Cart 2

pGDP2 vector (V006072)

Price Information

Cat No. Plasmid Name Availability Add to cart
V006072 pGDP2 In stock, 1 week for quality controls

Buy one, get one free!

Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.

Basic Vector Information

Vector Name:
pGDP2
Antibiotic Resistance:
Kanamycin
Length:
4591 bp
Type:
Cloning vector
Replication origin:
ori
Source/Author:
Stokes JM, MacNair CR, Ilyas B, French S, Cote J-P., Farha MA, Sieron AO, Coombes BK, Brown ED.

pGDP2 vector Map

Copy Sequence View Map
pGDP24591 bp600120018002400300036004200RBSProbable phosphatidylethanolamine transferase Mcr-16xHisrrnB T2 terminatorKanRoribomrop

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.

pGDP2 vector Sequence

LOCUS       V006072                 4591 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V006072
VERSION     V006072
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 4591)
  AUTHORS   Stokes JM, MacNair CR, Ilyas B, French S, Cote J-P., Farha MA,
            Sieron AO, Coombes BK, Brown ED.
  TITLE     An antiprotozoal drug overcomes acquired colistin resistance
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4591)
  AUTHORS   Stokes JM, MacNair CR, Ilyas B, French S, Cote J-P., Farha MA,
            Sieron AO, Coombes BK, Brown ED.
  TITLE     Direct Submission
  JOURNAL   Submitted (16-SEP-2016) Biochemistry and Biomedical Sciences,
            McMaster University, 1200 Main Street West, Hamilton, Ontario L8N
            3Z5, Canada
REFERENCE   3  (bases 1 to 4591)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4591)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing
            ##Assembly-Data-END##
            SGRef: number: 1; type: "Journal Article"; journalName:
            "Unpublished"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (16-SEP-2016) Biochemistry and Biomedical Sciences, McMaster
            University, 1200 Main Street West, Hamilton, Ontario L8N 3Z5,
            Canada"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4591
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             150..172
                     /label="RBS"
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             179..1801
                     /gene="mcr1"
                     /label="Probable phosphatidylethanolamine transferase
                     Mcr-1"
                     /note="Probable phosphatidylethanolamine transferase Mcr-1
                     from Escherichia coli. Accession#: A0A0R6L508"
     CDS             1811..1828
                     /label="6xHis"
                     /note="6xHis affinity tag"
     terminator      complement(1890..1917)
                     /label="rrnB T2 terminator"
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     CDS             complement(2011..2823)
                     /label="KanR"
                     /note="aminoglycoside phosphotransferase"
     rep_origin      2945..3533
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     misc_feature    complement(3719..3861)
                     /label="bom"
                     /note="basis of mobility region from pBR322"
     CDS             complement(3966..4154)
                     /label="rop"
                     /note="Rop protein, which maintains plasmids at low copy
                     number"