• 规格：20μl质粒
• 启动子：CMV IE94 promoter
• 原核抗性：氨苄青霉素Amp
• 克隆菌株：大肠杆菌Stbl3
• 表达宿主：斑马鱼
• 质粒简介：          pCS2-nCas9n-nanos 3’UTR是一个鱼类基因编辑Cas9表达质粒。
• 质粒图谱1： png/1-1P1061I046417.png

质粒序列： 点击查看序列

LOCUS       Exported                8906 bp ds-DNA     circular SYN 29-DEC-2017

DEFINITION  synthetic circular DNA

FEATURES             Location/Qualifiers

     source          1..8906

                     /organism="synthetic DNA construct"

                     /mol_type="other DNA"

     promoter        35..53

                     /label=SP6 promoter

                     /note="promoter for bacteriophage SP6 RNA polymerase"

     regulatory      99..108

                     /regulatory_class="other"

                     /note="vertebrate consensus sequence for strong initiation 

                     of translation (Kozak, 1987)"

     CDS             117..137

                     /codon_start=1

                     /product="nuclear localization signal of SV40 (simian virus

                     40) large T antigen"

                     /label=SV40 NLS

                     /translation="PKKKRKV"

     CDS             144..4244

                     /codon_start=1

                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 

                     pyogenes MGAS15252 Type II CRISPR/Cas system"

                     /label=Cas9

                     /note="generates RNA-guided double strand breaks in DNA"

                     /translation="MDKKYSIGLDIGTNSVGWAVITDDYKVPSKKFKVLGNTDRHSIKK

                     NLIGALLFGSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES

                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLADSTDKADLRLIYLALAHMIK

                     FRGHFLIEGDLNPDNSDVDKLFIQLVQIYNQLFEENPINASRVDAKAILSARLSKSRRL

                     ENLIAQLPGEKRNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ

                     IGDQYADLFLAAKNLSDAILLSDILRVNSEITKAPLSASMIKRYDEHHQDLTLLKALVR

                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL

                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG

                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY

                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD

                     SVEISGVEDRFNASLGAYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDRGMIEER

                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM

                     QLIHDDSLTFKEDIQKAQVSGQGHSLHEQIANLAGSPAIKKGILQTVKIVDELVKVMGH

                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL

                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFIKDDSIDNKVLTRSDKNRGKSDNVPS

                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV

                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN

                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE

                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE

                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT

                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL

                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA

                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD

                     ATLIHQSITGLYETRIDLSQLGGD"

     CDS             4260..4280

                     /codon_start=1

                     /product="nuclear localization signal of SV40 (simian virus

                     40) large T antigen"

                     /label=SV40 NLS

                     /translation="PKKKRKV"

     misc_feature    4317..4932

                     /label=nanos 3'UTR

     polyA_signal    4955..5089

                     /label=SV40 poly(A) signal

                     /note="SV40 polyadenylation signal"

     rep_origin      complement(5643..6231)

                     /direction=LEFT

                     /label=ori

                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 

                     replication"

     CDS             complement(6402..7262)

                     /codon_start=1

                     /gene="bla"

                     /product="beta-lactamase"

                     /label=AmpR

                     /note="confers resistance to ampicillin, carbenicillin, and

                     related antibiotics"

                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI

                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS

                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW

                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA

                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS

                     LIKHW"

     promoter        complement(7263..7367)

                     /gene="bla"

                     /label=AmpR promoter

     rep_origin      complement(7393..7848)

                     /direction=LEFT

                     /label=f1 ori

                     /note="f1 bacteriophage origin of replication; arrow 

                     indicates direction of (+) strand synthesis"

     promoter        7922..8906

                     /label=CMV IE94 promoter

                     /note="enhancer/promoter region of simian cytomegalovirus 

                     major immediate early transcription unit IE94"

