pRSFDuet-v2 RsTAL At4CL
(Plasmid #126524)

• Purpose: expresses TAL (Rhodobacter sphaeroides) and 4CL (Arabidopsis thaliana paralog 1) for PYP chromophore biosynthesis
• Depositing Lab: Andrew Woolley
• Publication: Brechun et al Biochemistry. 2019 Jun 11;58(23):2682-2694. doi: 10.1021/acs.biochem.9b00279. Epub 2019 May 30.

Backbone

• Vector backbone: pRSFDuet
• Backbone manufacturer: Novagen
• Backbone size w/o insert (bp): 3593
• Total vector size (bp): 7026
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert 1

• Gene/Insert name: tyrosine ammonia lyase
• Alt name: TAL
• Species: Rhodobacter sphaeroides
• Insert Size (bp): 1677
• GenBank ID: WP_011339422.1
• Promoter: T7
• Tag / Fusion Protein:
  • His (N terminal on insert)

Cloning Information for Gene/Insert 1

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

Gene/Insert 2

• Gene/Insert name: p-coumaroyl-CoA ligase
• Alt name: 4CL
• Alt name: pCL
• Species: A. thaliana (mustard weed)
• Insert Size (bp): 1686
• GenBank ID: U18675
• Entrez Gene: 4CL1 (a.k.a. AT1G51680, 4-COUMARATE:COA LIGASE, 4-COUMARATE:COA LIGASE 1, 4-coumarate:CoA ligase 1, 4CL.1, ARABIDOPSIS THALIANA 4-COUMARATE:COA LIGASE 1, AT4CL1, F19C24.11, F19C24_11)
• Promoter: T7

Cloning Information for Gene/Insert 2

• Cloning method: Gibson Cloning
• 5′ sequencing primer: custom
• 3′ sequencing primer: pET-RP

Resource Information

• Supplemental Documents:
  • pRSFDuet-v2_RsTAL_At4CL.gb

Terms and Licenses

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