pAc5.1-V5-His A

pAc5.1-V5-His A

载体基本信息

出品公司:	Ybscience
载体名称:	pAc5.1-V5-His A
质粒类型:	昆虫表达载体；果蝇表达载体
克隆方法:	多克隆位点，限制性内切酶
启动子:	P-AC5(Actin 5C)
载体大小:	5375 bp
5' 测序引物及序列:	AC5 Forward：5'd[ACACAAAGCCGCTCCATCAG]3'
3' 测序引物及序列:	--
载体标签:	V5 Epitope Tag, His Tag(C-末端)
载体抗性:	氨苄青霉素
真核筛选标记:	无
克隆菌株:	TOP10；DH5-T1R
宿主细胞（系）:	Sf9, High Five
备注:	pAc5.1-V5-His 载体可以与筛选载体pCoHygro或者pCoBlast共转染，建立稳表达细胞系。
瞬表达/稳表达:	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

 

The DES-Constitutive Kit provides the vector pAc5.1/V5-His for high-level constitutive expression of recombinant proteins and a choice of Blasticidin (pCoBlast) or hygromycin (pCoHygro) selection. The vector offers the strong, constitutive promoter from the Drosophila actin 5C gene. The pAc5.1/V5-His vector also offers the following features:

 Small size (5.4 kb) to improve DNA yields and increase subcloning efficiency
 C-terminal V5 epitope tag for rapid detection with Invitrogens Anti-V5 Antibody
 C-terminal 6xHis tag for simple purification of recombinant fusion proteins using nickel-chelating resin

To facilitate cloning, a set of three vectors-A, B, and C-is provided. Each vector has the multiple cloning site in a different reading frame relative to the coding sequence of the C-terminal tag.

 

