pSCBE3-HF

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pSCBE3HF载体基本信息

质粒名称: pSCBE3-HF
出品公司: Addgene
目录编号: 133967
存储实验室: Yuhui Sun

载体骨架基本信息
载体名称: pYH7
载体出品公司: Yuhui Sun
空载体大小: --
完整质粒大小: --
载体修饰: --
载体类型: CRISPR,合成生物学
筛选标记: 安普霉素用于链霉菌的筛选
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: HF-BE3
物种来源: Streptomyces
插入大小: --
基因突变: --
GenBank编号 --
启动子: --
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: pSCBE3-HF was a gift from Yuhui Sun (Addgene plasmid # 133967 ; http://n2t.net/addgene:133967 ; RRID:Addgene_133967)
参考文献部分: Base editing in Streptomyces with Cas9-deaminase fusions. Zhong Z, Guo J, Deng L, Chen L, Wang J, Li S, Xu W, Deng Z, Sun Y. BioRxiv 630137 10.1101/630137

序号

pSCBE3HF载体图谱质粒图谱和多克隆位点信息

pSCBE3-HF载体图谱

pSCBE3HF载体简介

pSCBE3HF载体序列

LOCUS       pSCBE3-HF              13610 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Expresses HF-BE3 CRISPR base editor and gRNA scaffold in 
            Streptomyces.
ACCESSION   .
VERSION     .
KEYWORDS    pSCBE3-HF
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13610)
  AUTHORS   Zhong Z, Guo J, Deng L, Chen L, Wang J, Li S, Xu W, Deng Z, Sun Y
  TITLE     Base editing in Streptomyces with Cas9-deaminase fusions
  JOURNAL   BioRxiv 630137
REFERENCE   2  (bases 1 to 13610)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..13610
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(250..270)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(250..268)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(297..315)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        383..487
                     /gene="bla"
                     /label=AmpR promoter
     CDS             488..1348
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(706..725)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1519..2107
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2008..2027
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2261..2278
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    2492..2626
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2516..2535)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2570..2589
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        5844..6145
                     /label=rpsL promoter
                     /note="promoter for the Xylanimonas cellulosilytica 
                     ribosomal protein S12 gene"
     CDS             6146..6832
                     /codon_start=1
                     /product="cytidine deaminase (C to U editing enzyme) from 
                     rat"
                     /label=APOBEC-1
                     /note="can use ssDNA as a substrate (Komor et al., 2016)"
                     /translation="MSSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWG
                     GRHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFL
                     SRYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSN
                     EAHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHI
                     LWATGLK"
     CDS             10994..11245
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     oriT            11608..11717
                     /note="incP origin of transfer"
     CDS             11750..12121
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     misc_RNA        12277..12352
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             complement(12866..56)
