pCMV-Tag 4A

pCMV-Tag 4A

载体基本信息

出品公司:	Ybscience
载体名称:	pCMV-Tag 4A
质粒类型:	哺乳动物表达载体
高拷贝/低拷贝:	高拷贝
克隆方法:	多克隆位点；限制性内切酶
启动子:	CMV
载体大小:	4319 bp
5' 测序引物及序列:	CMV Forward: CGCAAATGGGCGGTAGGCGTG
3' 测序引物及序列:	T7
载体标签:	FLAG tag (C-末端）
载体抗性:	卡那霉素
筛选标记:	Neomycin
克隆菌株:	XL1-Blue
宿主细胞（系）:	--
备注:	FLAG标签含8个氨基酸残基(DYKDDDDK)； pCMV-Tag 4 A,B,C的差别仅在于读码框（阅读框）处。
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

 

ADDITIONAL MATERIALS REQUIRED
T4 DNA ligase
Taq DNA polymerase
Taq DNA polymerase buffer
TE buffer
CIAP
Falcon 2059 polypropylene tubes (15-ml)
Water baths (37°C and 42°C)

XL1-Blue Genotype: recA1 endA1 gyrA96 thi-1 hsdR17 supE44 relA1 lac [F´ proAB lacIqZΔM15 Tn10 (Tetr)].
The XL1-Blue cells supplied with the pCMV-Tag vectors are not competent cells. Refer to Hanahan (1983) for a protocol for producing competent cells.

 

