Basic Vector Information
- Vector Name:
- pLV3-U6-MCS-shRNA
- Antibiotic Resistance:
- Ampicillin
- Length:
- 7983 bp
- Type:
- RNA interference
- Replication origin:
- ori
- Host:
- Mammalian cells, Lentivirus
- Promoter:
- RSV
- Growth Temperature:
- 37℃
pLV3-U6-MCS-shRNA 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 ℃.
pLV3-U6-MCS-shRNA vector Sequence
LOCUS 62056_15730 7983 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7983) AUTHORS . TITLE Direct Submission FEATURES Location/Qualifiers source 1..7983 /mol_type="other DNA" /organism="synthetic DNA construct" LTR 338..571 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 643..777 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" rep_origin 804..939 /label=SV40 ori /note="SV40 origin of replication" promoter complement(960..978) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(988..1004) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" rep_origin 1146..1601 /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" promoter 1627..1731 /label=AmpR promoter CDS 1732..2589 /label=AmpR /note="beta-lactamase" rep_origin 2763..3351 /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" protein_bind 3639..3660 /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." promoter 3675..3705 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3713..3729 /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 3737..3753 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" promoter 3774..3792 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" promoter 3820..4046 /label=RSV promoter /note="Rous sarcoma virus enhancer/promoter" LTR 4047..4227 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 4274..4399 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 4892..5125 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 5310..5354 /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 5503..5544 /note="Protein Tat from Human immunodeficiency virus type 1 group M subtype B (isolate WMJ22). Accession#: P12509" promoter 5652..5892 /label=U6 promoter /note="RNA polymerase III promoter for human U6 snRNA" misc_feature 7814..7931 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1"
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