载体序列

LOCUS       pAc5.1/V5-His A	5375 bp 	DNA	SYN
DEFINITION  pAc5.1/V5-His A
ACCESSION   
KEYWORDS    
SOURCE      
  ORGANISM  other sequences; artificial sequences; vectors.
FEATURES             Location/Qualifiers
     source          1..5375
                     /organism="pAc5.1/V5-His A"
                     /mol_type="other DNA"
     CDS             726..1454
                     /label="ORF frame 3"
                     /translation="MMPDHRLISASFAERSACAVVVFWNSFNFRTVRFDFSFLAYSKS
                     KVKPKKRDGNTKCGKTVVEGKGCGAAEGRVGRGVAGSVAGRDVTDVGATPLTMCACVY
                     YSCLATRRLFFSFYLALSSAISYACSTHRMLPCPLCVILARNRQLFKQRLVNENAKIN
                     KSTIWLLIAMCVQPTIATKATTQWLSLFHFLFASACEQDGTTGKRNYADKEQTKFWPK
                     NIKAPDTNAFAITIAIFNIVYEAV*"
     CDS             complement(1940..2416)
                     /label="ORF frame 2"
                     /translation="MLSALKSRSKLLPYMGMNCSPIGFSFTENFLRNWVFVLVCVLYI
                     AMKSVLPWVLSVAACDSIWGLLIFDTSAGYLYAVMVTVLLHIFSPTTHCMPKHTYVHV
                     FIVGLLVMSSDIKTLQLNNIHISLRLLLSALQIGYSGTFVQHTSSSRGITSRRKHT*"
     misc_feature    2525..2544
                     /label="AC5_primer"
                     /translation="MLSALKSRSKLLPYMGMNCSPIGFSFTENFLRNWVFVLVCVLYI
                     AMKSVLPWVLSVAACDSIWGLLIFDTSAGYLYAVMVTVLLHIFSPTTHCMPKHTYVHV
                     FIVGLLVMSSDIKTLQLNNIHISLRLLLSALQIGYSGTFVQHTSSSRGITSRRKHT*"
     misc_feature    complement(2751..2768)
                     /label="BGH_rev_primer"
                     /translation="MLSALKSRSKLLPYMGMNCSPIGFSFTENFLRNWVFVLVCVLYI
                     AMKSVLPWVLSVAACDSIWGLLIFDTSAGYLYAVMVTVLLHIFSPTTHCMPKHTYVHV
                     FIVGLLVMSSDIKTLQLNNIHISLRLLLSALQIGYSGTFVQHTSSSRGITSRRKHT*"
     terminator      complement(2811..2930)
                     /label="SV40_PA_terminator"
                     /translation="MLSALKSRSKLLPYMGMNCSPIGFSFTENFLRNWVFVLVCVLYI
                     AMKSVLPWVLSVAACDSIWGLLIFDTSAGYLYAVMVTVLLHIFSPTTHCMPKHTYVHV
                     FIVGLLVMSSDIKTLQLNNIHISLRLLLSALQIGYSGTFVQHTSSSRGITSRRKHT*"
     misc_feature    complement(2825..2844)
                     /label="EBV_rev_primer"
                     /translation="MLSALKSRSKLLPYMGMNCSPIGFSFTENFLRNWVFVLVCVLYI
                     AMKSVLPWVLSVAACDSIWGLLIFDTSAGYLYAVMVTVLLHIFSPTTHCMPKHTYVHV
                     FIVGLLVMSSDIKTLQLNNIHISLRLLLSALQIGYSGTFVQHTSSSRGITSRRKHT*"
     CDS             complement(2958..3587)
                     /label="ORF frame 1"
                     /translation="MLVRGAEPMEKRQQRGLFTVPGLLLAGKSRERSLKLSLMVVIYL
                     PGQHGLQRGHPDAAGSEKNHNGEGHPASRRERQQDVAQRVGRHAGDNGLLLAETFGGG
                     TSDEGLSEGVQDSEYRKRQADHRRAPAKAVLAENDPERCRHLSYELHDKEDSHKCGDD
