Islam and the Modren World
pET-32a-c(+) Vectors pET-32a DNA pET-32b DNA pET-32c DNA Cat. No. 69015-3 69016-3 69017-3
TB122 12/98 The pET-32 series is designed for cloning and high-level expression of peptide sequences fused with the 109aa Trx•Tag™ thioredoxin protein (1). Cloning sites are available for producing fusion proteins also containing cleavable His•Tag® and S•Tag™ sequences for detection and purification. Unique sites are shown on the circle map. Note that the sequence is numbered by the pBR322 convention, so the T7 expression region is reversed on the circle map. The cloning/expression region of the coding strand transcribed by T7 RNA polymerase is shown below. The f1 origin is oriented so that infection with helper phage will produce virions containing single-stranded DNA that corresponds to the coding strand. Therefore, single-stranded sequencing should be performed using the T7 terminator primer (Cat. No. 69337-3).
1. LaVallie, E.R., DiBlasio, E.A., Kovacic, S., Grant, K.L., Schendel, P.F. and McCoy, J.M. (1993) Bio/Technology 11, 187–193.
pET-32a(+) sequence landmarks
Ap (4 445 -53 02 )
T7 promoter T7 transcription start Trx•Tag coding sequence His•Tag coding sequence S•Tag coding sequence Multiple cloning sites (Nco I - Xho I) His•Tag coding sequence T7 terminator lacI coding sequence pBR322 origin bla coding sequence f1 origin
764-780 763 366-692 327-344 249-293 158-217 140-157 26-72 1171-2250 3684 4445-5302 5434-5889
Bpu1102 I(80)
Ava I(158) Xho I(158) Eag I(166) Not I(166) Hind III(173) Sal I(179) Sac I(190) EcoR I(192) BamH I(198) EcoR V(206) Nco I(212) Bgl II(241) Kpn I(238) Nsp V(268) Msc I(351) Rsr II(589)
Dra III(5658)
f1
Sca I(4995) Pvu I(4885)
-5889) (5434 gin ori
trxA (36 669 2
)
Xba I(729) SgrA I(840) Sph I(996) EcoN I(1056) ApaB I(1205)
The maps for pET-32b(+) and pET-32c(+) are the same as pET-32a(+) (shown) with the following exceptions: pET-32b(+) is a 5899bp plasmid; subtract 1bp from each site beyond BamH I at 198.
TB122 12/98 The pET-32 series is designed for cloning and high-level expression of peptide sequences fused with the 109aa Trx•Tag™ thioredoxin protein (1). Cloning sites are available for producing fusion proteins also containing cleavable His•Tag® and S•Tag™ sequences for detection and purification. Unique sites are shown on the circle map. Note that the sequence is numbered by the pBR322 convention, so the T7 expression region is reversed on the circle map. The cloning/expression region of the coding strand transcribed by T7 RNA polymerase is shown below. The f1 origin is oriented so that infection with helper phage will produce virions containing single-stranded DNA that corresponds to the coding strand. Therefore, single-stranded sequencing should be performed using the T7 terminator primer (Cat. No. 69337-3).
1. LaVallie, E.R., DiBlasio, E.A., Kovacic, S., Grant, K.L., Schendel, P.F. and McCoy, J.M. (1993) Bio/Technology 11, 187–193.
pET-32a(+) sequence landmarks
Ap (4 445 -53 02 )
T7 promoter T7 transcription start Trx•Tag coding sequence His•Tag coding sequence S•Tag coding sequence Multiple cloning sites (Nco I - Xho I) His•Tag coding sequence T7 terminator lacI coding sequence pBR322 origin bla coding sequence f1 origin
764-780 763 366-692 327-344 249-293 158-217 140-157 26-72 1171-2250 3684 4445-5302 5434-5889
Bpu1102 I(80)
Ava I(158) Xho I(158) Eag I(166) Not I(166) Hind III(173) Sal I(179) Sac I(190) EcoR I(192) BamH I(198) EcoR V(206) Nco I(212) Bgl II(241) Kpn I(238) Nsp V(268) Msc I(351) Rsr II(589)
Dra III(5658)
f1
Sca I(4995) Pvu I(4885)
-5889) (5434 gin ori
trxA (36 669 2
)
Xba I(729) SgrA I(840) Sph I(996) EcoN I(1056) ApaB I(1205)
The maps for pET-32b(+) and pET-32c(+) are the same as pET-32a(+) (shown) with the following exceptions: pET-32b(+) is a 5899bp plasmid; subtract 1bp from each site beyond BamH I at 198.