pBI-CMV5

pBI-CMV5

载体基本信息

出品公司:	Ybscience
载体名称:	pBI-CMV5
质粒类型:	哺乳动物细胞表达载体；双向表达载体；荧光素酶报告载体
高拷贝/低拷贝:	高拷贝
克隆方法:	限制性内切酶，多克隆位点
启动子:	miniCMV
载体大小:	3759 bp
5' 测序引物及序列:	--
3' 测序引物及序列:	--
载体标签:	--
载体抗性:	氨苄青霉素
筛选标记:	--
克隆菌株:	DH5α 等通用菌株
宿主细胞（系）:	常规细胞系（293、CV-1、CHO等）
备注:	哺乳动物细胞表达载体pBI-CMV5是双向表达载体；MetLuc是已知分子量最小的荧光素酶。
稳定性:	瞬表达
组成型/诱导型:	组成型
病毒/非病毒:	非病毒

 

载体质粒图谱和多克隆位点信息

载体简介

 

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.

 

载体序列

LOCUS       pBI-CMV5   3759 bp    DNA   circular            23-MAR-2009
COMMENT     Created by Clontech Laboratories Inc. http://www.clontech.com
COMMENT     Cat. No. 631757,632547
FEATURES             Location/Qualifiers
     misc_feature    complement(3691..3759)
                     /label=Minimal\CMV\Promoter\2
     misc_feature    675..862
                     /label=SV40\polyA\signal
     misc_feature    complement(2795..2982)
                     /label=SV40\polyA\signal
     misc_feature    1038..1637
                     /label=pUC\origin
     misc_feature    complement(1799..2659)
                     /label=Amp(R)
     misc_feature    64..473
                     /label=Enhancer
     misc_feature    474..599
                     /label=Minimal\CMV\Promoter\1
     misc_feature    complement(3017..3676)
                     /label=MetLuc
BASE COUNT      899 a       961 c       926 g       973 t 
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