pTJV1Sc-rpoB
(Plasmid #166978)

• Purpose: Generates ssDNA in vivo targeting E. coli rpoB for recombineering w/ Beta recombinase; temp. sensitive pSC101 ori and sgRNA for Cas9 counterselection; aada for spectinomycin resistance
• Depositing Lab: Christopher Reisch
• Publication: Ellington et al Synth Biol (Oxf). 2022 May 3;7(1):ysac007. doi: 10.1093/synbio/ysac007. eCollection 2022.

Backbone

• Vector backbone: pKDsgRNA
• Backbone manufacturer: Reisch et al Sci Rep. 2015 Oct 14;5:15096. doi: 10.10
• Backbone size w/o insert (bp): 3500
• Total vector size (bp): 5900
• Modifications to backbone: removed lambda exo, bet, and gam
• Vector type: Bacterial Expression, CRISPR, Synthetic Biology

Growth in Bacteria

• Bacterial Resistance(s): Spectinomycin, 50 μg/mL
• Growth Temperature: 30°C
• Growth Strain(s): DH5alpha
• Copy number: Low Copy

Gene/Insert 1

• Gene/Insert name: Ec86 Retron
• Insert Size (bp): 1200
• GenBank ID: KM923747.1
• Entrez Gene: ECD_00831 (a.k.a. ECD_00831)
• Promoter: J23101

Cloning Information for Gene/Insert 1

• Cloning method: Gibson Cloning
• 5′ sequencing primer: tccccgaaaagtgccacc
• 3′ sequencing primer: CAACCGGATCCTTAGGTCT

Gene/Insert 2

• Gene/Insert name: lambda bet
• Insert Size (bp): 800
• GenBank ID: KM923747.1
• Promoter: VanCC

Cloning Information for Gene/Insert 2

• Cloning method: Gibson Cloning
• 5′ sequencing primer: ATTGGATCCAATTGACAGC

Resource Information

• A portion of this plasmid was derived from a plasmid made by: Farzadfard et al Science. 2014 Nov 14;346(6211):1256272.

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry