pUC57-4eCOL2A1
(Plasmid #97211)

• Purpose: For cloning of a COL2A1-based, chondrogenesis-responsive promoter, and subcloning this insert into new vectors
• Depositing Lab: Ryan Porter
• Publication: De La Vega et al Tissue Eng Part C Methods. 2019 Feb 6. doi: 10.1089/ten.TEC.2018.0295.

Backbone

• Vector backbone: pUC57
• Backbone manufacturer: Genscript
• Backbone size w/o insert (bp): 2710
• Total vector size (bp): 3149
• Modifications to backbone: Protective bases were added to both sides of insert, allowing for subcloning into pUC57 by EcoRV
• Vector type: Vector is designed for cloning and generation of ExoIII deletions

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: 4 repeats of COL2A1 intronic enhancer (+2126/+2174) upstream of core promoter (-164/+37)
• Alt name: COL2A1
• Species: H. sapiens (human)
• Insert Size (bp): 407
• Entrez Gene: COL2A1 (a.k.a. ANFH, AOM, COL11A3, SEDC, STL1)
• Promoter: COL2A1 regulatory elements

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: SpeI (not destroyed)
• 3′ cloning site: BamHI (not destroyed)
• 5′ sequencing primer: TGTAAAACGACGGCCAGT
• 3′ sequencing primer: CAGGAAACAGCTATGAC

Resource Information

• A portion of this plasmid was derived from a plasmid made by: Insert synthesized by GenScript prior to subcloning into pUC57

Terms and Licenses

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

Depositor Comments

This construct can be used to help subclone the inserted chondrogenesis-specific promoter into various expression vectors.

An NheI site has been placed in between the 4 x enhancer repeats and the core COL2A1 promoter. This will allow those repeats to be removed, if desired (by SpeI and NheI digestion). It will also allow an additional set of repeats to be added (i.e., increasing the total to 8) after NheI digestion of the destination vector.