pTT3-CD109-bio-His vector (V016587)

Price Information

Cat No. Plasmid Name Availability Add to cart
V016587 pTT3-CD109-bio-His 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:
pTT3-CD109-bio-His
Antibiotic Resistance:
Ampicillin
Length:
10907 bp
Type:
Protein expression
Replication origin:
ori
Host:
Mammalian cells
Promoter:
CMV
Growth Strain(s):
DH5a
Growth Temperature:
37℃

pTT3-CD109-bio-His vector Map

pTT3-CD109-bio-His10907 bp50010001500200025003000350040004500500055006000650070007500800085009000950010000105006xHisbeta-globin poly(A) signalAmpR promoterAmpRoriCAP binding sitelac promoterCMV enhancerCMV promoterCD109 antigen

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.

pTT3-CD109-bio-His vector Sequence

LOCUS       V016587                10907 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V016587
VERSION     V016587
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 10907)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..10907
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             644..661
                     /label="6xHis"
                     /note="6xHis affinity tag"
     polyA_signal    713..768
                     /label="beta-globin poly(A) signal"
                     /note="rabbit beta-globin polyadenylation signal (Gil and
                     Proudfoot, 1987)"
     promoter        3090..3194
                     /label="AmpR promoter"
     CDS             3195..4052
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      4226..4814
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     protein_bind    5102..5123
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        5138..5168
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     enhancer        5487..5866
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5867..6070
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     CDS             join(6631..10907,1..13)
                     /gene="CD109"
                     /label="CD109 antigen"
                     /note="CD109 antigen from Homo sapiens. Accession#: Q6YHK3"