Basic Vector Information
- Vector Name:
- pGWB616
- Antibiotic Resistance:
- Streptomycin
- Length:
- 10856 bp
- Type:
- Gateway binary vector
- Replication origin:
- ori
- Host:
- Plants
- Source/Author:
- Nakamura S, Mano S, Tanaka Y, Ohnishi M, Nakamori C, Araki M, Niwa T, Nishimura M, Kaminaka H, Nakagawa T, Sato Y, Ishiguro S.
- Promoter:
- NOS
pGWB616 vector Map
pGWB616 vector Sequence
LOCUS 40924_23459 10856 bp DNA circular SYN 18-DEC-2018
DEFINITION Gateway binary vector pGWB616 DNA, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10856)
AUTHORS Nakamura S, Mano S, Tanaka Y, Ohnishi M, Nakamori C, Araki M, Niwa
T, Nishimura M, Kaminaka H, Nakagawa T, Sato Y, Ishiguro S.
TITLE Gateway binary vectors with the bialaphos resistance gene, bar, as a
selection marker for plant transformation
JOURNAL Biosci. Biotechnol. Biochem. 74 (6), 1315-1319 (2010)
PUBMED 20530878
REFERENCE 2 (bases 1 to 10856)
AUTHORS Tanaka Y, Nakamura S, Ishiguro 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 10856)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 10856)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Biosci.
Biotechnol. Biochem."; date: "2010"; volume: "74"; issue: "6";
pages: "1315-1319"
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..10856
/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 2065..2094
/codon_start=1
/label=Myc
/note="Myc (human c-Myc proto-oncogene) epitope tag"
/translation="EQKLISEEDL"
CDS 2101..2130
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 2146..2175
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 2182..2211
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
terminator 2242..2494
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
primer_bind complement(2528..2544)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 2552..2568
/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(2576..2606)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(2621..2642)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
terminator complement(2678..2930)
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
CDS complement(3093..3641)
/codon_start=1
/label=BlpR
/note="phosphinothricin acetyltransferase"
/translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
WQLDFSLPVPPRPVLPVTEI"
promoter complement(3693..3872)
/label=NOS promoter
/note="nopaline synthase promoter"
misc_feature complement(4378..4402)
/label=LB T-DNA repeat
/note="left border repeat from nopaline C58 T-DNA"
CDS 4923..5711
/codon_start=1
/label=SmR
/note="aminoglycoside adenylyltransferase (Murphy, 1985)"
/translation="MGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
rep_origin 5960..6548
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
misc_feature complement(6734..6874)
/label=bom
/note="basis of mobility region from pBR322"
rep_origin complement(7218..7412)
/direction=LEFT
/label=pVS1 oriV
/note="origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
CDS complement(7481..8551)
/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(8983..9609)
/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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