.1"
/translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
gene 1342..2151
/gene="kanR"
/label=kanR
regulatory 2152..2392
/regulatory_class="terminator"
/gene="TEF1"
gene 2152..2392
/gene="TEF1"
/label=TEF1
terminator 2157..2354
/label=TEF terminator
/note="Ashbya gossypii TEF terminator"
misc_feature 2393..5545
/label=derived from Cloning vector pUC119
/note="derived from Cloning vector pUC119"
primer_bind complement(2405..2421)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
rep_origin 2634..3089
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 3371..3475
/gene="bla"
/label=AmpR promoter
CDS 3476..4336
/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 4507..5095
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind 5383..5404
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 5419..5449
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5457..5473
/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 5481..5497
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
LOCUS Exported 5545 bp ds-DNA circular SYN 18-DEC-2018
DEFINITION S. pombe expression vector pLYS1K DNA, complete sequence.
ACCESSION AB364152
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5545)
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 5545)
AUTHORS Matsuyama A, Shirai A, Yoshida M.
TITLE Fission yeast integration vector pLYS1K
JOURNAL Published Only in Database (2007)
REFERENCE 3 (bases 1 to 5545)
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 5545)
AUTHORS .
TITLE Direct Submission
COMMENT Fission yeast expression vector.
FEATURES Location/Qualifiers
source 1..5545
/organism="Schizosaccharomyces pombe expression vector
pLYS1K"
/mol_type="other DNA"
/db_xref="taxon:477430"
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..956
/pseudo
/gene="lys1"
/label=derived from Schizosaccharomyces pombe
/note="derived from Schizosaccharomyces pombe"
CDS 242..956
/pseudo
/gene="lys1"
/label=lys1
/note="NotI site is created at the middle of this fragment
(613-620)."
gene 242..956
/pseudo
/gene="lys1"
/label=lys1
regulatory 959..1341
/regulatory_class="promoter"
/gene="TEF1"
gene 959..1341
/gene="TEF1"
/label=TEF1
gene 998..2354
/label=kanMX
/note="yeast selectable marker conferring kanamycin
resistance (Wach et al., 1994)"
promoter 998..1341
/label=TEF promoter
/note="Ashbya gossypii TEF promoter"
CDS 1342..2151
/codon_start=1
/gene="kanR"
/product="G418-resistance gene"
/label=kanR
/protein_id="BAF90991.1"
/translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
gene 1342..2151
/gene="kanR"
/label=kanR
regulatory 2152..2392
/regulatory_class="terminator"
/gene="TEF1"
gene 2152..2392
/gene="TEF1"
/label=TEF1
terminator 2157..2354
/label=TEF terminator
/note="Ashbya gossypii TEF terminator"
misc_feature 2393..5545
/label=derived from Cloning vector pUC119
/note="derived from Cloning vector pUC119"
primer_bind complement(2405..2421)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
rep_origin 2634..3089
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 3371..3475
/gene="bla"
/label=AmpR promoter
CDS 3476..4336
/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 4507..5095
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind 5383..5404
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 5419..5449
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5457..5473
/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 5481..5497
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"