pCas-Guide-EF1a-CD4

价格： ¥800 - 1200

品牌：Ybscience

货号：YB-10323

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载体基本信息

出品公司:	Ybscience
载体名称:	pCas-Guide-EF1a-CD4
质粒类型:	CRISPR/Cas9载体；基因敲除载体；哺乳动物细胞载体
高拷贝/低拷贝:	高拷贝
插入位点:	限制性内切酶，多克隆位点
启动子:	U6
载体大小:	11006 bp
5' 测序引物及序列:	5’ gatcgATGGGAGGTGGTATGGGAGGg 3’
3' 测序引物及序列:	5’ aaaacCCTCCCATACCACCTCCCATc 3’
载体标签:	Myc(C-ter）
载体抗性:	氨苄青霉素
筛选标记:	--
克隆菌株:	Stbl3
宿主细胞（系）:	哺乳动物细胞
备注:	--
稳定性:	瞬表达
组成型/诱导型:	组成型
病毒/非病毒:	非病毒

载体质粒图谱和多克隆位点信息

pCas-Guide-EF1a-CD4载体图谱

载体简介

pCas-Guide-EF1a-CD4载体特征All-in-one vectorExpresses human codon-optimized Cas9, C-terminal Myc-DDK taggedBamH I and BsmB I sites are used for genomic target sequence cloningExpresses gRNA containing the inserted target sequence and the gRNA scaffoldEF1a promoter driven CD4 to enrich transfected cellsCF3 primer (GE100008) can be used to sequence the target sequence after cloning

