pMac-L vector (V013013)

Price Information

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V013013 pMac-L 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:
pMac-L
Antibiotic Resistance:
Ampicillin
Length:
7934 bp
Type:
Protein expression, Antibody expression
Replication origin:
ori
Host:
Mammalian cells
Promoter:
CMV
Growth Strain(s):
DH5a
Growth Temperature:
37℃

pMac-L vector Map

pMac-L7934 bp30060090012001500180021002400270030003300360039004200450048005100540057006000630066006900720075007800SV40 poly(A) signaloriAmpRAmpR promoterT7Te terminatorSV40 promoterGlutamine synthetasesmall t intronSV40 NLSSV40 poly(A) signalCMV enhancerCMV promoterCMV intron AhIg-kappa-CLpoly(A) signal

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.

pMac-L vector Sequence

LOCUS       V013013                 7934 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V013013
VERSION     V013013
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 7934)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7934
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    110..231
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(500..1088)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(1262..2119)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(2120..2224)
                     /label="AmpR promoter"
     terminator      2393..2420
                     /label="T7Te terminator"
                     /note="phage T7 early transcription terminator"
     promoter        2505..2834
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     CDS             2873..3991
                     /gene="GLUL"
                     /label="Glutamine synthetase"
                     /note="Glutamine synthetase from Acomys cahirinus.
                     Accession#: Q9QY94"
     intron          4126..4191
                     /label="small t intron"
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             4321..4341
                     /label="SV40 NLS"
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
     polyA_signal    4766..4900
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     enhancer        5471..5850
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5851..6054
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     intron          6190..7016
                     /label="CMV intron A"
                     /note="human cytomegalovirus intron A (Chapman et al.,
                     1991)"
     CDS             7182..7499
                     /label="hIg-kappa-CL"
                     /note="Human immunoglobulin kappa light chain constant
                     region"
     polyA_signal    7712..7760
                     /label="poly(A) signal"
                     /note="synthetic polyadenylation signal"