https://eln.excellgen.com/index.php?action=history&feed=atom&title=PKA_Alpha_pLVX-Plus2 2026-10-18T06:19:03Z Revision history for this page on the wiki MediaWiki 1.44.0 https://eln.excellgen.com/index.php?title=PKA_Alpha_pLVX-Plus2&diff=792&oldid=prev 2011-05-03T21:18:12Z

<p></p> <p><b>New page</b></p><div>Clone PKA alpha into pLVX-Plus2&lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> subclone into pLVX-Plus2 from pMF8400&lt;br&gt;cut PKA-alpha in pMF8400 with Nde1 + BamH1 --&amp;gt; 1978 bp&lt;br&gt;cut pLVX-Plus2 with Nde+BamH1&amp;nbsp;<br /> <br /> CMV promoter with intron&lt;br&gt;CATATG CCAAGT ACGCCCCCTA TTGACGTCAA&lt;br&gt; 601 TGACGGTAAA TGGCCCGCCT GGCATTATGC CCAGTACATG ACCTTATGGG ACTTTCCTAC TTGGCAGTAC ATCTACGTAT TAGTCATCGC TATTACCATG <br /> <br /> 701 GTGATGCGGT TTTGGCAGTA CATCAATGGG CGTGGATAGC GGTTTGACTC ACGGGGATTT CCAAGTCTCC ACCCCATTGA CGTCAATGGG AGTTTGTTTT <br /> <br /> 801 GGCACCAAAA TCAACGGGAC TTTCCAAAAT GTCGTAACAA CTCCGCCCCA TTGACGCAAA TGGGCGGTAG GCGTGTACGG TGGGAGGTCT ATATAAGCAG <br /> <br /> 901 AGCTCGTTTA GTGAACCGTC AGATCGCCTG GAGACGCCAT CCACGCTGTT TTGACCTCCA TAGAAGACAC CGGGACCGAT CCAGCCTCCA TCGGCTCGCA <br /> <br /> 1001 TCTCTCCTTC ACGCGCCCGC CGCCCTACCT GAGGCCGCCA TCCACGCCGG TTGAGTCGCG TTCTGCCGCC TCCCGCCTGT GGTGCCTCCT GAACTGCGTC <br /> <br /> 1101 CGCCGTCTAG GTAAGTTTAA AGCTCAGGTC GAGACCGGGC CTTTGTCCGG CGCTCCCTTG GAGCCTACCT AGACTCAGCC GGCTCTCCAC GCTTTGCCTG&lt;br&gt; &lt;br&gt; &lt;br&gt; 1201 ACCCTGCTTG CTCAACTCTA GTTAACGGTG GAGGGCAGTG TAGTCTGAGC AGTACTCGTT GCTGCCGCGC GCGCCACCAG ACATAATAGC TGACAGACTA&lt;br&gt; IL2 secretory signal&lt;br&gt; 1301 ACAGACTGTT CCTTTCCATG GGTCTTTTCT GCAGTCACCG TCGTCGACAC GTGTGATCAG ATAGG GCC ACC ATGTACAGG ATGCAACTCC TGTCTTGCAT&lt;br&gt; &lt;br&gt; 1401 TGCACTAAGT CTTGCACTTG TCAC&lt;br&gt;EcoR1&lt;br&gt;GAATTC &lt;br&gt;6His&lt;br&gt; cac cac cac cat cac cac cac ACC GGT &lt;br&gt; ggcaacg ccgccgccgc caagaagggc agcgagcagg&lt;br&gt;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp; &amp;nbsp; &amp;nbsp;&amp;nbsp; EcoR1&lt;br&gt; 241 agagcgtgaa a gaattc tta gccaaagcca aagaagattt tcttaaaaaa tgggaaagtc&lt;br&gt; 301 ccgctcagaa cacagcccac ttggatcagt ttgaacgaat caagaccctc ggcacgggct&lt;br&gt; 361 ccttcgggcg ggtgatgctg gtgaaacaca aggagaccgg gaaccactat gccatgaaga&lt;br&gt; 421 tcctcgacaa acagaaggtg gtgaaactga aacagatcga acacaccctg aatgaaaagc&lt;br&gt; 481 gcatcctgca agctgtcaac tttccgttcc tcgtcaaact cgagttctcc ttcaaggaca&lt;br&gt; 541 actcaaactt atacatggtc atggagtacg tgcccggcgg ggagatgttc tcacacctac&lt;br&gt; 601 ggcggatcgg aaggttcagt gagccccatg cccgtttcta cgcggcccag atcgtcctga&lt;br&gt; 661 cctttgagta tctgcactcg ctggatctca tctacaggga cctgaagccg gagaatctgc&lt;br&gt; 721 tcattgacca gcagggctac attcaggtga cagacttcgg tttcgccaag cgcgtgaagg&lt;br&gt; 781 gccgcacttg gaccttgtgc ggcacccctg agtacctggc ccctgagatt atcctgagca&lt;br&gt; 841 aaggctacaa caaggccgtg gactggtggg ccctgggggt tcttatctat gaaatggccg&lt;br&gt; 901 ctggctaccc gcccttcttc gcagaccagc ccatccagat ctatgagaag atcgtctctg&lt;br&gt; 961 ggaaggtgcg cttcccttcc cacttcagct ctgacttgaa ggacctgctg cggaacctcc&lt;br&gt; 1021 tgcaggtaga tctcaccaag cgctttggga acctcaagaa tggggtcaac gatatcaaga&lt;br&gt; 1081 accacaagtg gtttgccaca actgactgga ttgccatcta ccagaggaag gtggaagctc&lt;br&gt; 1141 ccttcatacc aaagtttaaa ggccctgggg atacgagtaa ctttgacgac tatgaggaag&lt;br&gt; 1201 aagaaatccg ggtctccatc aatgagaagt gtggcaagga gttttctgag ttt&lt;br&gt;TCT AGA TGG AGC CAC CCG CAG TTC GAA AAG TAA &lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> BamH1 SfiI&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp; EcoR1 PmeI SfiI Xho1 &lt;br&gt; <br /> <br /> GGATCC GGCCGCTGCGGCC GAATTCGTTTAAAC GGCCAAACAGGCC TCGAG A&lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> Digest with EcoR1 --&amp;gt;1060bp fragment<br /> <br /> [[Image:PLVXPlus2-PKA-alpha_2011-5-3_digestion.JPG]] <br /> <br /> <br /> --[[User:Paul.xu|Paul.xu]] 16:18, 3 May 2011 (EST)<br /> [[Category:Cloning]]</div>

Paul.xu