pFastBac1-6×His-ADAR1-p150 vector (V014812)

Price Information

Cat No. Plasmid Name Availability Add to cart
V014812 pFastBac1-6×His-ADAR1-p150 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:
pFastBac1-6×His-ADAR1-p150
Antibiotic Resistance:
Ampicillin, Gentamycin
Length:
8470 bp
Type:
Protein expression
Replication origin:
ori
Host:
Insect cells
Promoter:
polyhedrin
5' Primer:
pFastbac-F
Growth Temperature:
37℃

pFastBac1-6×His-ADAR1-p150 vector Map

pFastBac1-6×His-ADAR1-p1508470 bp4008001200160020002400280032003600400044004800520056006000640068007200760080008400f1 oriAmpR promoterAmpRoriTn7RGmRPc promoterpolyhedrin promoter6xHisDouble-stranded RNA-specific adenosine deaminaseSV40 poly(A) signalTn7L

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.

pFastBac1-6×His-ADAR1-p150 vector Sequence

LOCUS       V014812                 8470 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V014812
VERSION     V014812
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8470)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8470
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      2..457
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        484..588
                     /label="AmpR promoter"
     CDS             589..1446
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      1620..2208
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     mobile_element  complement(2513..2737)
                     /label="Tn7R"
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(2807..3337)
                     /label="GmR"
                     /note="gentamycin acetyltransferase"
     promoter        complement(3526..3554)
                     /label="Pc promoter"
                     /note="class 1 integron promoter"
     promoter        3906..3997
                     /label="polyhedrin promoter"
                     /note="promoter for the baculovirus polyhedrin gene"
     CDS             4049..4066
                     /label="6xHis"
                     /note="6xHis affinity tag"
     CDS             4067..7744
                     /gene="ADAR"
                     /label="Double-stranded RNA-specific adenosine deaminase"
                     /note="Double-stranded RNA-specific adenosine deaminase
                     from Homo sapiens. Accession#: P55265"
     polyA_signal    7960..8094
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     mobile_element  complement(8123..8288)
                     /label="Tn7L"
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"