pDHtsk-gdpA-EGFP(156-238aa)-Linker2-GeneB-G418 vector (V013975)

Price Information

Cat No. Plasmid Name Availability Add to cart
V013975 pDHtsk-gdpA-EGFP(156-238aa)-Linker2-GeneB-G418 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:
pDHtsk-gdpA-EGFP(156-238aa)-Linker2-GeneB-G418
Antibiotic Resistance:
Kanamycin
Length:
11746 bp
Type:
Protein expression
Replication origin:
ori
Host:
Hyphal fungi
Selection Marker:
Hyg
Promoter:
NOS
Growth Strain(s):
DH5a
Growth Temperature:
37℃

pDHtsk-gdpA-EGFP(156-238aa)-Linker2-GeneB-G418 vector Map

pDHtsk-gdpA-EGFP(156-238aa)-Linker2-GeneB-G41811746 bp50010001500200025003000350040004500500055006000650070007500800085009000950010000105001100011500T7 promoterKS primerVC155Mitogen-activated protein kinase mpkBT3 promoterNOS promoterNeoR/KanRNOS terminatorT3 promoterlac promoterCAP binding siteRB T-DNA repeatpVS1 StaApVS1 RepApVS1 oriVbomoriKanRLB T-DNA repeatCaMV poly(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.

pDHtsk-gdpA-EGFP(156-238aa)-Linker2-GeneB-G418 vector Sequence

LOCUS       V013975                11746 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V013975
VERSION     V013975
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 11746)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..11746
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        191..209
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(958..974)
                     /label="KS primer"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     CDS             992..1240
                     /label="VC155"
                     /note="C-terminal fragment of mVenus for use in bimolecular
                     fluorescence complementation (BiFC) (Kodama and Hu, 2010)"
     CDS             1268..2302
                     /gene="mpkB"
                     /label="Mitogen-activated protein kinase mpkB"
                     /note="Mitogen-activated protein kinase mpkB from
                     Aspergillus fumigatus (strain CBS 144.89 / FGSC A1163 /
                     CEA10). Accession#: B0Y8W7"
     promoter        2316..2334
                     /label="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2676..2859
                     /label="NOS promoter"
                     /note="nopaline synthase promoter"
     CDS             2883..3671
                     /label="NeoR/KanR"
                     /note="aminoglycoside phosphotransferase from Tn5"
     terminator      4064..4316
                     /label="NOS terminator"
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(4662..4680)
                     /label="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        complement(4749..4780)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4795..4816)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     misc_feature    5184..5208
                     /label="RB T-DNA repeat"
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             6508..7134
                     /label="pVS1 StaA"
                     /note="stability protein from the Pseudomonas plasmid pVS1
                     (Heeb et al., 2000)"
     CDS             7571..8635
                     /label="pVS1 RepA"
                     /note="replication protein from the Pseudomonas plasmid
                     pVS1 (Heeb et al., 2000)"
     rep_origin      8704..8898
                     /label="pVS1 oriV"
                     /note="origin of replication for the Pseudomonas plasmid
                     pVS1 (Heeb et al., 2000)"
     misc_feature    9242..9382
                     /label="bom"
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(9568..10156)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(10246..11037)
                     /label="KanR"
                     /note="aminoglycoside phosphotransferase"
     misc_feature    11462..11486
                     /label="LB T-DNA repeat"
                     /note="left border repeat from nopaline C58 T-DNA"
     polyA_signal    complement(11564..11738)
                     /label="CaMV poly(A) signal"
                     /note="cauliflower mosaic virus polyadenylation signal"