载体序列

LOCUS       Exported File          11006 bp ds-DNA    circular SYN 23-七月-2016DEFINITION  Mammalian vector for co-expressing a guide RNA (gRNA) together with             Cas9 and CD4, for genome editing using the S. pyogenes CRISPR/Cas9             system.ACCESSION   .VERSION     .KEYWORDS    pCas-Guide-EF1a-CD4SOURCE      synthetic DNA construct  ORGANISM  synthetic DNA constructREFERENCE   1  (bases 1 to 11006)  AUTHORS   OriGene  TITLE     Direct Submission  JOURNAL   Exported 2016-07-23 from SnapGene Viewer 1.5.3            http://www.snapgene.comFEATURES             Location/Qualifiers     source          1..11006                     /organism="synthetic DNA construct"                     /lab_host="Mammalian Cells"                     /mol_type="other DNA"     promoter        90..330                     /note="U6"                     /note="RNA polymerase III promoter for human U6 snRNA"                     /note="color: #000000; direction: RIGHT"     misc_RNA        384..459                     /note="gRNA scaffold"                     /note="guide RNA scaffold for the Streptococcus pyogenes                      CRISPR/Cas9 system"                     /note="color: #00ccff; direction: RIGHT"     terminator      460..465                     /note="polIII terminator"                     /note="RNA polymerase III transcription terminator"                     /note="color: #ffffff"     promoter        480..1658                     /note="EF-1-alpha promoter"                     /note="strong constitutive promoter for human elongation                      factor EF-1-alpha"                     /note="color: #000000; direction: RIGHT"     intron          711..1649                     /note="EF-1-alpha intron A"                     /note="intron upstream of the start codon of human                      EF-1-alpha"                     /note="color: #------"     CDS             1679..2950                     /codon_start=1                     /product="truncated human CD4 cell surface glycoprotein                      lacking the cytoplasmic tail"                     /note="CD4 (truncated)"                     /note="This feature has 2 segments:                     ???1:?1679?..?1753?/?#ff6600?/?signal?peptide                     ???2:?1754?..?2950?/?#ff6600                     Cleavage site after base 1753"                     /translation="MNRGVPFRHLLLVLQLALLPAATQGKKVVLGKKGDTVELTCTASQ                     KKSIQFHWKNSNQIKILGNQGSFLTKGPSKLNDRADSRRSLWDQGNFPLIIKNLKIEDS                     DTYICEVEDQKEEVQLLVFGLTANSDTHLLQGQSLTLTLESPPGSSPSVQCRSPRGKNI                     QGGKTLSVSQLELQDSGTWTCTVLQNQKKVEFKIDIVVLAFQKASSIVYKKEGEQVEFS                     FPLAFTVEKLTGSGELWWQAERASSSKSWITFDLKNKEVSVKRVTQDPKLQMGKKLPLH                     LTLPQALPQYAGSGNLTLALEAKTGKLHQEVNLVVMRATQLQKNLTCEVWGPTSPKLML                     SLKLENKEAKVSKREKAVWVLNPEAGMWQCLLSDSGQVLLESNIKVLPTWSTPVQPMAL                     IVLGGVAGLLLFIGLGIFFCVRCRH"     enhancer        3560..3863                     /note="CMV enhancer"                     /note="human cytomegalovirus immediate early enhancer"                     /note="color: #000000"     promoter        3864..4067                     /note="CMV promoter"                     /note="human cytomegalovirus (CMV) immediate early                      promoter"                     /note="color: #000000; direction: RIGHT"     promoter        4093..4111                     /note="T7 promoter"                     /note="promoter for bacteriophage T7 RNA polymerase"                     /note="color: #ffffff; direction: RIGHT"     CDS             4170..8273                     /codon_start=1                     /locus_tag="                     "                     /product="Cas9 (Csn1) endonuclease from the?Streptococcus                      pyogenes?Type II CRISPR/Cas system"                     /note="Cas9"                     /note="generates RNA-guided double strand breaks in DNA"                     /note="color: #993366"                     /protein_id="                     "                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL                     DATLIHQSITGLYETRIDLSQLGGD"     CDS             8274..8294                     /codon_start=1                     /locus_tag="                     "                     /product="nuclear localization signal of SV40 large T                      antigen"                     /note="SV40 NLS"                     /note="                     "                     /note="color: #993366"                     /protein_id="                     "                     /translation="PKKKRKV"     CDS             8301..8321                     /codon_start=1                     /locus_tag="                     "                     /product="nuclear localization signal of SV40 large T                      antigen"                     /note="SV40 NLS"                     /note="                     "                     /note="color: #993366"                     /protein_id="                     "                     /translation="PKKKRKV"     CDS             8343..8372                     /codon_start=1                     /product="Myc (human c-Myc oncogene) epitope tag"                     /note="Myc"                     /note="color: #cc99b2"                     /translation="EQKLISEEDL"     CDS             8391..8414                     /codon_start=1                     /product="FLAG(R) epitope tag, followed by an enterokinase                      cleavage site"                     /note="FLAG"                     /note="color: #cc99b2"                     /translation="DYKDDDDK"     polyA_signal    8461..9083                     /note="hGH poly(A) signal"                     /note="human growth hormone polyadenylation signal"                     /note="color: #a6acb3"     rep_origin      complement(9204..9792)                     /direction=LEFT                     /note="ori"                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of                      replication"                     /note="color: #ffff00"     CDS             complement(9967..10827)                     /codon_start=1                     /gene="bla"                     /product="beta-lactamase"                     /note="AmpR"                     /note="confers resistance to ampicillin, carbenicillin, and                     related antibiotics"                     /note="This feature has 2 segments:                     ???1:??9967?..?10758?/?#ccffcc                     ???2:?10759?..?10827?/?#ccffcc?/?signal?sequence                     Cleavage site after base 10758"                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS                     LIKHW"ORIGIN        1 gaattcccca gtggaaagac gcgcaggcaa aacgcaccac gtgacggagc gtgaccgcgc       61 gccgagcgcg cgccaaggtc gggcaggaag agggcctatt tcccatgatt ccttcatatt      121 tgcatatacg atacaaggct gttagagaga taattagaat taatttgact gtaaacacaa      181 agatattagt acaaaatacg tgacgtagaa agtaataatt tcttgggtag tttgcagttt      241 taaaattatg ttttaaaatg gactatcata tgcttaccgt aacttgaaag tatttcgatt      301 tcttgggttt atatatcttg tggaaaggac gcgggatcca ctggaccagg cagcagcgtc      361 agaagacttt tttggaacgt ctcgttttag agctagaaat agcaagttaa aataaggcta      421 gtccgttatc aacttgaaaa agtggcaccg agtcggtgct ttttttggtg tacacgtgag      481 gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg      541 gaggggtcgg caattgaacc ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg      601 atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag      661 tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag gtaagtgccg      721 tgtgtggttc ccgcgggcct ggcctcttta cgggttatgg cccttgcgtg ccttgaatta      781 cttccacctg gctgcagtac gtgattcttg atcccgagct tcgggttgga agtgggtggg      841 agagttcgag gccttgcgct taaggagccc cttcgcctcg tgcttgagtt gaggcctggc      901 ctgggcgctg gggccgccgc gtgcgaatct ggtggcacct tcgcgcctgt ctcgctgctt      961 tcgataagtc tctagccatt taaaattttt gatgacctgc tgcgacgctt tttttctggc     1021 aagatagtct tgtaaatgcg ggccaagatc tgcacactgg tatttcggtt