pacAd5 CMVK- NpA

$228.00

  • Model: PVT11032
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pacAd5 CMVK- NpA

PVT11032    2ug

 

pacAd5 CMVK- NpA Information

Promoter: CMV promoter

Replicon: pBR322 ori

Terminator: SV40 poly (A) signal

Plasmid size: 6215bp

Prokaryotic resistance: ampicillin

Clone strain: Escherichia coli Stbl3

Culture conditions: 37 C, aerobic, LB

 

pacAd5 CMVK- NpA Description

PacAd5 CMVK-NpA plasmid is an adenovirus plasmid, and CMV promoter drives the expression of target genes.Recombinant adenoviruses have tremendous potential in both research and therapeutic applications. There are numerous advantages they provide when introducing genetic material into host cells. The permissive host cell range is very wide. The virus has been used to infect many mammalian cell types (both replicative and non-replicative) for high expression of the recombinant protein. Recombinant adenoviruses are especially useful for gene transfer and protein expression in cell lines that have low transfection efficiency with liposome. After entering cells, the virus remains epichromosomal (i.e. does not integrate into the host chromosome so does not activate or inactivate host genes). Recently, recombinant adenoviruses have been used to deliver RNAi into cells. Two methods have traditionally been used to generate recombinant adenoviruses. The first involves homologous recombination of a shuttle vector containing gene of interest and an adenoviral backbone plasmid vector (restricted in E1/E3) in an adenovirus packaging cell line. The isolation of recombinant adenovirus by this method involves performing multiple plaque isolations to avoid wild-type virus and is extremely laborious and time consuming. The second method, pAdEasy system, employs the homologous recombination machinery in E. coli, a recombinant adenovirus is produced by a doublerecombination event between cotransformed adenoviral backbone plasmid vector and a shuttle vector carrying the gene of interest. For the pAdEasy method, the system is high fidelity, but inefficient and requires the screening of many bacterial colonies. This results in a significant time commitment even before transfection of recombinant DNA into E1-expressing cells such as HEK293 cells.

 

pacAd5 CMVK- NpA Multiple cloning site

pacAd5-CMVK-NpA-vector

 

 

pacAd5 CMVK- NpA Sequence

LOCUS       Exported                6215 bp ds-DNA     circular SYN 05-NOV-2017
DEFINITION  synthetic circular DNA
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6215)
            
FEATURES             Location/Qualifiers
     source          1..6215
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_signal     1..353
                     /label=Ad5
     repeat_region   16..118
                     /label=ITR
                     /note="inverted terminal repeat of human adenovirus 
                     serotype 5"
     enhancer        385..688
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        689..892
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     misc_feature    913..1002
                     /label=MCS
     polyA_signal    1014..1148
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    1437..3901
                     /label=AD5
     rep_origin      complement(4388..4976)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5147..6007)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6008..6112)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aattaattaa gctagcatca tcaataatat accttatttt ggattgaagc caatatgata
       61 atgagggggt ggagtttgtg acgtggcgcg gggcgtggga acggggcggg tgacgtagta
      121 gtgtggcgga agtgtgatgt tgcaagtgtg gcggaacaca tgtaagcgac ggatgtggca
      181 aaagtgacgt ttttggtgtg cgccggtgta cacaggaagt gacaattttc gcgcggtttt
      241 aggcggatgt tgtagtaaat ttgggcgtaa ccgagtaaga tttggccatt ttcgcgggaa
      301 aactgaataa gaggaagtga aatctgaata attttgtgtt actcatagcg cgtaatattt
      361 gtctagggag atcagcctgc aggtcgttac ataacttacg gtaaatggcc cgcctggctg
      421 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
      481 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
      541 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
      601 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
      661 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg
      721 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
      781 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt
      841 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctcgtttagt
      901 gaaccgtcag ttggtaccgt ttaaactcga ggtcgacggt atcgataagc ttgatatcga
      961 attcctgcag cccgggggat ccactagttc tagagcggcc gccacagcgg ggagatccag
     1021 acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat
     1081 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
     1141 aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg
     1201 aggtttttta aagcaagtaa aacctctaca aatgtggtat ggctgattat gatccggctg
     1261 cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     1321 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     1381 tgttggcggg tgtcggggcg cagccatgag gtcgactcta gtccccgcgg tggcagatct
     1441 ggaaggtgct gaggtacgat gagacccgca ccaggtgcag accctgcgag tgtggcggta
     1501 aacatattag gaaccagcct gtgatgctgg atgtgaccga ggagctgagg cccgatcact
     1561 tggtgctggc ctgcacccgc gctgagtttg gctctagcga tgaagataca gattgaggta
     1621 ctgaaatgtg tgggcgtggc ttaagggtgg gaaagaatat ataaggtggg ggtcttatgt
     1681 agttttgtat ctgttttgca gcagccgccg ccgccatgag caccaactcg tttgatggaa
     1741 gcattgtgag ctcatatttg acaacgcgca tgcccccatg ggccggggtg cgtcagaatg
     1801 tgatgggctc cagcattgat ggtcgccccg tcctgcccgc aaactctact accttgacct
     1861 acgagaccgt gtctggaacg ccgttggaga ctgcagcctc cgccgccgct tcagccgctg
     1921 cagccaccgc ccgcgggatt gtgactgact ttgctttcct gagcccgctt gcaagcagtg
     1981 cagcttcccg ttcatccgcc cgcgatgaca agttgacggc tcttttggca caattggatt
     2041 ctttgacccg ggaacttaat gtcgtttctc agcagctgtt ggatctgcgc cagcaggttt
     2101 ctgccctgaa ggcttcctcc cctcccaatg cggtttaaaa cataaataaa aaaccagact
     2161 ctgtttggat ttggatcaag caagtgtctt gctgtcttta tttaggggtt ttgcgcgcgc
     2221 ggtaggcccg ggaccagcgg tctcggtcgt tgagggtcct gtgtattttt tccaggacgt
     2281 ggtaaaggtg actctggatg ttcagataca tgggcataag cccgtctctg gggtggaggt
     2341 agcaccactg cagagcttca tgctgcgggg tggtgttgta gatgatccag tcgtagcagg
     2401 agcgctgggc gtggtgccta aaaatgtctt tcagtagcaa gctgattgcc aggggcaggc
     2461 ccttggtgta agtgtttaca aagcggttaa gctgggatgg gtgcatacgt ggggatatga
     2521 gatgcatctt ggactgtatt tttaggttgg ctatgttccc agccatatcc ctccggggat
     2581 tcatgttgtg cagaaccacc agcacagtgt atccggtgca cttgggaaat ttgtcatgta
     2641 gcttagaagg aaatgcgtgg aagaacttgg agacgccctt gtgacctcca agattttcca
     2701 tgcattcgtc cataatgatg gcaatgggcc cacgggcggc ggcctgggcg aagatatttc
     2761 tgggatcact aacgtcatag ttgtgttcca ggatgagatc gtcataggcc atttttacaa
     2821 agcgcgggcg gagggtgcca gactgcggta taatggttcc atccggccca ggggcgtagt
     2881 taccctcaca gatttgcatt tcccacgctt tgagttcaga tggggggatc atgtctacct
     2941 gcggggcgat gaagaaaacg gtttccgggg taggggagat cagctgggaa gaaagcaggt
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