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Addgene

pBAC-lacZ
(Plasmid #13422)

Ordering

This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 13422 Standard format: Plasmid sent in bacteria as agar stab 1 $85
Cloning Grade DNA 13422-DNA.cg 2 µg of cloning grade DNA in Tris buffer 1 $105

Backbone

  • Vector backbone
    pBAC-lacZ
  • Backbone size (bp) 11151
  • Vector type
    Bacterial Expression, Cre/Lox

Growth in Bacteria

  • Bacterial Resistance(s)
    Chloramphenicol, 25 μg/mL
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Copy number
    Low Copy

Gene/Insert

  • Gene/Insert name
    None

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site BsaI (not destroyed)
  • 3′ cloning site BsaI (not destroyed)
  • 5′ sequencing primer CGC CAG GGT TTT CCC AGT CAC GAC
  • (Common Sequencing Primers)

Resource Information

Terms and Licenses

  • Academic/Nonprofit Terms
  • Industry Terms
    • Not Available to Industry
Trademarks:
  • Zeocin® is an InvivoGen trademark.

Depositor Comments

The complete assembled sequence for this plasmid may not be accurate. Addgene suggests sequence verifying the construct before starting your experiments.

Bacterial cell-based two-hybrid (B2H) reporter vector. Use 12.5 ug/ml chloramphenicol for growth.

The pBAC-lacZ plasmid is a mini-F' that can be replicated in standard E. coli strains, but because it is maintained as a single copy episome, it gives low DNA yields. pBAC-lacZ also contains a second, higher copy number origin of replication (oriV) that is only active in the presence of a trans-acting factor encoded by the trfA gene. Transformax EPI300 cells express trfA from an
inducible promoter that we have found is inducible with arabinose. When the trfA gene is induced in Transformax EPI300 cells, the copy number of pBAC-lacZ plasmid is increased in the cells and reasonable yields of plasmid can be obtained using a standard miniprep procedure.

Further notes on propagating this plasmid: grow an overnight culture (without arabinose) and then the next morning subculture 1:10 into medium that has a final concentration of arabinose of 1 mM. You then grow this culture for 5 to 6 hours at 37 C and then harvest the DNA.

Information for Cloning Grade DNA (Catalog # 13422-DNA.cg) ( Back to top)

Purpose

Cloning grade DNA is suitable for use in PCR, cloning reactions, or transformation into E. coli. The purity and amount is not suitable for direct transfections.

Delivery

  • Amount 2 µg
  • Guaranteed Concentration 100 ng/µl +/- 5 ng/µl
  • Pricing $105 USD
  • Storage DNA can be stored at 4℃ (short term) or -20℃ (long term).

Terms and Licenses

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

Quality Control

Addgene has verified this plasmid using Next Generation Sequencing. Results are available here

How to cite this plasmid ( Back to top)

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:

    pBAC-lacZ was a gift from Keith Joung (Addgene plasmid # 13422 ; http://n2t.net/addgene:13422 ; RRID:Addgene_13422)
  • For your References section:

    Standardized reagents and protocols for engineering zinc finger nucleases by modular assembly. Wright DA, Thibodeau-Beganny S, Sander JD, Winfrey RJ, Hirsh AS, Eichtinger M, Fu F, Porteus MH, Dobbs D, Voytas DF, Joung JK. Nat Protoc. 2006;1(3):1637-52. doi: 10.1038/nprot.2006.259 10.1038/nprot.2006.259 PubMed 17406455