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