Basic Vector Information
- Vector Name:
- pGWB459
- Antibiotic Resistance:
- Streptomycin
- Length:
- 11558 bp
- Type:
- Gateway binary vector
- Replication origin:
- ori
- Host:
- Plants
- Source/Author:
- Tanaka Y, Nakamura S, Nakagawa T.
- Promoter:
- NOS
pGWB459 vector Map
pGWB459 vector Sequence
LOCUS 40924_23049 11558 bp DNA circular SYN 18-DEC-2018
DEFINITION Gateway binary vector pGWB459 DNA, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 11558)
AUTHORS Tanaka Y, Nakamura S, Nakagawa T.
TITLE Development of the tagRFP Gateway Binary Vectors
JOURNAL Published Only in Database (2010)
REFERENCE 2 (bases 1 to 11558)
AUTHORS Tanaka Y, Nakamura S, Nakagawa T.
TITLE Direct Submission
JOURNAL Submitted (22-JAN-2010) Contact:Tsuyoshi Nakagawa Shimane
University, Center for Integrated Research in Science; 1060
Nishikawatsu, Matsue, Shimane 690-8504, Japan
REFERENCE 3 (bases 1 to 11558)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 11558)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Published
Only in Database (2010)"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(22-JAN-2010) Contact:Tsuyoshi Nakagawa Shimane University, Center
for Integrated Research in Science; 1060 Nishikawatsu, Matsue,
Shimane 690-8504, Japan"
COMMENT SGRef: number: 3; type: "Journal Article"
COMMENT constructed using pPZP221.
FEATURES Location/Qualifiers
source 1..11558
/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 338..462
/label=attR1
/note="recombination site for the Gateway(R) LR reaction"
promoter 499..529
/label=lac UV5 promoter
/note="E. coli lac promoter with an 'up' mutation"
CDS 583..1239
/codon_start=1
/label=CmR
/note="chloramphenicol acetyltransferase"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
CDS 1584..1886
/codon_start=1
/label=ccdB
/note="CcdB, a bacterial toxin that poisons DNA gyrase"
/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
protein_bind complement(1930..2054)
/label=attR2
/note="recombination site for the Gateway(R) LR reaction"
CDS 2065..2775
/codon_start=1
/label=TagRFP
/note="monomeric derivative of red fluorescent protein from
Entacmaea quadricolor (Merzlyak et al., 2007)"
/translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
RIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRSD
MALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAV
ARYCDLPSKLGHKLN"
terminator 2800..3052
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
primer_bind complement(3086..3102)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 3110..3126
/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(3134..3164)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(3179..3200)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
terminator complement(3234..3486)
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
CDS complement(3879..4667)
/codon_start=1
/label=NeoR/KanR
/note="aminoglycoside phosphotransferase from Tn5"
/translation="IEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRPV
LFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLSS
HLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQG
LAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIAL
ATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
promoter complement(4691..4874)
/label=NOS promoter
/note="nopaline synthase promoter"
misc_feature complement(5080..5104)
/label=LB T-DNA repeat
/note="left border repeat from nopaline C58 T-DNA"
CDS 5625..6413
/codon_start=1
/label=SmR
/note="aminoglycoside adenylyltransferase (Murphy, 1985)"
/translation="MGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
rep_origin 6662..7250
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
misc_feature complement(7436..7576)
/label=bom
/note="basis of mobility region from pBR322"
rep_origin complement(7920..8114)
/direction=LEFT
/label=pVS1 oriV
/note="origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
CDS complement(8183..9253)
/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(9685..10311)
/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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