pUDP004

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联系方式:I47-825O-882O

相关技术服务:质粒构建    基因合成    质粒大提

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载体基本信息

质粒名称: pUDP004
出品公司: Addgene
目录编号: 101165
存储实验室: Jean-Marc Daran

载体骨架基本信息
载体名称: not applicable
载体出品公司: --
空载体大小: --
完整质粒大小: --
载体修饰: --
载体类型: 酵母表达,CRISPR
筛选标记: amdS(赋予乙酰胺生长的乙酰胺酶基因)
载体抗性: 氨苄霉素
生长温度: 37 ℃
宿主菌: DH5a
拷贝数: 高拷贝

插入基因信息
插入基因名称: --
物种来源: --
插入大小: --
基因突变: --
GenBank编号 --
启动子: ScTDH3
标签/融合蛋白: --
克隆方法: Gibson 克隆
5’克隆位点: --
3’克隆位点: --
5’测序引物 --
3’测序引物 --

文献引用方法
材料和方法部分: pUDP004 was a gift from Jean-Marc Daran (Addgene plasmid # 101165 ; http://n2t.net/addgene:101165 ; RRID:Addgene_101165)
参考文献部分: CRISPR-Cas9 mediated gene deletions in lager yeast Saccharomyces pastorianus. de Vries ARG, de Groot PA, van den Broek M, Daran JG. Microb Cell Fact. 2017 Dec 5;16(1):222. doi: 10.1186/s12934-017-0835-1. 10.1186/s12934-017-0835-1 [pii] PubMed 29207996

