Basic Vector Information
35SCoreIN-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 ℃.
35SCoreIN-pFLEV vector Sequence
LOCUS 40924_50 4027 bp DNA circular SYN 17-DEC-2018
DEFINITION Cloning vector 35SCoreIN-pFLEV, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4027)
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 4027)
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 4027)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 4027)
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..4027
/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 447..493
/label=minimal CaMV 35S promoter
/note="minimal 35S promoter from cauliflower mosaic virus"
CDS 747..1463
/codon_start=1
/label=EGFP
/note="enhanced GFP"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITHGMDELYK"
terminator 1499..1751
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
primer_bind complement(1806..1822)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(1830..1846)
/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(1854..1884)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(1899..1920)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(2208..2796)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(2970..3827)
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(3828..3932)
/label=AmpR promoter
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