pBI-CMV5

pBI-CMV5

pBI-CMV5载体基本信息

pBI-CMV5载体质粒图谱和多克隆位点信息

pBI-CMV5载体描述

pBI-CMV5 is a mammalian bidirectional expression vector designed to constitutively express a protein of interest and Metridia luciferase (MetLuc), a secreted reporter protein that can be easily detected in the medium surrounding the cells.

Metridia luciferase is a naturally secreted luciferase from the marine copepod Metridia longa. The human codon-optimized Metridia luciferase gene encodes a 219 amino acid (24 kDa) polypeptide that includes a 17 amino acid N-terminal signal peptide necessary for secretion (1). Metridia luciferase is expressed and secreted into the surrounding medium, where it is easily detected by the addition of a chemiluminescent substrate.

Protein expression is driven by one of two constitutively active, minimal human cytomegalovirus promoters: PminCMV1 (located upstream of the multiple cloning site [MCS]), drives the expression of the protein of interest, and PminCMV2 drives the expression of MetLuc. To allow propagation and selection in E. coli, the vector contains a ColE1 origin of replication and an ampicillin resistance gene (Ampr).

The pBI-CMV5 vector, available as part of the Bidirectional Secreted Luciferase System (Cat. No. 631757), is designed to constitutively express a protein of interest and Metridia luciferase. The gene of interest must contain an initiation codon and a stop codon.

pBI-CMV5 can be transfected into mammalian cells using any standard transfection method. Cells expressing the secreted Metridia luciferase can be detected by adding chemiluminescent substrate (provided) to a sample of the cell medium.

Propagation in E. coli

 Recommended host strain: DH5α? and other general purpose strains.

 Selectable marker: plasmid confers resistance to ampicillin (100 μg/ml) in E. coli hosts.

 E. coli replication origin: ColE1

 Copy number: low

 Plasmid incompatibility group: pMB1/ColE1

References

1. Markova, S.V. et al. (2004) J. Bio. Chem. 279(5):3212-3217.

Note: The vector sequence was compiled from information in the sequence databases, published literature, and other sources, together with partial sequences obtained by Clontech. This vector has not been completely sequenced.

