Biomedical Engineering Reference
In-Depth Information
Table 3
Vectors designed for co-expression in
E. coli
E. coli
co-expression
compatibility
Vector
linearization
sites
Origin
(copy no.)
Vector
backbone
Vector
description
Vector
Tag
Resistance
Forward extension
Reverse extension
pOPINCDE
POI-C-HIS
CloDF13
(20-40)
Str
pCDFDuet-1 T7lacO,
lacI
pOPINs,
pOPINRSs
NcoI PmeI
Kozak(pET/TriEx)
Inffwd
KH6-pTriExInfrev
AGGAGATATA
CCATG
GTGATGGTGAT
GTTT
pOPINCDF
N-HIS-3C-POI CloDF13
(20-40)
Str
pCDFDuet-1 T7lacO,
lacI
pOPINs,
pOPINRSs
KpnI HinDIII
Opti3CInffwd
AAGTTCTGTTT
CAGGGCCCG
In-Fusion™ 3
site
ATGGTCTAGA
AAGCTTTA
′
pOPINCDJ
N-HIS-GST-
3C-POI
CloDF13
(20-40)
Str
pCDFDuet-1 T7lacO,
lacI
pOPINs,
pOPINRSs
KpnI HinDIII
Opti3CInffwd
AAGTTCTGTT
TCAGGGCCCG
In-Fusion™ 3
site
ATGGTCTAGA
AAGCTTTA
′
pOPINCDM
N-HIS-MBP-
3C-POI
CloDF13
(20-40)
Str
pCDFDuet-1 T7lacO,
lacI
pOPINs,
pOPINRSs
KpnI HinDIII
Opti3CInffwd
AAGTTCTGTT
TCAGGGCCCG
In-Fusion™ 3
site
ATGGTCTAGA
AAGCTTTA
′
pOPINRSE
POI-C-HIS
RSF1030
(>100)
Kan
pRSFDuet-1
T7lacO,
lacI
pOPINs,
pOPINCDs
NcoI PmeI
Kozak(pET/TriEx)
Inffwd
KH6-pTriExInfrev
AGGAGATATA
CCATG
GTGATGGTGAT
GTTT
pOPINRSF
N-HIS-3C-POI RSF1030
(>100)
Kan
pRSFDuet-1
T7lacO,
lacI
pOPINs,
pOPINCDs
KpnI HinDIII
Opti3CInffwd AAG
TTCTGTTTCAG
GGCCCG
In-Fusion™ 3
′
site
ATGGTCTAGA
AAGCTTTA
pOPINRSJ
N-HIS-GST-
3C-POI
RSF1030
(>100)
Kan
pRSFDuet-1
T7lacO,
lacI
pOPINs
KpnI HinDIII
Opti3CInffwd
AAGTTCTGTT
TCAGGGCCCG
In-Fusion™ 3
′
site
ATGGTCTAGA
AAGCTTTA
pOPINCDs
The vectors use the same homology extensions as either pOPINE (black) or pOPINF (bold) allowing the same PCR products to be used for either the POPINF or pOPINE vector families.
If untagged protein is required the target sequence is cloned in a pOPINE family member with a stop codon at its carboxy-terminus.
POI
protein of interest,
N-HIS
N-terminal His
6
tag,
C-His
C-terminal His
6
tag,
3C
Rhinovirus 3C protease site,
GST
glutathione-S-transferase,
MBP
maltose binding protein.