                     /codon_start=1
                     /gene="aac(3)IV"
                     /product="aminoglycoside 3-N-acetyltransferase type IV     
                               "
                     /label=ApmR
                     /note="confers resistance to apramycin (Paget and Davies, 
                     1996)"
                     /translation="MSSAVECNVVQYEWRKAELIGQLLNLGVTPGGVLLVHSSFRSVRP
                     LEDGPLGLIEALRAALGPGGTLVMPSWSGLDDEPFDPATSPVTPDLGVVSDTFWRLPNV
                     KRSAHPFAFAAAGPQAEQIISDPLPLPPHSPASPVARVHELDGQVLLLGVGHDANTTLH
                     LAELMAKVPYGVPRHCTILQDGKLVRVDYLENDHCCERFALADRWLKEKSLQKEGPVGH
                     AFARLIRSRDIVATALGQLGRDPLIFLHPPEAGCEECDAARQSIG"
ORIGIN
        1 agctcggctt ttcgccattc gtattgcacg acattgcact ccaccgctga tgacatcagt
       61 cgatcatagc acgatcaacg gcactgttgc aaatagtcgg tggtgataaa cttatcatcc
      121 ccttttgctg atggagctgc acatgaaccc attcaaaggc cggcattttc agcgtgacat
      181 cattctgtgg gccgtacgct ggtactgcaa atacggcatc agttaccgtg agctgcaggt
      241 cgacggatcc ctttagtgag ggttaattgc ggccgcgaat tcttgaagac gaaagggcct
      301 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
      361 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
      421 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
      481 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
      541 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
      601 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
      661 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
      721 attatcccgt gttgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
      781 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
      841 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
      901 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
      961 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     1021 cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     1081 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     1141 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     1201 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     1261 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     1321 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     1381 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     1441 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     1501 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     1561 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     1621 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
     1681 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     1741 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     1801 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     1861 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     1921 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     1981 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     2041 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     2101 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     2161 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     2221 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     2281 agcggaagag cgctgacttc cgcgtttcca gactttacga aacacggaaa ccgaagacca
     2341 ttcatgttgt tgctcaggtc gcagacgttt tgcagcagca gtcgcttcac gttcgctcgc
