Basic Vector Information
- Vector Name:
- pEGP2×35S-nYFP-NOS
- Antibiotic Resistance:
- Kanamycin
- Length:
- 10629 bp
- Type:
- Bioluminescence resonance energy transfer (BRET)
- Replication origin:
- ori
- Host:
- Plants
- Selection Marker:
- Hyg
- Promoter:
- CaMV 35S (enhanced)
- Growth Strain(s):
- DH5a
- Growth Temperature:
- 37℃
pEGP2×35S-nYFP-NOS 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 ℃.
pEGP2×35S-nYFP-NOS vector Sequence
LOCUS 62056_9115 10629 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10629) AUTHORS . TITLE Direct Submission FEATURES Location/Qualifiers source 1..10629 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 1..678 /label=CaMV 35S promoter (enhanced) /note="cauliflower mosaic virus 35S promoter with a duplicated enhancer region" CDS 746..1768 /codon_start=1 /label=HygR /note="aminoglycoside phosphotransferase from E. coli" /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK K" polyA_signal 1811..1985 /label=CaMV poly(A) signal /note="cauliflower mosaic virus polyadenylation signal" misc_feature complement(2063..2087) /label=LB T-DNA repeat /note="left border repeat from nopaline C58 T-DNA" CDS 2512..3303 /codon_start=1 /label=KanR /note="aminoglycoside phosphotransferase" /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF" rep_origin 3393..3981 /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" misc_feature complement(4167..4307) /label=bom /note="basis of mobility region from pBR322" rep_origin complement(4651..4845) /direction=LEFT /label=pVS1 oriV /note="origin of replication for the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" CDS complement(4910..5974) /codon_start=1 /label=pVS1 RepA /note="replication protein from the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" /translation="GRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESWQA AADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLSKR DRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKPGR VFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGEAL ISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYRLA RRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPILV MRYRNLIEGEASAGS" CDS complement(6411..7037) /codon_start=1 /label=pVS1 StaA /note="stability protein from the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI" misc_feature complement(8337..8361) /label=RB T-DNA repeat /note="right border repeat from nopaline C58 T-DNA" promoter 9008..9352 /label=CaMV 35S promoter /note="strong constitutive promoter from cauliflower mosaic virus" CDS 9494..10012 /codon_start=1 /label=VN173 /note="N-terminal fragment of mVenus for use in bimolecular fluorescence complementation (BiFC) (Kodama and Hu, 2010)" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIE" terminator 10029..10281 /label=NOS terminator /note="nopaline synthase terminator and poly(A) signal" promoter complement(10373..10403) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind complement(10418..10439) /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP."
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