pC003 - LshC2C2 locus into pACYC184 for spacer cloning

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载体基本信息

质粒名称: pC003 - LshC2C2 locus into pACYC184 for spacer cloning 
出品公司: Addgene
目录编号: 79152
存储实验室: Feng Zhang

载体骨架基本信息
载体名称: pACYC184
载体出品公司: --
载体抗性: 氯霉素
空载体大小: 4723 bp
完整质粒大小: 10444 bp
载体修饰: --
载体类型: --
筛选标记: --
高拷贝/低拷贝: 低拷贝
生长条件: 37 ℃

插入基因信息
插入基因名称: LshC2c2 locus
物种来源: Leptotrichia shahii
插入大小: --
基因突变: --
GenBank编号 --
启动子 --
标签/融合蛋白: --
克隆方法: --
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: pC003 - LshC2C2 locus into pACYC184 for spacer cloning was a gift from Feng Zhang (Addgene plasmid # 79152 ; http://n2t.net/addgene:79152 ; RRID:Addgene_79152)
参考文献部分: C2c2 is a single-component programmable RNA-guided RNA-targeting CRISPR effector. Abudayyeh OO, Gootenberg JS, Konermann S, Joung J, Slaymaker IM, Cox DB, Shmakov S, Makarova KS, Semenova E, Minakhin L, Severinov K, Regev A, Lander ES, Koonin EV, Zhang F. Science. 2016 Jun 2. pii: aaf5573. 10.1126/science.aaf5573 PubMed 27256883

载体图谱质粒图谱和多克隆位点信息

pC003 - LshC2C2 locus into pACYC184 for spacer cloning质粒图谱

载体简介

载体序列

LOCUS       pC003 - LshC2C2 locus into pACYC184 for spacer cloning               10392 bp ds-DNA     circular SYN 06-AUG-2018
DEFINITION  LshC2c2 locus in pACYC184 with BsaI sites for spacer cloning.
ACCESSION   .
VERSION     .
KEYWORDS    pC003 - LshC2C2 locus into pACYC184 for spacer cloning
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10392)
  AUTHORS   Abudayyeh OO, Gootenberg JS, Konermann S, Joung J, Slaymaker IM, Cox
            DB, Shmakov S, Makarova KS, Semenova E, Minakhin L, Severinov K, 
            Regev A, Lander ES, Koonin EV, Zhang F
  TITLE     C2c2 is a single-component programmable RNA-guided RNA-targeting 
            CRISPR effector.
  JOURNAL   Science. 2016 Jun 2. pii: aaf5573.
  PUBMED    27256883
REFERENCE   2  (bases 1 to 10392)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 6, 2018 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..10392
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             complement(1475..2665)
                     /codon_start=1
                     /gene="tet"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSIIGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     primer_bind     2294..2313
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             3062..7231
                     /codon_start=1
                     /product="type VI CRISPR effector from Leptotrichia shahii 
                     with RNA-guided RNAse activity (Abudayyeh et al., 2016)"
                     /label=LshC2c2
                     /translation="MGNLFGHKRWYEVRDKKDFKIKRKVKVKRNYDGNKYILNINENNN
                     KEKIDNNKFIRKYINYKKNDNILKEFTRKFHAGNILFKLKGKEGIIRIENNDDFLETEE
                     VVLYIEAYGKSEKLKALGITKKKIIDEAIRQGITKDDKKIEIKRQENEEEIEIDIRDEY
                     TNKTLNDCSIILRIIENDELETKKSIYEIFKNINMSLYKIIEKIIENETEKVFENRYYE
                     EHLREKLLKDDKIDVILTNFMEIREKIKSNLEILGFVKFYLNVGGDKKKSKNKKMLVEK
