pDsRed-Express2

价格:2500元

联系方式:I47-825O-882O

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载体基本信息

出品公司: TAKARA
载体名称: pDsRed-Express2
质粒类型: 大肠杆菌表达载体;cDNA克隆载体
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: lac
载体大小: 3311 bp
5' 测序引物及序列: --
3' 测序引物及序列: --
载体标签: DsRed-Express2
载体抗性: 氨苄青霉素(Ampicillin)
克隆菌株: DH5alpha
表达菌株: 原核细胞
备注: pDsRed-Express2载体是原核表达载体,也用作DsRed-Express2的cDNA克隆载体;
DsRed-Express2具有细胞毒性低的特点,适用于体内荧光标记;
在DsRed-Express2上下游分别有一个多克隆位点区(MCS),如果将目的基因片段插入到5‘端MCS区,表达的产物则不是DsRed-Express2融合蛋白。
产品目录号: 632535
稳定性: 稳表达
组成型/诱导型: --
病毒/非病毒: 非病毒

载体质粒图谱和多克隆位点信息

pDsRed-Express2 载体图谱



pDsRed-Express2 多克隆位点

pDsRed-Express2 载体特征

载体简介

pDsRed-Express2载体描述

pDsRed-Express2 is a prokaryotic expression vector that encodes DsRed-Express2, a variant of the Discosoma sp. red fluorescent protein, DsRed (1). DsRed-Express2 retains the fast maturation and high photostability characteristic of its predecessor, DsRed-Express (2), and has been engineered (through additional amino acid substitutions) for increased solubility and reduced cytotoxicity (3). Although it most likely forms the same tetrameric structure as wild-type DsRed, DsRed-Express2 displays a greatly reduced tendency to aggregate; this results in minimal cytotoxicity, which makes DsRed-Express2 much better suited for in vivo applications involving sensitive cells, such as primary or stem cells. DsRed-Express2 also exhibits extremely low residual green fluorescence, which allows cells expressing the protein to be effectively separated from other fluorescently labeled cell populations by flow cytometry.
In pDsRed-Express2, the DsRed-Express2 coding sequence is flanked by separate and distinct multiple cloning sites (i.e., the 5' MCS and 3' MCS) that make it easy to excise the gene for use in other cloning applications. In E. coli, DsRed-Express2 is expressed from the lac promoter (Plac) as a fusion with several amino acids, including the first five amino acids of the LacZ protein. Note, however, that if the DsRed-Express2 coding sequence is excised using a restriction site in the 5' MCS, the protein will no longer be expressed as a fusion (as it is when it is expressed from the lac promoter). A Kozak consensus sequence is located immediately upstream of the DsRed-Express2 coding sequence to enhance translational efficiency in eukaryotic cells (4).The entire DsRed-Express2 expression cassette in pDsRed-Express2 is supported by a pUC19 backbone, which contains a high-copy number origin of replication and an ampicillin resistance gene (Ampr) for propagation and selection in E. coli.

pDsRed-Express2 is primarily intended to serve as a source of DsRed-Express2 cDNA. The flanking MCS regions make it possible to excise the DsRed-Express2 coding sequence and insert it into other vector systems. The vector can also be used to express DsRed-Express2 in bacteria. Cells expressing DsRed-Express2 (excitation and emission maxima: 554 nm and 591 nm, respectively) can be detected by either fluorescence microscopy or flow cytometry 8–12 hours after transfection.
For Western analysis, DsRed-Express2 can be detected with either the Living Colors DsRed Polyclonal Anti-body (Cat. No. 632496) or the Living Colors DsRed Monoclonal Antibody (Cat. Nos. 632392 and 632393).

pDsRed-Express2载体含有以下元件:
 Plac (lac Promoter): 95–178
CAP binding site: 111–124
–35 region: 143–148; –10 region: 167–172
lac operator: 179–199
 lacZ-DsRed-Express2 fusion expressed in E. coli
Ribosome binding site: 206–209
Start codon (ATG): 217–219; Stop codon 961–963
 5' MCS (5' multiple cloning site): 234–292
 DsRed-Express2 (Discosoma sp. red fluorescent protein variant)
Kozak consensus translation initiation site: 282–292
Start codon (ATG): 289–291; Stop codon: 964–966
 3' MCS (3' multiple cloning site): 966–1065
 Ampr (Ampicillin resistance gene): 1511–2371
 pUC origin of replication: 2519–3161

Propagation in E. coli
 Recommended host strain: DH5α
 Selectable marker: plasmid confers resistance to ampicillin (50 μg/ml) in E. coli hosts.
 E. coli replication origin: pUC
 Copy number: high
 Plasmid incompatibility group: pMB1/ColE1

Excitation and emission maxima of DsRed-Express2
 Excitation maximum = 554 nm
 Emission maximum = 591 nm

