Basic Vector Information
pnCGW 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 ℃.
pnCGW vector Sequence
LOCUS 40924_33073 6516 bp DNA circular SYN 18-DEC-2018 DEFINITION Gateway vector pnCGW DNA, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6516) 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 6516) 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 6516) TITLE Direct Submission REFERENCE 4 (bases 1 to 6516) 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..6516 /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" CDS 1111..1629 /codon_start=1 /label=VN173 /note="N-terminal fragment of mVenus for use in bimolecular fluorescence complementation (BiFC) (Kodama and Hu, 2010)" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK ANFKIRHNIE" protein_bind 1642..1766 /label=attR1 /note="recombination site for the Gateway(R) LR reaction" promoter 1803..1833 /label=lac UV5 promoter /note="E. coli lac promoter with an 'up' mutation" CDS 1887..2543 /codon_start=1 /label=CmR /note="chloramphenicol acetyltransferase" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" CDS 2888..3190 /codon_start=1 /label=ccdB /note="CcdB, a bacterial toxin that poisons DNA gyrase" /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI" protein_bind complement(3234..3358) /label=attR2 /note="recombination site for the Gateway(R) LR reaction" terminator 3384..3636 /label=NOS terminator /note="nopaline synthase terminator and poly(A) signal" primer_bind complement(3645..3661) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" rep_origin 3874..4329 /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" promoter 4611..4715 /label=AmpR promoter CDS 4716..5573 /codon_start=1 /label=AmpR /note="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" rep_origin 5747..6335 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication"
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