pBI-CMV5载体序列

ORIGIN

        1 CTCGAGTAGT TATTAATAGT AATCAATTAC GGGGTCATTA GTTCATAGCC CATATATGGA

       61 GTTCCGCGTT ACATAACTTA CGGTAAATGG CCCGCCTGGC TGACCGCCCA ACGACCCCCG

      121 CCCATTGACG TCAATAATGA CGTATGTTCC CATAGTAACG CCAATAGGGA CTTTCCATTG

      181 ACGTCAATGG GTGGAGTATT TACGGTAAAC TGCCCACTTG GCAGTACATC AAGTGTATCA

      241 TATGCCAAGT ACGCCCCCTA TTGACGTCAA TGACGGTAAA TGGCCCGCCT GGCATTATGC

      301 CCAGTACATG ACCTTATGGG ACTTTCCTAC TTGGCAGTAC ATCTACGTAT TAGTCATCGC

      361 TATTACCATG GTGATGCGGT TTTGGCAGTA CATCAATGGG CGTGGATAGC GGTTTGACTC

      421 ACGGGGATTT CCAAGTCTCC ACCCCATTGA CGTCAATGGG AGTTTGTTTT GGCACCAAAA

      481 TCAACGGGAC TTTCCAAAAT GTCGTAACAA CTCCGCCCCA TTGACGCAAA TGGGCGGTAG

      541 GCGTGTACGG TGGGAGGTCT ATATAAGCAG AGCTGGTTTA GTGAACCGTC AGATCCGCTA

      601 GGGATCCTCT AGTCAGCTGA CGCGTGCTAG CGCGGCCGCA TCGATAAGCT TGTCGACGAT

      661 ATCTCCAGAG GATCATAATC AGCCATACCA CATTTGTAGA GGTTTTACTT GCTTTAAAAA

      721 ACCTCCCACA CCTCCCCCTG AACCTGAAAC ATAAAATGAA TGCAATTGTT GTTGTTAACT

      781 TGTTTATTGC AGCTTATAAT GGTTACAAAT AAAGCAATAG CATCACAAAT TTCACAAATA

      841 AAGCATTTTT TTCACTGCCC CGAGCTTCCT CGCTCACTGA CTCGCTGCGC TCGGTCGTTC

      901 GGCTGCGGCG AGCGGTATCA GCTCACTCAA AGGCGGTAAT ACGGTTATCC ACAGAATCAG

      961 GGGATAACGC AGGAAAGAAC ATGTGAGCAA AAGGCCAGCA AAAGGCCAGG AACCGTAAAA

     1021 AGGCCGCGTT GCTGGCGTTT TTCCATAGGC TCCGCCCCCC TGACGAGCAT CACAAAAATC

     1081 GACGCTCAAG TCAGAGGTGG CGAAACCCGA CAGGACTATA AAGATACCAG GCGTTTCCCC

     1141 CTGGAAGCTC CCTCGTGCGC TCTCCTGTTC CGACCCTGCC GCTTACCGGA TACCTGTCCG

     1201 CCTTTCTCCC TTCGGGAAGC GTGGCGCTTT CTCATAGCTC ACGCTGTAGG TATCTCAGTT

     1261 CGGTGTAGGT CGTTCGCTCC AAGCTGGGCT GTGTGCACGA ACCCCCCGTT CAGCCCGACC

     1321 GCTGCGCCTT ATCCGGTAAC TATCGTCTTG AGTCCAACCC GGTAAGACAC GACTTATCGC

     1381 CACTGGCAGC AGCCACTGGT AACAGGATTA GCAGAGCGAG GTATGTAGGC GGTGCTACAG

     1441 AGTTCTTGAA GTGGTGGCCT AACTACGGCT ACACTAGAAG AACAGTATTT GGTATCTGCG

     1501 CTCTGCTGAA GCCAGTTACC TTCGGAAAAA GAGTTGGTAG CTCTTGATCC GGCAAACAAA

     1561 CCACCGCTGG TAGCGGTGGT TTTTTTGTTT GCAAGCAGCA GATTACGCGC AGAAAAAAAG

     1621 GATCTCAAGA AGATCCTTTG ATCTTTTCTA CGGGGTCTGA CGCTCAGTGG AACGAAAACT

     1681 CACGTTAAGG GATTTTGGTC ATGAGATTAT CAAAAAGGAT CTTCACCTAG ATCCTTTTAA

     1741 ATTAAAAATG AAGTTTTAAA TCAATCTAAA GTATATATGA GTAAACTTGG TCTGACAGTT

     1801 ACCAATGCTT AATCAGTGAG GCACCTATCT CAGCGATCTG TCTATTTCGT TCATCCATAG