载体序列

LOCUS       pCMV-Tag 4A	4319 bp 	DNA	SYN
DEFINITION  pCMV-Tag 4A
ACCESSION   
KEYWORDS    
SOURCE      
  ORGANISM  other sequences; artificial sequences; vectors.
FEATURES             Location/Qualifiers
     source          1..4319
                     /organism="pCMV-Tag 4A"
                     /mol_type="other DNA"
     promoter        16..568
                     /label="CMV_immearly_promoter"
     misc_feature    71..358
                     /label="CAG_enhancer"
     misc_feature    525..545
                     /label="CMV_fwd_primer"
     promoter        526..595
                     /label="CMV_promoter"
     promoter        620..639
                     /label="T3_promoter"
     misc_feature    complement(726..742)
                     /label="pBluescriptKS_primer"
     promoter        complement(821..839)
                     /label="T7_promoter"
     misc_feature    896..955
                     /label="SV40_3_splice"
     terminator      1006..1234
                     /label="SV40_PA_terminator"
     misc_feature    1204..1223
                     /label="EBV_rev_primer"
     rep_origin      complement(1373..1679)
                     /label="f1_origin"
     promoter        1758..1786
                     /label="AmpR_promoter"
     misc_feature    complement(1852..1872)
                     /label="pBABE_3_primer"
     misc_feature    complement(1858..2073)
                     /label="SV40_enhancer"
     promoter        1870..2138
                     /label="SV40_promoter"
     rep_origin      2037..2114
                     /label="SV40_origin"
     misc_feature    2099..2118
                     /label="SV40pro_F_primer"
     CDS             2221..3015
                     /label="ORF frame 1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR
                     PVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL
                     LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRY
                     QDIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
     gene            2224..3012
                     /label="NeoR/KanR (variant)"
                     /gene="NeoR/KanR (variant)"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR
                     PVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL
                     LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRY
                     QDIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
     CDS             complement(2530..3066)
                     /label="ORF frame 3"
                     /translation="MAGWASLGRSFRTPESRSEELVKKAIEGDALRIGSGDTVKHEEA
                     VSPFAAKFFSNITGSQRYVLIAVRHTQPATVDESRKAAIFHHDIRQAGIAMGHDEILA
                     VGHARLEPGEQFGWREPLMFFVQIILIDKTGFHPSTCSLDAMFRLVVEWAGSRIKRMQ
                     PPHCISHDGYFLGRSKVR*"
     terminator      3190..3459
                     /label="TK_PA_terminator"
                     /translation="MAGWASLGRSFRTPESRSEELVKKAIEGDALRIGSGDTVKHEEA
                     VSPFAAKFFSNITGSQRYVLIAVRHTQPATVDESRKAAIFHHDIRQAGIAMGHDEILA
                     VGHARLEPGEQFGWREPLMFFVQIILIDKTGFHPSTCSLDAMFRLVVEWAGSRIKRMQ
                     PPHCISHDGYFLGRSKVR*"
     rep_origin      3607..4226
                     /label="pBR322_origin"
                     /translation="MAGWASLGRSFRTPESRSEELVKKAIEGDALRIGSGDTVKHEEA
                     VSPFAAKFFSNITGSQRYVLIAVRHTQPATVDESRKAAIFHHDIRQAGIAMGHDEILA
                     VGHARLEPGEQFGWREPLMFFVQIILIDKTGFHPSTCSLDAMFRLVVEWAGSRIKRMQ
                     PPHCISHDGYFLGRSKVR*"
ORIGIN
    1 ATGCATTAGT 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 GCGATTACGC CAAGCTCGAA ATTAACCCTC ACTAAAGGGA ACAAAAGCTG GAGCTCCACC
  661 GCGGTGGCGG CCGCTCTAGC CCGGGCGGAT CCCCCGGGCT GCAGGAATTC GATATCAAGC
  721 TTATCGATAC CGTCGACCTC GAGGATTACA AGGATGACGA CGATAAGTAG GGCCCGGTAC
  781 CTTAATTAAT TAAGGTACCA GGTAAGTGTA CCCAATTCGC CCTATAGTGA GTCGTATTAC
  841 AATTCACTCG ATCGCCCTTC CCAACAGTTG CGCAGCCTGA ATGGCGAATG GAGATCCAAT
  901 TTTTAAGTGT ATAATGTGTT AAACTACTGA TTCTAATTGT TTGTGTATTT TAGATTCACA
