Vectors: Protein A fusion
Number and Name
Cloning Sites
Marker
Promoter
5' UTR
3' UTR
Comments
5
pRIT2T
C - EcoRI, XcyI, SmaI, BamHI, SalI, PstI
Amp
lambda +
PSQ
-fusion vector
11
pSPRIT3
N - XhoI, PvuII, HindIII
SP6 +
-ClaI/Klenow gives in frame TGA -> creates NruI site
17
pRI4
N - EcoRI, NcoI, BglII, BamHI, XbaI, SalI, XhoI, PvuII
no promoter
EDG
-PrA fusion vector with lots of N-term sites
-M13 ori
-GC clamp
27
pSPRIT5
N - XhoI, PvuII, HindIII, EcoRI, NcoI, BglII, BamHI, XbaI, SalI, XhoI, PvuII, HindIII
SP6 -
T7 +
-PrA fusion vector
45
pSPRIT5S
N - XhoI, PvuII, HindIII, EcoRI, NcoI, BglII, BamHI, XbaI, SalI, XhoI, PvuII
PLA
-PrA with in frame stop codon
53
pSPRIT6
N - NcoI, PvuII, HindIII
PKQ
-PrA vector with NcoI linker
57
pGEMSVPA
N - HindIII, BglII, NcoI, ApaI, NcoI, PvuII
C - EcoRI, NruI, EcoRV, AvaI, SmaI, KpnI, SacI, EcoRI
T7 -
UTK
ESQ
-high expression vector with PrA (5' UTK and Kozaks)
118
pAOFAD
C - NcoI, XbaI, BamHI, XcyI, SmaI, AvaI, EcoRI
-adenovirus epitope not in frame
143
pAEPI19
C - EcoRI, XcyI, SmaI, AvaI, BamHI, NcoI
-UT-AD
-19 kDa protein from adenovirus C-term PrA fusion
197
pSPTACPrA
V - NcoI, ApaI, SphI, PstI, SalI, XbaI, BamHI, XbaI, BamHI, XbaI, SalI, XhoI, PvuII, HindIII
Tac +
glo
-over expression
-same as #184 but has PvuI linker in EcoRV site
303
PRIT2TEK
c - XcyI, SmaI, BamHI, SalI, PstI
-PrA affinity tail
-EcoRI polylinker shifts frame of polylinker
318
pSP57FB
N - HindIII, BglII, NcoI, PvuII, HindIII
C - EcoRI, BamHI, EcoRV, AvaI, SmaI, KpnI, SacI, EcoRI
-#273 with BamHI linker
331
pSPTACPrAdeltaBam+EcoRI
N - SphI, PstI, SalI, XbaI, EcoRI, XbaI, SalI, XhoI, PvuII, HindIII
-EcoRI replaces BamHI site
669
pSPTACPrAlacIq
N - ApaI, SphI, PstI, SalI, XbaI, BamHI, XbaI, SalI, XhoI, PvuII, HindIII
-contains lacIq to regulate Tac promoter
-PrA out of frame -> use for fusion of PrA to c-term of your protein
676
pSPUTKL2PrA
N - BglII, SphI, SacII, AflII, PstI, SacI, NcoI, PvuII, HindIII
-oligo added 5' to PrA sequence to allow further cloning
684
pSPTACPrAN
N - PvuI, HindIII, NcoI
-PrA in frame
768
PERPrA
N - NcoI, ApaI, PvuII
T7+
-lacI
-lac operator
-T7 protein expression system
773
pDOC56PrA
C - XbaI, SalI, PstI, SphI, HindIII
Lac +
lambda -
-f1 ori
872
lacIqStpTi
N - NcoI, ApaI, SphI, PstI, SalI, XbaI, BamHI, XbaI, SalI, XhoI, PvuII, HindIII
transcription stop sequence
-669 with T1 termination sequence (bacterial) after PrA
-lacIq
-PrA no longer has appropriate stop codon
1093
pPrAdMfeI
N - HindIII, SacI, BglI, AflII, NcoI, MfeI
UTS
-stop codon at beginning of PrA
1120
pPrAMfe2
N - HindIII, SacI, BglII, AflII, NcoI, MfeI
-MfeI site in frame in PrA
1232
FL22
N - NcoI, StyI, Eco1301, DsaI
-SP6 promoter screwed up
1255
PBAD24PrA-Arac
C - XbaI, AccI, SalI, BspMI, PstI, SphI, HindIII
AraBC +
-use SURE cells to transform
-use DH5alpha for over expression