Clone: EvNO00025965 ← Click Here To Order

Clone ID:EvNO00025965
Is Verified:U
Status:AVAILABLE
Source:J. Kamil in Coen lab (BCMP/HMS)
Description:Vector with kanR gene insert NTAP tag ORF; useful for two-step Red recombination BAC mutagenesis approach.
Comments:Synthetic affinity tag ORF was designed in order to eliminate repeated elements present in the native CTAP described in PubMed 10504710
Type:No insert
Verification Method:
Distribution:Academic and non-profit labs
Special MTA:

Related Identifiers:

Original Clone ID pSP72 syn-ProteinC NTAP AphAI
SYNONYM pSP72 syn-Protein C NTAP AphA
SYNONYM pSP72-syn-Protein_C-NTAP-AphAI
GenBank Accession EU040032
GI 155197413

Vector Information:

Vector Name:pNTAP-TSR
Type:bacterial plasmid
Description:Vector for BAC mutagenesis via Red recombination (kanR excised upon I-SceI and Rec E/T induction) with disrupted affinity tag (synthetic ORF eliminates nucleotide repeats present in wild-type); kanamycin resistance; restriction enzyme cloning.
Properties:BAC mutagenesis, I-SceI site, Red recombination, bacterial expression, multiple cloning site, mutagenesis vector, with tag/fusion/marker
Comments:PTP N-terminal TAP tag (see PMID 16278461) added via synthetic construct that does not have direct DNA repeats present in original construct and is codon optimized for human
Map:pSP72-PTP-NTAP-ISceI.pdf
Sequence:pSP72-syn-ProteinC-NTAP-AphAI.txt
Size (bp):3968
Form:dsDNA

Host Information:

Host Strain Is Used In Distribution Description
EB5-alpha T1, T5 phage resistant Y EB5-alpha host as supplied by Edge BioSystems

Antibiotic Selections:

Host Type Marker
bacterial kanamycin

Recommended Growth Condition:

Host Type Selection Condition Growth Condition Comments
bacterial 50ug/mL kanamycin Growth with the single antibiotic in LB at 37 degrees is recommended. Conditions for Gateway-type vectors in recombined (with insert) form and other kanamycin resistant vectors.

Authors:

Author Name Author Type
Donald Coen Adademic researcher, clone donor's PI
Jeremy Kamil Academic researcher, clone donor

Publications:

PMID Title
16526409 Two-step red-mediated recombination for versatile high-efficiency markerless DNA manipulation in Escherichia coli
16278461 Highly efficient tandem affinity purification of trypanosome protein complexes based on a novel epitope combination
17634236 Human cytomegalovirus protein kinase UL97 forms a complex with the tegument phosphoprotein pp65.