                     ILNINVDLTVEDIADFVIKELEFWNITKRIEKVKKVNNEFLEKRRNRTYIKSYVLLDKH
                     EKFKIERENKKDKIVKFFVENIKNNSIKEKIEKILAEFKIDELIKKLEKELKKGNCDTE
                     IFGIFKKHYKVNFDSKKFSKKSDEEKELYKIIYRYLKGRIEKILVNEQKVRLKKMEKIE
                     IEKILNESILSEKILKRVKQYTLEHIMYLGKLRHNDIDMTTVNTDDFSRLHAKEELDLE
                     LITFFASTNMELNKIFSRENINNDENIDFFGGDREKNYVLDKKILNSKIKIIRDLDFID
                     NKNNITNNFIRKFTKIGTNERNRILHAISKERDLQGTQDDYNKVINIIQNLKISDEEVS
                     KALNLDVVFKDKKNIITKINDIKISEENNNDIKYLPSFSKVLPEILNLYRNNPKNEPFD
                     TIETEKIVLNALIYVNKELYKKLILEDDLEENESKNIFLQELKKTLGNIDEIDENIIEN
                     YYKNAQISASKGNNKAIKKYQKKVIECYIGYLRKNYEELFDFSDFKMNIQEIKKQIKDI
                     NDNKTYERITVKTSDKTIVINDDFEYIISIFALLNSNAVINKIRNRFFATSVWLNTSEY
                     QNIIDILDEIMQLNTLRNECITENWNLNLEEFIQKMKEIEKDFDDFKIQTKKEIFNNYY
                     EDIKNNILTEFKDDINGCDVLEKKLEKIVIFDDETKFEIDKKSNILQDEQRKLSNINKK
                     DLKKKVDQYIKDKDQEIKSKILCRIIFNSDFLKKYKKEIDNLIEDMESENENKFQEIYY
                     PKERKNELYIYKKNLFLNIGNPNFDKIYGLISNDIKMADAKFLFNIDGKNIRKNKISEI
                     DAILKNLNDKLNGYSKEYKEKYIKKLKENDDFFAKNIQNKNYKSFEKDYNRVSEYKKIR
                     DLVEFNYLNKIESYLIDINWKLAIQMARFERDMHYIVNGLRELGIIKLSGYNTGISRAY
                     PKRNGSDGFYTTTAYYKFFDEESYKKFEKICYGFGIDLSENSEINKPENESIRNYISHF
                     YIVRNPFADYSIAEQIDRVSNLLSYSTRYNNSTYASVFEVFKKDVNLDYDELKKKFKLI
                     GNNDILERLMKPKKVSVLELESYNSDYIKNLIIELLTKIENTNDTL"
     rep_origin      9000..9545
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     9446..9465
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9662..9679
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        10071..10173
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             10174..441
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(10240..10259)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 cgtatggcaa tgaaagacgg tgagctggtg atatgggata gtgttcaccc ttgttacacc
       61 gttttccatg agcaaactga aacgttttca tcgctctgga gtgaatacca cgacgatttc
      121 cggcagtttc tacacatata ttcgcaagat gtggcgtgtt acggtgaaaa cctggcctat
      181 ttccctaaag ggtttattga gaatatgttt ttcgtctcag ccaatccctg ggtgagtttc
      241 accagttttg atttaaacgt ggccaatatg gacaacttct tcgcccccgt tttcaccatg
      301 ggcaaatatt atacgcaagg cgacaaggtg ctgatgccgc tggcgattca ggttcatcat
      361 gccgtttgtg atggcttcca tgtcggcaga atgcttaatg aattacaaca gtactgcgat
      421 gagtggcagg gcggggcgta atttttttaa ggcagttatt ggtgccctta aacgcctggt
      481 tgctacgcct gaataagtga taataagcgg atgaatggca gaaattcgaa agcaaattcg
      541 acccggtcgt cggttcaggg cagggtcgtt aaatagccgc ttatgtctat tgctggttta
      601 ccggtttatt gactaccgga agcagtgtga ccgtgtgctt ctcaaatgcc tgaggccagt
      661 ttgctcaggc tctccccgtg gaggtaataa ttgacgatat gatcatttat tctgcctccc
      721 agagcctgat aaaaacggtt agcgcttcgt taatacagat gtaggtgttc cacagggtag
      781 ccagcagcat cctgcgatgc agatccggaa cataatggtg cagggcgctt gtttcggcgt
