Basic Vector Information
- Vector Name:
- pGWB528
- Antibiotic Resistance:
- Streptomycin
- Length:
- 11696 bp
- Type:
- Gateway binary vector
- Replication origin:
- ori
- Source/Author:
- 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.
pGWB528 vector Map
pGWB528 vector Sequence
LOCUS 40924_23244 11696 bp DNA circular SYN 18-DEC-2018
DEFINITION Gateway binary vector pGWB528 DNA, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 11696)
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 11696)
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 11696)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 11696)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Biosci.
Biotechnol. Biochem. (2007) In press"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(21-FEB-2007) Tsuyoshi Nakagawa, Shimane University, Center for
Integrated Research in Science"; volume: " 1060 Nishikawatsu,
Matsue, Shimane 690-8504, Japan
(E-mail:tnakagaw@life.shimane-u.ac.jp, Tel:81-852-32-6595, Fax";
pages: "81-852-32-6109"
COMMENT SGRef: number: 3; type: "Journal Article"
COMMENT constructed using pPZP221.
FEATURES Location/Qualifiers
source 1..11696
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_feature complement(54..78)
/label=RB T-DNA repeat
/note="right border repeat from nopaline C58 T-DNA"
primer_bind 281..297
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 323..447
/label=attR1
/note="recombination site for the Gateway(R) LR reaction"
promoter 484..514
/label=lac UV5 promoter
/note="E. coli lac promoter with an 'up' mutation"
CDS 568..1224
/codon_start=1
/label=CmR
/note="chloramphenicol acetyltransferase"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
CDS 1569..1871
/codon_start=1
/label=ccdB
/note="CcdB, a bacterial toxin that poisons DNA gyrase"
/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
protein_bind complement(1915..2039)
/label=attR2
/note="recombination site for the Gateway(R) LR reaction"
CDS 2050..2127
/codon_start=1
/label=CBP
/note="calmodulin-binding peptide"
/translation="KRRWKKNFIAVSAANRFKKISSSGAL"
CDS 2155..2175
/codon_start=1
/label=TEV site
/note="tobacco etch virus (TEV) protease recognition and
cleavage site"
/translation="ENLYFQG"
CDS 2215..2388
/codon_start=1
/label=ProtA
/note="IgG-binding unit of Staphylococcus aureus protein A"
/translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
AEAKKLNDAQAPK"
CDS 2392..2562
/codon_start=1
/product="IgG-binding unit of Staphylococcus aureus protein
A"
/label=ProtA
/translation="DNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLLA
EAKKLNGAQAPK"
terminator 2617..2869
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
primer_bind complement(2903..2919)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 2927..2943
/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(2951..2981)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(2996..3017)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
terminator complement(3053..3305)
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
CDS complement(3459..4481)
/codon_start=1
/label=HygR
/note="aminoglycoside phosphotransferase from E. coli"
/translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGAFSESLTYCISRRAQGVTLQDLP
ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
E"
promoter complement(4533..4712)
/label=NOS promoter
/note="nopaline synthase promoter"
misc_feature complement(5218..5242)
/label=LB T-DNA repeat
/note="left border repeat from nopaline C58 T-DNA"
CDS 5763..6551
/codon_start=1
/label=SmR
/note="aminoglycoside adenylyltransferase (Murphy, 1985)"
/translation="MGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
rep_origin 6800..7388
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
misc_feature complement(7574..7714)
/label=bom
/note="basis of mobility region from pBR322"
rep_origin complement(8058..8252)
/direction=LEFT
/label=pVS1 oriV
/note="origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
CDS complement(8321..9391)
/codon_start=1
/label=pVS1 RepA
/note="replication protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
/translation="VSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
LVMRYRNLIEGEASAGS"
CDS complement(9823..10449)
/codon_start=1
/label=pVS1 StaA
/note="stability protein from the Pseudomonas plasmid pVS1
(Heeb et al., 2000)"
/translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
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