Recruited polymerase expression - pCMV-T7-EcKlenow-CC(N5) (LM2696)
(Plasmid
#208969)
-
PurposeRecruited EcKlenow DNA polymerase (-exo) with a C-terminal N5 coiled coil (CC) domain, expressed from CMV or T7 promoters.
-
Depositing Lab
-
Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
---|---|---|---|---|---|
Plasmid | 208969 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
-
Vector backbonepCMV-T7-SpCas9-P2A-EGFP (RTW3027)
-
Vector typeMammalian Expression, CRISPR ; in vitro transcription; T7 promoter
Growth in Bacteria
-
Bacterial Resistance(s)Ampicillin, 100 μg/mL
-
Growth Temperature37°C
-
Growth Strain(s)DH5alpha
-
Copy numberHigh Copy
Gene/Insert
-
Gene/Insert nameEcKlenow-BPNLS-CC(N5)
-
Alt nameLM2696
-
SpeciesSynthetic
-
MutationEcKlenow(-exo;D355A/E357A)
- Promoter CMV and T7
-
Tag
/ Fusion Protein
- BPNLS (C terminal on insert)
Cloning Information
- Cloning method Gibson Cloning
- 5′ sequencing primer oBK6928-CCAAGTCTCCACCCCATTGACG
- 3′ sequencing primer oBK219-GGGAGTGGCACCTTCCAGGGTC (Common Sequencing Primers)
Resource Information
-
Supplemental Documents
Terms and Licenses
-
Academic/Nonprofit Terms
-
Industry Terms
- Not Available to Industry
Trademarks:
- Zeocin® is an InvivoGen trademark.
Depositor Comments
Please visit https://doi.org/10.1101/2023.09.12.557440 for bioRxiv preprint.
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:
Recruited polymerase expression - pCMV-T7-EcKlenow-CC(N5) (LM2696) was a gift from Benjamin Kleinstiver (Addgene plasmid # 208969 ; http://n2t.net/addgene:208969 ; RRID:Addgene_208969) -
For your References section:
Click editing enables programmable genome writing using DNA polymerases and HUH endonucleases. Ferreira da Silva J, Tou CJ, King EM, Eller ML, Rufino-Ramos D, Ma L, Cromwell CR, Metovic J, Benning FMC, Chao LH, Eichler FS, Kleinstiver BP. Nat Biotechnol. 2024 Jul 22. doi: 10.1038/s41587-024-02324-x. 10.1038/s41587-024-02324-x PubMed 39039307