P0383 pSilent-1

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P0383 pSilent-1 20μl "启动子: TrpC
复制子: pUC ori，F1 ori
终止子: TRPC
质粒分类: 丝状真菌载体；RNAi基因沉默干扰质粒
质粒大小: 7056bp
原核抗性: 氨苄青霉素Ampicillin
筛选标记: 潮霉素Hygromycin
克隆菌株: DH5α
培养条件: 37℃，有氧，LB
表达宿主: 丝状真菌
5'测序引物: 根据序列设计
3'测序引物: 根据序列设计
质粒简介
pSilent-1质粒是一种用于真菌RNAi基因沉默干扰的载体，其多克隆位点区域的酶切位点有XhoI,SnaBI,HindIII （MCS1）和BglII,SphI,StuI,KpnI,ApaI（MCS2）等。

质粒图谱

质粒序列
Gene silencing vector pSilent-1 DNA, complete sequence
GenBank: AB303070.2
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 LOCUS       AB303070                7056 bp    DNA     circular SYN 11-DEC-2008
DEFINITION  Gene silencing vector pSilent-1 DNA, complete sequence.
ACCESSION   AB303070
VERSION     AB303070.2  GI:217272796
KEYWORDS    .
SOURCE      Gene silencing vector pSilent-1
  ORGANISM  Gene silencing vector pSilent-1
            other sequences; artificial sequences; vectors.
REFERENCE   1
  AUTHORS   Nakayashiki,H., Hanada,S., Nguyen,B.Q., Kadotani,N., Tosa,Y. and
            Mayama,S.
  TITLE     RNA silencing as a tool for exploring gene function in ascomycete
            fungi
  JOURNAL   Fungal Genet. Biol. 42 (4), 275-283 (2005)
   PUBMED   15749047
REFERENCE   2  (bases 1 to 7056)
  AUTHORS   Nakayashiki,H.
  TITLE     Direct Submission
  JOURNAL   Submitted (09-MAY-2007) Contact:Hitoshi Nakayashiki Kobe
            University, Graduate school of agricultural science; 1 Rokko-dai
            Nada, Kobe 657-8501, Japan
COMMENT     On Dec 10, 2008 this sequence version replaced gi:146455412.
FEATURES             Location/Qualifiers
     source          1..7056
                     /organism=""Gene silencing vector pSilent-1""
                     /mol_type=""other DNA""
                     /db_xref=""taxon:442632""
                     /note=""Gene silencing vector for ascomysete fungi""
     CDS             129..989
                     /note=""unnamed protein product; Ampicillin resistance
                     gene""
                     /codon_start=1
                     /transl_table=11
                     /protein_id=""BAF62257.1""
                     /db_xref=""GI:146455413""
                     /translation=""MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW""
     regulatory      2226..2592
                     /regulatory_class=""promoter""
                     /note=""Aspergillus nidulans TrpC gene""
     CDS             2593..3618
                     /note=""unnamed protein product; Hygromycin resistance
                     gene""
                     /codon_start=1
                     /transl_table=11
                     /protein_id=""BAF62258.1""
                     /db_xref=""GI:146455414""
                     /translation=""MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQD
                     LPETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVY
                     HWQTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWS
                     EAMFGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQ
                     SLVDGNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPS
                     TRPRAKE""
     regulatory      3638..4123
                     /regulatory_class=""terminator""
                     /note=""Aspergillus nidulans TrpC gene""
     regulatory      4162..5447
                     /regulatory_class=""promoter""
                     /note=""Aspergillus nidulans TrpC gene""
     misc_feature    5451..5476
                     /note=""multi cloning site""
     intron          5491..5624
                     /note=""Magnaporthe grisea cutinase gene""
     misc_feature    5637..5673
                     /note=""multi cloning site""
     regulatory      5675..6404
                     /regulatory_class=""terminator""
                     /note=""Aspergillus nidulans TrpC gene""
ORIGIN    
        1 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
       61 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
      121 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
      181 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
      241 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
      301 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
      361 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
      421 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
      481 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
      541 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
      601 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
      661 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
      721 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