载体图谱质粒图谱和多克隆位点信息

pUDP004载体图谱

载体简介

载体序列

LOCUS       pUDP004               10664 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  E. coli/S. cerevisiae shuttle vector carrying amd S marker and 
            Spcas9D147Y P411T allowing cloning of ribozyme flanked g-RNA for 
            Cas9 editing (HH-gRNA-HDV).
ACCESSION   .
VERSION     .
KEYWORDS    pUDP004
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10664)
  AUTHORS   de Vries ARG, de Groot PA, van den Broek M, Daran JG
  TITLE     CRISPR-Cas9 mediated gene deletions in lager yeast Saccharomyces 
            pastorianus.
  JOURNAL   Microb Cell Fact. 2017 Dec 5;16(1):222. doi: 
            10.1186/s12934-017-0835-1.
  PUBMED    29207996
REFERENCE   2  (bases 1 to 10664)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..10664
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             2574..2597
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2601..2621
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2622..6722
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTYKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSITHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             6726..6746
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        7665..8331
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase"
     primer_bind     8201..8224
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     primer_bind     complement(8393..8411)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             complement(8845..9705)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGTRVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     9468..9487
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9706..9810)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      9913..10501
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10402..10421
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gacatggagg cccagaatac cctccttgac agtcttgacg tgcgcagctc aggggcatga
       61 tgtgactgtc gcccgtacat ttagcccata catccccatg tataatcatt tgcatccata
      121 cattttgatg gccgcacggc gcgaagcaaa aattacggct cctcgctgca gacctgcgag
      181 cagggaaacg ctcccctcac agacgcgttg aattgtcccc acgccgcgcc cctgtagaga
      241 aatataaaag gttaggattt gccactgagg ttcttctttc atatacttcc ttttaaaatc
      301 ttgctaggat acagttctca catcacatcc gaacataaac aaggatccat gccacaatct
      361 tgggaagaat tggctgctga caagagagct agattggcta agactatccc agacgaatgg
      421 aaggttcaaa ctttgccagc tgaagactct gttatcgact tcccaaagaa gtctggtatc
      481 ttgtctgaag ctgaattgaa gatcactgaa gcttctgctg ctgacttggt ttctaagttg
      541 gctgctggtg aattgacttc tgttgaagtt actttggctt tctgtaagag agctgctatc
      601 gctcaacaat tgactaactg tgctcacgaa ttcttcccag acgctgcttt ggctcaagct
      661 agagaattgg acgaatacta cgctaagcac aaaagacctg ttggtccatt gcacggtttg
      721 ccaatctctt tgaaggacca attgagagtt aagggttacg aaacttctat gggttacatc
      781 tcttggttga acaagtacga cgaaggtgac tctgttttga ctactatgtt gagaaaggct
      841 ggtgctgttt tctacgttaa gacttctgtt ccacaaactt tgatggtttg tgaaactgtt
      901 aacaacatca tcggtagaac tgttaaccca agaaacaaga actggtcttg tggtggttct
      961 tctggtggtg aaggtgctat cgttggtatc agaggtggtg ttatcggtgt tggtactgac
     1021 atcggtggtt ctatcagagt tccagctgct ttcaacttct tgtacggttt aagaccatct
     1081 cacggtagat tgccatacgc taagatggct aactctatgg aaggtcaaga aactgtacac
     1141 tctgttgttg gtccaatcac tcactctgtt gaagacttga gattgttcac taagtctgtt
     1201 ttgggtcaag aaccatggaa gtacgactct aaggttatcc caatgccatg gagacaatct
     1261 gaatctgaca tcatcgcttc taagatcaag aacggtggtt tgaacatcgg ttactacaac
     1321 ttcgacggta acgttttgcc acacccacca atcttgagag gtgttgaaac tactgttgct
     1381 gctttggcta aggctggtca cactgttact ccatggactc catacaagca cgacttcggt
     1441 cacgacttga tctctcacat ctacgctgct gacggttctg ctgacgttat gagagacatc
