Vector Name:
pMAPLe3
Synonyms:
''
Sequencing Primer:
Forward:
T7
Reverse:
T7 terminator
Description:
Bacterial expression vector with a T7 promoter, N-terminal MBP and C-terminal 6xHis tag-TEV protease site; constitutively expresses TEV protease to produce C-terminal His-tagged protein; contains the ccdB death cassette; kanamycin resistance in bacteria; ligation independent cloning (LIC).
Comments:
Requires special host strain E. coli DB3.1 for growth
Size (bp):
8205
Parent Vector:
None
Properties:
bacterial expression, ligation independent cloning (LIC), with tag/fusion/marker
Author Name:
David Eisenberg
Mark Arbing
Integrated Center for Structure and Function Innovation
PSI
Mark Arbing
Integrated Center for Structure and Function Innovation
PSI
Publications:
None
Vector Map:
Vector Sequence:
Sequence:
Vector Features:
Type
Name
Description
Start
End
bacterial operon
lac operator
Lac operator CDS
6034
6058
bacterial origin
ColE
ColE1 pBR322 origin
2084
2084
death cassette
ccdB
ccdB death cassette
7239
8012
gene fragment
TEV cds
TEV protease cds (TEV protease is constitutively expressed from this plasmid allowing in vivo proteolysis of the expressed fusion protien so that only the target with His6 tag is purified)
3402
4432
phage origin
f1
f1 phage origin
12
467
primer
T7 term primer
T7 term reverse sequencing primer
8122
8140
promoter
T7
T7 promoter
6015
6031
protease cleavage site
TEV
TEV protease clevage site
7215
7235
repressor protein gene
LacI
LacI repressor gene
4549
5628
restriction site
ScaI
two ScaI sites from 7233-7238 and 8013-8018
0
0
ribosome binding site
RBS
ribosomal binding site
6068
6094
selectable marker
KanR
kanamycin resistance gene
563
1375
tag
MBP
N-terminal MBP tag
6102
7205
tag
His
C-terminal 6xHis tag
8019
8036
trxn termination sequence
T7 term
T7 transcription termination
8134
8180