      841 gggtatggtg gcaggccccg tggccggggg actgttgggc gctgccggca cctgtcctac
      901 gagttgcatg ataaagaaga cagtcataag tgcggcgacg atagtcatgc cccgcgccca
      961 ccggaaggag ctaccggcag cggtgcggac tgttgtaact cagaataaga aatgaggccg
     1021 ctcatggcgt tgactctcag tcatagtatc gtggtatcac cggttggttc cactctctgt
     1081 tgcgggcaac ttcagcagca cgtaggggac ttccgcgttt ccagacttta cgaaacacgg
     1141 aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt
     1201 cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc ccgccagcct
     1261 agccgggtcc tcaacgacag gagcacgatc atgcgcaccc gtggccagga cccaacgctg
     1321 cccgagatgc gccgcgtgcg gctgctggag atggcggacg cgatggatat gttctgccaa
     1381 gggttggttt gcgcattcac agttctccgc aagaattgat tggctccaat tcttggagtg
     1441 gtgaatccgt tagcgaggtg ccgccggctt ccattcaggt cgaggtggcc cggctccatg
     1501 caccgcgacg caacgcgggg aggcagacaa ggtatagggc ggcgcctaca atccatgcca
     1561 acccgttcca tgtgctcgcc gaggcggcat aaatcgccgt gacgatcagc ggtccaatga
     1621 tcgaagttag gctggtaaga gccgcgagcg atccttgaag ctgtccctga tggtcgtcat
     1681 ctacctgcct ggacagcatg gcctgcaacg cgggcatccc gatgccgccg gaagcgagaa
     1741 gaatcataat ggggaaggcc atccagcctc gcgtcgcgaa cgccagcaag acgtagccca
     1801 gcgcgtcggc cgccatgccg gcgataatgg cctgcttctc gccgaaacgt ttggtggcgg
     1861 gaccagtgac gaaggcttga gcgagggcgt gcaagattcc gaataccgca agcgacaggc
     1921 cgatcatcgt cgcgctccag cgaaagcggt cctcgccgaa aatgacccag agcgctgccg
     1981 gcacctgtcc tacgagttgc atgataaaga agacagtcat aagtgcggcg acgatagtca
     2041 tgccccgcgc ccaccggaag gagctgactg ggttgaaggc tctcaagggc atcggtcgac
     2101 gctctccctt atgcgactcc tgcattagga agcagcccag tagtaggttg aggccgttga
     2161 gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc gcccaacagt cccccggcca
     2221 cggggcctgc caccataccc acgccgaaac aagcgctcat gagcccgaag tggcgagccc
     2281 gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct gtggcgccgg
     2341 tgatgccggc cacgatgcgt ccggcgtaga ggatccacag gacgggtgtg gtcgccatga
     2401 tcgcgtagtc gatagtggct ccaagtagcg aagcgagcag gactgggcgg cggccaaagc
     2461 ggtcggacag tgctccgaga acgggtgcgc atagaaattg catcaacgca tatagcgcta
     2521 gcagcacgcc atagtgactg gcgatgctgt cggaatggac gatatcccgc aagaggcccg
     2581 gcagtaccgg cataaccaag cctatgccta cagcatccag ggtgacggtg ccgaggatga
     2641 cgatgagcgc attgttagat ttcatacacg gtgcctgact gcgttagcaa tttaactgtg
     2701 ataaactacc gcattaaagc tcgtttaaca gctgggaaaa tgtggaattt gaaacaaatt
     2761 atcaggatga aagatttatt gatttggtaa cagaaggcag aacatttaga acaggttcaa
     2821 atgggaaaat taagttggat ctaaaggcga aagaaggatt gattttacga aaagtaagga
     2881 tagatgggga ttatgagtaa atagaagtag ggagctatgt ttgacggctc cttatttttt
     2941 tgagatttaa tattgatgtg gtatactaaa aatggaaaat tgtatttttg attagaaaga
     3001 tgtaaaattg atttaattta aaaatatttt attagattaa agtagaaacg aggagattgg
     3061 aatgggaaat ttatttggac ataagagatg gtatgaagtt cgtgataaaa aagattttaa
     3121 aataaaacgg aaagtaaaag ttaaaaggaa ctatgatgga aacaaataca ttttaaatat
     3181 taatgaaaat aataataaag aaaaaattga taataataaa tttattagaa aatatattaa