     2401 gtatcggtga ttcattctgc taaccagtaa ggcaaccccg ccagcctagc cgggtcctca
     2461 acgacaggag cacgatcatg cgcacccgtc agatccagac atgataagat acattgatga
     2521 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     2581 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttcccg cacgatcagc
     2641 gtcccgccga ccaggagcag tgcggccagc actgccgcta acgcctggtc ctggtcccgg
     2701 tcctggtggt gcatcagtcc tccccgtgat cacttcggca cccaccgtag tgatcacccc
     2761 cgacagcgga tcaaggggtt tgcgggtccc ggtcggcgcc gggcggggga ggcaggagcc
     2821 gccgacgctg cctctgggac gggccggacg gcagggggac cggcggccgg gcgagctgca
     2881 gccgggggtc cggcagggcc ggagcgggcg gaaccgtgct ctgacctgcg gcccgagttt
     2941 cgtcacgtga cggaatggaa ggctgctgca tttcgtcacg tgacgtatct cggcgagcga
     3001 ctgccgacgc cacggcggac acgatcgcct cgcgctggcg ccgggcctcg tacgcccgct
     3061 ggcggcagga gcggcggcag tagtcccggc tccggccgac gccggattgc ttgatctccg
     3121 agccgcacca ggcgcagagc ttcgcgccgt cggcgtccct gggggtggtg gtgctcatgg
     3181 ccgacgaccg tacgcggcac gtctcgtagc gaggcgagtc gggcgcgagg taccgcctgc
     3241 acgaagtgcc ggcggggccg accccgggcg agtaatccca ggattactcc cgcggcttcg
     3301 accccggccg ccgtcgccgc gtacgtcacc gacccccgcc gtacgtcacc gggatgacgt
     3361 acggcggggg ggagcgagtt agtgcgaagt gggcccactt gcgagccggg cgatgtgccg
     3421 ggcggcccgc tcctggcggt cgtcggcgtc gtcgtcctgg tcgtcgtcct gctctcgccg
     3481 tcggcgtgca gttgcttcct cgcggcgctg ggcgagggcg gcgagcatgt cggcgtacgc
     3541 ctcggccacc tcccccgccg tgagcaccac cactgtgtcg gccgcgtcgg ccagcgccag
     3601 gacctcccgc acccgttcgc ccacggccgc cgaatcctcg ttgccgtcct tgccttcggc
     3661 ggcccgggtc gcctcgaggt cgagggcgcg gcgggtgacc gcgtgccatc cgtcctcggt
     3721 cacggcgacc ccggcccgca gctccccgcc gtcggcgtcg gccgccagga gcagatcgag
     3781 gtcgtcggcc tcggtgtcgc cgccgtcgag cccgagcatc tgccgcaggt agcgggtcca
     3841 ttcgatggcc cggcgtcccc gggttgcccg ctcgtactcg tgccagcgcg agaggttcca
     3901 ctccagcgag ccgaccccgg cggcgtcgtc ctcggtcatg ccgccggtca ggtccccgat
     3961 ccgtccgagg agttcgaacg gggcgacgtt cccgccggtc gccgtcttga ggtcggcgcg
     4021 ggcgagttcg agggcgggcg ccttcccgtc ctgggtcttg gcgatgtact cggcgaggtc
     4081 gttggcgtcg cgctcggtct ccagccgctt gaagtcgacg ccgtgccggt cgtcgggcgt
     4141 gaaggcgggg ttgaccttgc gcagggcggc ggtccacacg gaccgccagt gcccctgcca
     4201 ctcgtcgagc gcggcgccgg tcggctcgaa ggtggcgacg atctgcttcg cggaccgctc
     4261 cccctcggtc cggccgccga ccaggacgat cgcgtggatg tgcgggtgcc agccgttgat
     4321 ctgccccacg gtgacttcgg tcgcgcggat catgccgacg tacccgatcc ggtctcggat
     4381 gccctcgcgg tcggcggccc ggtgcccgtc cttggcccgg cgtccggccc acgtgccgcc
     4441 cgtgatcagt cgctggtagg cgcccggccg ccgggggctg tccggcgtct tccgggtgcc
     4501 ctggagggcg tccatgaggt ccgcgagccg gtccgtgtgc ccatggcggg ccgtgaaggt
     4561 gaccaggtag gcggtccccc cgcgcttgat ccactcgacc acggcggcgg tgatctcctc
     4621 ggcccgcttg tgccggatcg tggcggcgca gaccgggcag agccagatcc gcccgcaccg
     4681 catcaggccc aggaccacgg acgttccggc cgccgtctgg gcgacgatca cgccggaggc
     4741 agggtccatc agggcgcggc cgcagccctt gcacgcggcg tccccgctga tccgccacag
     4801 cgtccggcgg cggctgtacc gggcggcttt ccgcagtcgg gcagcgtcgc tccgcgacgt
     4861 gcttcctact tccgagaggc tgtcgcctct cgggctctcc ccatccaccc cgtccggaga
     4921 aaccgcaggt cggaggggtg cgggaaactc tgttgtttct ttcccaaggt gttcgctttt
     4981 gcctcgggcg gcatctcgcg tcacacgcgc gatcgcccgc ttcgctgcca tccggcagcg
     5041 gtctgagcag tagatacgcg gccgtttgcc cggtgtgtgg gcaattgcgg tcccgcagtg
