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Browse > Keith Joung > pBAC-lacZ
Plasmid 13422: pBAC-lacZ
Gene/insert name: none
Insert size (bp): Unknown
Vector backbone: pBAC-lacZ  (Search Vector Database)
Type of vector: Bacterial expression,Cre/Lox
Backbone size (bp): 11151
Cloning site 5': BsaI
Site destroyed during cloning: No
Cloning site 3': BsaI
Site destroyed during cloning: No
5' Sequencing primer: CGC CAG GGT TTT CCC AGT CAC GAC  (List of Sequencing Primers)
Bacteria resistance: Chloramphenicol
High or low copy: High Copy
Grow in standard E. coli @ 37C: Yes
Sequence:View sequence
Author's Map:View map
Plasmid Provided In:Transformax EPI300
Principal Investigator:Keith Joung
Terms and Licenses:MTA

Comments: 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.

Addgene has sequenced a portion of this plasmid for verification. Click here for the sequencing result.

Click on map to enlarge

Selected features
rrnB_T1_terminator 44 - 87
rrnB_T2_terminator 219 - 246
lac_promoter 436 - 465
M13_pUC_rev_primer 476 - 498
M13_reverse_primer 497 - 515
M13_forward20_primer 564 - 548
M13_pUC_fwd_primer 579 - 557
Orf frame 3 507 - 3602
lacZ 555 - 3599
trpA_terminator 3623 - 3650
loxP 3702 - 3669
cosN 3914 - 4003
Orf frame 1 5895 - 4924
Orf frame 3 7070 - 5895
Orf frame 2 8404 - 7649
Orf frame 3 10365 - 11024
CAT/CamR 10365 - 11024
Unique restriction sites

NotI 2
HindIII 539
AatII 1165
ClaI 1366
ApaI 4832
SpeI 5074
KpnI 6593
StuI 8626
SacII 9320

Please acknowledge the principal investigator if you use this plasmid in a publication.

Also, please include the text "Addgene plasmid 13422" in your Materials and Methods section. This information allows Addgene to create a link from the plasmid page to your publication.

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