     1861 TTGCCTGACT CCCCGTCGTG TAGATAACTA CGATACGGGA GGGCTTACCA TCTGGCCCCA

     1921 GTGCTGCAAT GATACCGCGA GACCCACGCT CACCGGCTCC AGATTTATCA GCAATAAACC

     1981 AGCCAGCCGG AAGGGCCGAG CGCAGAAGTG GTCCTGCAAC TTTATCCGCC TCCATCCAGT

     2041 CTATTAATTG TTGCCGGGAA GCTAGAGTAA GTAGTTCGCC AGTTAATAGT TTGCGCAACG

     2101 TTGTTGCCAT TGCTACAGGC ATCGTGGTGT CACGCTCGTC GTTTGGTATG GCTTCATTCA

     2161 GCTCCGGTTC CCAACGATCA AGGCGAGTTA CATGATCCCC CATGTTGTGC AAAAAAGCGG

     2221 TTAGCTCCTT CGGTCCTCCG ATCGTTGTCA GAAGTAAGTT GGCCGCAGTG TTATCACTCA

     2281 TGGTTATGGC AGCACTGCAT AATTCTCTTA CTGTCATGCC ATCCGTAAGA TGCTTTTCTG

     2341 TGACTGGTGA GTACTCAACC AAGTCATTCT GAGAATAGTG TATGCGGCGA CCGAGTTGCT

     2401 CTTGCCCGGC GTCAACACGG GATAATACCG CGCCACATAG CAGAACTTTA AAAGTGCTCA

     2461 TCATTGGAAA ACGTTCTTCG GGGCGAAAAC TCTCAAGGAT CTTACCGCTG TTGAGATCCA

     2521 GTTCGATGTA ACCCACTCGT GCACCCAACT GATCTTCAGC ATCTTTTACT TTCACCAGCG

     2581 TTTCTGGGTG AGCAAAAACA GGAAGGCAAA ATGCCGCAAA AAAGGGAATA AGGGCGACAC

     2641 GGAAATGTTG AATACTCATA CTCTTCCTTT TTCAATATTA TTGAAGCATT TATCAGGGTT

     2701 ATTGTCTCAT GAGCGGATAC ATATTTGAAT GTATTTAGAA AAATAAACAA ATAGGGGTTC

     2761 CGCGCACATT TCCCCGAAAA GTGCCACCTG ACGTCGGCAG TGAAAAAAAT GCTTTATTTG

     2821 TGAAATTTGT GATGCTATTG CTTTATTTGT AACCATTATA AGCTGCAATA AACAAGTTAA

     2881 CAACAACAAT TGCATTCATT TTATGTTTCA GGTTCAGGGG GAGGTGTGGG AGGTTTTTTA

     2941 AAGCAAGTAA AACCTCTACA AATGTGGTAT GGCTGATTAT GATCCTCTAG ACTGCAGCCT

     3001 CAGGAGATCT GGGCCCTCAC CTGTCGCCGG CCATGCCCTT GATGTTGTGC ACCTCTTTCT

     3061 GGATCTTGTC GGCGAAGCTG GCGCAGCGGT CGGGCAGCCA CTTCTTCAGC AGCTCGCTGC

     3121 ACTTCACGTT GGCCAGGCCC TTCAGGCAGC CGGTGGTGCA GCTGGCGCAT CTGTCCACCT

     3181 GGGCGATGAA CTGTTCCATG GGGGCCATTT CTTTGAAGCC GCTGATCTCG GGGATGTCCA

     3241 CGATGGCGCC CACGATGCCG GCCTGGCCGG TTTTCTTGTC GCCGCCGTAG TCGTGGCACC

     3301 TGCCGGGGAT GTACACCTTC ATCTTGGCGG TGCACTTGAT CTTGCTCAGG CAGATCAGGC

     3361 AGCCCCGGGT GCAGCCGGCC TTGAAGGCGT TGGCTTCCAT TTCCATGATG ACGGCCAGGG

     3421 GCAGCTTCTT GCCGGGCATC TTGCCCCGGT TGGCGTCGGT CTCGGTGGCC ACGAAGTTGC

     3481 TGGCCCGGTC GGTGTCGGCG ATGTCGGCCT TGATCATCAC TTCCATGGTC TCCCAGATGG

     3541 TGAACAGGTC GGTTTCCAGG TTGGTGATGC CGAACTTGCC TTCCAGGCCC ACGATGTCGA

     3601 TGTTGGGGTC GAACTCGGTG CTCTTGGCCT GCACCAGGGC GCTGAACACC AGGGTGAACA

     3661 CCACCTTGAT GTCCATGGTG AATTCTCCAG GCGATCTGAC GGTTCACTAA ACGAGCTCTG

     3721 CTTATATAGG CCTCCCACCG TACACGCCAC CTCGACATA

//

其他哺乳动物表达载体：