     3241 ttacaaaaaa aatgataata ttcttaaaga atttacaaga aaatttcatg caggaaatat
     3301 tctatttaag ttgaaaggta aagaaggaat cataagaata gaaaataatg atgatttttt
     3361 ggaaacagaa gaagttgtat tatatataga agcatatgga aaatctgaga aattgaaagc
     3421 attggggatt acaaaaaaga aaataataga tgaagcaatt aggcaaggaa taactaaaga
     3481 tgataaaaaa atagaaataa agagacaaga gaatgaagaa gaaatagaga tagatatcag
     3541 agatgagtat actaataaaa cactaaatga ttgttcaata atattaagaa taatagaaaa
     3601 tgatgaatta gaaacaaaaa aatcaattta cgaaattttt aaaaatatta acatgagttt
     3661 atataagatt atagaaaaaa taattgaaaa tgaaacagaa aaagtatttg aaaatcgata
     3721 ttatgaagaa catttaagag aaaaattatt gaaagatgat aaaatagatg ttattttgac
     3781 taattttatg gaaataagag aaaaaataaa aagtaattta gaaattctgg gttttgtaaa
     3841 attttatctt aatgttggtg gtgataaaaa gaaatctaaa aataaaaaaa tgcttgttga
     3901 aaaaatttta aatataaatg tagatttaac agtagaagat atagctgatt ttgttataaa
     3961 agagttagaa ttttggaata ttactaaaag aattgaaaaa gtaaaaaaag ttaataatga
     4021 atttcttgaa aagagaagaa atagaacata tataaaatct tatgtactat tagataagca
     4081 cgaaaaattt aaaatagaaa gagaaaataa aaaagataaa attgtaaaat tttttgtaga
     4141 aaatataaaa aataatagta taaaagaaaa aatagaaaaa attttagcag aatttaaaat
     4201 agatgaatta ataaaaaaat tagaaaaaga actaaaaaaa ggaaattgtg atacagaaat
     4261 ttttggaata tttaaaaaac attataaagt taattttgat tcgaaaaaat tttcaaagaa
     4321 atctgatgaa gaaaaagaac tatataaaat catatatcga tatttgaaag gaagaattga
     4381 aaaaatattg gtaaatgaac aaaaagtcag attaaagaaa atggaaaaaa tagaaataga
     4441 aaaaatttta aatgaaagta tattatctga aaaaatttta aaaagagtta agcaatatac
     4501 gttagagcat attatgtatt tagggaaatt aagacataat gatatagata tgacaacagt
     4561 aaatactgat gatttttcta gacttcacgc taaagaggaa ttggatttag aattgattac
     4621 tttttttgct tctaccaata tggaattaaa taagattttt tctagagaaa atataaataa
     4681 tgatgaaaat atagatttct ttggaggaga tcgagaaaaa aattacgtac ttgataaaaa
     4741 aattttaaat tcaaaaataa aaataataag agacttagat tttatagata ataaaaataa
     4801 cattacaaat aattttatta ggaaatttac aaagatagga acaaatgaaa gaaacaggat
     4861 attacatgcg attagcaagg aaagagattt acaaggaacg caagatgact ataataaagt
     4921 cataaatata attcaaaatt taaaaatatc agatgaagaa gtgtcaaaag cattaaattt
     4981 agatgtagta tttaaagata aaaaaaatat aattactaaa ataaatgata ttaaaatttc
     5041 agaagaaaat aataatgata ttaaatattt accatccttt tctaaagtac ttcctgaaat
     5101 attaaattta tataggaata atcctaagaa tgaaccattt gatactatag aaacagaaaa
     5161 aatagtatta aatgctttaa tatatgtgaa taaagaattg tataagaaat taattttaga
     5221 agatgattta gaagaaaatg aaagtaagaa tatattttta caagaattaa agaaaacttt
     5281 aggaaacatt gatgaaatag atgaaaatat tatagaaaat tattataaaa atgctcagat
     5341 ttctgcttca aaaggtaata ataaagctat taaaaaatat cagaaaaaag taatagaatg
     5401 ttatattgga tatttgagaa aaaattatga ggaacttttt gatttttcag attttaaaat
     5461 gaatattcaa gaaattaaga aacaaataaa agacattaat gataataaaa cttacgaaag
     5521 aataactgta aagactagtg ataaaactat agtaataaat gatgattttg aatatataat
     5581 ttcaatattt gcactgttaa atagtaatgc ggttataaat aaaattagaa atagattttt