     5101 gcagcggggc ccggcgggcc gatctggcaa tgcctcggca tcgctccgta ctctgggcac
     5161 gagcaacgtt cctgtctcgc ccggctaagg ggcgcgagtc tgggagcgga cgggtcggag
     5221 gtgcgaagtc cggcccgttg ctctttggtc tggtgggaat cctggcacca atcgggccag
     5281 aggttccctc cgccactccc gacgcccctt ggggctggtg tgacttggag ggccgaagag
     5341 agccccgccg gtgatccggc ggggctttga cgtgcggtca gtgcgtgtgt cggcgagcga
     5401 tggccacgag gccctggaag ccgagcggtc cggcgaagtc ggcccagtcg caaccgggct
     5461 cagcgcagtg ggcggaccag ccaccgccgt tggggtcctg gaccaggttc acggtcccct
     5521 cggtcaggcg tccgtcgaag tcggtcatgg tcggtctcct ggtgggtggg ggcggggcgc
     5581 cagcacgaag tgccggcgcc ccgcgggggt tggtcgggtc aggcgccgaa ccggcgggcg
     5641 gcggcggcga ccaggccgtc ggcggcggcc atggcgcggt cgcggtcggt ggtgagggcg
     5701 gtgcggtcgg cggcggccca gtcctgctct cacgcaacgt ctacgtggac gctcagggcg
     5761 acacgatcga ggtcggggag tccgtcgctc cggcgggccg ctggcgcgtc caccgggact
     5821 gatcaaggcg aatacttcat atggccctgc aggcggaagt caggtagaca cgacttccgc
     5881 tagtccttgc aaggtctgct gacgtgaggc ggggcggtcg tttttgaccg ccctgccttc
     5941 gtcatgtagg ctcgctcgct gtgcctggcg tgtcatcaga cgcccaggtc ccggtgccgt
     6001 gaggcccggg ccatcgagcc ggtggtacgt ggctgcggtc cccttgtgag ggctgcgcgc
     6061 cgtgtgctgt ccggcgcgca cagccttgaa tccacccgcg ggggccggcc ggtctccgtg
     6121 agctcgagta gacgacggag acgtaatgtc gtccgagacc ggcccggtgg ccgtggaccc
     6181 gaccctgcgt cgccgcatcg agccgcacga gttcgaggtg ttcttcgacc cgcgggagct
     6241 gcgcaaggag acctgcctgc tgtacgagat caactggggc ggccggcact cgatctggcg
     6301 ccacacctcg cagaacacca acaagcacgt cgaggtgaac ttcatcgaga agttcaccac
     6361 cgagcggtac ttctgcccga acacccgctg ctcgatcacc tggttcctgt cctggtcccc
     6421 gtgcggcgag tgctcccggg cgatcaccga gttcctgtcc cgctacccgc acgtcaccct
     6481 gttcatctac atcgcccggc tgtaccacca cgccgacccg cggaaccggc agggcctgcg
     6541 cgacctgatc tcctccggcg tgaccatcca gatcatgacc gagcaggagt ccggctactg
     6601 ctggcggaac ttcgtcaact actcgccgtc caacgaggcc cactggccgc ggtacccgca
     6661 cctgtgggtg cgcctgtacg tcctggagct gtactgcatc atcctgggcc tgccgccgtg
     6721 cctgaacatc ctgcgtcgca agcagccgca gctgaccttc ttcaccatcg ccctgcagtc
     6781 ctgccactac cagcgcctgc cgccgcacat cctgtgggcg accggcctga agtccggctc
     6841 ggagaccccg ggcacctcgg agtccgccac cccggagtcc gacaagaagt acagcatcgg
     6901 cctggccatc ggcaccaaca gcgtgggctg ggcggtcatc accgacgagt acaaggtccc
     6961 ctccaagaag ttcaaggtcc tgggcaacac cgaccggcac tcgatcaaga agaacctgat
     7021 cggcgccctg ctcttcgaca gcggcgagac cgccgaggcg acccgcctga agcggaccgc
     7081 gcgccggcgc tacacccggc gcaagaaccg catctgctac ctccaggaaa tcttctccaa
     7141 cgagatggcc aaggtcgacg actcgttctt ccaccggctc gaggagagct tcctggtgga
     7201 ggaggacaag aagcacgagc gccacccgat cttcggcaac atcgtcgacg aggtggccta
     7261 ccacgagaag taccccacca tctaccacct ccgcaagaag ctggtcgact cgaccgacaa
     7321 ggcggacctg cggctcatct acctggccct cgcgcacatg atcaagttcc gcggccactt
     7381 cctcatcgag ggcgacctga acccggacaa ctccgacgtc gacaagctct tcatccagct
     7441 ggtgcagacc tacaaccagc tgttcgagga gaaccccatc aacgccagcg gcgtcgacgc
     7501 caaggcgatc ctctccgcgc gcctgagcaa gtcccggcgc ctggagaacc tcatcgccca
     7561 gctgccgggc gagaagaaga acggcctctt cggcaacctg atcgcgctgt cgctcggcct
     7621 gacccccaac ttcaagagca acttcgacct ggccgaggac gcgaagctcc agctgtccaa
     7681 ggacacctac gacgacgacc tggacaacct gctcgcccag atcggcgacc agtacgcgga
     7741 cctcttcctg gccgcgaaga acctctcgga cgccatcctg ctcagcgaca tcctgcgggt
