Price Information
Cat No. | Plasmid Name | Availability | Add to cart |
---|---|---|---|
V016883 | pCCLc-MND-B2705-SABR-Backbone | In stock, 1 week for quality controls |
Buy one, get one free! |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
- Vector Name:
- pCCLc-MND-B2705-SABR-Backbone
- Antibiotic Resistance:
- Ampicillin
- Length:
- 10900 bp
- Type:
- Protein expression
- Replication origin:
- ori
- Host:
- Mammalian cells, Lentivirus
- Growth Strain(s):
- Stbl3
- Growth Temperature:
- 37℃
pCCLc-MND-B2705-SABR-Backbone vector Map
Plasmid 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 ℃.
6. The concentration of plasmid re-measurement sometimes differs from the nominal value, which may be due to the position of the lyophilized plasmid in the tube, the efficiency of the re-dissolution, the measurement bias, and adsorption on the wall of the tube, therefore, it is recommended to transform and extract the plasmid before using it
General Plasmid Transform Protocol
1. Take one 100μl of the competent cells and thaw it on ice for 10min, add 2μl of plasmid, then ice bath for 30min, then heat-shock it at 42℃ for 60s, do not stir, and then ice bath for 2min.
2. Add 900μl of LB liquid medium without antibiotics, and incubate at 37℃ for 45min (30℃ for 1-1.5 hours) with 180rpm shaking.
3. Centrifuge at 6000rpm for 5min, leave only 100μl of supernatant to resuspend the bacterial precipitate and spread it onto the target plasmid-resistant LB plate.
4. Invert the plate and incubate at 37℃ for 14h, or at 30℃ for 20h.
5. Pick a single colony into LB liquid medium, add the corresponding antibiotics, incubate at 220rpm for 14h, and extract the plasmid according to the experimental needs and the instructions of the plasmid extraction kit.
pCCLc-MND-B2705-SABR-Backbone vector Sequence
LOCUS V016883 10900 bp DNA circular SYN 01-JAN-1980 DEFINITION Exported. ACCESSION V016883 VERSION V016883 KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct . REFERENCE 1 (bases 1 to 10900) AUTHORS . TITLE Direct Submission FEATURES Location/Qualifiers source 1..10900 /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" promoter 136..154 /label="T3 promoter" /note="promoter for bacteriophage T3 RNA polymerase" enhancer 255..634 /label="CMV enhancer" /note="human cytomegalovirus immediate early enhancer" promoter 635..838 /label="CMV promoter" /note="human cytomegalovirus (CMV) immediate early promoter" LTR 853..1033 /label="5' LTR (truncated)" /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 1080..1205 /label="HIV-1 Psi" /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1698..1931 /label="RRE" /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 2116..2160 /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 2309..2350 /note="Protein Tat from Human immunodeficiency virus type 1 group M subtype B (isolate WMJ22). Accession#: P12509" /label="Protein Tat" misc_feature 2457..2574 /label="cPPT/CTS" /note="central polypurine tract and central termination sequence of HIV-1" promoter 2620..3157 /label="MNDU3" /note="The U3 region from the myeloproliferative sarcoma virus (MPSV) LTR with the negative control region (NCR) removed" sig_peptide 3177..3254 /label="hGH signal sequence" /note="human growth hormone signal sequence" CDS 7038..7103 /label="F2A" /note="2A peptide from foot-and-mouth disease virus polyprotein" LTR 7600..7833 /label="3' LTR (Delta-U3)" /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 7905..8039 /label="SV40 poly(A) signal" /note="SV40 polyadenylation signal" rep_origin 8066..8201 /label="SV40 ori" /note="SV40 origin of replication" promoter complement(8222..8240) /label="T7 promoter" /note="promoter for bacteriophage T7 RNA polymerase" rep_origin 8408..8863 /label="f1 ori" /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" promoter 8889..8993 /label="AmpR promoter" CDS 8994..9851 /label="AmpR" /note="beta-lactamase" rep_origin 10025..10613 /direction=RIGHT /label="ori" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication"