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.
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
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