pThermoCas9_ctrl vector (V007240)

MSDS

Basic Vector Information

Vector Name:: pThermoCas9_ctrl

Antibiotic Resistance:: Chloramphenicol

Length:: 7467 bp

Type:: Bacterial Expression

Replication origin:: ori

Promoter:: B. coagulans DSM 1 pta promoter

Cloning Method:: Gibson Cloning

5' Primer:: ggatcccatgacgctagtatccagctggGTCATAGTTCCCCTGAGAT

3' Primer:: CCCTCCCATGCACAATAG

pThermoCas9_ctrl vector Vector Map

Plasmid Resuspension 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 ℃.

pThermoCas9_ctrl vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       pThermoCas9_ctrl        7467 bp DNA     circular SYN 13-MAY-2021
DEFINITION  Expresses ThermoCas9 and its sgRNA module.
ACCESSION   .
VERSION     .
KEYWORDS    pThermoCas9_ctrl.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7467)
  AUTHORS   Mougiakos I, Mohanraju P, Bosma EF, Vrouwe V, Finger Bou M, 
            Naduthodi MIS, Gussak A, Brinkman RBL, van Kranenburg R, van der 
            Oost J
  TITLE     Characterizing a thermostable Cas9 for bacterial genome editing and 
            silencing.
  JOURNAL   Nat Commun. 2017 Nov 21;8(1):1647. doi: 10.1038/s41467-017-01591-4.
  PUBMED    29162801
REFERENCE   2  (bases 1 to 7467)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 7467)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat
            Commun."; date: "2017-11-21"; pages: "
            10.1038/s41467-017-01591-4"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7467
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(46..62)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(70..86)
                     /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(94..124)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(139..160)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(293..940)
                     /gene="cat"
                     /label=cat
                     /note="Chloramphenicol acetyltransferase from
                     Staphylococcus aureus. Accession#: P00485"
     CDS             complement(1050..2051)
                     /label=repB
                     /note="RepB replication protein"
     primer_bind     complement(2572..2589)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2743..3331)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"

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