| 出品公司: | Clontech |
|---|---|
| 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。 |
| 产品目录号: | 631757 |
| 稳定性: | 瞬表达 |
| 组成型/诱导型: | 组成型 |
| 病毒/非病毒: | 非病毒 |
买家导航
载体基本信息
载体图谱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
//
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