  961 GTCCCAAGGC TCATTTCAGG CCCCTCAGTC CTCACAGTCT GTTCATGATC ATAATCAGCC
 1021 ATACCACATT TGTAGAGGTT TTACTTGCTT TAAAAAACCT CCCACACCTC CCCCTGAACC
 1081 TGAAACATAA AATGAATGCA ATTGTTGTTG TTAACTTGTT TATTGCAGCT TATAATGGTT
 1141 ACAAATAAAG CAATAGCATC ACAAATTTCA CAAATAAAGC ATTTTTTTCA CTGCATTCTA
 1201 GTTGTGGTTT GTCCAAACTC ATCAATGTAT CTTAACGCGT AAATTGTAAG CGTTAATATT
 1261 TTGTTAAAAT TCGCGTTAAA TTTTTGTTAA ATCAGCTCAT TTTTTAACCA ATAGGCCGAA
 1321 ATCGGCAAAA TCCCTTATAA ATCAAAAGAA TAGACCGAGA TAGGGTTGAG TGTTGTTCCA
 1381 GTTTGGAACA AGAGTCCACT ATTAAAGAAC GTGGACTCCA ACGTCAAAGG GCGAAAAACC
 1441 GTCTATCAGG GCGATGGCCC ACTACGTGAA CCATCACCCT AATCAAGTTT TTTGGGGTCG
 1501 AGGTGCCGTA AAGCACTAAA TCGGAACCCT AAAGGGAGCC CCCGATTTAG AGCTTGACGG
 1561 GGAAAGCCGG CGAACGTGGC GAGAAAGGAA GGGAAGAAAG CGAAAGGAGC GGGCGCTAGG
 1621 GCGCTGGCAA GTGTAGCGGT CACGCTGCGC GTAACCACCA CACCCGCCGC GCTTAATGCG
 1681 CCGCTACAGG GCGCGTCAGG TGGCACTTTT CGGGGAAATG TGCGCGGAAC CCCTATTTGT
 1741 TTATTTTTCT AAATACATTC AAATATGTAT CCGCTCATGA GACAATAACC CTGATAAATG
 1801 CTTCAATAAT ATTGAAAAAG GAAGAATCCT GAGGCGGAAA GAACCAGCTG TGGAATGTGT
 1861 GTCAGTTAGG GTGTGGAAAG TCCCCAGGCT CCCCAGCAGG CAGAAGTATG CAAAGCATGC
 1921 ATCTCAATTA GTCAGCAACC AGGTGTGGAA AGTCCCCAGG CTCCCCAGCA GGCAGAAGTA
 1981 TGCAAAGCAT GCATCTCAAT TAGTCAGCAA CCATAGTCCC GCCCCTAACT CCGCCCATCC
 2041 CGCCCCTAAC TCCGCCCAGT TCCGCCCATT CTCCGCCCCA TGGCTGACTA ATTTTTTTTA
 2101 TTTATGCAGA GGCCGAGGCC GCCTCGGCCT CTGAGCTATT CCAGAAGTAG TGAGGAGGCT
 2161 TTTTTGGAGG CCTAGGCTTT TGCAAAGATC GATCAAGAGA CAGGATGAGG ATCGTTTCGC
 2221 ATGATTGAAC AAGATGGATT GCACGCAGGT TCTCCGGCCG CTTGGGTGGA GAGGCTATTC
 2281 GGCTATGACT GGGCACAACA GACAATCGGC TGCTCTGATG CCGCCGTGTT CCGGCTGTCA
 2341 GCGCAGGGGC GCCCGGTTCT TTTTGTCAAG ACCGACCTGT CCGGTGCCCT GAATGAACTG
 2401 CAAGACGAGG CAGCGCGGCT ATCGTGGCTG GCCACGACGG GCGTTCCTTG CGCAGCTGTG
 2461 CTCGACGTTG TCACTGAAGC GGGAAGGGAC TGGCTGCTAT TGGGCGAAGT GCCGGGGCAG
 2521 GATCTCCTGT CATCTCACCT TGCTCCTGCC GAGAAAGTAT CCATCATGGC TGATGCAATG
 2581 CGGCGGCTGC ATACGCTTGA TCCGGCTACC TGCCCATTCG ACCACCAAGC GAAACATCGC
 2641 ATCGAGCGAG CACGTACTCG GATGGAAGCC GGTCTTGTCG ATCAGGATGA TCTGGACGAA
 2701 GAACATCAGG GGCTCGCGCC AGCCGAACTG TTCGCCAGGC TCAAGGCGAG CATGCCCGAC
 2761 GGCGAGGATC TCGTCGTGAC CCATGGCGAT GCCTGCTTGC CGAATATCAT GGTGGAAAAT
 2821 GGCCGCTTTT CTGGATTCAT CGACTGTGGC CGGCTGGGTG TGGCGGACCG CTATCAGGAC
 2881 ATAGCGTTGG CTACCCGTGA TATTGCTGAA GAACTTGGCG GCGAATGGGC TGACCGCTTC
 2941 CTCGTGCTTT ACGGTATCGC CGCTCCCGAT TCGCAGCGCA TCGCCTTCTA TCGCCTTCTT
 3001 GACGAGTTCT TCTGAGCGGG ACTCTGGGGT TCGAAATGAC CGACCAAGCG ACGCCCAACC
 3061 TGCCATCACG AGATTTCGAT TCCACCGCCG CCTTCTATGA AAGGTTGGGC TTCGGAATCG
 3121 TTTTCCGGGA CGCCGGCTGG ATGATCCTCC AGCGCGGGGA TCTCATGCTG GAGTTCTTCG
 3181 CCCACCCTAG GGGGAGGCTA ACTGAAACAC GGAAGGAGAC AATACCGGAA GGAACCCGCG
 3241 CTATGACGGC AATAAAAAGA CAGAATAAAA CGCACGGTGT TGGGTCGTTT GTTCATAAAC
 3301 GCGGGGTTCG GTCCCAGGGC TGGCACTCTG TCGATACCCC ACCGAGACCC CATTGGGGCC
 3361 AATACGCCCG CGTTTCTTCC TTTTCCCCAC CCCACCCCCC AAGTTCGGGT GAAGGCCCAG
 3421 GGCTCGCAGC CAACGTCGGG GCGGCAGGCC CTGCCATAGC CTCAGGTTAC TCATATATAC
 3481 TTTAGATTGA TTTAAAACTT CATTTTTAAT TTAAAAGGAT CTAGGTGAAG ATCCTTTTTG
 3541 ATAATCTCAT GACCAAAATC CCTTAACGTG AGTTTTCGTT CCACTGAGCG TCAGACCCCG
 3601 TAGAAAAGAT CAAAGGATCT TCTTGAGATC CTTTTTTTCT GCGCGTAATC TGCTGCTTGC
 3661 AAACAAAAAA ACCACCGCTA CCAGCGGTGG TTTGTTTGCC GGATCAAGAG CTACCAACTC
 3721 TTTTTCCGAA GGTAACTGGC TTCAGCAGAG CGCAGATACC AAATACTGTC CTTCTAGTGT
 3781 AGCCGTAGTT AGGCCACCAC TTCAAGAACT CTGTAGCACC GCCTACATAC CTCGCTCTGC
 3841 TAATCCTGTT ACCAGTGGCT GCTGCCAGTG GCGATAAGTC GTGTCTTACC GGGTTGGACT
 3901 CAAGACGATA GTTACCGGAT AAGGCGCAGC GGTCGGGCTG AACGGGGGGT TCGTGCACAC
 3961 AGCCCAGCTT GGAGCGAACG ACCTACACCG AACTGAGATA CCTACAGCGT GAGCTATGAG
 4021 AAAGCGCCAC GCTTCCCGAA GGGAGAAAGG CGGACAGGTA TCCGGTAAGC GGCAGGGTCG
 4081 GAACAGGAGA GCGCACGAGG GAGCTTCCAG GGGGAAACGC CTGGTATCTT TATAGTCCTG
 4141 TCGGGTTTCG CCACCTCTGA CTTGAGCGTC GATTTTTGTG ATGCTCGTCA GGGGGGCGGA
 4201 GCCTATGGAA AAACGCCAGC AACGCGGCCT TTTTACGGTT CCTGGCCTTT TGCTGGCCTT
 4261 TTGCTCACAT GTTCTTTCCT GCGTTATCCC CTGATTCTGT GGATAACCGT ATTACCGCC