     1501 tctgcttctg gtgaaccagc tatcccaaac atcaaggact tgttgaaccc aaacatcaag
     1561 gctgttaaca tgaacgaatt gtgggacact cacttgcaaa agtggaacta ccaaatggaa
     1621 tacttggaaa agtggagaga agctgaagaa aaggctggta aggaattgga cgctatcatc
     1681 gctccaatca ctccaactgc tgctgttaga cacgaccaat tcagatacta cggttacgct
     1741 tctgttatca acttgttgga cttcacttct gttgttgttc cagttacttt cgctgacaag
     1801 aacatcgaca agaagaacga atctttcaag gctgtttctg aattggacgc tttggttcaa
     1861 gaagaatacg acccagaagc ttaccacggt gctccagttg ctgttcaagt tatcggtaga
     1921 agattgtctg aagaaagaac tttggctatc gctgaagaag ttggtaagtt gttgggtaac
     1981 gttgttactc cataactcga ggacaataaa aagattcttg ttttcaagaa cttgtcattt
     2041 gtatagtttt tttatattgt agttgttcta ttttaatcaa atgttagcgt gatttatatt
     2101 ttttttcgcc tcgacatcat ctgcccagat gcgaagttaa gtgcgcagaa agtaatatca
     2161 tgcgtcaatc gtatgtgaat gctggtcgct atactggagc tctcgagaac cctcaccttt
     2221 cgagaggacg atgcccgtgt ctaaatgatt cgaccagcct aagaatgttc aacaatctat
     2281 aatcagtcca tagtcaacaa gagccctcaa agttgatatc gtttagtgat ctccctcgag
     2341 atgagcggag gatggttgct ttttggggag ccctaagtgg tacgtgcggc gagcgggatg
     2401 tttccccatt aggcaaagga cgaccgggta acccctcgag aaaaaaaatt tttcgccgct
     2461 aatctggtgt tatataaagc tccccctgct cctggaattt ttttccttgt caactcacac
     2521 cggaaatcaa ggcatttcat tctgagtagt tcttaaaaac ataatcaaca atggattata
     2581 aagatgacga tgacaaacct ccaaaaaaga agagaaaggt cgataagaaa tactcaatag
     2641 gcttagatat cggcacaaat agcgtcggat gggcggtgat cactgatgaa tataaggttc
     2701 cgtctaaaaa gttcaaggtt ctgggaaata cagaccgcca cagtatcaaa aaaaatctta
     2761 taggggctct tttatttgac agtggagaga cagcggaagc gactcgtctc aaacggacag
     2821 ctcgtagaag gtatacacgt cggaagaatc gtatttgtta tctacaggag attttttcaa
     2881 atgagatggc gaaagtagat gatagtttct ttcatcgact tgaagagtct tttttggtgg
     2941 aagaagacaa gaagcatgaa cgtcatccta tttttggaaa tatagtagat gaagttgctt
     3001 atcatgagaa atatccaact atctatcatc tgcgaaaaaa attggtagat tctacttata
     3061 aagcggattt gcgcttaatc tatttggcct tagcgcatat gattaagttt cgtggtcatt
     3121 ttttgattga gggagattta aatcctgata atagtgatgt ggacaaacta tttatccagt
     3181 tggtacaaac ctacaatcaa ttatttgaag aaaaccctat taacgcaagt ggagtagatg
     3241 ctaaagcgat tctttctgca cgattgagta aatcaagacg attagaaaat ctcattgctc
     3301 agctccccgg tgagaagaaa aatggcttat ttgggaatct cattgctttg tcattgggtt
     3361 tgacccctaa ttttaaatca aattttgatt tggcagaaga tgctaaatta cagctttcaa
     3421 aagatactta cgatgatgat ttagataatt tattggcgca aattggagat caatatgctg
     3481 atttgttttt ggcagctaag aatttatcag atgctatttt actttcagat atcctaagag
     3541 taaatactga aataactaag gctcccctat cagcttcaat gattaaacgc tacgatgaac
     3601 atcatcaaga cttgactctt ttaaaagctt tagttcgaca acaacttcca gaaaagtata
     3661 aagaaatctt ttttgatcaa tcaaaaaacg gatatgcagg ttatattgat gggggagcta
     3721 gccaagaaga attttataaa tttatcaaac caattttaga aaaaatggat ggtactgagg
     3781 aattattggt gaaactaaat cgtgaagatt tgctgcgcaa gcaacggacc tttgacaacg
     3841 gctctattac ccatcaaatt cacttgggtg agctgcatgc tattttgaga agacaagaag
     3901 acttttatcc atttttaaaa gacaatcgtg agaagattga aaaaatcttg acttttcgaa
     3961 ttccttatta tgttggtcca ttggcgcgtg gcaatagtcg ttttgcatgg atgactcgga
     4021 agtctgaaga aacaattacc ccatggaatt ttgaagaagt tgtcgataaa ggtgcttcag
     4081 ctcaatcatt tattgaacgc atgacaaact ttgataaaaa tcttccaaat gaaaaagtac
     4141 taccaaaaca tagtttgctt tatgagtatt ttacggttta taacgaattg acaaaggtca
     4201 aatatgttac tgaaggaatg cgaaaaccag catttctttc aggtgaacag aagaaagcca
     4261 ttgttgattt actcttcaaa acaaatcgaa aagtaaccgt taagcaatta aaagaagatt
     4321 atttcaaaaa aatagaatgt tttgatagtg ttgaaatttc aggagttgaa gatagattta
     4381 atgcttcatt aggtacctac catgatttgc taaaaattat taaagataaa gattttttgg