载体序列

LOCUS       pDsRed-Express2	3311 bp 	DNA	SYN
DEFINITION  pDsRed-Express2
ACCESSION   
KEYWORDS    
SOURCE      
  ORGANISM  other sequences; artificial sequences; vectors.
FEATURES             Location/Qualifiers
     source          1..3311
                     /organism="pDsRed-Express2"
                     /mol_type="other DNA"
     promoter        143..172
                     /label="lac_promoter"
     misc_feature    186..208
                     /label="M13_pUC_rev_primer"
     promoter        207..225
                     /label="M13_reverse_primer"
     CDS             217..966
                     /label="ORF frame 1"
                     /translation="MTMITPSLHACRSTLEDPRVPVATMDSTENVIKPFMRFKVHMEG
                     SVNGHEFEIEGEGEGKPYEGTQTAKLQVTKGGPLPFAWDILSPQFQYGSKVYVKHPAD
                     IPDYKKLSFPEGFKWERVMNFEDGGVVTVTQDSSLQDGTFIYHVKFIGVNFPSDGPVM
                     QKKTLGWEPSTERLYPRDGVLKGEIHKALKLKGGGHYLVEFKSIYMAKKPVKLPGYYY
                     VDSKLDITSHNEDYTVVEQYERAEARHHLFQ*"
     gene            292..963
                     /label="E2_Crimson (variant)"
                     /gene="E2_Crimson (variant)"
                     /translation="MTMITPSLHACRSTLEDPRVPVATMDSTENVIKPFMRFKVHMEG
                     SVNGHEFEIEGEGEGKPYEGTQTAKLQVTKGGPLPFAWDILSPQFQYGSKVYVKHPAD
                     IPDYKKLSFPEGFKWERVMNFEDGGVVTVTQDSSLQDGTFIYHVKFIGVNFPSDGPVM
                     QKKTLGWEPSTERLYPRDGVLKGEIHKALKLKGGGHYLVEFKSIYMAKKPVKLPGYYY
                     VDSKLDITSHNEDYTVVEQYERAEARHHLFQ*"
     misc_feature    complement(469..489)
                     /label="dsRed1_N_primer"
                     /translation="MTMITPSLHACRSTLEDPRVPVATMDSTENVIKPFMRFKVHMEG
                     SVNGHEFEIEGEGEGKPYEGTQTAKLQVTKGGPLPFAWDILSPQFQYGSKVYVKHPAD
                     IPDYKKLSFPEGFKWERVMNFEDGGVVTVTQDSSLQDGTFIYHVKFIGVNFPSDGPVM
                     QKKTLGWEPSTERLYPRDGVLKGEIHKALKLKGGGHYLVEFKSIYMAKKPVKLPGYYY
                     VDSKLDITSHNEDYTVVEQYERAEARHHLFQ*"
     misc_feature    881..904
                     /label="dsRed1_C_primer"
                     /translation="MTMITPSLHACRSTLEDPRVPVATMDSTENVIKPFMRFKVHMEG
                     SVNGHEFEIEGEGEGKPYEGTQTAKLQVTKGGPLPFAWDILSPQFQYGSKVYVKHPAD
                     IPDYKKLSFPEGFKWERVMNFEDGGVVTVTQDSSLQDGTFIYHVKFIGVNFPSDGPVM
                     QKKTLGWEPSTERLYPRDGVLKGEIHKALKLKGGGHYLVEFKSIYMAKKPVKLPGYYY
                     VDSKLDITSHNEDYTVVEQYERAEARHHLFQ*"
     misc_feature    complement(1101..1123)
                     /label="pGEX_3_primer"
                     /translation="MTMITPSLHACRSTLEDPRVPVATMDSTENVIKPFMRFKVHMEG
                     SVNGHEFEIEGEGEGKPYEGTQTAKLQVTKGGPLPFAWDILSPQFQYGSKVYVKHPAD
