Clone: EvNO00025963
| Clone ID: | EvNO00025963 |
| Is Verified: | U |
| Status: | AVAILABLE |
| Source: | J. Kamil in Coen lab (BCMP/HMS) |
| Description: | Vector with kanR gene inside CTAP tag ORF (synthetic CTAP designed to avoid repeats, optimize for human codon usage); useful for two-step Red recombination BAC mutagenesis. |
| Comments: | |
| Type: | No insert |
| Verification Method: | |
| Distribution: | Academic and non-profit labs |
| Special MTA: |
Related Identifiers:
| Original Clone ID | pSP72 syn-CTAP AphAI |
| SYNONYM | pSP72-syn-CTAP-AphAI |
Vector Information:
| Vector Name: | pSP72 syn-CTAP AphAI |
| 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 form); kanamycin resistance; restriction enzyme cloning. |
| Properties: | BAC mutagenesis, I-SceI site, Red recombination, in vitro transcription, multiple cloning site, mutagenesis vector, with tag/fusion/marker |
| Comments: | See pSP72-syn-CTAP for map without the kanR (AphAI) insert and I-SceI site that make this useful for BAC mutagenesis |
| Map: | pSP72-syn-CTAP-AphAI.pdf |
| Sequence: | pSP72-syn-CTAP-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 |
| 10504710 | A generic protein purification method for protein complex characterization and proteome exploration |