     5641 tgcaacatca gtttggttaa atacttcaga atatcaaaat attatagata ttttagatga
     5701 aataatgcaa ttaaatactt tgagaaatga atgtataact gaaaattgga atcttaattt
     5761 agaagaattt attcaaaaaa tgaaagagat agaaaaagac tttgatgatt ttaaaataca
     5821 aactaaaaaa gaaattttta ataattatta tgaggatata aaaaataata tattaactga
     5881 atttaaagat gatattaatg gttgtgatgt tttagagaaa aaattagaaa aaattgtaat
     5941 ttttgatgat gaaactaaat ttgagataga taaaaaatcg aatatattac aagatgaaca
     6001 aagaaagtta tcaaacataa ataaaaagga tttgaaaaaa aaggttgatc agtatataaa
     6061 agataaagat caggaaataa agagtaaaat attgtgtaga ataatattta attctgattt
     6121 tttaaaaaaa tacaaaaaag aaatagataa tttaatagaa gatatggaat cagagaatga
     6181 aaataaattt caagaaatat attatcctaa agaacgtaag aatgaattat atatttataa
     6241 aaagaattta tttttaaata ttggaaatcc aaattttgat aaaatttatg gattaatttc
     6301 aaatgatatt aaaatggctg atgcaaaatt tttatttaat attgatggta aaaatattag
     6361 aaaaaataaa atatctgaaa ttgatgcaat attgaaaaat ctaaatgata aactaaatgg
     6421 atattctaaa gaatacaaag agaagtatat aaaaaaattg aaagaaaatg atgacttttt
     6481 tgcaaaaaat atacaaaata aaaactataa atcatttgaa aaagattata atagagtttc
     6541 tgaatacaaa aaaattagag atttggtaga atttaattat ttgaataaaa tagaaagtta
     6601 tttgatagat ataaattgga aacttgctat tcaaatggct agatttgaaa gagatatgca
     6661 ctatattgta aatgggctaa gagaattagg aattattaag ttaagtggct acaatactgg
     6721 aataagtaga gcttatccta agcgtaatgg atctgatggc ttttatacaa caacagcata
     6781 ctataaattt tttgatgaag aatcttataa aaaatttgag aaaatatgtt atggatttgg
     6841 aattgatttg tctgaaaatt ctgaaattaa taagccggaa aatgaaagta ttagaaacta
     6901 tatttcacat ttctatattg taagaaatcc gtttgctgat tacagtattg cagaacaaat
     6961 tgatagagtt tctaatttat tatcatatag tacacgttat aataattcaa cttatgcaag
     7021 tgtatttgaa gtatttaaaa aagatgtgaa tttagattat gatgaattaa agaaaaaatt
     7081 taaacttatc ggtaataatg atatacttga aagattaatg aaacctaaaa aagtttctgt
     7141 tttagaatta gaaagttata attctgatta tattaaaaac ttgataatag aacttttaac
     7201 aaaaatagaa aatacgaatg atacgttata agaatgaaat ttataaggag tatgattaaa
     7261 atgcccaata taattcacat aacaagggct gatgatttgt caatatcgaa taatcagctt
     7321 ataatggttg atgaagacaa taatgatgag aaaaataaga tttctttgaa tgatatatct
     7381 gcgatagtta ttgaaaactg ctattgtaag attagtgcga ttttgcagtt gaggctggtg
     7441 gaaaataaca ttccgattat tatttgcaac gagaaacatc agcctgaaat tcattcgctg
     7501 ggattattta atcattttca agtgacttta cgaataaatg agcagattga gtgggaaaaa
     7561 gagaaaaagg aaaaattgtg gagcaggata gtggagaata agattgaaaa tcaaagagca
     7621 ttgttagagt atcttgaaaa aagtgatatt tccattgaaa gactgaaaac atataaggaa
     7681 aacttgaaaa aagatgatgc gagtgcagaa catcaggaag caatagcttc aaggatatat
     7741 tttcaggaat tgtatagtaa tagttttaaa agatttgatg aagatggagt gaattctgca
     7801 cttaattatg gatatatgat tttaagagct ataatatcat caaagattgt agcaaaagga
     7861 tttcatccaa gtttaggatt acatcataag tcacagttta atgcctataa tttttctgat
     7921 gatataatag aggtttttcg tcctatggtg gattatcttg tttatatgta caaggatatt
     7981 ttgaatgaag tgaaattaag taaggaaata aggcaaaaaa tacttcttgt agcacagcag
