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product summary
company name :
Addgene
product type :
cDNA
product name :
pPV454
catalog :
42922
citations: 1
product information
Catalog Number :
42922
Product Name :
pPV454
article :
| doi | 10.1371/journal.pone.0038587 |
| id | 6336 |
| pubmed_id | 22701676 |
bacterial resistance :
Ampicillin and Kanamycin
cloning :
| backbone | pCR4-TOPO | |
| backbone_mutation | ||
| backbone_origin | Invitrogen | |
| backbone_size | 3956 | |
| promoter | ||
| sequencing_primer_3 | ||
| sequencing_primer_5 | ||
| vector_types |
|
growth notes :
Ceage1p-cDNA-R GAGGGGCTTCGTCCCATAGACATcttaaaatcttaaaatgaaagcttg Ceage1-Cat-F CCTCCGTGGAAATGAAGAGCACatcaagatcatcacccgacaag Ceage1-Cat-R cttgtcgggtgatgatcttgatGTGCTCTTCATTTCCACGGAGG Ceage1-unc54-F CCACGCAGTCAAACACTACTGAgataagagctccgcatcggccg Ceage1-unc54-R cggccgatgcggagctcttatcTCAGTAGTGTTTGACTGCGTGG
caagctttcattttaagattttaagATGTCTATGGGACG
AAGCCCCTC
The construct Ce-age-1p::Ce-age-1(3,549bp)::Ce-unc-54t, contained in plasmid pPV454, was assembled by overlap extension PCR. An 873 bp segment of Ce-age-1 5 region was amplified from genomic DNA and fused to 2,376 bp of the 5 region of Ce-age-1, amplified from cDNA, using the overlapping primers Ceage1p-cDNA-F and -R (below). This was subsequently fused to the remainder of the Ce-age-1 cDNA with the plasmid Pdpy-30-age-1, a gift from Yuichi Iino (University of Tokyo, Tokyo, Japan), as template and using the overlapping primers Ceage1-Cat-F and -R (below). Finally, the promoter and full-length 3,549 bp Ce-age-1 cDNA were fused to 765 bp of the Ce-unc-54 3 region (terminator), amplified from Pdpy-30-age-1 template, using the overlapping primers Ceage1-unc54-F and -R (below). The PCR product was T/A cloned into the pCR4-TOPO vector (Life Technologies) and the insert fully sequenced. Ceage1p-cDNA-F
caagctttcattttaagattttaagATGTCTATGGGACG
AAGCCCCTC
The construct Ce-age-1p::Ce-age-1(3,549bp)::Ce-unc-54t, contained in plasmid pPV454, was assembled by overlap extension PCR. An 873 bp segment of Ce-age-1 5 region was amplified from genomic DNA and fused to 2,376 bp of the 5 region of Ce-age-1, amplified from cDNA, using the overlapping primers Ceage1p-cDNA-F and -R (below). This was subsequently fused to the remainder of the Ce-age-1 cDNA with the plasmid Pdpy-30-age-1, a gift from Yuichi Iino (University of Tokyo, Tokyo, Japan), as template and using the overlapping primers Ceage1-Cat-F and -R (below). Finally, the promoter and full-length 3,549 bp Ce-age-1 cDNA were fused to 765 bp of the Ce-unc-54 3 region (terminator), amplified from Pdpy-30-age-1 template, using the overlapping primers Ceage1-unc54-F and -R (below). The PCR product was T/A cloned into the pCR4-TOPO vector (Life Technologies) and the insert fully sequenced. Ceage1p-cDNA-F
origin :
37
pi :
|
plasmid copy :
Ce-age-1 cDNA and Ce-unc-54 3' region (terminator) were amplified from the plasmid Pdpy-30-age-1 from Yuichi Iino, University of Tokyo, Tokyo, Japan.
resistance markers :
| 298 |
tags :
Unknown
terms :
| Neomycin (select with G418) |
company information
Addgene
490 Arsenal Way, Suite 100
Watertown, MA 02472
Watertown, MA 02472
info@addgene.org
https://www.addgene.org617.225.9000
headquarters: USA
questions and comments