Basic Vector Information
- Vector Name:
- pLCN DSB Repair Reporter (DRR)
- Antibiotic Resistance:
- Ampicillin
- Length:
- 8444 bp
- Type:
- Lentiviral
- Replication origin:
- ori
- Selection Marker:
- Neomycin (select with G418)
- Copy Number:
- High Copy
- Promoter:
- RSV
- 5' Primer:
- CGC AAA TGG GCG GTA GGC GTG
pLCN DSB Repair Reporter (DRR) 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 ℃.
pLCN DSB Repair Reporter (DRR) vector Sequence
LOCUS pLCN_DSB_Repair 8444 bp DNA circular SYN 13-MAY-2021
DEFINITION DSB Repair Reporter (DRR). Expresses GFP after cut of two inverted
ISce1 sites and repair by NHEJ..
ACCESSION .
VERSION .
KEYWORDS pLCN DSB Repair Reporter (DRR).
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8444)
AUTHORS Arnoult N, Correia A, Ma J, Merlo A, Garcia-Gomez S, Maric M,
Tognetti M, Benner CW, Boulton SJ, Saghatelian A, Karlseder J
TITLE Regulation of DNA repair pathway choice in S and G2 phases by the
NHEJ inhibitor CYREN.
JOURNAL Nature. 2017 Sep 20;549(7673):548-552. doi: 10.1038/nature24023.
PUBMED 28959974
REFERENCE 2 (bases 1 to 8444)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 8444)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; doi: "10"; journalName:
"Nature"; date: "2017-09-20- 20"; volume: "549"; issue: "7673";
pages: "548-552"
COMMENT SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..8444
/mol_type="other DNA"
/organism="synthetic DNA construct"
protein_bind 1..22
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 37..67
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 75..91
/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)."
primer_bind 99..115
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
promoter 136..154
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 182..408
/label=RSV promoter
/note="Rous sarcoma virus enhancer/promoter"
LTR 409..589
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 636..761
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1254..1487
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 1672..1716
/label=gp41 peptide
/note="antigenic peptide corresponding to amino acids 655
to 669 of the HIV envelope protein gp41 (Lutje Hulsik et
al., 2013)"
CDS 1865..1906
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
misc_feature 1983..2099
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
enhancer 2122..2425
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 2426..2629
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 2626..2650
/label=LNCX
/note="Human CMV promoter, forward primer"
CDS 2687..3487
/label=NeoR/KanR
/note="aminoglycoside phosphotransferase from Tn5"
CDS 3488..3541
/label=T2A
/note="2A peptide from Thosea asigna virus capsid protein"
CDS 3584..3610
/label=HA
/note="HA (human influenza hemagglutinin) epitope tag"
CDS 3630..4346
/label=EGFP
/note="enhanced GFP"
misc_feature 4458..5046
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
LTR 5144..5377
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 5449..5583
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
rep_origin 5610..5745
/label=SV40 ori
/note="SV40 origin of replication"
promoter complement(5766..5784)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(5794..5810)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
rep_origin 5952..6407
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 6433..6537
/label=AmpR promoter
CDS 6538..7395
/label=AmpR
/note="beta-lactamase"
rep_origin 7569..8157
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 8311..8328
/label=L4440
/note="L4440 vector, forward primer"
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