Basic Vector Information
pnRGW vector Vector Map
Plasmid Resuspension Protocol:
1. Centrifuge at 5,000×g for 5 min.
2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.
3. Close the tube and incubate for 10 minutes at room temperature.
4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.
5.Store the plasmid at -20 ℃.
pnRGW vector Sequence
LOCUS 40924_33497 6489 bp DNA circular SYN 18-DEC-2018
DEFINITION Gateway vector pnRGW DNA, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6489)
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 6489)
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 6489)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 6489)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "PLoS ONE";
date: "2016"; volume: "11"; issue: "8"; pages: "E0160717"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(10-JUL-2013) Contact:Shoji Mano National Institute for Basic
Biology, Department of Cell Biology; 38 Nishigonaka, Myodaiji,
Okazaki, Aichi 444-8585, Japan"
COMMENT SGRef: number: 3; type: "Journal Article"
COMMENT These colnes were obtained at our laboratory.
FEATURES Location/Qualifiers
source 1..6489
/mol_type="other DNA"
/organism="synthetic DNA construct"
protein_bind 107..128
/label=CAP binding site
/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
/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"
promoter 745..1090
/label=CaMV 35S promoter
/note="strong constitutive promoter from cauliflower mosaic
virus"
misc_feature 1111..1605
/label=nmrfp1
/note="nmrfp1"
CDS 1114..1143
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
protein_bind 1615..1739
/label=attR1
/note="recombination site for the Gateway(R) LR reaction"
promoter 1776..1806
/label=lac UV5 promoter
/note="E. coli lac promoter with an 'up' mutation"
CDS 1860..2516
/codon_start=1
/label=CmR
/note="chloramphenicol acetyltransferase"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
CDS 2861..3163
/codon_start=1
/label=ccdB
/note="CcdB, a bacterial toxin that poisons DNA gyrase"
/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
protein_bind complement(3207..3331)
/label=attR2
/note="recombination site for the Gateway(R) LR reaction"
terminator 3357..3609
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
primer_bind complement(3618..3634)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
rep_origin 3847..4302
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 4584..4688
/label=AmpR promoter
CDS 4689..5546
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 5720..6308
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
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