Basic Vector Information
L071 RRI-Green 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 ℃.
L071 RRI-Green vector Sequence
LOCUS 40924_1519 8254 bp DNA circular SYN 17-DEC-2018
DEFINITION Cloning vector L071 RRI-Green, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8254)
AUTHORS Micklem DR, Lorens JB.
TITLE Retroviral vectors for expression of shRNAs
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 8254)
AUTHORS Micklem DR, Lorens JB.
TITLE Direct Submission
JOURNAL Submitted (24-JAN-2008) Biomedicine, University of Bergen, Jonas
Lies vei 91, Bergen 5009, Norway
REFERENCE 3 (bases 1 to 8254)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 8254)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(24-JAN-2008) Biomedicine, University of Bergen, Jonas Lies vei 91,
Bergen 5009, Norway"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..8254
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 63..366
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 367..570
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
LTR 571..746
/label=5' LTR (truncated)
/note="truncated long terminal repeat from Moloney murine
sarcoma virus"
CDS 1240..1656
/label=gag (truncated)
/note="truncated Moloney murine leukemia virus (MMLV) gag
gene lacking the start codon"
misc_feature 1666..2044
/label=pol region
/note="Moloney murine leukemia virus (MMLV) pol region
containing the splice acceptor site"
polyA_signal complement(2063..2270)
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
misc_feature complement(2901..2985)
/label=FMDV 2A self-cleaving linker region
/note="FMDV 2A self-cleaving linker region"
CDS complement(2992..3708)
/label=EGFP
/note="enhanced GFP"
promoter complement(3734..4896)
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
promoter 5105..5346
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
misc_feature 5355..5745
/note="stuffer fragment; lost during cloning of shRNAs
between unique BglII and AarI sites"
misc_feature 5355
/label=U6 promoter-dependent transcription initiation
/note="U6 promoter-dependent transcription initiation"
misc_feature 5802..5869
/label=R region
/note="R region"
protein_bind complement(6055..6071)
/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(6079..6109)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(6124..6145)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(6433..7021)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(7198..8055)
/label=AmpR
/note="beta-lactamase"
promoter complement(8056..8160)
/label=AmpR promoter
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