ORIGIN

        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag

       61 ctacttgttc tttttgcagg atccgatcca atgtcgacgc caccatggct tctccaccta

      121 agaagaagag aaaggtggga agcatggata agaagtatag catcggcctg gatattggaa

      181 ctaactccgt gggttgggca gtgattacag acgactacaa ggtccctagc aagaaattta

      241 aggtgctggg taacaccgac aggcacagca tcaagaaaaa tctgattgga gccctgctgt

      301 tcggttctgg agagactgcc gaagcaacac gcctgaaaag aacagcaaga aggcgctata

      361 ccagaaggaa gaatagaatc tgttacctgc aggagatttt ctctaacgaa atggctaagg

      421 tggacgattc attctttcat aggctggagg aaagtttcct ggtcgaggaa gataagaaac

      481 acgagcgcca tcctatcttt ggaaacattg tggacgaggt cgcctatcac gaaaaatacc

      541 caaccatcta tcatctgcgc aagaaactgg ctgactctac tgataaagcc gacctgagac

      601 tgatctatct ggctctggcc cacatgatta agttcagggg tcattttctg atcgagggcg

      661 atctgaaccc cgacaattcc gatgtggaca agctgttcat ccagctggtc cagatttaca

      721 atcagctgtt tgaggaaaac cctattaatg cttccagagt ggacgcaaaa gctatcctgt

      781 cagccaggct gtccaagtca cgcagactgg agaacctgat tgcacagctg cccggagaaa

      841 agaggaacgg tctgtttgga aatctgatcg ctctgagtct gggcctgact cctaacttca

      901 aaagcaattt tgatctggct gaggacgcca aactgcagct gtcaaaggac acatatgacg

      961 atgacctgga taacctgctg gcacagatcg gagatcagta cgctgacctg ttcctggctg

     1021 ccaaaaatct gtccgacgca atcctgctgt cagatattct gagagtgaac agcgagatta

     1081 caaaagctcc tctgagtgcc agcatgatca agagatatga cgagcaccat caggatctga

     1141 ccctgctgaa ggctctggtc aggcagcagc tgccagagaa gtacaaggaa attttctttg

     1201 atcagtccaa gaacggctac gccggttata tcgacggagg cgcatcacag gaggaattct

     1261 acaagtttat caaacctatt ctggagaaga tggacggaac tgaggaactg ctggtgaaac

     1321 tgaatagaga ggacctgctg aggaagcagc gcacatttga taacggttcc atcccacacc

     1381 agattcatct gggagagctg cacgctatcc tgaggcgcca ggaagacttc tacccctttc

     1441 tgaaagataa ccgcgagaag atcgaaaaaa ttctgacctt cagaatccct tactatgtgg

     1501 gtccactggc tcgcggaaac agcagatttg cctggatgac tcgcaaatcc gaggaaacca

     1561 ttactccttg gaacttcgag gaagtggtcg ataagggcgc ctctgcacag tccttcatcg

     1621 agagaatgac taattttgac aaaaacctgc ccaatgagaa agtgctgcct aagcactccc

     1681 tgctgtacga gtatttcact gtctataacg aactgacaaa ggtgaaatac gtcaccgagg

     1741 gcatgagaaa gccagccttc ctgtcaggag agcagaagaa agcaatcgtg gatctgctgt

     1801 ttaaaaccaa caggaaagtg actgtcaagc agctgaagga ggactacttc aagaaaattg

     1861 aatgcttcga ttccgtggag atcagcggag tcgaagacag atttaacgca agcctgggcg

     1921 cttaccacga tctgctgaag atcattaagg ataaagactt cctggacaac gaggaaaatg

     1981 aggatatcct ggaagacatt gtgctgacac tgaccctgtt tgaggacaga ggaatgatcg

     2041 aggaaagact gaaaacctat gctcatctgt tcgatgacaa ggtgatgaaa cagctgaaga

     2101 gaaggcgcta cactggctgg ggtagactga gcaggaagct gatcaacggc attagggata

     2161 aacagtcagg aaagacaatc ctggactttc tgaaaagtga tggcttcgcc aaccgcaatt

     2221 ttatgcagct gattcacgat gacagtctga ccttcaaaga ggacatccag aaggctcagg

     2281 tgtctggaca gggccactcc ctgcatgagc agattgcaaa cctggctgga agcccagcca

     2341 tcaagaaagg cattctgcag acagtgaaaa tcgtcgatga gctggtgaaa gtcatgggcc

     2401 ataagcccga aaacatcgtg attgagatgg ctcgcgaaaa tcagacaacc cagaagggtc

     2461 agaagaacag tagagagagg atgaaaagaa tcgaggaagg cattaaggag ctgggtagcc

     2521 agatcctgaa agagcaccca gtggaaaaca cacagctgca gaatgagaag ctgtatctgt

     2581 actatctgca gaatggaaga gatatgtacg tggaccagga gctggatatt aacaggctgt

     2641 ctgattacga cgtggatcat atcgtccccc agagtttcat caaagatgac agcattgaca

     2701 acaaggtgct gaccaggtcc gacaaaaaca gaggaaaatc agataatgtc cctagtgagg

     2761 aagtggtcaa gaaaatgaag aactactgga gacagctgct gaatgccaaa ctgatcactc

     2821 agaggaagtt tgataacctg acaaaagcag agcgcggtgg actgtcagaa ctggacaaag

     2881 ctggattcat caagaggcag ctggtggaaa cacgccagat cactaaacac gtcgcacaga

     2941 ttctggatag tcgcatgaac acaaagtacg atgagaatga caaactgatc agagaagtga

     3001 aggtcattac cctgaagagt aaactggtca gcgactttag gaaagatttc cagttttata

     3061 aggtccgcga gattaacaat tatcaccatg cccatgacgc atacctgaac gccgtggtcg

     3121 gtaccgcact gatcaagaaa tacccaaaac tggagagcga attcgtgtac ggagactata

     3181 aggtgtacga tgtcagaaaa atgatcgcca agtccgagca ggaaattgga aaagctactg

     3241 ccaagtattt cttttactca aacatcatga atttctttaa gacagagatc accctggcca

     3301 atggagaaat ccgcaaaagg cccctgattg agacaaacgg agagacaggc gaaatcgtgt

     3361 gggacaaagg cagagatttt gcaaccgtga ggaaggtcct gagcatgcct caagtgaata

     3421 tcgtcaagaa aactgaggtg cagacaggcg gtttctcaaa agaaagtatt ctgccaaaac

     3481 gcaactctga taagctgatc gctagaaaga aagactggga ccctaagaag tatggaggct

  &nb