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Functional Genomics & RNAi

Non-Selection Vector

The pACD4 TargeTron Non-Selection Vector Set (TV0010) is a set of four linearized vectors intended for use with the TargeTron Gene Knockout System (TA0100) when the use of a selectable marker is not desired. Each vector in the pACD4 vector set is similar to the pACD4K-C TargeTron vector with the main difference being they do not have a kanamycin RAM marker to select for chromosomal insertion. The pACD4 vector set also differs from the pACD4K-C TargeTron vector in that each vector in this set contains either an A, C, G or T base in the δ+1 position. This base variation facilitates proper base pairing of group II intron precursor RNA for different target site designs resulting in more efficient precursor RNA splicing to form active TargeTron RNPs.1

pACD4 Vector Sequence

Technical Bulletin

Vector Map

Data

Ordering Information

TargeTron Vector

References

  1. Perutka, J. et al., J. Biol. Biol., 336 (2), 421-439 (2004)

 


Related Products
TargeTron Gene Knockout System




TargeTron® Vector pACD4K-C-loxP Vector Map


Feature Position
p15A ori 585-1497
T7 promoter 1741-1762
5' exon (IBS) 1802-1826
Intron RNA 1827-2741
EBS2 (exon binding sequence 2) 2049-2053
EBS1d 2102-2111
delta +1 position 2742
3' exon 2742-2751
LtrA ORF 2988-4787
T1/T2 transcriptional terminator 4920-5190
chloramphenicol (Cm) resistance 5861-219

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TargeTron® Non-Selection Data


M=Marker, 50-2000 bp

1=bcr intron (204 bp)

2=lax Z intron (266 bp)

3=yec E intron (318 bp)

4=lac Z intron (266 bp)

Two genes, bcr and lacZ, were sequentially knocked out with vector pACD4-C (no RAM selection) in E. coli BL21 (DE3). In a second experiment, yecE and lacZ were sequentially knocked out using vector pACD4-C (no RAM selection) in E. coli BL 21(DE3). To confirm insertions, colony PCR was performed using an intron specific primer and a gene specific primer that spanned the 3' intron-gene junctions.




TargeTron is a registered trademark of InGex, LLC.



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TargeTron® Gene Knockout System