Plasmids for tagging genes in situ

This page is dedicated to the very useful in-situ C-terminal tagging vectors developed in Thomas Seebeck's lab (A vector series for rapid PCR-mediated C-terminal in situ tagging of Trypanosoma brucei genes (2006) Oberholzer M, Morand S, Kunz S & Seebeck T. Mol Biochem Parasitol 145:117-20. DOI PMID) and variants that we have constructed including a selection in which we have incorporated a floxed positive-negative selection cassette so the markers can be re-used and there will be minimal disruption of the 3' UTR after cre-mediated marker elimination (one 34-bp lox sequence will remain). Please contact Michael Oberholzer for the original vectors and addgene for our variants. Some columns in the table are for our lab information only. Please inform me if any of the many links on this page yield the wrong item!


Find George Cross Lab Plasmids
To ensure continued availability of our plasmids, we have deposited the major ones with Addgene. Please use the link to the left to check availability and to request them.
  
Lab Ref Systematic Name * Tag Marker Source Local Availability Bacterial Stock SeqBuilder file GenBank file
  pBSIIKS+ -      
1 pMOTag2 - PUR Oberholzer -
2 pMOTag3 - NEO Oberholzer Aug-07 No    
3 pMOTag4 - HYG Oberholzer Aug-07
4 pMOTag5 - BLE Oberholzer -
5 pMotag2H 3x HA PUR Cross (LEK) Yes
6 pMOTag3H 3x HA NEO Oberholzer - Yes    
7 pMOTag4H 3x HA HYG Oberholzer - Yes
8 pMOTag5H 3x HA BLE Cross (LEK) May 2008 Yes
9 pMOTag4M 3x c-Myc HYG Oberholzer -
10 pMOTag33M 3x c-Myc ** NEO Oberholzer Aug 2007 No    
11 pMOTag43M 3x c-Myc HYG Oberholzer Aug 2007 No
12 pMOTag53M 3x c-Myc BLE Cross (LMF) May 2008 Yes
13 pMOTag4F FLAG HYG Oberholzer Aug 2007 Yes    
14 pMOTag3S Strep-tagII NEO Oberholzer -
15 pMOTag4S Strep-tagII HYG Oberholzer -
16 pMOTag3G GFP NEO Oberholzer -
17 pMOTag4G GFP HYG Oberholzer -
18 pMOTag4YH eYFP - 3x HA HYG Oberholzer -
19 pMOTag2T TY1 PUR Cross (LMF) May 2008 Yes
20 pMOTag4HM 3x HA & 3x c-Myc HYG Cross (JW) Feb 2008  
Floxed Vectors              
21 pMOTag2T-crelox-HYG TY1 HYG Cross (JL) May 2009  
22 pMOTag2T-crelox-PUR TY1 PUR Cross (JL) May 2009  
23 pMOTag4H-crelox-HYG 3X HA HYG Cross (JL) May 2009  
24 pMOTag4H-crelox-PUR 3X HA PUR Cross (JL) May 2009  
25 pMOTag4HM-crelox-HYG 3X HA & 3X C-Myc HYG Cross (JL) May 2009  
26 pMOTag4HM-crelox-PUR 3X HA & 3X C-Myc PUR Cross (JL) May 2009  
27 pMOTag43M-crelox-HYG 3X C-Myc HYG Cross (JL) May 2009  
28 pMOTag43M-crelox-PUR 3X C-Myc PUR Cross (JL) May 2009  
                 
Notes
* Restriction sites within the drug-resistance genes may differ from those stated in the paper, in our experience.
** The Seebeck lab (and others personally) have reported that the 3x c-myc tag with antibody 9E10 (Santa Cruz Biotechnology) gives the most sensitive detection, with no background on immunoblots of procyclic or bloodstream forms.
 
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