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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
References
- 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
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| 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
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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)
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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
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