pCCLc-MND-B2705-SABR-Backbone vector (V016883)

Price Information

Cat No. Plasmid Name Availability Add to cart
V016883 pCCLc-MND-B2705-SABR-Backbone In stock, 1 week for quality controls

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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

pCCLc-MND-B2705-SABR-Backbone10900 bp5001000150020002500300035004000450050005500600065007000750080008500900095001000010500CAP binding sitelac promoterT3 promoterCMV enhancerCMV promoter5' LTR (truncated)HIV-1 PsiRREgp41 peptideProtein TatcPPT/CTSMNDU3hGH signal sequenceF2A3' LTR (Delta-U3)SV40 poly(A) signalSV40 oriT7 promoterf1 oriAmpR promoterAmpRori

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"