     8041 aaggtattat ttaacaataa gaaatatgat tttttccaag cggtagacta ttatttggat
     8101 agtatacgaa attattttgt aaaagatgaa gaagtgataa ttccgacatt gagcgtgatg
     8161 gactatgagt attgagtatt ttggtctgct tatgtatgat tttcctatgc agacagatgt
     8221 tgagagattt gaatatagga cttttaggaa aaaattgata gaaaagggat attatcagct
     8281 tcaaaaatcg gtatatataa tgaggtcgaa aacaaaagaa aaaatcgaac tggcagaaaa
     8341 acagttatca atgctcgctc cagaaaattc atcaatccgt tccattatcc tgacagaaga
     8401 tcagtttcaa aaatgaaggt acttgcagga gaagtgacaa tgggagaaaa aatttcacaa
     8461 aacaaaatat tggaattttg aaaaatcaaa aaatatggta taatagtata gggtggttct
     8521 atattattaa taaagagaaa aataggattg aaagtattga aaataaagac aaaatagaaa
     8581 aattagaaat aaatatctca tcagcaagaa attaggatgg gatatagacc accccaatat
     8641 cgaaggggac taaaacggag accgagattg gtctcggata tagaccaccc caatatcgaa
     8701 ggggactaaa actctaaaga attatctatt ctgtctttta aattgatata gaccacccca
     8761 atatcgaaaa gtgatattta ataaaaataa cttctgagtg agaataaaat ttcaattctt
     8821 gctcattttt tattgttttt tgaatatggt tgcttggttg ttctgaaacg aaaatttttg
     8881 gagatgtttt aaatttttag gttgaaaaaa aataagaatt atactataat aaataattat
     8941 gcgattaggc taactagagc ctaatttaaa tattttatct gattaataag atgatcttct
     9001 tgagatcgtt ttggtctgcg cgtaatctct tgctctgaaa acgaaaaaac cgccttgcag
     9061 ggcggttttt cgaaggttct ctgagctacc aactctttga accgaggtaa ctggcttgga
     9121 ggagcgcagt caccaaaact tgtcctttca gtttagcctt aaccggcgca tgacttcaag
     9181 actaactcct ctaaatcaat taccagtggc tgctgccagt ggtgcttttg catgtctttc
     9241 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcggact gaacgggggg
     9301 ttcgtgcata cagtccagct tggagcgaac tgcctacccg gaactgagtg tcaggcgtgg
     9361 aatgagacaa acgcggccat aacagcggaa tgacaccggt aaaccgaaag gcaggaacag
     9421 gagagcgcac gagggagccg ccagggggaa acgcctggta tctttatagt cctgtcgggt
     9481 ttcgccacca ctgatttgag cgtcagattt cgtgatgctt gtcagggggg cggagcctat
     9541 ggaaaaacgg ctttgccgcg gccctctcac ttccctgtta agtatcttcc tggcatcttc
     9601 caggaaatct ccgccccgtt cgtaagccat ttccgctcgc cgcagtcgaa cgaccgagcg
     9661 tagcgagtca gtgagcgagg aagcggaata tatcctgtat cacatattct gctgacgcac
     9721 cggtgcagcc ttttttctcc tgccacatga agcacttcac tgacaccctc atcagtgcca
     9781 acatagtaag ccagtataca ctccgctagc gctgatgtcc ggcggtgctt ttgccgttac
     9841 gcaccacccc gtcagtagct gaacaggagg gacagctgat agaaacagaa gccactggag
     9901 cacctcaaaa acaccatcat acactaaatc agtaagttgg cagcatcacc cgacgcactt
     9961 tgcgccgaat aaatacctgt gacggaagat cacttcgcag aataaataaa tcctggtgtc
    10021 cctgttgata ccgggaagcc ctgggccaac ttttggcgaa aatgagacgt tgatcggcac
    10081 gtaagaggtt ccaactttca ccataatgaa ataagatcac taccgggcgt attttttgag
    10141 ttatcgagat tttcaggagc taaggaagct aaaatggaga aaaaaatcac tggatatacc
    10201 accgttgata tatcccaatg gcatcgtaaa gaacattttg aggcatttca gtcagttgct
    10261 caatgtacct ataaccagac cgttcagctg gatattacgg cctttttaaa gaccgtaaag
    10321 aaaaataagc acaagtttta tccggccttt attcacattc ttgcccgcct gatgaatgct
    10381 catccggaat tc
//

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