     4441 ataatgaaga aaatgaagat atcttagagg atattgtttt aacattgacc ttatttgaag
     4501 atagggagat gattgaggaa agacttaaaa catatgctca cctctttgat gataaggtga
     4561 tgaaacagct taaacgtcgc cgttatactg gttggggacg tttgtctcga aaattgatta
     4621 atggtattag ggataagcaa tctggcaaaa caatattaga ttttttgaaa tcagatggtt
     4681 ttgccaatcg caattttatg cagctgatcc atgatgatag tttgacattt aaagaagaca
     4741 ttcaaaaagc acaagtgtct ggacaaggcg atagtttaca tgaacatatt gcaaatttag
     4801 ctggtagccc tgctattaaa aaaggtattt tacagactgt aaaagttgtt gatgaattgg
     4861 tcaaagtaat ggggcggcat aagccagaaa atatcgttat tgaaatggca cgtgaaaatc
     4921 agacaactca aaagggccag aaaaattcgc gagagcgtat gaaacgaatc gaagaaggta
     4981 tcaaagaatt aggaagtcag attcttaaag agcatcctgt tgaaaatact caattgcaaa
     5041 atgaaaagct ctatctctat tatctccaaa atggaagaga catgtatgtg gaccaagaat
     5101 tagatattaa tcgtttaagt gattatgatg tcgatcacat tgttccacaa agtttcctta
     5161 aagacgattc aatagacaat aaggtcttaa cgcgttctga taaaaatcgt ggtaaatcgg
     5221 ataacgttcc aagtgaagaa gtagtcaaaa agatgaaaaa ctattggaga caacttctaa
     5281 acgccaagtt aatcactcaa cgtaagtttg ataatttaac gaaagctgaa cgtggaggtt
     5341 tgagtgaact tgataaagct ggttttatca aacgccaatt ggttgaaact cgccaaatca
     5401 ctaagcatgt ggcacaaatt ttggatagtc gcatgaatac taaatacgat gaaaatgata
     5461 aacttattcg agaggttaaa gtgattacct taaaatctaa attagtttct gacttccgaa
     5521 aagatttcca attctataaa gtacgtgaga ttaacaatta ccatcatgcc catgatgcgt
     5581 atctaaatgc cgtcgttgga actgctttga ttaagaaata tccaaaactt gaatcggagt
     5641 ttgtctatgg tgattataaa gtttatgatg ttcgtaaaat gattgctaag tctgagcaag
     5701 aaataggcaa agcaaccgca aaatatttct tttactctaa tatcatgaac ttcttcaaaa
     5761 cagaaattac acttgcaaat ggagagattc gcaaacgccc tctaatcgaa actaatgggg
     5821 aaactggaga aattgtctgg gataaagggc gagattttgc cacagtgcgc aaagtattgt
     5881 ccatgcccca agtcaatatt gtcaagaaaa cagaagtaca gacaggcgga ttctccaagg
     5941 agtcaatttt accaaaaaga aattcggaca agcttattgc tcgtaaaaaa gactgggatc
     6001 caaaaaaata tggtggtttt gatagtccaa cggtagctta ttcagtccta gtggttgcta
     6061 aggtggaaaa agggaaatcg aagaagttaa aatccgttaa agagttacta gggatcacaa
     6121 ttatggaaag aagttccttt gaaaaaaatc cgattgactt tttagaagct aaaggatata
     6181 aggaagttaa aaaagactta atcattaaac tacctaaata tagtcttttt gagttagaaa
     6241 acggtcgtaa acggatgctg gctagtgccg gagaattaca aaaaggaaat gagctggctc
     6301 tgccaagcaa atatgtgaat tttttatatt tagctagtca ttatgaaaag ttgaagggta
     6361 gtccagaaga taacgaacaa aaacaattgt ttgtggagca gcataagcat tatttagatg
     6421 agattattga gcaaatcagt gaattttcta agcgtgttat tttagcagat gccaatttag
     6481 ataaagttct tagtgcatat aacaaacata gagacaaacc aatacgtgaa caagcagaaa
     6541 atattattca tttatttacg ttgacgaatc ttggagctcc cgctgctttt aaatattttg
     6601 atacaacaat tgatcgtaaa cgatatacgt ctacaaaaga agttttagat gccactctta
     6661 tccatcaatc catcactggt ctttatgaaa cacgcattga tttgagtcag ctaggaggtg
     6721 accctccaaa aaagaagaga aaggtctgag cggttttgta taactaaata atattggaaa
     6781 ctaaatacga atacccaaat tttttatcta aattttgccg aaagattaaa atctgcagag
     6841 atatccgaaa caggtaaatg gatgtttcaa tccctgtagt cagtcaggaa cccatattat
     6901 attacagtat tagtcgccgc ttaggcacgc ctttaattag caaaatcaaa ccttaagtgc
     6961 atatgccgta taagggaaac tcaaagaact ggcatcgcaa aaatgaaaac atacgttgaa
     7021 actacggcaa aggattggtc agatcgcttc atacagggaa agttcggcat caacatcttt
     7081 ggataatatc agaatgagaa agaacagata cgcagtacgt tttttggtga gctctttgca
     7141 cttctttagt tctttccatc aatatcagtt ttttaaactt ttaggactaa aagtgatgtt
     7201 taacttcaaa atgtttaaaa ttttgttctt cccgacgttc attaagaata ctaatacact
     7261 ttaataatta ttttaatatt ttgttctaaa taatgacttt taattaaaaa agataaaata
     7321 taaaaacatc ataataactc accagaggtt aagaacaaaa aaacaaatta gatatctgct
     7381 aatccaatat agttaaatca atctttcctt ggtataatgg gtatattaca tatatttcaa