tttggggccg     1081 cgggcggcga cggggcccgt gcgtcccagc gcacatgttc ggcgaggcgg ggcctgcgag     1141 cgcggccacc gagaatcgga cgggggtagt ctcaagctgg ccggcctgct ctggtgcctg     1201 gcctcgcgcc gccgtgtatc gccccgccct gggcggcaag gctggcccgg tcggcaccag     1261 ttgcgtgagc ggaaagatgg ccgcttcccg gccctgctgc agggagctca aaatggagga     1321 cgcggcgctc gggagagcgg gcgggtgagt cacccacaca aaggaaaagg gcctttccgt     1381 cctcagccgt cgcttcatgt gactccacgg agtaccgggc gccgtccagg cacctcgatt     1441 agttctcgag cttttggagt acgtcgtctt taggttgggg ggaggggttt tatgcgatgg     1501 agtttcccca cactgagtgg gtggagactg aagttaggcc agcttggcac ttgatgtaat     1561 tctccttgga atttgccctt tttgagtttg gatcttggtt cattctcaag cctcagacag     1621 tggttcaaag tttttttctt ccatttcagg tgtcgtgact atagggcggc cggacgtgat     1681 gaaccgggga gtccctttta ggcacttgct tctggtgctg caactggcgc tcctcccagc     1741 agccactcag ggaaagaaag tggtgctggg caaaaaaggg gatacagtgg aactgacctg     1801 tacagcttcc cagaagaaga gcatacaatt ccactggaaa aactccaacc agataaagat     1861 tctgggaaat cagggctcct tcttaactaa aggtccatcc aagctgaatg atcgcgctga     1921 ctcaagaaga agcctttggg accaaggaaa ctttcccctg atcatcaaga atcttaagat     1981 agaagactca gatacttaca tctgtgaagt ggaggaccag aaggaggagg tgcaattgct     2041 agtgttcgga ttgactgcca actctgacac ccacctgctt caggggcaga gcctgaccct     2101 gaccttggag agcccccctg gtagtagccc ctcagtgcaa tgtaggagtc caaggggtaa     2161 aaacatacag ggggggaaga ccctctccgt gtctcagctg gagctccagg atagtggcac     2221 ctggacatgc actgtcttgc agaaccagaa gaaggtggag ttcaaaatag acatcgtggt     2281 gctagctttc cagaaggcct ccagcatagt ctataagaaa gagggggaac aggtggagtt     2341 ctccttccca ctcgccttta cagttgaaaa gctgacgggc agtggcgagc tgtggtggca     2401 ggcggagagg gcttcctcct ccaagtcttg gatcaccttt gacctgaaga acaaggaagt     2461 gtctgtaaaa cgggttaccc aggaccctaa gctccagatg ggcaagaagc tcccgctcca     2521 cctcaccctg ccccaggcct tgcctcagta tgctggctct ggaaacctca ccctggccct     2581 tgaagcgaaa acaggaaagt tgcatcagga agtgaacctg gtggtgatga gagccactca     2641 gctccagaaa aatttgacct gtgaggtgtg gggacccacc tcccctaagc tgatgctgag     2701 cttgaaactg gagaacaagg aggcaaaggt ctcgaagcgg gagaaggcgg tgtgggtgct     2761 gaaccctgag gcggggatgt ggcagtgtct gctgagtgac tcgggacagg tcctgctgga     2821 atccaacatc aaggttctgc ccacatggtc caccccggtg cagccaatgg ccctgattgt     2881 gctggggggc gtcgccggcc tcctgctttt cattgggcta ggcatcttct tctgtgtcag     2941 gtgccggcac accggttagg atatcagatc cccgggatgc agaaattgat gatctattaa     3001 acaataaaga tgtccactaa aatggaagtt tttcctgtca tactttgtta agaagggtga     3061 gaacagagta cctacatttt gaatggaagg attggagcta cgggggtggg ggtggggtgg     3121 gattagataa atgcctgctc tttactgaag gctctttact attgctttat gataatgttt     3181 catagttgga tatcataatt taaacaagca aaaccaaatt aagggccagc tcattcctcc     3241 cactcatgat ctatagatct atagatctct cgtgggatca ttgtttttct cttgattccc     3301 actttgtggt tctaagtact gtggtttcca aatgtgtcag tttcatagcc tgaagaacga     3361 gatcagcagc ctctgttcca catacacttc attctcagta ttgttttgcc aagttctaat     3421 tccatcagaa gctggtcgag atccggaacc cttaatataa cttcgtataa tgtatgctat     3481 acgaagttat taggtccact agttattaat agtaatcaat tacggggtca ttagttcata     3541 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc     3601 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag     3661 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac     3721 atcaagtgta tcatatgcca agtccgcccc ctattgacgt caatgacggt aaatggcccg     3781 cctggcatta tgcccagtac atgaccttac gggactttcc tacttggcag tacatctacg     3841 tattagtcat cgctattacc atggtgatgc ggttttggca gtacaccaat gggcgtggat     3901 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt     3961 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa taaccccgcc ccgttgacgc     4021 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctcgt ttagtgaacc     4081 gtcagaattt tgtaatacga ctcactatag ggcggccggg aattcgtcga ctggaaccgg     4141 taccgaggag atctgccgcc gcgatcgcca tggataagaa atactcaata ggactggata     4201 ttggcacaaa tagcgtcgga tgggctgtga tcactgatga atataaggtt ccttctaaaa     4261 agttcaaggt tctgggaaat acagaccgcc acagtatcaa aaaaaatctt