Bonomi M, Muller EGD., Pellarin R, Kim SJ, Russel D, Ramsden R, Sundin BA, Davis TN, Sali A.
pTAF18 载体载体图谱
pTAF18 载体载体序列
LOCUS Exported 6476 bp ds-DNA circular SYN 18-DEC-2018
DEFINITION Yeast integrative vector pTAF18, complete sequence.
ACCESSION KF177463
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6476)
AUTHORS Bonomi M, Muller EGD., Pellarin R, Kim SJ, Russel D, Ramsden R,
Sundin BA, Davis TN, Sali A.
TITLE Protein complex structures from Bayesian modeling of in vivo FRET
data
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 6476)
AUTHORS Bonomi M, Muller EGD., Pellarin R, Kim SJ, Russel D, Ramsden R,
Sundin BA, Davis TN, Sali A.
TITLE Direct Submission
JOURNAL Submitted (31-MAY-2013) Biochemistry, University of Washington, 1705
NE Pacific St, Seattle, WA 98195-7350, USA
REFERENCE 3 (bases 1 to 6476)
AUTHORS .
TITLE Direct Submission
COMMENT ##Assembly-Data-START##
Assembly Method :: CLC Main Workbench v. 6
Sequencing Technology :: Sanger dideoxy sequencing; Genbank
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..6476
/organism="Yeast integrative vector pTAF18"
/mol_type="other DNA"
/db_xref="taxon:1400783"
promoter 1..19
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
regulatory 34..433
/regulatory_class="promoter"
/note="TEF"
CDS 460..834
/codon_start=1
/product="NLS-TAF18"
/label=NLS-TAF18
/note="TAF18 with a nuclear localization signal; one of two
subunits of the human TBP-associated factor
hTAF(II)/hTAT(II)18 heterodimer"
/protein_id="AGW21635.1"
/translation="MVPKKKRKVLIDPKKKRKVDPKKKRKVLINLFSKELRCMMYGFGD
DQNPYTESVDILEDLVIEFITEMTHKAMSIGRQGRVQVEDIVFLIRKDPRKFARVKDLL
TMNEELKRARKAFDEANYGS"
CDS 466..486
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 496..516
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 520..540
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
primer_bind complement(843..859)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
terminator 908..1097
/gene="S. cerevisiae CYC1"
/label=CYC1 terminator
/note="transcription terminator for CYC1"
promoter complement(1120..1138)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(1159..1175)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 1183..1199
/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(1207..1237)
/label=lac promoter
/note="promoter for the E. coli lac operon"
rep_origin 1505..2172
/label=pUC
/note="pUC"
rep_origin complement(1561..2149)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(2320..3180)
/codon_start=1
/product="AmpR"
/label=AmpR
/note="ampicillin resistance protein"
/protein_id="AGW21636.1"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(3181..3285)
/gene="bla"
/label=AmpR promoter
CDS complement(4043..5149)
/codon_start=1
/product="LEU2"
/label=LEU2
/protein_id="AGW21637.1"
/translation="MSAPMSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDF
ENHLIGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKEL
QLYANLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDS
EQYTVPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLK
VQHQLIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDK
NTAFGLYEPCHGSAPDLPKNKVDPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAG
IRTGDLGGSNSTTEVGDAVAEEVKKILA"
CDS complement(4043..5137)
/codon_start=1
/gene="S. cerevisiae LEU2"
/product="3-isopropylmalate dehydrogenase, required for
leucine biosynthesis"
/label=LEU2
/note="yeast auxotrophic marker"
/translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
IGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
GLYEPCHGSAPDLPKNKVDPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
DLGGSNSTTEVGDAVAEEVKKILA"
promoter complement(5150..5554)
/gene="S. cerevisiae LEU2"
/label=LEU2 promoter
rep_origin complement(5854..6309)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind 6454..6470
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
Bonomi M, Muller EGD., Pellarin R, Kim SJ, Russel D, Ramsden R, Sundin BA, Davis TN, Sali A.
pTAF18 载体载体图谱
pTAF18 载体载体序列
LOCUS Exported 6476 bp ds-DNA circular SYN 18-DEC-2018
DEFINITION Yeast integrative vector pTAF18, complete sequence.
ACCESSION KF177463
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6476)
AUTHORS Bonomi M, Muller EGD., Pellarin R, Kim SJ, Russel D, Ramsden R,
Sundin BA, Davis TN, Sali A.
TITLE Protein complex structures from Bayesian modeling of in vivo FRET
data
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 6476)
AUTHORS Bonomi M, Muller EGD., Pellarin R, Kim SJ, Russel D, Ramsden R,
Sundin BA, Davis TN, Sali A.
TITLE Direct Submission
JOURNAL Submitted (31-MAY-2013) Biochemistry, University of Washington, 1705
NE Pacific St, Seattle, WA 98195-7350, USA
REFERENCE 3 (bases 1 to 6476)
AUTHORS .
TITLE Direct Submission
COMMENT ##Assembly-Data-START##
Assembly Method :: CLC Main Workbench v. 6
Sequencing Technology :: Sanger dideoxy sequencing; Genbank
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..6476
/organism="Yeast integrative vector pTAF18"
/mol_type="other DNA"
/db_xref="taxon:1400783"
promoter 1..19
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
regulatory 34..433
/regulatory_class="promoter"
/note="TEF"
CDS 460..834
/codon_start=1
/product="NLS-TAF18"
/label=NLS-TAF18
/note="TAF18 with a nuclear localization signal; one of two
subunits of the human TBP-associated factor
hTAF(II)/hTAT(II)18 heterodimer"
/protein_id="AGW21635.1"
/translation="MVPKKKRKVLIDPKKKRKVDPKKKRKVLINLFSKELRCMMYGFGD
DQNPYTESVDILEDLVIEFITEMTHKAMSIGRQGRVQVEDIVFLIRKDPRKFARVKDLL
TMNEELKRARKAFDEANYGS"
CDS 466..486
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 496..516
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 520..540
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
primer_bind complement(843..859)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
terminator 908..1097
/gene="S. cerevisiae CYC1"
/label=CYC1 terminator
/note="transcription terminator for CYC1"
promoter complement(1120..1138)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(1159..1175)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 1183..1199
/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(1207..1237)
/label=lac promoter
/note="promoter for the E. coli lac operon"
rep_origin 1505..2172
/label=pUC
/note="pUC"
rep_origin complement(1561..2149)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(2320..3180)
/codon_start=1
/product="AmpR"
/label=AmpR
/note="ampicillin resistance protein"
/protein_id="AGW21636.1"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(3181..3285)
/gene="bla"
/label=AmpR promoter
CDS complement(4043..5149)
/codon_start=1
/product="LEU2"
/label=LEU2
/protein_id="AGW21637.1"
/translation="MSAPMSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDF
ENHLIGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKEL
QLYANLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDS
EQYTVPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLK
VQHQLIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDK
NTAFGLYEPCHGSAPDLPKNKVDPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAG
IRTGDLGGSNSTTEVGDAVAEEVKKILA"
CDS complement(4043..5137)
/codon_start=1
/gene="S. cerevisiae LEU2"
/product="3-isopropylmalate dehydrogenase, required for
leucine biosynthesis"
/label=LEU2
/note="yeast auxotrophic marker"
/translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
IGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
GLYEPCHGSAPDLPKNKVDPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
DLGGSNSTTEVGDAVAEEVKKILA"
promoter complement(5150..5554)
/gene="S. cerevisiae LEU2"
/label=LEU2 promoter
rep_origin complement(5854..6309)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind 6454..6470
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"