                     SHAPRPPEGADWVEGSQGHRSTDHNQPYHICRGFTCFKKPPTPPPEPET*"
     CDS             3036..3638
                     /label="ORF frame 3"
                     /translation="MISRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIMQLVGQVPA
                     ALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAIIAGMAADA
                     LGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSLAALTSQ
                     QKARNRKKAALLAFFHRLRPPDEHHKNRRSSQRWRNPTGL*"
     rep_origin      complement(3542..4161)
                     /label="pBR322_origin"
                     /translation="MISRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIMQLVGQVPA
                     ALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAIIAGMAADA
                     LGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSLAALTSQ
                     QKARNRKKAALLAFFHRLRPPDEHHKNRRSSQRWRNPTGL*"
     gene            complement(4316..5176)
                     /label="Ampicillin"
                     /gene="Ampicillin"
                     /translation="MISRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIMQLVGQVPA
                     ALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAIIAGMAADA
                     LGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSLAALTSQ
                     QKARNRKKAALLAFFHRLRPPDEHHKNRRSSQRWRNPTGL*"
     CDS             complement(4316..5176)
                     /label="ORF frame 2"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW*"
     promoter        complement(5218..5246)
                     /label="AmpR_promoter"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW*"
ORIGIN
    1 CTTCAAGGAA TTAATTCTAT ATTCTAAAAA CACAAATGAT ACTTCTAAAA AAAATCATGA
   61 ATGGCATCAA CTCTGAATCA AATCTTTGCA GATGCACCTA CTTCTCATTT CCACTGTCAC
  121 ATCATTTTTC CAGATCTCGC TGCCTGTTAT GTGGCCCACA AACCAAGACA CGTTTTATGG
  181 CCATTAAAGC TGGCTGATCG TCGCCAAACA CCAAATACAT ATCAATATGT ACATTCGAGA
  241 AAGAAGCGAT CAAAGAAGCG TCTTCGGGCG AGTAGGAGAA TGCGGAGGAG AAGGAGAACG
  301 AGCTGATCTA GTATCTCTCC ACAATCCAAT GCCAACTGAC CAACTGGCCA TATTCGGAGC
  361 AATTTGAAGC CAATTTCCAT CGCCTGGCGA TCGCTCCATT CTTGGCTATA TGTTTTTCAC
  421 CGTTCCCGGG GCCATTTTCA AAGACTCGTC GGTAAGATAA GATTGTGTCA CTCGCTGTCT
  481 CTCTTCATTT GTCGAAGAAT GCTGAGGAAT TTCGCGATGA CGTCGGCGAG TATTTTGAAG
  541 AATGAGAATA ATTTGTATTT ATACGAAAAT CAGTTAGTGG AATTTTCTAC AAAAACATGT
  601 TATCTATAGA TAATTTTGTT GCAAAATATG TTGACTATGA CAAAGATTGT ATGTATATAC
  661 CTTTAATGTA TTCTCATTTT CTTATGTATT TATAATGGCA ATGATGATAC TGATGATATT
  721 TTAAGATGAT GCCAGACCAC AGGCTGATTT CTGCGTCTTT TGCCGAACGC AGTGCATGTG
  781 CGGTTGTTGT TTTTTGGAAT AGTTTCAATT TTCGGACTGT CCGCTTTGAT TTCAGTTTCT
