PurposemTurquoise2^SEC^2xHA vector with ccdB sites for cloning homology arms
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||73343||Standard format: Plasmid sent in bacteria as agar stab||1||$75|
This material is available to academics and nonprofits only.
Vector backbonepUC19 (modified)
- Backbone size w/o insert (bp) 2600
- Total vector size (bp) 10500
Modifications to backboneAddition of ccdB markers to facilitate homology arm cloning
Vector typeWorm Expression, Cre/Lox, CRISPR
Selectable markersHygromycin ; sqt-1(d) (worm phenotypic marker)
Growth in Bacteria
Growth Strain(s)NEB Turbo
Growth instructionsThe dual ccdB sites in this vector may make it prone to recombination. It is recommended to pick several single clones and test by restriction digestion before use. This construct should be maintained as a purified plasmid stock in addition to a bacterial stock in case there is a need to re-transform.
SpeciesC. elegans (nematode), Synthetic
Insert Size (bp)6600
/ Fusion Proteins
- C. elegans codon-optimized mTurquoise2
- Cloning method Gibson Cloning
- 5′ sequencing primer M13 Forward (tgtaaaacgacggccagt)
- 3′ sequencing primer M13 Reverse (caggaaacagctatgaccatg) (Common Sequencing Primers)
pDD315 is a derivative of our published vectors pDD282-285 (Dickinson et al. Genetics 2015). It has a 2xHA tag in place of 3xFlag, and Lox511I sites in place of LoxP. These features allow pDD315 to be used in a genetic background that has already been modified using a green or red FP-SEC vector, without conflicts between epitope tags and Lox sites.
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.
For your Materials & Methods section:pDD315 (mTurquoise2^SEC^2xHA) was a gift from Bob Goldstein (Addgene plasmid # 73343 ; http://n2t.net/addgene:73343 ; RRID:Addgene_73343)