pAdVAntage哺乳辅助质粒

基本信息

启动子:	Lac
复制子:	pUC ori
质粒分类:	哺乳细胞，蛋白过表达载体
质粒大小:	4392bp
原核抗性:	Amp
克隆菌株:	DH5α
培养条件:	37℃，有氧 LB
表达宿主:	哺乳细胞
诱导方式:	无须诱导，瞬时表达
5'测序引物:	M13R:CAGGAAACAGCTATGACC
3'测序引物:	根据序列设计引物

质粒简介

       The use of pAdVAntage vectors were transfected into mammalian cells can promote the initiation of translation to enhance the instantaneous protein expression in many cell types.
       Transfection of mammalian cells with expression vector often leads to the expression of the target protein. The double stranded RNA (dsRNA), which is produced in the transfection process, is thought to activate the dsRNA- activation inhibitor (DAI), which is one of the many kinds of enzymes involved in the host cell antiviral defense system. DAI phosphorylation of translation initiation factor eIF-2, termination of translation and generation of protein. However, with the pAdVAntage vector were transfected into DAI in the process of translation inhibition by RNA polymerase III generated adeno-associated virus type I (RNA VAI RNA) offset. RNA DAI combined with VAI to prevent its activation, so as to complete the translation and protein expression.

质粒图谱

质粒序列

LOCUS       Exported                4392 bp ds-DNA     circular SYN 15-JUL-2016
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pAdVAntage
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4392)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, August 30, 2016 from SnapGene Viewer 3.1.4
FEATURES             Location/Qualifiers
     source          1..4392
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(1743..1759)
                     /note="M13 rev"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1767..1783
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="lac operator"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(1791..1821)
                     /note="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    1836..1857
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding site"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(2145..2733)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2904..3764)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3765..3869)
                     /gene="bla"
                     /note="AmpR promoter"
     primer_bind     4343..4359
                     /note="M13 fwd"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
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 gctttctcat
     2341 agctcacgct 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 agaagaacag 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