                     IPDYKKLSFPEGFKWERVMNFEDGGVVTVTQDSSLQDGTFIYHVKFIGVNFPSDGPVM
                     QKKTLGWEPSTERLYPRDGVLKGEIHKALKLKGGGHYLVEFKSIYMAKKPVKLPGYYY
                     VDSKLDITSHNEDYTVVEQYERAEARHHLFQ*"
     promoter        1441..1469
                     /label="AmpR_promoter"
                     /translation="MTMITPSLHACRSTLEDPRVPVATMDSTENVIKPFMRFKVHMEG
                     SVNGHEFEIEGEGEGKPYEGTQTAKLQVTKGGPLPFAWDILSPQFQYGSKVYVKHPAD
                     IPDYKKLSFPEGFKWERVMNFEDGGVVTVTQDSSLQDGTFIYHVKFIGVNFPSDGPVM
                     QKKTLGWEPSTERLYPRDGVLKGEIHKALKLKGGGHYLVEFKSIYMAKKPVKLPGYYY
                     VDSKLDITSHNEDYTVVEQYERAEARHHLFQ*"
     gene            1511..2371
                     /label="Ampicillin"
                     /gene="Ampicillin"
                     /translation="MTMITPSLHACRSTLEDPRVPVATMDSTENVIKPFMRFKVHMEG
                     SVNGHEFEIEGEGEGKPYEGTQTAKLQVTKGGPLPFAWDILSPQFQYGSKVYVKHPAD
                     IPDYKKLSFPEGFKWERVMNFEDGGVVTVTQDSSLQDGTFIYHVKFIGVNFPSDGPVM
                     QKKTLGWEPSTERLYPRDGVLKGEIHKALKLKGGGHYLVEFKSIYMAKKPVKLPGYYY
                     VDSKLDITSHNEDYTVVEQYERAEARHHLFQ*"
     CDS             1511..2371
                     /label="ORF frame 2"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW*"
     rep_origin      2526..3145
                     /label="pBR322_origin"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW*"
ORIGIN
    1 AGCGCCCAAT ACGCAAACCG CCTCTCCCCG CGCGTTGGCC GATTCATTAA TGCAGCTGGC
   61 ACGACAGGTT TCCCGACTGG AAAGCGGGCA GTGAGCGCAA CGCAATTAAT GTGAGTTAGC
  121 TCACTCATTA GGCACCCCAG GCTTTACACT TTATGCTTCC GGCTCGTATG TTGTGTGGAA
  181 TTGTGAGCGG ATAACAATTT CACACAGGAA ACAGCTATGA CCATGATTAC GCCAAGCTTG
  241 CATGCCTGCA GGTCGACTCT AGAGGATCCC CGGGTACCGG TCGCCACCAT GGATAGCACT
  301 GAGAACGTCA TCAAGCCCTT CATGCGCTTC AAGGTGCACA TGGAGGGCTC CGTGAACGGC
  361 CACGAGTTCG AGATCGAGGG CGAGGGCGAG GGCAAGCCCT ACGAGGGCAC CCAGACCGCC
  421 AAGCTGCAGG TGACCAAGGG CGGCCCCCTG CCCTTCGCCT GGGACATCCT GTCCCCCCAG
  481 TTCCAGTACG GCTCCAAGGT GTACGTGAAG CACCCCGCCG ACATCCCCGA CTACAAGAAG
  541 CTGTCCTTCC CCGAGGGCTT CAAGTGGGAG CGCGTGATGA ACTTCGAGGA CGGCGGCGTG
  601 GTGACCGTGA CCCAGGACTC CTCCCTGCAG GACGGCACCT TCATCTACCA CGTGAAGTTC
  661 ATCGGCGTGA ACTTCCCCTC CGACGGCCCC GTAATGCAGA AGAAGACTCT GGGCTGGGAG
  721 CCCTCCACCG AGCGCCTGTA CCCCCGCGAC GGCGTGCTGA AGGGCGAGAT CCACAAGGCG
  781 CTGAAGCTGA AGGGCGGCGG CCACTACCTG GTGGAGTTCA AGTCAATCTA CATGGCCAAG
  841 AAGCCCGTGA AGCTGCCCGG CTACTACTAC GTGGACTCCA AGCTGGACAT CACCTCCCAC
  901 AACGAGGACT ACACCGTGGT GGAGCAGTAC GAGCGCGCCG AGGCCCGCCA CCACCTGTTC
  961 CAGTAGCGGC CGCGACTCTA GAATTCCAAC TGAGCGCCGG TCGCTACCAT TACCAACTTG
 1021 TCTGGTGTCA AAAATAATAG GCCTACTAGT CGGCCGTACG GGCCCTTTCG TCTCGCGCGT
