2012 Promega catalogue
2012 Promega catalogue
2012 Promega catalogue
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Life<br />
Science<br />
Catalog<br />
<strong>2012</strong><br />
Worldwide Contact List<br />
Section<br />
Contents<br />
Table of<br />
Contents<br />
Cell Signaling<br />
276<br />
Reporter Vector and Luciferase Sequencing<br />
Primers<br />
Product Size Cat.# Price ($)<br />
RVprimer3 (clockwise) 2 µg E4481 126.00<br />
RVprimer4 (counterclockwise) 2 µg E4491 126.00<br />
GLprimer1 (clockwise) 2 µg E1651 126.00<br />
GLprimer2 (counterclockwise) 2 µg E1661 126.00<br />
For Research Use Only. Not for Use in Diagnostic Procedures.<br />
Description: The Reporter Vector (RV) Sequencing Primers are designed<br />
for use with the pGL3 and pGL4 Luciferase Vectors, Chroma-Luc Vectors<br />
and pCAT ® 3 Reporter Vectors. RVprimer3 binds upstream of the luc+, luc2 or<br />
CAT gene, and sequencing runs clockwise across the multiple cloning region.<br />
RVprimer4 binds downstream of the luc+, luc2 or CAT polyadenylation region<br />
in the Promoter and Basic Vectors and downstream of the SV40 enhancer<br />
region of the Enhancer and Control Vectors. Both primers can be used to<br />
sequence double-stranded templates, but only RVprimer4 can be used to<br />
sequence single-stranded templates.<br />
RVprimer3<br />
The GLprimer1 sequences clockwise across the cloning sites upstream of the<br />
luciferase gene in the pGL2 Vectors. GLprimer2 sequences counterclockwise<br />
across the cloning sites upstream of the luciferase gene in pGL2 or pGL3 Vectors.<br />
Both GLprimers can be used to sequence double-stranded DNA, but only<br />
the GLprimer2 can be used to sequence single-stranded DNA.<br />
Primer Sequences<br />
• GLprimer1: 5´-d(TGTATCTTATGGTACTGTAACTG)-3´<br />
• GLprimer2: 5´-d(CTTTATGTTTTTGGCGTCTTCCA)-3´<br />
• RVprimer3: 5´-d(CTAGCAAAATAGGCTGTCCC)-3´<br />
• RVprimer4: 5´-d(GACGATAGTCATGCCCCGCG)-3´<br />
Storage Conditions: Store at –20°C. The primers are supplied dried.<br />
Reporter Vector Sequencing Primer Information.<br />
GLprimer2 RVprimer3 RVprimer4<br />
Sequences from luc ORF into Sequences from upstream of Sequences from downstream of<br />
multiple cloning region. Will multiple cloning region into reporter ORF and poly adenylation<br />
sequence through SV40 promoter multiple cloning region. sequences into SalI, BamHI multiple<br />
if present. cloning region, which is intended<br />
for cloning enhancer elements.<br />
pGL3 Vectors<br />
pCAT ® 3 Vectors<br />
Chroma-Luc (Click Beetle)<br />
Vectors (pCBR, pCBG68, pCBG99)<br />
pGL4 Vectors<br />
5′ . . . CTAGCAAAATAGGCTGTCCCCAGTGCAAGTGCAGGTGCCAGAACATTTCTCTATCGATA<br />
GGTACCGAGCTCTTACGCGTGCTAGCCCGGGCTCGAGATCTGCGATCTAAGTAAGCTTGG . . .<br />
KpnI<br />
Acc65I<br />
SacI MluI<br />
NheI<br />
XmaI<br />
SmaI<br />
CATTCCGGTACTGTTGGTAAAGCCACCATGGAAGACGCCAAAAACATAAAG . . . (1892bp) . . . GGATCCGTCGAC<br />
NcoI<br />
XhoI<br />
luc+ Coding Region<br />
Start<br />
SV40<br />
Promoter<br />
BglII HindIII<br />
RVprimer4<br />
SV40<br />
Enhancer<br />
GLprimer2 BamHI SalI<br />
CGATGCCCTTGAGAGCCTTCAACCCAGTCAGCTCCTTCCGGTGGGCGCGGGGCATGACTATCGTC . . . 3′<br />
pGL3 Vector multiple cloning region. The upstream and downstream cloning sites and the locations of the sequencing primers, RVprimer3, GLprimer2 and<br />
RVprimer4, are shown. The arrows above the primers indicate the direction of sequencing. The positions of the promoter (in pGL3-Promoter and pGL3-Control)<br />
and the enhancer (in pGL3-Enhancer and pGL3-Control) are shown as insertions into the sequence of pGL3-Basic (note that the promoter replaces four bases of<br />
pGL3-Basic). The sequence shown is of the ssDNA produced using the f1 origin.<br />
For complete and up-to-date product information visit: www.promega.com/catalog<br />
9490LA<br />
0756MA08_4A
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