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Detailed Vector Information: pSP72 syn-CTAP AphAI

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Explanation of Terms


Gene: Gene Symbol:  No insert
Gene Name:  No insert
Original Clone ID: pSP72 syn-CTAP AphAI
Type: No insert
Vector Name: pSP72 syn-CTAP AphAI              
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: None
Publications: PMID: 16526409
Title: Two-step red-mediated recombination for versatile high-efficiency markerless DNA manipulation in Escherichia coli
PMID: 10504710
Title: A generic protein purification method for protein complex characterization and proteome exploration
Authors: Donald Coen
Jeremy Kamil
EvNO00025963

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AVAILABLE
CPD only
Special MTA: None

 

 

 Recommended Growth Condition:

Distributed in bacterial strain : EB5-alpha T1, T5 phage resistant
Antibiotic Selection: Host Type:  bacterial    Marker:  kanamycin
Bacterial Selection Condition:  50ug/mL kanamycin
Growth Condition:  Growth with the single antibiotic in LB at 37 degrees is recommended.
Comments:  Conditions for Gateway-type vectors in recombined (with insert) form and other kanamycin resistant vectors.
       DyNA Vector Map

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Vector Name: pSP72 syn-CTAP AphAI

pSP72syn-CTAPAphAI in advanced viewed

Synonyms: pSP72-PTP-AphAI, pSP72-PTP-KanR
Sequencing Primer: Forward:  Unknown
Reverse:  Unknown
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.
Comments: See pSP72-syn-CTAP for map without the kanR (AphAI) insert and I-SceI site that make this useful for BAC mutagenesis
Size (bp): 3968
Parent Vector: None
Properties: BAC mutagenesis, I-SceI site, Red recombination, in vitro transcription, multiple cloning site, with tag/fusion/marker
Author Name:
Publications: PMID: 16526409
Title: Two-step red-mediated recombination for versatile high-efficiency markerless DNA manipulation in Escherichia coli

PMID: 10504710
Title: A generic protein purification method for protein complex characterization and proteome exploration

Vector Map:         Vector Sequence:
Sequence:

 



Vector Features:

Type Name Description Start Position End Position
MCS I-SceI site I-SceI endonuclease cleavage site 263 280
MCS I-SceI site I-SceI endonuclease cleavage site 263 280
MCS MCS MCS (XbaI, BamHI, KpnI, EcoRI, BglII) 1547 1591
bacterial origin ColE1 ori ColE1-type origin of replication 1864 2546
binding site IgG 1 IgG bind site 61 234
binding site IgG 2 IgG bind site 1273 1458
primer site T7 T7 primer bind site/promoter 1605 1624
recombination site recomb site 2 50 bp recombination substrate (Red strategy) 1251 1300
recombination site recomb site 1 50 bp recombination substrate (Red strategy) 213 262
selectable marker AmpR ampicillin resistance gene 2644 3303
selectable marker KanR AphAI kanamycin resistance gene 263 1250
tag Protein C Protein C fragment (affinity tag fragment) 1510 1545

 

Original Clone ID: pSP72 syn-CTAP AphAI
SYNONYM: pSP72-syn-CTAP-AphAI
Species Specific ID: None
Target GenBank: None