ataggggctc     4321 ttctgtttga cagtggagag acagccgaag ctactagact caaacggaca gctaggagaa     4381 ggtatacaag acggaagaat aggatttgtt atctccagga gattttttca aatgagatgg     4441 ccaaagtgga tgatagtttc tttcatagac ttgaagagtc ttttttggtg gaagaagaca     4501 agaagcatga aagacatcct atttttggaa atatagtgga tgaagttgct tatcacgaga     4561 aatatccaac tatctatcat ctgagaaaaa aattggtgga ttctactgat aaagccgatt     4621 tgcgcctgat ctatttggcc ctggcccaca tgattaagtt tagaggtcat tttttgattg     4681 agggcgatct gaatcctgat aatagtgatg tggacaaact gtttatccag ttggtgcaaa     4741 cctacaatca actgtttgaa gaaaacccta ttaacgcaag tggagtggat gctaaagcca     4801 ttctttctgc aagattgagt aaatcaagaa gactggaaaa tctcattgct cagctccccg     4861 gtgagaagaa aaatggcctg tttgggaatc tcattgcttt gtcattgggt ttgaccccta     4921 attttaaatc aaattttgat ttggcagaag atgctaaact ccagctttca aaagatactt     4981 acgatgatga tctggataat ctgttggctc aaattggaga tcaatatgct gatttgtttt     5041 tggcagctaa gaatctgtca gatgctattc tgctttcaga catcctgaga gtgaatactg     5101 aaataactaa ggctcccctg tcagcttcaa tgattaaacg ctacgatgaa catcatcaag     5161 acttgactct tctgaaagcc ctggttagac aacaacttcc agaaaagtat aaagaaatct     5221 tttttgatca atcaaaaaac ggatatgcag gttatattga tggcggcgca agccaagaag     5281 aattttataa atttatcaaa ccaattctgg aaaaaatgga tggtactgag gaactgttgg     5341 tgaaactgaa tagagaagat ttgctgcgca agcaacggac ctttgacaac ggctctattc     5401 cccatcaaat tcacttgggt gagctgcatg ctattttgag aagacaagaa gacttttatc     5461 catttctgaa agacaataga gagaagattg aaaaaatctt gacttttagg attccttatt     5521 atgttggtcc attggccaga ggcaatagta ggtttgcatg gatgactcgg aagtctgaag     5581 aaacaattac cccatggaat tttgaagaag ttgtcgataa aggtgcttca gctcaatcat     5641 ttattgaacg catgacaaac tttgataaaa atcttccaaa tgaaaaagtg ctgccaaaac     5701 atagtttgct ttatgagtat tttaccgttt ataacgaatt gacaaaggtc aaatatgtta     5761 ctgaaggaat gagaaaacca gcatttcttt caggtgaaca gaagaaagcc attgttgatc     5821 tgctcttcaa aacaaatagg aaagtgaccg ttaagcaact gaaagaagat tatttcaaaa     5881 aaatagaatg ttttgatagt gttgaaattt caggagttga agatagattt aatgcttcac     5941 tgggtacata ccatgatttg ctgaaaatta ttaaagataa agattttttg gataatgaag     6001 aaaatgaaga catcctggag gatattgttc tgacattgac cctgtttgaa gatagggaga     6061 tgattgagga aagacttaaa acatacgctc acctctttga tgataaggtg atgaaacagc     6121 ttaaaagacg cagatatact ggttggggaa ggttgtccag aaaattgatt aatggtatta     6181 gggataagca atctggcaaa acaatactgg attttttgaa atcagatggt tttgccaatc     6241 gcaattttat gcagctcatc catgatgata gtttgacatt taaagaagac atccaaaaag     6301 cacaagtgtc tggacaaggc gatagtctgc atgaacatat tgcaaatctg gctggtagcc     6361 ctgctattaa aaaaggtatt ctccagactg tgaaagttgt tgatgaattg gtcaaagtga     6421 tggggcggca taagccagaa aatatcgtta ttgaaatggc aagagaaaat cagacaactc     6481 aaaagggcca gaaaaattcc agagagagga tgaaaagaat cgaagaaggt atcaaagaac     6541 tgggaagtca gattcttaaa gagcatcctg ttgaaaatac tcaattgcaa aatgaaaagc     6601 tctatctcta ttatctccaa aatggaagag atatgtatgt ggaccaagaa ctggatatta     6661 ataggctgag tgattatgat gtcgatcaca ttgttccaca aagtttcctt aaagacgatt     6721 caatagacaa taaggtcctg accaggtctg ataaaaatag aggtaaatcc gataacgttc     6781 caagtgaaga agtggtcaaa aagatgaaaa actattggag acaacttctg aacgccaagc     6841 tgatcactca aaggaagttt gataatctga ccaaagctga aagaggaggt ttgagtgaac     6901 ttgataaagc tggttttatc aaacgccaat tggttgaaac tcgccaaatc actaagcatg     6961 tggcacaaat tttggatagt cgcatgaata ctaaatacga tgaaaatgat aaacttatta     7021 gagaggttaa agtgattacc ctgaaatcta aactggtttc tgacttcaga aaagatttcc     7081 aattctataa agtgagagag attaacaatt accatcatgc ccatgatgcc tatctgaatg     7141 ccgtcgttgg aactgctttg attaagaaat atccaaaact tgaaagcgag tttgtctatg     7201 gtgattataa agtttatgat gttaggaaaa tgattgctaa gtctgagcaa gaaataggca     7261 aagcaaccgc aaagtatttc ttttactcta atatcatgaa cttcttcaaa acagaaatta     7321 cacttgcaaa tggagagatt cgcaaacgcc ctctgatcga aactaatggg gaaactggag     7381 aaattgtctg ggataaaggg agagattttg ccacagtgcg caaagtgttg tccatgcccc     7441 aagtcaatat cgtcaagaaa acagaagtgc agacaggcgg attctctaag gagtcaattc     7501 tgccaaaaag aaattccgac aagctgattg ctaggaaaaa