Basic Vector Information
35SCoreTIF1-pFLEV 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 ℃.
35SCoreTIF1-pFLEV vector Sequence
LOCUS 40924_60 3964 bp DNA circular SYN 17-DEC-2018 DEFINITION Cloning vector 35SCoreTIF1-pFLEV, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3964) AUTHORS Grant TN, De La Torre CM, Zhang N, Finer JJ. TITLE Synthetic introns help identify sequences in the 5' UTR intron of the Glycine max polyubiquitin (Gmubi) promoter that give increased promoter activity JOURNAL Planta (2017) In press PUBMED 28070655 REFERENCE 2 (bases 1 to 3964) AUTHORS Finer J, Grant T, De La Torre C, Zhang N. TITLE Direct Submission JOURNAL Submitted (02-SEP-2016) Horticulture and Crop Science, Ohio State University, 1680 Madison Ave, Wooster, OH 44691, USA REFERENCE 3 (bases 1 to 3964) TITLE Direct Submission REFERENCE 4 (bases 1 to 3964) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Planta (2017) In press" COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted (02-SEP-2016) Horticulture and Crop Science, Ohio State University, 1680 Madison Ave, Wooster, OH 44691, USA" COMMENT SGRef: number: 3; type: "Journal Article" COMMENT ##Assembly-Data-START## Sequencing Technology :: Sanger dideoxy sequencing ##Assembly-Data-END## FEATURES Location/Qualifiers source 1..3964 /mol_type="other DNA" /organism="synthetic DNA construct" primer_bind 379..395 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" promoter 619..665 /label=minimal CaMV 35S promoter /note="minimal 35S promoter from cauliflower mosaic virus" CDS 684..1400 /codon_start=1 /label=EGFP /note="enhanced GFP" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITHGMDELYK" terminator 1436..1688 /label=NOS terminator /note="nopaline synthase terminator and poly(A) signal" primer_bind complement(1743..1759) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind complement(1767..1783) /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)." promoter complement(1791..1821) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind complement(1836..1857) /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(2145..2733) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(2907..3764) /codon_start=1 /label=AmpR /note="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(3765..3869) /label=AmpR promoter
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