Basic Vector Information
- Vector Name:
- pEGFPpA-ACN
- Antibiotic Resistance:
- Ampicillin
- Length:
- 7904 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Wu S, Ying G, Wu Q, Capecchi MR.
- Promoter:
- T7
pEGFPpA-ACN 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 ℃.
pEGFPpA-ACN vector Sequence
LOCUS 40924_17259 7904 bp DNA circular SYN 17-DEC-2018
DEFINITION Cloning vector pEGFPpA-ACN, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7904)
AUTHORS Wu S, Ying G, Wu Q, Capecchi MR.
TITLE A protocol for constructing gene targeting vectors: generating
knockout mice for the cadherin family and beyond
JOURNAL Nat Protoc 3 (6), 1056-1076 (2008)
PUBMED 18546598
REFERENCE 2 (bases 1 to 7904)
AUTHORS Wu S, Capecchi MR.
TITLE Direct Submission
JOURNAL Submitted (29-FEB-2008) Howard Hughes Medical Institute, University
of Utah, 15N 2030E, Salt Lake City, UT 84112, USA
REFERENCE 3 (bases 1 to 7904)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 7904)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat
Protoc"; date: "2008"; volume: "3"; issue: "6"; pages: "1056-1076"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(29-FEB-2008) Howard Hughes Medical Institute, University of Utah,
15N 2030E, Salt Lake City, UT 84112, USA"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..7904
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 1..19
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
CDS 44..64
/label=SV40 NLS
/note="nuclear localization signal of SV40 (simian virus
40) large T antigen"
CDS 62..775
/label=hrGFP
/note="humanized Renilla green fluorescent protein"
CDS 813..1526
/label=EGFP
/note="enhanced GFP"
polyA_signal 1652..1773
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
protein_bind 1799..1832
/label=loxP
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
polyA_signal 1859..1907
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
regulatory complement(1859..1906)
/label=HSV thymidine kinase polyA signal
/note="HSV thymidine kinase polyA signal"
/regulatory_class="polyA_signal_sequence"
CDS complement(1917..2717)
/label=NeoR/KanR
/note="aminoglycoside phosphotransferase from Tn5"
regulatory complement(2734..3440)
/label=RNA polymerase II large subunit promoter
/note="RNA polymerase II large subunit promoter"
/regulatory_class="promoter"
polyA_signal complement(3456..3504)
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
intron complement(3854..3919)
/label=small t intron
/note="SV40 (simian virus 40) small t antigen intron"
CDS complement(4745..4765)
/label=SV40 NLS
/note="nuclear localization signal of SV40 (simian virus
40) large T antigen"
CDS complement(4789..4800)
/label=WELQut site
/note="WELQut protease recognition and cleavage site"
misc_recomb 5475..5508
/label=loxP site
/note="loxP site"
protein_bind complement(5475..5508)
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
promoter complement(5559..5577)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
rep_origin complement(5835..6423)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(6597..7454)
/label=AmpR
/note="beta-lactamase"
promoter complement(7455..7559)
/label=AmpR promoter
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