  841 TGGCTTATTC AAAAAGCAAA GTAAAGCCAA AAAAGCGAGA TGGCAATACC AAATGCGGCA
  901 AAACGGTAGT GGAAGGAAAG GGGTGCGGGG CAGCGGAAGG AAGGGTGGGG CGGGGCGTGG
  961 CGGGGTCTGT GGCTGGGCGC GACGTCACCG ACGTTGGAGC CACTCCTTTG ACCATGTGTG
 1021 CGTGTGTGTA TTATTCGTGT CTCGCCACTC GCCGGTTGTT TTTTTCTTTT TATCTCGCTC
 1081 TCTCTAGCGC CATCTCGTAC GCATGCTCAA CGCACCGCAT GTTGCCGTGT CCTTTATGCG
 1141 TCATTTTGGC TCGAAATAGG CAATTATTTA AACAAAGATT AGTCAACGAA AACGCTAAAA
 1201 TAAATAAGTC TACAATATGG TTACTTATTG CCATGTGTGT GCAGCCAACG ATAGCAACAA
 1261 AAGCAACAAC ACAGTGGCTT TCCCTCTTTC ACTTTTTGTT TGCAAGCGCG TGCGAGCAAG
 1321 ACGGCACGAC CGGCAAACGC AATTACGCTG ACAAAGAGCA GACGAAGTTT TGGCCGAAAA
 1381 ACATCAAGGC GCCTGATACG AATGCATTTG CAATAACAAT TGCGATATTT AATATTGTTT
 1441 ATGAAGCTGT TTGACTTCAA AACACACAAA AAAAAAAATA AAACAAATTA TTTGAAAGAG
 1501 AATTAGGAAT CGGACAGCTT ATCGTTACGG GCTAACAGCA CACCGAGACG AAATAGCTTA
 1561 CCTGACGTCA CAGCCTCTGG AAGAACTGCC GCCAAGCAGA GAGAGAGAGA AAAAGAGGGA
 1621 GAGCAGCTTA GACCGCATGT GCTTGTGTGT GAGGCGTCTC TCTCTTCGTC TCCTGTTTGC
 1681 GCAAACGCAT AGACTGCACT GAGAAAATCG ATTACCTATT TTTTATGAAT GAATATTTGC
 1741 ACTATTACTA TTCAAAACTA TTAAGATAGC AATCACATTC AATAGCCAAA TACTATACCA
 1801 CCTGAGCGAT GCAACGAAAT GATCAATTTG AGCAAAAATG CTGCATATTT AGGACGGCAT
 1861 CATTATAGAA ATGCTTCTTG CTGTGTACTT TTCTCTCGTC TGGCAGCTGT TTCGCCGTTA
 1921 TTGTTAAAAC CGGCTTAAGT TAGGTGTGTT TTCTACGACT AGTGATGCCC CTACTAGAAG
 1981 ATGTGTGTTG CACAAATGTC CCTGAATAAC CAATTTGAAG TGCAGATAGC AGTAAACGTA
 2041 AGCTAATATG AATATTATTT AACTGTAATG TTTTAATATC GCTGGACATT ACTAATAAAC
 2101 CCACTATAAA CACATGTACA TATGTATGTT TTGGCATACA ATGAGTAGTT GGGGAAAAAA
 2161 TGTGTAAAAG CACCGTGACC ATCACAGCAT AAAGATAACC AGCTGAAGTA TCGAATATGA
 2221 GTAACCCCCA AATTGAATCA CATGCCGCAA CTGATAGGAC CCATGGAAGT ACACTCTTCA
 2281 TGGCGATATA CAAGACACAC ACAAGCACGA ACACCCAGTT GCGGAGGAAA TTCTCCGTAA
 2341 ATGAAAACCC AATCGGCGAA CAATTCATAC CCATATATGG TAAAAGTTTT GAACGCGACT
 2401 TGAGAGCGGA GAGCATTGCG GCTGATAAGG TTTTAGCGCT AAGCGGGCTT TATAAAACGG
 2461 GCTGCGGGAC CAGTTTTCAT ATCACTACCG TTTGAGTTCT TGTGCTGTGT GGATACTCCT
 2521 CCCGACACAA AGCCGCTCCA TCAGCCAGCA GTCGTCTAAT CCAGAGACCC CGGATCGGGG
 2581 TACCTACTAG TCCAGTGTGG TGGAATTCTG CAGATATCCA GCACAGTGGC GGCCGCTCGA
 2641 GTCTAGAGGG CCCTTCGAAG GTAAGCCTAT CCCTAACCCT CTCCTCGGTC TCGATTCTAC
 2701 GCGTACCGGT CATCATCACC ATCACCATTG AGTTTAAACC CGCTGATCAG CCTCGACTGT
 2761 GCCTTCTAAG GCCTGAGCTC GCTGATCAGC CTCGATCGAG GATCCAGACA TGATAAGATA
 2821 CATTGATGAG TTTGGACAAA CCACAACTAG AATGCAGTGA AAAAAATGCT TTATTTGTGA
 2881 AATTTGTGAT GCTATTGCTT TATTTGTAAC CATTATAAGC TGCAATAAAC AAGTTAACAA
 2941 CAACAATTGC ATTCATTTTA TGTTTCAGGT TCAGGGGGAG GTGTGGGAGG TTTTTTAAAG
 3001 CAAGTAAAAC CTCTACAAAT GTGGTATGGC TGATTATGAT CAGTCGACCG ATGCCCTTGA
 3061 GAGCCTTCAA CCCAGTCAGC TCCTTCCGGT GGGCGCGGGG CATGACTATC GTCGCCGCAC
