pCMV-HA-C

The pCMV-HA Mammalian Expression Vector expresses proteins containing an N-terminal hemagglutinin (HA) epitope tag. The HA epitope tag is well-characterized and highly immunoreactive. High-level expression in mammalian cells is driven from the human cytomegalovirus immediate early promoter/enhancer (PCMV IE). The vector contains an intron (SV40 splice donor/splice acceptor); the epitope tag; an MCS; and a polyadenylation signal from SV40. This vectors also possesses the ampicillin resistance gene for selection in E. coli.

载体用途

To create a fusion of a gene of interest and the HA tag, insert the gene into the MCS in frame with the HA coding sequence. The resulting HA-tagged proteins can be identified with the HA-Tag Polyclonal Antibody (IgG), provided with this vector, or another antibody raised against the HA tag. The epitope tag is also useful for facilitating purification of the protein, identifying associated proteins, characterizing new proteins by immunoprecipitation, and determining subcellular localization. The MCS in this vector is compatible with the MCSs in Clontech's Matchmaker? Two-Hybrid System Vectors. Compatibility with System 3 Vectors is noted in the MCS diagram. Consult the Vector Information Packet provided with any Matchmaker vector for complete information.

After obtaining putative positive clones in your Matchmaker two-hybrid screen, use the pCMV-Myc and pCMV-HA Vectors to verify the interactions identified in yeast directly in mammalian cells. To accomplish this, subclone the selected inserts into the pCMV-Myc Vector and the "bait" insert into the pCMV-HA Vector. Alternatively, clone the "bait" insert into pCMV-Myc and the library inserts into pCMV-HA. To confirm predicted interactions in vivo via coimmunoprecipitation, cotransfect pCMV-Myc with the pCMV-HA Vector into mammalian cells and immunoprecipitate using the c-Myc Monoclonal or HA-Tag Polyclonal Antibody provided with the vectors.

