Kamigaki A, Nito K, Hikino K, Goto-Yamada S, Nishimura M, Nakagawa T, Mano S.
pcRGW 载体载体图谱
pcRGW 载体载体序列
LOCUS Exported 6237 bp ds-DNA circular SYN 17-DEC-2018
DEFINITION Gateway vector pcRGW DNA, complete sequence.
ACCESSION AB830537
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6237)
AUTHORS Kamigaki A, Nito K, Hikino K, Goto-Yamada S, Nishimura M, Nakagawa
T, Mano S.
TITLE Gateway Vectors for Simultaneous Detection of Multiple
Protein-Protein Interactions in Plant Cells Using Bimolecular
Fluorescence Complementation
JOURNAL PLoS ONE 11 (8), E0160717 (2016)
PUBMED 27490375
REFERENCE 2 (bases 1 to 6237)
AUTHORS Mano S, Nakagawa T.
TITLE Direct Submission
JOURNAL Submitted (10-JUL-2013) Contact:Shoji Mano National Institute for
Basic Biology, Department of Cell Biology; 38 Nishigonaka, Myodaiji,
Okazaki, Aichi 444-8585, Japan
REFERENCE 3 (bases 1 to 6237)
AUTHORS .
TITLE Direct Submission
COMMENT These colnes were obtained at our laboratory.
FEATURES Location/Qualifiers
source 1..6237
/organism="Gateway vector pcRGW"
/mol_type="other DNA"
/db_xref="taxon:1356410"
protein_bind 107..128
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 143..173
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 181..197
/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 205..221
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
regulatory 258..1091
/regulatory_class="promoter"
/note="35S promoter"
promoter 745..1090
/label=CaMV 35S promoter
/note="strong constitutive promoter from cauliflower mosaic
virus"
misc_feature 1111..1353
/label=cmrfp1
/note="cmrfp1"
CDS 1114..1140
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
misc_feature complement(1363..1487)
/label=attR1
/note="attR1"
protein_bind 1363..1487
/gene="mutant version of attR"
/label=attR1
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
promoter 1524..1554
/label=lac UV5 promoter
/note="E. coli lac promoter with an ""up"" mutation"
CDS 1608..2267
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=cat
/protein_id="BAV44452.1"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
gene 1608..2267
/gene="cat"
/label=cat
CDS 2609..2914
/codon_start=1
/gene="ccdB"
/label=ccdB
/protein_id="BAV44453.1"
/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
gene 2609..2914
/gene="ccdB"
/label=ccdB
misc_feature 2955..3079
/label=attR2
/note="attR2"
protein_bind complement(2955..3079)
/gene="mutant version of attR"
/label=attR2
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
regulatory 3090..3365
/regulatory_class="terminator"
/note="NOS terminator"
terminator 3105..3357
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
primer_bind complement(3366..3382)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
rep_origin 3595..4050
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 4332..4436
/gene="bla"
/label=AmpR promoter
CDS 4437..5297
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=bla
/protein_id="BAV44454.1"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
gene 4437..5297
/gene="bla"
/label=bla
rep_origin complement(5458..6058)
/direction=LEFT
/label=colE1 ori
/note="colE1 ori"
rep_origin 5468..6056
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
Kamigaki A, Nito K, Hikino K, Goto-Yamada S, Nishimura M, Nakagawa T, Mano S.
pcRGW 载体载体图谱
pcRGW 载体载体序列
LOCUS Exported 6237 bp ds-DNA circular SYN 17-DEC-2018
DEFINITION Gateway vector pcRGW DNA, complete sequence.
ACCESSION AB830537
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6237)
AUTHORS Kamigaki A, Nito K, Hikino K, Goto-Yamada S, Nishimura M, Nakagawa
T, Mano S.
TITLE Gateway Vectors for Simultaneous Detection of Multiple
Protein-Protein Interactions in Plant Cells Using Bimolecular
Fluorescence Complementation
JOURNAL PLoS ONE 11 (8), E0160717 (2016)
PUBMED 27490375
REFERENCE 2 (bases 1 to 6237)
AUTHORS Mano S, Nakagawa T.
TITLE Direct Submission
JOURNAL Submitted (10-JUL-2013) Contact:Shoji Mano National Institute for
Basic Biology, Department of Cell Biology; 38 Nishigonaka, Myodaiji,
Okazaki, Aichi 444-8585, Japan
REFERENCE 3 (bases 1 to 6237)
AUTHORS .
TITLE Direct Submission
COMMENT These colnes were obtained at our laboratory.
FEATURES Location/Qualifiers
source 1..6237
/organism="Gateway vector pcRGW"
/mol_type="other DNA"
/db_xref="taxon:1356410"
protein_bind 107..128
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 143..173
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 181..197
/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 205..221
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
regulatory 258..1091
/regulatory_class="promoter"
/note="35S promoter"
promoter 745..1090
/label=CaMV 35S promoter
/note="strong constitutive promoter from cauliflower mosaic
virus"
misc_feature 1111..1353
/label=cmrfp1
/note="cmrfp1"
CDS 1114..1140
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
misc_feature complement(1363..1487)
/label=attR1
/note="attR1"
protein_bind 1363..1487
/gene="mutant version of attR"
/label=attR1
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
promoter 1524..1554
/label=lac UV5 promoter
/note="E. coli lac promoter with an ""up"" mutation"
CDS 1608..2267
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=cat
/protein_id="BAV44452.1"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
gene 1608..2267
/gene="cat"
/label=cat
CDS 2609..2914
/codon_start=1
/gene="ccdB"
/label=ccdB
/protein_id="BAV44453.1"
/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
gene 2609..2914
/gene="ccdB"
/label=ccdB
misc_feature 2955..3079
/label=attR2
/note="attR2"
protein_bind complement(2955..3079)
/gene="mutant version of attR"
/label=attR2
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
regulatory 3090..3365
/regulatory_class="terminator"
/note="NOS terminator"
terminator 3105..3357
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
primer_bind complement(3366..3382)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
rep_origin 3595..4050
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 4332..4436
/gene="bla"
/label=AmpR promoter
CDS 4437..5297
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=bla
/protein_id="BAV44454.1"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
gene 4437..5297
/gene="bla"
/label=bla
rep_origin complement(5458..6058)
/direction=LEFT
/label=colE1 ori
/note="colE1 ori"
rep_origin 5468..6056
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"