Vector: pSP72 syn-CTAP AphAI

Number of plasmids with this backbone: 1
Name: pSP72 syn-CTAP AphAI
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.
Synonyms: pSP72-PTP-KanR, pSP72-PTP-AphAI
Type: bacterial plasmid
Form: dsDNA
Size (bp):: 3968
Properties: BAC mutagenesis, I-SceI site, Red recombination, in vitro transcription, multiple cloning site, mutagenesis vector, with tag/fusion/marker
Vector Map: pSP72-syn-CTAP-AphAI.pdf
Vector Sequence: pSP72-syn-CTAP-AphAI.txt
Comments: See pSP72-syn-CTAP for map without the kanR (AphAI) insert and I-SceI site that make this useful for BAC mutagenesis

Features:

Type Name Description Start Position End Position
tag IgG site 1 IgG bind site 61 234
recombination site recomb site 1 50 bp recombination substrate (Red strategy) 213 262
tag IgG site 2 IgG bind site (disrupted) 235 262
MCS I-SceI site I-SceI endonuclease cleavage site 263 280
selectable marker kanR AphAI kanamycin resistance gene 263 1250
MCS I-SceI site I-SceI endonuclease cleavage site 263 280
tag IgG site 1 IgG bind site (disrupted) 1251 1272
recombination site recomb site 2 50 bp recombination substrate (Red strategy) 1251 1300
tag IgG site 2 IgG bind site 1273 1458
tag Protein C Protein C fragment (affinity tag fragment) 1510 1545
MCS MCS MCS (XbaI, BamHI, KpnI, EcoRI, BglII) 1547 1591
primer site T7 T7 primer bind site/promoter 1605 1624
bacterial origin ColE1 ori ColE1-type origin of replication 1864 2546
selectable marker ampR ampicillin resistance gene 2644 3303

Authors:

Author Name Author Type Creation Date

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

Parent Vector:

Name Comments