| 载体基本信息 |
| 载体名称 |
pGWB405 |
载体抗性 |
Spectinomycin/Streptomycin |
| 载体长度 |
12414 bp |
载体类型 |
Gateway binary vector |
| 载体来源 |
Nakagawa T, Suzuki T, Murata S, Nakamura S, Hino T, Maeo K, Tabata R, Kawai T, Tanaka K, Niwa Y, Watanabe Y, Nakamura K, Kimura T, Ishiguro S. |
pGWB405 载体载体图谱
pGWB405 载体载体序列
LOCUS Exported 12414 bp ds-DNA circular SYN 18-DEC-2018
DEFINITION Gateway binary vector pGWB405 DNA, complete sequence.
ACCESSION AB294429
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 12414)
AUTHORS Nakagawa T, Suzuki T, Murata S, Nakamura S, Hino T, Maeo K, Tabata
R, Kawai T, Tanaka K, Niwa Y, Watanabe Y, Nakamura K, Kimura T,
Ishiguro S.
TITLE Improved Gateway Binary Vectors: High-Performance Vectors for
Creation of Fusion Constructs in Transgenic Analysis of Plants
JOURNAL Biosci. Biotechnol. Biochem. (2007) In press
PUBMED 17690442
REFERENCE 2 (bases 1 to 12414)
AUTHORS Nakagawa T, Ishiguro S.
TITLE Direct Submission
JOURNAL Submitted (21-FEB-2007) Tsuyoshi Nakagawa, Shimane University,
Center for Integrated Research in Science; 1060 Nishikawatsu,
Matsue, Shimane 690-8504, Japan
(E-mail:tnakagaw@life.shimane-u.ac.jp, Tel:81-852-32-6595,
Fax:81-852-32-6109)
REFERENCE 3 (bases 1 to 12414)
AUTHORS .
TITLE Direct Submission
COMMENT constructed using pPZP221.
FEATURES Location/Qualifiers
source 1..12414
/organism="Gateway binary vector pGWB405"
/mol_type="other DNA"
/db_xref="taxon:426956"
misc_feature complement(54..78)
/label=T-DNA right border
/note="T-DNA right border"
primer_bind 281..297
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
regulatory 325..1159
/regulatory_class="promoter"
/note="35S promoter"
promoter 812..1157
/label=CaMV 35S promoter
/note="strong constitutive promoter from cauliflower mosaic
virus"
misc_feature complement(1179..1303)
/label=attR1
/note="attR1"
protein_bind 1179..1303
/gene="mutant version of attR"
/label=attR1
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
promoter 1340..1370
/label=lac UV5 promoter
/note="E. coli lac promoter with an ""up"" mutation"
CDS 1424..2083
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=cat
/protein_id="BAF47429.1"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
gene 1424..2083
/gene="cat"
/label=cat
CDS 2425..2730
/codon_start=1
/gene="ccdB"
/label=ccdB
/protein_id="BAF47430.1"
/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
VPRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
gene 2425..2730
/gene="ccdB"
/label=ccdB
misc_feature 2771..2895
/label=attR2
/note="attR2"
protein_bind complement(2771..2895)
/gene="mutant version of attR"
/label=attR2
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
CDS 2912..3631
/codon_start=1
/gene="gfp"
/product="synthetic green fluorescent protein"
/label=gfp
/protein_id="BAF47431.1"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITHGMDELYK"
gene 2912..3631
/gene="gfp"
/label=gfp
regulatory 3640..4049
/regulatory_class="terminator"
/note="NOS terminator"
terminator 3656..3908
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
primer_bind complement(3942..3958)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 3966..3982
/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(3990..4020)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4035..4056
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
regulatory complement(4083..4342)
/regulatory_class="terminator"
/note="NOS terminator"
terminator 4090..4342
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
CDS complement(4732..5526)
/codon_start=1
/gene="nptII"
/product="neomycin phosphotransferase II"
/label=nptII
/protein_id="BAF47432.1"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
gene complement(4732..5526)
/gene="nptII"
/label=nptII
regulatory complement(5539..5845)
/regulatory_class="promoter"
/note="NOS promoter"
promoter complement(5547..5730)
/function="NOS promoter"
/label=NOS promoter
/note="nopaline synthase promoter"
promoter complement(5583..5762)
/label=NOS promoter
/note="nopaline synthase promoter"
misc_feature complement(5936..5960)
/label=T-DNA left border
/note="T-DNA left border"
CDS 6481..7272
/codon_start=1
/gene="aadA"
/product="aminoglycoside adenylyltransferase (Murphy,
1985)"
/label=SmR
/note="confers resistance to spectinomycin and
streptomycin"
/translation="MGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
rep_origin 7518..8106
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
misc_feature 8292..8432
/label=bom
/note="basis of mobility region from pBR322"
rep_origin 8776..8970
/label=pVS1 oriV
/note="origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
CDS complement(9036..10109)
/codon_start=1
/product="replication protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
/label=pVS1 RepA
/translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
LVMRYRNLIEGEASAGS"
CDS complement(10538..11167)
/codon_start=1
/product="stability protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
/label=pVS1 StaA
/translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"