Co-transfection of mammalian cells with the pAdVAntage™ Vector enhances transient protein expression in a variety of cell types by increasing translation initiation. Transfection of mammalian cells with an expression vector often results in suboptimal expression of the protein of interest. Double-stranded RNA (dsRNA) generated during transfection is thought to activate the dsRNA-activated inhibitor (DAI), one of several enzymes involved in the host cell's antiviral defense system. Activated DAI phosphorylates the translation initiation factor eIF-2, halting translation and therefore protein production (1, reviewed in reference 2; see Figure 1). However, inhibition of translation by DAI can be overcome with the adenoviral VAI RNA (Virus Associated I RNA) that is produced by RNA polymerase III following co-transfection with the pAdVAntage™ Vector. The VAI RNA binds to DAI, preventing its activation, thereby allowing translation and protein expression (3,4, reviewed in reference 2; see Figure 1). The pAdVAntage™ Vector contains base pairs 9,831 to 11,555 of the adenovirus type 2 genome on a 1724bp Sal I-Hind III fragment which encodes the virus-associated RNA genes, VAI and VAII.

 back Return to this vector's summary.


LOCUS       CVU47294     4392 bp    DNA             SYN       26-FEB-1996
DEFINITION  Cloning vector pAdVantage adenovirus VAI RNA gene, adenovirus
            VAII RNA gene, and beta-lactamase (AmpR) gene, complete cds.
ACCESSION   U47294
NID         g1200457
KEYWORDS    .
SOURCE      Cloning vector pAdVantage.
  ORGANISM  Cloning vector pAdVantage
            artificial sequence; cloning vectors.
REFERENCE   1  (bases 1 to 4392)
  AUTHORS   Groskreutz,D.J. and Schenborn,E.T.
  TITLE     Increased Gene Expression in Mammalian cell lines~using
            pAdVAntage(TM) DNA as a Co-transfectant
  JOURNAL   Promega Notes 48, 8-12 (1994)
REFERENCE   2  (bases 1 to 4392)
  AUTHORS   Groskreutz,D.J. and Schenborn,E.T.
  TITLE     Direct Submission
  JOURNAL   Submitted (26-JAN-1996) D.J. Groskreutz, R&D, Promega Corporation,
            5445 East Cheryl Parkway, Madison, WI 53711, USA
COMMENT
            NCBI gi: 1200457
FEATURES               Location/Qualifiers
     source          1..4392
                     /organism="Cloning vector pAdVantage"
     misc_feature    1..1725
                     /note="adenovirus DNA insert"
     misc_RNA        780..939
                     /product="adenovirus VAI RNA"
     misc_RNA        1036..1198
                     /product="adenovirus VAII RNA"
     CDS             complement(2904..3764)
                     /gene="AmpR"
                     /note="ampicillin resistance;  NCBI gi: 1200458"
                     /codon_start=1
                     /product="beta-lactamase"
                     /db_xref="PID:g1200458"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW"
BASE COUNT      972 a   1177 c   1266 g    977 t
ORIGIN      
        1 gtcgacagaa gcaccatgtc cttgggtccg gcctgctgaa tgcgcaggcg gtcggccatg
       61 ccccaggctt cgttttgaca tcggcgcagg tctttgtagt agtcttgcat gagcctttct
      121 accggcactt cttcttctcc ttcctcttgt cctgcatctc ttgcatctat cgctacggcg
      181 gcggcggagt ttggccgtag gtggcgccct cttcctccca tgcgtgtgac cccgaagccc
      241 ctcatcggct gaagcagggc caggtcggcg acaacgcgct cggctaatat ggcctgctgc
      301 acctgcgtga gggtagactg gaagtcatcc atgtccacaa agcggtggta tgcgcccgtg
      361 ttgatggtgt aagtgcagtt ggccataacg gaccagttaa cggtctggtg acccggctgc
      421 gagagctcgg tgtacctgag acgcgagtaa gcccttgagt caaagacgta gtcgttgcaa
      481 gtccgcacca ggtactgata tcccaccaaa aagtgcggcg gcggctggcg gtagaggggc
      541 cagcgtaggg tggccggggc tccgggggcg aggtcttcca acataaggcg atgatatccg
      601 tagatgtacc tggacatcca ggtgatgccg gcggcggtgg tggaggcgcg cggaaagtcg
      661 cggacgcggt tccagatgtt gcgcagcggc aaaaagtgct ccatggtcgg gacgctctgg
      721 ccggtgaggc gtgcgcagtc gttgacgctc tagaccgtgc aaaaggagag cctgtaagcg
      781 ggcactcttc cgtggtctgg tggataaatt cgcaagggta tcatggcgga cgaccggggt
      841 tcgaaccccg gatccggccg tccgccgtga tccatgcggt taccgcccgc gtgtcgaacc
      901 caggtgtgcg acgtcagaca acgggggagc gctccttttg gcttccttcc aggcgcggcg
      961 gctgctgcgc tagctttttt ggccactggc cgcgcgcggc gtaagcggtt aggctggaaa
     1021 gcgaaagcat taagtggctc gctccctgta gccggagggt tattttccaa gggttgagtc
     1081 gcaggacccc cggttcgagt ctcgggccgg ccggactgcg gcgaacgggg gtttgcctcc
     1141 ccgtcatgca agaccccgct tgcaaattcc tccggaaaca gggacgagcc ccttttttgc
     1201 ttttcccaga tgcatccggt gctgcggcag atgcgccccc ctcctcagca gcggcaagag
     1261 caagagcagc ggcagacatg cagggcaccc tccccttctc ctaccgcgtc aggaggggca
     1321 acatccgcgg ctgacgcggc ggcagatggt gattacgaac ccccgcggcg ccgggcccgg
     1381 cactacctgg acttggagga gggcgagggc ctggcgcggc taggagcgcc ctctcctgag
     1441 cgacacccaa gggtgcagct gaagcgtgac acgcgcgagg cgtacgtgcc gcggcagaac
     1501 ctgtttcgcg accgcgaggg agaggagccc gaggagatgc gggatcgaaa gttccacgca
     1561 gggcgcgagt tgcggcatgg cctgaaccgc gagcggttgc tgcgcgagga ggactttgag
     1621 cccgacgcgc ggaccgggat tagtcccgcg cgcgcacacg tggcggccgc cgacctggta
     1681 accgcgtacg agcagacggt gaaccaggag attaactttc aaaaaagctt ggcgtaatca
     1741 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     1801 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     1861 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     1921 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     1981 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     2041 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     2101 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     2161 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     2221 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     2281 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa
     2341 tgctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     2401 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     2461 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     2521 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     2581 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     2641 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     2701 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     2761 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     2821 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     2881 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     2941 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     3001 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     3061 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     3121 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     3181 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     3241 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     3301 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     3361 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     3421 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     3481 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     3541 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     3601 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     3661 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     3721 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     3781 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     3841 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     3901 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt
     3961 cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg
     4021 gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg
     4081 ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga
     4141 gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg
     4201 cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
     4261 ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca
     4321 gggttttccc agtcacgacg ttgtaaaacg acggccagtg aattcgagct cggtacccgg
     4381 ggatcctcta ga
//