agactgggac ccaaaaaaat     7561 atggtggttt tgatagtcca accgtggctt attcagtcct ggtggttgct aaggtggaaa     7621 aagggaaatc caagaagctg aaatccgtta aagagctgct ggggatcaca attatggaaa     7681 gaagttcctt tgaaaaaaat cccattgact ttctggaagc taaaggatat aaggaagtta     7741 aaaaagacct gatcattaaa ctgcctaaat atagtctttt tgagctggaa aacggtagga     7801 aacggatgct ggctagtgcc ggagaactgc aaaaaggaaa tgagctggct ctgccaagca     7861 aatatgtgaa ttttctgtat ctggctagtc attatgaaaa gttgaagggt agtccagaag     7921 ataacgaaca aaaacaattg tttgtggagc agcataagca ttatctggat gagattattg     7981 agcaaatcag tgaattttct aagagagtta ttctggcaga tgccaatctg gataaagttc     8041 ttagtgcata taacaaacat agagacaaac caataagaga acaagcagaa aatatcattc     8101 atctgtttac cttgaccaat cttggagcac ccgctgcttt taaatacttt gatacaacaa     8161 ttgataggaa aagatatacc tctacaaaag aagttctgga tgccactctt atccatcaat     8221 ccatcactgg tctttatgaa acacgcattg atttgagtca gctgggaggt gaccccaaga     8281 aaaaacgcaa ggtggaagat cctaagaaaa agcggaaagt ggacacgcgt acgcggccgc     8341 tcgagcagaa actcatctca gaagaggatc tggcagcaaa tgatatcctg gattacaagg     8401 atgacgacga taaggtttaa acggccggcc gcggtcatag ctgtttcctg aacagatccc     8461 gggtggcatc cctgtgaccc ctccccagtg cctctcctgg ccctggaagt tgccactcca     8521 gtgcccacca gccttgtcct aataaaatta agttgcatca ttttgtctga ctaggtgtcc     8581 ttctataata ttatggggtg gaggggggtg gtatggagca aggggcaagt tgggaagaca     8641 acctgtaggg cctgcggggt ctattgggaa ccaagctgga gtgcagtggc acaatcttgg     8701 ctcactgcaa tctccgcctc ctgggttcaa gcgattctcc tgcctcagcc tcccgagttg     8761 ttgggattcc aggcatgcat gaccaggctc agctaatttt tgtttttttg gtagaggcgg     8821 ggtttcacca tattggccag gctggtctcc aactcctaat ctcaggtgat ctacccacct     8881 tggcctccca aattgctggg attacaggcg tgaaccactg ctcccttccc tgtccttctg     8941 attttaaaat aactatacca gcaggaggac gtccagacac agcataggct acctggccat     9001 gcccaaccgg tgggacattt gagttgcttg cttggcactg tcctctcatg cgttgggtcc     9061 actcagtaga tgcctgttga attgggtacg cggccagcgg cgagcggtat cagctcactc     9121 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtccgt     9181 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa     9241 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt     9301 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg     9361 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc     9421 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc     9481 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta     9541 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct     9601 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc     9661 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa     9721 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa     9781 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgac     9841 gcgtaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat     9901 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc     9961 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc    10021 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc    10081 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc    10141 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc    10201 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt    10261 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc    10321 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa    10381 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt    10441 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg    10501 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc    10561 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa    10621 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt    10681 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt    10741 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag    10801 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta    10861 tcagggttat tgtctcatga tgatattatt ttatcttgtg caatgtaaca tcagagattt    10921 tgagacacgg gccagagctg ccaggaaaca gctatgacca tgtaatacga ctcactatag    10981 gggatatcag ctggatggca gttaac