 1081 TTCGGTGATG ACGGTGAAAA CCTCTGACAC ATGCAGCTCC CGGAGACGGT CACAGCTTGT
 1141 CTGTAAGCGG ATGCCGGGAG CAGACAAGCC CGTCAGGGCG CGTCAGCGGG TGTTGGCGGG
 1201 TGTCGGGGCT GGCTTAACTA TGCGGCATCA GAGCAGATTG TACTGAGAGT GCACCATATG
 1261 CGGTGTGAAA TACCGCACAG ATGCGTAAGG AGAAAATACC GCATCAGGCG GCCTTAAGGG
 1321 CCTCGTGATA CGCCTATTTT TATAGGTTAA TGTCATGATA ATAATGGTTT CTTAGACGTC
 1381 AGGTGGCACT TTTCGGGGAA ATGTGCGCGG AACCCCTATT TGTTTATTTT TCTAAATACA
 1441 TTCAAATATG TATCCGCTCA TGAGACAATA ACCCTGATAA ATGCTTCAAT AATATTGAAA
 1501 AAGGAAGAGT ATGAGTATTC AACATTTCCG TGTCGCCCTT ATTCCCTTTT TTGCGGCATT
 1561 TTGCCTTCCT GTTTTTGCTC ACCCAGAAAC GCTGGTGAAA GTAAAAGATG CTGAAGATCA
 1621 GTTGGGTGCA CGAGTGGGTT ACATCGAACT GGATCTCAAC AGCGGTAAGA TCCTTGAGAG
 1681 TTTTCGCCCC GAAGAACGTT TTCCAATGAT GAGCACTTTT AAAGTTCTGC TATGTGGCGC
 1741 GGTATTATCC CGTATTGACG CCGGGCAAGA GCAACTCGGT CGCCGCATAC ACTATTCTCA
 1801 GAATGACTTG GTTGAGTACT CACCAGTCAC AGAAAAGCAT CTTACGGATG GCATGACAGT
 1861 AAGAGAATTA TGCAGTGCTG CCATAACCAT GAGTGATAAC ACTGCGGCCA ACTTACTTCT
 1921 GACAACGATC GGAGGACCGA AGGAGCTAAC CGCTTTTTTG CACAACATGG GGGATCATGT
 1981 AACTCGCCTT GATCGTTGGG AACCGGAGCT GAATGAAGCC ATACCAAACG ACGAGCGTGA
 2041 CACCACGATG CCTGTAGCAA TGGCAACAAC GTTGCGCAAA CTATTAACTG GCGAACTACT
 2101 TACTCTAGCT TCCCGGCAAC AATTAATAGA CTGGATGGAG GCGGATAAAG TTGCAGGACC
 2161 ACTTCTGCGC TCGGCCCTTC CGGCTGGCTG GTTTATTGCT GATAAATCTG GAGCCGGTGA
 2221 GCGTGGGTCT CGCGGTATCA TTGCAGCACT GGGGCCAGAT GGTAAGCCCT CCCGTATCGT
 2281 AGTTATCTAC ACGACGGGGA GTCAGGCAAC TATGGATGAA CGAAATAGAC AGATCGCTGA
 2341 GATAGGTGCC TCACTGATTA AGCATTGGTA ACTGTCAGAC CAAGTTTACT CATATATACT
 2401 TTAGATTGAT TTAAAACTTC ATTTTTAATT TAAAAGGATC TAGGTGAAGA TCCTTTTTGA
 2461 TAATCTCATG ACCAAAATCC CTTAACGTGA GTTTTCGTTC CACTGAGCGT CAGACCCCGT
 2521 AGAAAAGATC AAAGGATCTT CTTGAGATCC TTTTTTTCTG CGCGTAATCT GCTGCTTGCA
 2581 AACAAAAAAA CCACCGCTAC CAGCGGTGGT TTGTTTGCCG GATCAAGAGC TACCAACTCT
 2641 TTTTCCGAAG GTAACTGGCT TCAGCAGAGC GCAGATACCA AATACTGTTC TTCTAGTGTA
 2701 GCCGTAGTTA GGCCACCACT TCAAGAACTC TGTAGCACCG CCTACATACC TCGCTCTGCT
 2761 AATCCTGTTA CCAGTGGCTG CTGCCAGTGG CGATAAGTCG TGTCTTACCG GGTTGGACTC
 2821 AAGACGATAG TTACCGGATA AGGCGCAGCG GTCGGGCTGA ACGGGGGGTT CGTGCACACA
 2881 GCCCAGCTTG GAGCGAACGA CCTACACCGA ACTGAGATAC CTACAGCGTG AGCTATGAGA
 2941 AAGCGCCACG CTTCCCGAAG GGAGAAAGGC GGACAGGTAT CCGGTAAGCG GCAGGGTCGG
 3001 AACAGGAGAG CGCACGAGGG AGCTTCCAGG GGGAAACGCC TGGTATCTTT ATAGTCCTGT
 3061 CGGGTTTCGC CACCTCTGAC TTGAGCGTCG ATTTTTGTGA TGCTCGTCAG GGGGGCGGAG
 3121 CCTATGGAAA AACGCCAGCA ACGCGGCCTT TTTACGGTTC CTGGCCTTTT GCTGGCCTTT
 3181 TGCTCACATG TTCTTTCCTG CGTTATCCCC TGATTCTGTG GATAACCGTA TTACCGCCTT
 3241 TGAGTGAGCT GATACCGCTC GCCGCAGCCG AACGACCGAG CGCAGCGAGT CAGTGAGCGA
 3301 GGAAGCGGAA G
//

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