      781 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
      841 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
      901 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
      961 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     1021 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     1081 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     1141 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     1201 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     1261 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
     1321 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     1381 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     1441 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     1501 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     1561 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     1621 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     1681 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     1741 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     1801 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     1861 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     1921 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     1981 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
     2041 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
     2101 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg
     2161 ccaagcgcgc aattaaccct cactaaaggg aacaaaagct ggagctccac cgcggtggcg
     2221 gccgcgacgt taactgatat tgaaggagca ctttttgggc ttggctggag ctagtggagg
     2281 tcaacaatga atgcctattt tggtttagtc gtccaggcgg tgagcacaaa atttgtgtcg
     2341 tttgacaaga tggttcattt aggcaactgg tcagatcagc cccacttgta gcagtagcgg
     2401 cggcgctcga agtgtgactc ttattagcag acaggaacga ggacattatt atcatctgct
     2461 gcttggtgca cgataacttg gtgcgtttgt caagcaaggt aagtgaacga cccggtcata
     2521 ccttcttaag ttcgcccttc ctccctttat ttcagattca atctgactta cctattctac
     2581 ccaagcatcg atatgaaaaa gcctgaactc accgcgacgt ctgtcgagaa gtttctgatc
     2641 gaaaagttcg acagcgtctc cgacctgatg cagctctcgg agggcgaaga atctcgtgct
     2701 ttcagcttcg atgtaggagg gcgtggatat gtcctgcggg taaatagctg cgccgatggt
     2761 ttctacaaag atcgttatgt ttatcggcac tttgcatcgg ccgcgctccc gattccggaa
     2821 gtgcttgaca ttggggaatt cagcgagagc ctgacctatt gcatctcccg ccgtgcacag
     2881 ggtgtcacgt tgcaagacct gcctgaaacc gaactgcccg ctgttctgca gccggtcgcg
     2941 gaggccatgg atgcgatcgc tgcggccgat cttagccaga cgagcgggtt cggcccattc
     3001 ggaccgcaag gaatcggtca atacactaca tggcgtgatt tcatatgcgc gattgctgat
     3061 ccccatgtgt atcactggca aactgtgatg gacgacaccg tcagtgcgtc cgtcgcgcag
     3121 gctctcgatg agctgatgct ttgggccgag gactgccccg aagtccggca cctcgtgcac
     3181 gcggatttcg gctccaacaa tgtcctgacg gacaatggcc gcataacagc ggtcattgac
     3241 tggagcgagg cgatgttcgg ggattcccaa tacgaggtcg ccaacatctt cttctggagg
     3301 ccgtggttgg cttgtatgga gcagcagacg cgctacttcg agcggaggca tccggagctt
     3361 gcaggatcgc cgcggctccg ggcgtatatg ctccgcattg gtcttgacca actctatcag
     3421 agcttggttg acggcaattt cgatgatgca gcttgggcgc agggtcgatg cgacgcaatc
     3481 gtccgatccg gagccgggac tgtcgggcgt acacaaatcg cccgcagaag cgcggccgtc
     3541 tggaccgatg gctgtgtaga agtactcgcc gatagtggaa accgacgccc cagcactcgt
     3601 ccgagggcaa aggaatagag tagatgccga ccggatcgat ccacttaacg ttactgaaat
     3661 catcaaacag cttgacgaat ctggatataa gatcgttggt gtcgatgtca gctccggagt
     3721 tgagacaaat ggtgttcagg atctcgataa gatacgttca tttgtccaag cagcaaagag
     3781 tgccttctag tgatttaata gctccatgtc aacaagaata aaacgcgttt cgggtttacc
     3841 tcttccagat acagctcatc tgcaatgcat taatgcattg gacctcgcaa ccctagtacg
     3901 cccttcaggc tccggcgaag cagaagaata gcttagcaga gtctattttc attttcggga
     3961 gacgagatca agcagatcaa cggtcgtcaa gagacctacg agactgagga atccgctctt
     4021 ggctccacgc gactatatat ttgtctctaa ttgtactttg acatgctcct cttctttact
     4081 ctgatagctt gactatgaaa attccgtcac cagcccctgg gttgcggccg ctctagaact
     4141 agtggatccc ccgggctgca ggaattcatg ccagttgttc ccagtgatct tcgtttcgaa
     4201 gatggacact cccaatttgt gcaagttatt cggcctacct ggctgtggcc gaggcgcgtt
     4261 atcatgaccg tcgctgttca aagataaggc gagaagtttg cgggctgtct tgacgatatg
     4321 gcttcgttca gacagatata gttcccggag tcgcaggcgt ctattcttct ccgaaacaaa
     4381 ctcggctgca ctgtttccat caccgggtct ggcgttgagg atgtcagcga aactcggccc
     4441 ggcaagtgac acccgaaaag tatcgactcc ggctgcccgt ttcaagctag tggcttcctc
     4501 atcagcgagt cggccaagga gacgtgaagc aggacgggtt tgccattcca agaccgtgat
     4561 ccgaagcgcg ttgatttcat caatcccagc cttttcgctc aaccaaagag catcggcttt
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