Clone: EvNO00025965

Clone ID:EvNO00025965
Is Verified:U
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
Size (bp):3968

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.


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


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.