0001 GAGTTCGAGC TTGCATGCCT GCAGGTCGTT ACATAACTTA CGGTAAATGG CCCGCCTGGC 
0061 TGACCGCCCA ACGACCCCCG CCCATTGACG TCAATAATGA CGTATGTTCC CATAGTAACG 
0121 CCAATAGGGA CTTTCCATTG ACGTCAATGG GTGGAGTATT TACGGTAAAC TGCCCACTTG 
0181 GCAGTACATC AAGTGTATCA TATGCCAAGT ACGCCCCCTA TTGACGTCAA TGACGGTAAA 
0241 TGGCCCGCCT GGCATTATGC CCAGTACATG ACCTTATGGG ACTTTCCTAC TTGGCAGTAC 
0301 ATCTACGTAT TAGTCATCGC TATTACCATG GTGATGCGGT TTTGGCAGTA CATCAATGGG 
0361 CGTGGATAGC GGTTTGACTC ACGGGGATTT CCAAGTCTCC ACCCCATTGA CGTCAATGGG 
0421 AGTTTGTTTT GGCACCAAAA TCAACGGGAC TTTCCAAAAT GTCGTAACAA CTCCGCCCCA 
0481 TTGACGCAAA TGGGCGGTAG GCGTGTACGG TGGGAGGTCT ATATAAGCAG AGCTCGTTTA 
0541 GTGAACCGTC AGATCGCCTG GAGACGCCAT CCACGCTGTT TTGACCTCCA TAGAAGACAC 
0601 CGGGACCGAT CCAGCCTCCG GACTCTAGAG GATCCGGTAC TAGAGGAACT GAAAAACCAG 
0661 AAAGTTAACT GGTAAGTTTA GTCTTTTTGT CTTTTATTTC AGGTCCCGGA TCCGGTGGTG 
0721 GTGCAAATCA AAGAACTGCT CCTCAGTGGA TGTTGCCTTT ACTTCTAGGC CTGTACGGAA 
0781 GTGTTACTTC TGCTCTAAAA GCTGCGGAAT TGTACCCGCG GGCCCAGGCC CGAATTCGGT 
0841 CGACCGAGAT CTCTCGAGGT ACCGGCTACC CATACGATGT TCCAGATTAC GCTTAAGCGG 
0901 CCGCGGGGAT CCAGACATGA TAAGATACAT TGATGAGTTT GGACAAACCA CAACTAGAAT 
0961 GCAGTGAAAA AAATGCTTTA TTTGTGAAAT TTGTGATGCT ATTGCTTTAT TTGTAACCAT 
1021 TATAAGCTGC AATAAACAAG TTAACAACAA CAATTGCATT CATTTTATGT TTCAGGTTCA 
1081 GGGGGAGGTG TGGGAGGTTT TTTCGGATCC TCTAGAGTCG ATCTGCAGGC ATGCTAGCTT 
1141 GGCGTAATCA TGGTCATAGC TGTTTCCTGT GTGAAATTGT TATCCGCTCA CAATTCCACA 
1201 CAACATACGA GCCGGAAGCA TAAAGTGTAA AGCCTGGGGT GCCTAATGAG TGAGCTAACT 
1261 CACATTAATT GCGTTGCGCT CACTGCCCGC TTTCCAGTCG GGAAACCTGT CGTGCCAGCT 
1321 GCATTAATGA ATCGGCCAAC GCGCGGGGAG AGGCGGTTTG CGTATTGGGC GCTCTTCCGC 
1381 TTCCTCGCTC ACTGACTCGC TGCGCTCGGT CGTTCGGCTG CGGCGAGCGG TATCAGCTCA 
1441 CTCAAAGGCG GTAATACGGT TATCCACAGA ATCAGGGGAT AACGCAGGAA AGAACATGTG 
1501 AGCAAAAGGC CAGCAAAAGG CCAGGAACCG TAAAAAGGCC GCGTTGCTGG CGTTTTTCCA 
1561 TAGGCTCCGC CCCCCTGACG AGCATCACAA AAATCGACGC TCAAGTCAGA GGTGGCGAAA 
1621 CCCGACAGGA CTATAAAGAT ACCAGGCGTT TCCCCCTGGA AGCTCCCTCG TGCGCTCTCC 
1681 TGTTCCGACC CTGCCGCTTA CCGGATACCT GTCCGCCTTT CTCCCTTCGG GAAGCGTGGC 
1741 GCTTTCTCAT AGCTCACGCT GTAGGTATCT CAGTTCGGTG TAGGTCGTTC GCTCCAAGCT 
1801 GGGCTGTGTG CACGAACCCC CCGTTCAGCC CGACCGCTGC GCCTTATCCG GTAACTATCG 
1861 TCTTGAGTCC AACCCGGTAA GACACGACTT ATCGCCACTG GCAGCAGCCA CTGGTAACAG 
1921 GATTAGCAGA GCGAGGTATG TAGGCGGTGC TACAGAGTTC TTGAAGTGGT GGCCTAACTA 
1981 CGGCTACACT AGAAGGACAG TATTTGGTAT CTGCGCTCTG CTGAAGCCAG TTACCTTCGG 
2041 AAAAAGAGTT GGTAGCTCTT GATCCGGCAA ACAAACCACC GCTGGTAGCG GTGGTTTTTT 
2101 TGTTTGCAAG CAGCAGATTA CGCGCAGAAA AAAAGGATCT CAAGAAGATC CTTTGATCTT 
2161 TTCTACGGGG TCTGACGCTC AGTGGAACGA AAACTCACGT TAAGGGATTT TGGTCATGAG 
2221 ATTATCAAAA AGGATCTTCA CCTAGATCCT TTTAAATTAA AAATGAAGTT TTAAATCAAT 
2281 CTAAAGTATA TATGAGTAAA CTTGGTCTGA CAGTTACCAA TGCTTAATCA GTGAGGCACC 
2341 TATCTCAGCG ATCTGTCTAT TTCGTTCATC CATAGTTGCC TGACTCCCCG TCGTGTAGAT 
2401 AACTACGATA CGGGAGGGCT TACCATCTGG CCCCAGTGCT GCAATGATAC CGCGAGACCC 
2461 ACGCTCACCG GCTCCAGATT TATCAGCAAT AAACCAGCCA GCCGGAAGGG CCGAGCGCAG 
2521 AAGTGGTCCT GCAACTTTAT CCGCCTCCAT CCAGTCTATT AATTGTTGCC GGGAAGCTAG 
2581 AGTAAGTAGT TCGCCAGTTA ATAGTTTGCG CAACGTTGTT GCCATTGCTA CAGGCATCGT 
2641 GGTGTCACGC TCGTCGTTTG GTATGGCTTC ATTCAGCTCC GGTTCCCAAC GATCAAGGCG 
2701 AGTTACATGA TCCCCCATGT TGTGCAAAAA AGCGGTTAGC TCCTTCGGTC CTCCGATCGT 
2761 TGTCAGAAGT AAGTTGGCCG CAGTGTTATC ACTCATGGTT ATGGCAGCAC TGCATAATTC 
2821 TCTTACTGTC ATGCCATCCG TAAGATGCTT TTCTGTGACT GGTGAGTACT CAACCAAGTC 
2881 ATTCTGAGAA TAGTGTATGC GGCGACCGAG TTGCTCTTGC CCGGCGTCAA TACGGGATAA 
2941 TACCGCGCCA CATAGCAGAA CTTTAAAAGT GCTCATCATT GGAAAACGTT CTTCGGGGCG 
3001 AAAACTCTCA AGGATCTTAC CGCTGTTGAG ATCCAGTTCG ATGTAACCCA CTCGTGCACC 
3061 CAACTGATCT TCAGCATCTT TTACTTTCAC CAGCGTTTCT GGGTGAGCAA AAACAGGAAG 
3121 GCAAAATGCC GCAAAAAAGG GAATAAGGGC GACACGGAAA TGTTGAATAC TCATACTCTT 
3181 CCTTTTTCAA TATTATTGAA GCATTTATCA GGGTTATTGT CTCATGAGCG GATACATATT 
3241 TGAATGTATT TAGAAAAATA AACAAATAGG GGTTCCGCGC ACATTTCCCC GAAAAGTGCC 
3301 ACCTGACGTC TAAGAAACCA TTATTATCAT GACATTAACC TATAAAAATA GGCGTATCAC 
3361 GAGGCCCTTT CGTCTCGCGC GTTTCGGTGA TGACGGTGAA AACCTCTGAC ACATGCAGCT 
3421 CCCGGAGACG GTCACAGCTT GTCTGTAAGC GGATGCCGGG AGCAGACAAG CCCGTCAGGG 
3481 CGCGTCAGCG GGTGTTGGCG GGTGTCGGGG CTGGCTTAAC TATGCGGCAT CAGAGCAGAT 
3541 TGTACTGAGA GTGCACCATA TGCGGTGTGA AATACCGCAC AGATGCGTAA GGAGAAAATA 
3601 CCGCATCAGG CGCCATTCGC CATTCAGGCT GCGCAACTGT TGGGAAGGGC GATCGGTGCG 
3661 GGCCTCTTCG CTATTACGCC AGCTGGCGAA AGGGGGATGT GCTGCAAGGC GATTAAGTTG 
3721 GGTAACGCCA GGGTTTTCCC AGTCACGACG TTGTAAAACG ACGGCCAGT