     7441 ggaccgacac tcctaccaaa tatctaaaat ttaccatatt aacataacat gtatataaac
     7501 gtcaaatcat aatcagcact aacgtctcac ggatcgtata tgccgtagcg acaatctaag
     7561 aactatgcga ggacacgcta gggccaaggc aaaaagattc cttgattacg taagggagtt
     7621 agaatcattt tgaataaaaa acacgctttt tcagttcgag tttatcatta tcaatactgc
     7681 catttcaaag aatacgtaaa taattaatag tagtgatttt cctaacttta tttagtcaaa
     7741 aaattagcct tttaattctg ctgtaacccg tacatgccca aaataggggg cgggttacac
     7801 agaatatata acatcgtagg tgtctgggtg aacagtttat tcttggcatc cactaaatat
     7861 aatggagccc gctttttaag ctggcatcca gaaaaaaaaa gaatcccagc accaaaatat
     7921 tgttttcttc accaaccatc agttcatagg tccattctct tagcgcaact acagagaaca
     7981 ggggcacaaa caggcaaaaa acgggcacaa cctcaatgga gtgatgcaac ctgcctggag
     8041 taaatgatga cacaaggcaa ttgacccacg catgtatcta tctcattttc ttacaccttc
     8101 tattaccttc tgctctctct gatttggaaa aagctgaaaa aaaaggttga aaccagttcc
     8161 ctgaaattat tcccctactt gactaataag tatataaaga cggtaggtat tgattgtaat
     8221 tctgtaaatc tatttcttaa acttcttaaa ttctactttt atagttagtc ttttttttag
     8281 ttttaaaaca ccaagaactt agtttcgaat aaacacacat aaacaaacaa acgagacctg
     8341 cggaggtctc cacaggcccc ttttcctttg tcgatatcat gtaattagtt atgtcacgct
     8401 tacattcacg ccctcccccc acatccgctc taaccgaaaa ggaaggagtt agacaacctg
     8461 aagtctaggt ccctatttat ttttttatag ttatgttagt attaagaacg ttatttatat
     8521 ttcaaatttt tctttttttt ctgtacaaac gcgtgtacgc atgtaacatt atactgaaaa
     8581 ccttgcttga gaaggttttg ggacaagctt acgtctcacg gatcgtatat gccgtagcga
     8641 caatctaaga actatgcgag gacacgctag cagctgtatt cacgtagacg gataggtata
     8701 gccagacatc agcagcatac ttcgggaacc gtaggccacc gagcggccgc gattatcaaa
     8761 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     8821 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     8881 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     8941 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgggacc cacgctcacc
     9001 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     9061 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     9121 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     9181 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     9241 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     9301 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     9361 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     9421 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     9481 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     9541 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgtac ccaactgatc
     9601 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     9661 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     9721 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     9781 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgtcat
     9841 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     9901 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     9961 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
    10021 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
    10081 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
    10141 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
    10201 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
    10261 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
    10321 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
    10381 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcc
    10441 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
    10501 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
    10561 gttctttcct gcgttatccc ctgattctgt ggataaccgt gcggactata tgtgaaggca
    10621 tggctatggc acggcagaca ttccgccaga tcatcaatag gcac
//

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