Number and Name

Cloning Sites

Marker

Promoter

5' UTR

3' UTR

Comments

5

pRIT2T

C - EcoRI, XcyI, SmaI, BamHI, SalI, PstI

Amp

lambda +

PSQ

PSQ

-fusion vector

11

pSPRIT3

N - XhoI, PvuII, HindIII

Amp

SP6 +

PSQ

PSQ

-ClaI/Klenow gives in frame TGA -> creates NruI site

17

pRI4

N - EcoRI, NcoI, BglII, BamHI, XbaI, SalI, XhoI, PvuII

Amp

no promoter

PSQ

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

Amp

SP6 -

T7 +

PSQ

PSQ

-PrA fusion vector

45

pSPRIT5S

N - XhoI, PvuII, HindIII, EcoRI, NcoI, BglII, BamHI, XbaI, SalI, XhoI, PvuII

Amp

SP6 -

T7 +

PSQ

PLA

-PrA with in frame stop codon

53

pSPRIT6

N - NcoI, PvuII, HindIII

Amp

SP6 -

T7 +

PKQ

EDG

-PrA vector with NcoI linker

57

pGEMSVPA

N - HindIII, BglII, NcoI, ApaI, NcoI, PvuII

C - EcoRI, NruI, EcoRV, AvaI, SmaI, KpnI, SacI, EcoRI

Amp

SP6 +

T7 -

UTK

ESQ

-high expression vector with PrA (5' UTK and Kozaks)

118

pAOFAD

C - NcoI, XbaI, BamHI, XcyI, SmaI, AvaI, EcoRI

Amp

lambda +

PSQ

PSQ

-adenovirus epitope not in frame

143

pAEPI19

C - EcoRI, XcyI, SmaI, AvaI, BamHI, NcoI

Amp

lambda +

PSQ

PSQ

-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

Amp

SP6 +

Tac +

T7 -

glo

EDG

-over expression

-same as #184 but has PvuI linker in EcoRV site

303

PRIT2TEK

c - XcyI, SmaI, BamHI, SalI, PstI

Amp

lambda +

PSQ

PSQ

-over expression

-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

Amp

SP6 +

T7 -

UTK

PSQ

-#273 with BamHI linker

331

pSPTACPrAdeltaBam+EcoRI

N - SphI, PstI, SalI, XbaI, EcoRI, XbaI, SalI, XhoI, PvuII, HindIII

Amp

SP6 +

Tac +

T7 -

glo

PSQ

-over expression

-EcoRI replaces BamHI site

669

pSPTACPrAlacIq

N - ApaI, SphI, PstI, SalI, XbaI, BamHI, XbaI, SalI, XhoI, PvuII, HindIII

Amp

SP6 +

Tac +

glo

PSQ

-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

Amp

SP6 +

T7 -

UTK

EDG

-oligo added 5' to PrA sequence to allow further cloning

684

pSPTACPrAN

N - PvuI, HindIII, NcoI

Amp

SP6 +

Tac +

T7 -

PSQ

-contains lacIq to regulate Tac promoter

-PrA in frame

768

PERPrA

N - NcoI, ApaI, PvuII

Amp

T7+

PSQ

PSQ

-lacI

-lac operator

-T7 protein expression system

773

pDOC56PrA

C - XbaI, SalI, PstI, SphI, HindIII

Amp

Lac +

lambda -

PSQ

PSQ

-f1 ori

872

pSPTACPrA

lacIqStpTi

N - NcoI, ApaI, SphI, PstI, SalI, XbaI, BamHI, XbaI, SalI, XhoI, PvuII, HindIII

Amp

SP6 +

Tac +

glo

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

Amp

SP6 +

T7 -

UTS

PSQ

-stop codon at beginning of PrA

1120

pPrAMfe2

N - HindIII, SacI, BglII, AflII, NcoI, MfeI

Amp

SP6 +

T7 -

UTS

PSQ

-MfeI site in frame in PrA

1232

FL22

N - NcoI, StyI, Eco1301, DsaI

Amp

T7 +

SP6 +

Tac +

UTK

PSQ

-over expression

-SP6 promoter screwed up

1255

PBAD24PrA-Arac

C - XbaI, AccI, SalI, BspMI, PstI, SphI, HindIII

Amp

AraBC +

PSQ

PSQ

-use SURE cells to transform

-use DH5alpha for over expression