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pcDNA-GalNAc-T2-GFP-ESCargo(FTV) vector (V017052)

Price Information

Cat No. Plasmid Name Availability Add to cart
V017052 pcDNA-GalNAc-T2-GFP-ESCargo(FTV) 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:
pcDNA-GalNAc-T2-GFP-ESCargo(FTV)
Antibiotic Resistance:
Ampicillin
Length:
10051 bp
Type:
Protein expression
Replication origin:
ori
Host:
Mammalian cells
Selection Marker:
Hyg
Promoter:
EF-1α
Growth Strain(s):
DH5a
Growth Temperature:
37℃

pcDNA-GalNAc-T2-GFP-ESCargo(FTV) vector Map

Copy Sequence View Map
pcDNA-GalNAc-T2-GFP-ESCargo(FTV)10051 bp50010001500200025003000350040004500500055006000650070007500800085009000950010000EF-1-alpha promoterPolypeptide N-acetylgalactosaminyltransferase 2msGFP2bGH poly(A) signalCMV enhancerCMV promoterDsRed-Express2bGH poly(A) signalFRTHygRSV40 poly(A) signallac promoterCAP binding siteoriAmpRAmpR promoter

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.

pcDNA-GalNAc-T2-GFP-ESCargo(FTV) vector Sequence

LOCUS       V017052                10051 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V017052
VERSION     V017052
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 10051)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..10051
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        51..1213
                     /label="EF-1-alpha promoter"
                     /note="strong constitutive promoter for human elongation
                     factor EF-1-alpha"
     CDS             1233..2945
                     /gene="GALNT2"
                     /label="Polypeptide N-acetylgalactosaminyltransferase 2"
                     /note="Polypeptide N-acetylgalactosaminyltransferase 2 from
                     Homo sapiens. Accession#: Q10471"
     CDS             2964..3671
                     /label="msGFP2"
                     /note="photostable monomeric superfolder GFP (Valbuena et
                     al., 2020)"
     polyA_signal    3701..3812
                     /label="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     enhancer        4019..4398
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4399..4602
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     CDS             4890..5561
                     /label="DsRed-Express2"
                     /note="noncytotoxic tetrameric variant of DsRed fluorescent
                     protein (Strack et al., 2008)"
     polyA_signal    6028..6252
                     /label="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    6536..6583
                     /label="FRT"
                     /note="FLP-mediated recombination occurs in the 8-bp core
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             6591..7610
                     /label="HygR"
                     /note="aminoglycoside phosphotransferase from E. coli"
     polyA_signal    7743..7876
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     promoter        complement(7961..7991)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(8006..8027)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(8315..8903)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(9077..9934)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(9935..10039)
                     /label="AmpR promoter"