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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 con-vention, 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. T
he f1 origin is oriented so that infection with helper phage will produce virions containing single-stranded DNA that corre-sponds 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.
(996)(1056)(1205)
(1521)(1535)(1702)
(1730)(1732)(1932)
AlwN I Bsa I (4576)
pfaPst I Pvu I Sca I TB122 12/98vb
数据库编程实例
pET-32a(+) sequence landmarks
T7 promoter
764-780T7 transcription start 763Trx•Tag coding sequence 366-692His•Tag coding sequence 327-344S•Tag coding sequence 249-293Multiple cloning sites (Nco I - Xho I)
158-217His•Tag coding sequence 140-157T7 terminator
26-72lacI coding sequence 1171-2250pBR322 origin
3684
bla coding sequence 4445-5302f1 origin
5434-5889
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 Bam H I at 198. pET-32c(+) is a 5901bp plasmid; add 1bp to each site
beyond Bam H I at 198 except for Eco R V,which cuts at 209.
pET-32a-c(+) Vectors
Eam1105 I Cat. No.pET-32a DNA 69015-3pET-32b DNA 69016-3pET-32c DNA
tiammao69017-3
pET-32a(+) Restriction Sites
TB122 12/98
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