pTOR vector (V002568)

Price Information

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V002568 pTOR In stock, instant shipping

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

Basic Vector Information

Vector Name:
pTOR
Antibiotic Resistance:
Ampicillin
Length:
6531 bp
Type:
Cloning vector
Replication origin:
ori
Source/Author:
Belbahri L, Torche S, Mauch F.
Promoter:
lac

pTOR vector Map

pTOR6531 bp300600900120015001800210024002700300033003600390042004500480051005400570060006300f1 oriM13 fwdT7 promoter5' Bremia lactucae Ham34 promoterKS primerSK primer3' Bremia lactucae Ham34 terminator5' Bremia lactucae Hsp70 promoterNeoR/KanR3' Bremia lactucae Hsp70 terminatorT3 promoterM13 revlac operatorlac 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.

pTOR vector Sequence

LOCUS       Exported                6531 bp DNA     circular SYN 31-AUG-2024
DEFINITION  Cloning vector pTOR, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6531)
  AUTHORS   Belbahri L, Torche S, Mauch F.
  TITLE     Versatile cloning vectors for Phytophthora transformation
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6531)
  AUTHORS   Belbahri L, Torche S, Mauch F.
  TITLE     Direct Submission
  JOURNAL   Submitted (31-OCT-2007) Laboratory of Applied Genetics, School of 
            Engineering of Lullier, 150 Route de Presinge, Jussy 1254, 
            Switzerland
REFERENCE   3  (bases 1 to 6531)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6531)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 6531)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (31-OCT-2007) Laboratory of Applied Genetics, School of Engineering
            of Lullier, 150 Route de Presinge, Jussy 1254, Switzerland"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     SGRef: number: 4; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6531
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      4..459
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1011..1575
                     /label=5' Bremia lactucae Ham34 promoter
                     /note="5' Bremia lactucae Ham34 promoter"
                     /regulatory_class="promoter"
     primer_bind     1588..1604
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1638..1654)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      1689..2209
                     /label=3' Bremia lactucae Ham34 terminator
                     /note="3' Bremia lactucae Ham34 terminator"
                     /regulatory_class="terminator"
     regulatory      2217..2800
                     /label=5' Bremia lactucae Hsp70 promoter
                     /note="5' Bremia lactucae Hsp70 promoter"
                     /regulatory_class="promoter"
     CDS             2816..3607
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     regulatory      3797..4016
                     /label=3' Bremia lactucae Hsp70 terminator
                     /note="3' Bremia lactucae Hsp70 terminator"
                     /regulatory_class="terminator"
     promoter        complement(4343..4361)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4382..4398)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4406..4422)
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(4430..4460)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4475..4496)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4784..5372)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5546..6403)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(6404..6508)
                     /label=AmpR promoter