 3121 TTATGACTGT CTTCTTTATC ATGCAACTCG TAGGACAGGT GCCGGCAGCG CTCTGGGTCA
 3181 TTTTCGGCGA GGACCGCTTT CGCTGGAGCG CGACGATGAT CGGCCTGTCG CTTGCGGTAT
 3241 TCGGAATCTT GCACGCCCTC GCTCAAGCCT TCGTCACTGG TCCCGCCACC AAACGTTTCG
 3301 GCGAGAAGCA GGCCATTATC GCCGGCATGG CGGCCGACGC GCTGGGCTAC GTCTTGCTGG
 3361 CGTTCGCGAC GCGAGGCTGG ATGGCCTTCC CCATTATGAT TCTTCTCGCT TCCGGCGGCA
 3421 TCGGGATGCC CGCGTTGCAG GCCATGCTGT CCAGGCAGGT AGATGACGAC CATCAGGGAC
 3481 AGCTTCAAGG ATCGCTCGCG GCTCTTACCA GCCAGCAAAA GGCCAGGAAC CGTAAAAAGG
 3541 CCGCGTTGCT GGCGTTTTTC CATAGGCTCC GCCCCCCTGA CGAGCATCAC AAAAATCGAC
 3601 GCTCAAGTCA GAGGTGGCGA AACCCGACAG GACTATAAAG ATACCAGGCG TTTCCCCCTG
 3661 GAAGCTCCCT CGTGCGCTCT CCTGTTCCGA CCCTGCCGCT TACCGGATAC CTGTCCGCCT
 3721 TTCTCCCTTC GGGAAGCGTG GCGCTTTCTC AATGCTCACG CTGTAGGTAT CTCAGTTCGG
 3781 TGTAGGTCGT TCGCTCCAAG CTGGGCTGTG TGCACGAACC CCCCGTTCAG CCCGACCGCT
 3841 GCGCCTTATC CGGTAACTAT CGTCTTGAGT CCAACCCGGT AAGACACGAC TTATCGCCAC
 3901 TGGCAGCAGC CACTGGTAAC AGGATTAGCA GAGCGAGGTA TGTAGGCGGT GCTACAGAGT
 3961 TCTTGAAGTG GTGGCCTAAC TACGGCTACA CTAGAAGGAC AGTATTTGGT ATCTGCGCTC
 4021 TGCTGAAGCC AGTTACCTTC GGAAAAAGAG TTGGTAGCTC TTGATCCGGC AAACAAACCA
 4081 CCGCTGGTAG CGGTGGTTTT TTTGTTTGCA AGCAGCAGAT TACGCGCAGA AAAAAAGGAT
 4141 CTCAAGAAGA TCCTTTGATC TTTTCTACGG GGTCTGACGC TCAGTGGAAC GAAAACTCAC
 4201 GTTAAGGGAT TTTGGTCATG AGATTATCAA AAAGGATCTT CACCTAGATC CTTTTAAATT
 4261 AAAAATGAAG TTTTAAATCA ATCTAAAGTA TATATGAGTA AACTTGGTCT GACAGTTACC
 4321 AATGCTTAAT CAGTGAGGCA CCTATCTCAG CGATCTGTCT ATTTCGTTCA TCCATAGTTG
 4381 CCTGACTCCC CGTCGTGTAG ATAACTACGA TACGGGAGGG CTTACCATCT GGCCCCAGTG
 4441 CTGCAATGAT ACCGCGAGAC CCACGCTCAC CGGCTCCAGA TTTATCAGCA ATAAACCAGC
 4501 CAGCCGGAAG GGCCGAGCGC AGAAGTGGTC CTGCAACTTT ATCCGCCTCC ATCCAGTCTA
 4561 TTAATTGTTG CCGGGAAGCT AGAGTAAGTA GTTCGCCAGT TAATAGTTTG CGCAACGTTG
 4621 TTGCCATTGC TGCAGGCATC GTGGTGTCAC GCTCGTCGTT TGGTATGGCT TCATTCAGCT
 4681 CCGGTTCCCA ACGATCAAGG CGAGTTACAT GATCCCCCAT GTTGTGCAAA AAAGCGGTTA
 4741 GCTCCTTCGG TCCTCCGATC GTTGTCAGAA GTAAGTTGGC CGCAGTGTTA TCACTCATGG
 4801 TTATGGCAGC ACTGCATAAT TCTCTTACTG TCATGCCATC CGTAAGATGC TTTTCTGTGA
 4861 CTGGTGAGTA CTCAACCAAG TCATTCTGAG AATAGTGTAT GCGGCGACCG AGTTGCTCTT
 4921 GCCCGGCGTC AACACGGGAT AATACCGCGC CACATAGCAG AACTTTAAAA GTGCTCATCA
 4981 TTGGAAAACG TTCTTCGGGG CGAAAACTCT CAAGGATCTT ACCGCTGTTG AGATCCAGTT
 5041 CGATGTAACC CACTCGTGCA CCCAACTGAT CTTCAGCATC TTTTACTTTC ACCAGCGTTT
 5101 CTGGGTGAGC AAAAACAGGA AGGCAAAATG CCGCAAAAAA GGGAATAAGG GCGACACGGA
 5161 AATGTTGAAT ACTCATACTC TTCCTTTTTC AATATTATTG AAGCATTTAT CAGGGTTATT
 5221 GTCTCATGAG CGGATACATA TTTGAATGTA TTTAGAAAAA TAAACAAATA GGGGTTCCGC
 5281 GCACATTTCC CCGAAAAGTG CCACCTGACG TCTAAGAAAC CATTATTATC ATGACATTAA
 5341 CCTATAAAAA TAGGCGTATC ACGAGGCCCT TTCGT