Bonomi M, Muller EGD., Pellarin R, Kim SJ, Russel D, Ramsden R, Sundin BA, Davis TN, Sali A.
pBS47 载体载体图谱
pBS47 载体载体序列
LOCUS Exported 6628 bp ds-DNA circular SYN 17-DEC-2018
DEFINITION Yeast integrative vector pBS47, complete sequence.
ACCESSION KF177457
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6628)
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JOURNAL Unpublished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 6628)
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 6628)
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..6628
/organism="Yeast integrative vector pBS47"
/mol_type="other DNA"
/db_xref="taxon:1400777"
regulatory 2..401
/regulatory_class="promoter"
/note="TEF"
CDS 428..1990
/codon_start=1
/product="NLS-YFP-Pro10-CFP"
/label=NLS-YFP-Pro10-CFP
/note="YFP and CFP linked with 10 tandem prolines and a
nuclear localization signal"
/protein_id="AGW21617.1"
/translation="MVPKKKRKVLIDPKKKRKVDPKKKRKVLINMVSKGEELFTGVVPI
LVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTFGYGLKCFAR
YPDHMKRHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKE
DGNILGHKLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDG
PVLLPDNHYLSYQSALSKDPNEKRDHMVLLEFVTAAGITHGMDELYKGAPPPPPPPPPP
GASKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWP
TLVTTLTWGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGD
TLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIKAHFKIRHNIEDGSV
QLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITHGMDE
LYK"
CDS 434..454
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 464..484
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 488..508
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
primer_bind complement(2094..2110)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
terminator 2159..2348
/gene="S. cerevisiae CYC1"
/label=CYC1 terminator
/note="transcription terminator for CYC1"
promoter complement(2371..2389)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(2410..2426)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 2434..2450
/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(2458..2488)
/label=lac promoter
/note="promoter for the E. coli lac operon"
rep_origin 2756..3423
/label=pUC
/note="pUC"
rep_origin complement(2812..3400)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(3571..4431)
/codon_start=1
/product="AmpR"
/label=AmpR
/note="ampicillin resistance protein"
/protein_id="AGW21618.1"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFFHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(4432..4536)
/gene="bla"
/label=AmpR promoter
promoter 4826..5041
/gene="S. cerevisiae URA3"
/label=URA3 promoter
CDS 5042..5845
/codon_start=1
/product="URA3"
/label=URA3
/protein_id="AGW21619.1"
/translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
rep_origin complement(5974..6429)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind 6574..6590
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 6597..6615
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
Bonomi M, Muller EGD., Pellarin R, Kim SJ, Russel D, Ramsden R, Sundin BA, Davis TN, Sali A.
pBS47 载体载体图谱
pBS47 载体载体序列
LOCUS Exported 6628 bp ds-DNA circular SYN 17-DEC-2018
DEFINITION Yeast integrative vector pBS47, complete sequence.
ACCESSION KF177457
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6628)
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 6628)
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 6628)
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..6628
/organism="Yeast integrative vector pBS47"
/mol_type="other DNA"
/db_xref="taxon:1400777"
regulatory 2..401
/regulatory_class="promoter"
/note="TEF"
CDS 428..1990
/codon_start=1
/product="NLS-YFP-Pro10-CFP"
/label=NLS-YFP-Pro10-CFP
/note="YFP and CFP linked with 10 tandem prolines and a
nuclear localization signal"
/protein_id="AGW21617.1"
/translation="MVPKKKRKVLIDPKKKRKVDPKKKRKVLINMVSKGEELFTGVVPI
LVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTFGYGLKCFAR
YPDHMKRHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKE
DGNILGHKLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDG
PVLLPDNHYLSYQSALSKDPNEKRDHMVLLEFVTAAGITHGMDELYKGAPPPPPPPPPP
GASKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWP
TLVTTLTWGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGD
TLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIKAHFKIRHNIEDGSV
QLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITHGMDE
LYK"
CDS 434..454
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 464..484
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 488..508
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
primer_bind complement(2094..2110)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
terminator 2159..2348
/gene="S. cerevisiae CYC1"
/label=CYC1 terminator
/note="transcription terminator for CYC1"
promoter complement(2371..2389)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(2410..2426)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 2434..2450
/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(2458..2488)
/label=lac promoter
/note="promoter for the E. coli lac operon"
rep_origin 2756..3423
/label=pUC
/note="pUC"
rep_origin complement(2812..3400)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(3571..4431)
/codon_start=1
/product="AmpR"
/label=AmpR
/note="ampicillin resistance protein"
/protein_id="AGW21618.1"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFFHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(4432..4536)
/gene="bla"
/label=AmpR promoter
promoter 4826..5041
/gene="S. cerevisiae URA3"
/label=URA3 promoter
CDS 5042..5845
/codon_start=1
/product="URA3"
/label=URA3
/protein_id="AGW21619.1"
/translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
rep_origin complement(5974..6429)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind 6574..6590
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 6597..6615
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"