     7801 caacaccgag atcaccaagg ccccgctgtc ggcgagcatg atcaagcggt acgacgagca
     7861 ccaccaggac ctgaccctgc tcaaggccct cgtgcgccag cagctgcccg agaagtacaa
     7921 ggaaatcttc ttcgaccagt ccaagaacgg ctacgccggc tacatcgacg gcggcgcgtc
     7981 gcaggaggag ttctacaagt tcatcaagcc gatcctggag aagatggacg gcaccgagga
     8041 gctgctcgtc aagctgaacc gcgaggacct gctccgcaag cagcggacct tcgacaacgg
     8101 ctccatcccg caccagatcc acctgggcga gctgcacgcc atcctccggc gccaggagga
     8161 cttctacccc ttcctgaagg acaaccgcga gaagatcgag aagatcctga ccttccgcat
     8221 cccgtactac gtcggccccc tggcccgcgg caactcccgg ttcgcgtgga tgacccggaa
     8281 gtcggaggag accatcaccc cgtggaactt cgaggaggtc gtggacaagg gcgcctccgc
     8341 ccagtcgttc atcgagcgca tgaccgcctt cgacaagaac ctcccgaacg agaaggtcct
     8401 gcccaagcac tccctgctct acgagtactt caccgtgtac aacgagctga ccaaggtcaa
     8461 gtacgtgacc gagggcatgc ggaagccggc cttcctgtcg ggcgagcaga agaaggcgat
     8521 cgtcgacctg ctcttcaaga ccaaccgcaa ggtcaccgtg aagcagctga aggaggacta
     8581 cttcaagaag atcgagtgct tcgactccgt cgagatcagc ggcgtggagg accgcttcaa
     8641 cgcctccctg ggcacctacc acgacctgct caagatcatc aaggacaagg acttcctcga
     8701 caacgaggag aacgaggaca tcctggagga catcgtcctc accctgaccc tcttcgagga
     8761 ccgcgagatg atcgaggagc ggctcaagac ctacgcccac ctgttcgacg acaaggtgat
     8821 gaagcagctg aagcggcgcc ggtacaccgg ctggggcgcc ctctcccgga agctgatcaa
     8881 cggcatccgg gacaagcaga gcggcaagac catcctggac ttcctcaagt ccgacggctt
     8941 cgccaaccgc aacttcatgg cgctcatcca cgacgacagc ctgaccttca aggaggacat
     9001 ccagaaggcc caggtctcgg gccagggcga cagcctccac gagcacatcg ccaacctggc
     9061 gggctccccg gcgatcaaga agggcatcct ccagaccgtc aaggtcgtgg acgagctggt
     9121 caaggtgatg ggccgccaca agcccgagaa catcgtgatc gagatggccc gggagaacca
     9181 gaccacccag aagggccaga agaactcgcg cgagcggatg aagcggatcg aggagggcat
     9241 caaggagctg ggcagccaga tcctgaagga gcacccggtc gagaacaccc agctccagaa
     9301 cgagaagctg tacctctact acctccagaa cggccgcgac atgtacgtgg accaggagct
     9361 ggacatcaac cggctgtccg actacgacgt cgaccacatc gtgccgcagt ccttcctgaa
     9421 ggacgactcg atcgacaaca aggtcctgac ccgctcggac aagaaccggg gcaagtccga
     9481 caacgtgccc tcggaggagg tcgtgaagaa gatgaagaac tactggcgcc agctgctcaa
     9541 cgccaagctc atcacccagc gcaagttcga caacctgacc aaggccgagc ggggcggcct
     9601 gagcgagctg gacaaggcgg gcttcatcaa gcgccagctg gtcgagaccc gggcgatcac
     9661 caagcacgtg gcccagatcc tggactcccg gatgaacacc aagtacgacg agaacgacaa
     9721 gctgatccgc gaggtcaagg tgatcaccct caagagcaag ctggtctccg acttccgcaa
     9781 ggacttccag ttctacaagg tccgggagat caacaactac caccacgccc acgacgcgta
     9841 cctgaacgcc gtcgtgggca ccgcgctgat caagaagtac ccgaagctgg agtccgagtt
     9901 cgtctacggc gactacaagg tctacgacgt gcgcaagatg atcgccaaga gcgagcagga
     9961 gatcggcaag gccaccgcga agtacttctt ctactccaac atcatgaact tcttcaagac
    10021 cgagatcacc ctggccaacg gcgagatccg caagcggccc ctgatcgaga ccaacggcga
    10081 gaccggcgag atcgtctggg acaagggccg cgacttcgcc accgtccgga aggtgctgtc
    10141 gatgccgcag gtcaacatcg tgaagaagac cgaggtgcag accggcggct tcagcaagga
    10201 gtccatcctc cccaagcgca acagcgacaa gctgatcgcc cggaagaagg actgggaccc
    10261 gaagaagtac ggcggcttcg acagccccac cgtcgcctac tccgtgctgg tcgtggcgaa
    10321 ggtcgagaag ggcaagagca agaagctgaa gtccgtgaag gagctgctcg gcatcaccat
    10381 catggagcgc tcctcgttcg agaagaaccc gatcgacttc ctggaggcca agggctacaa
    10441 ggaggtcaag aaggacctca tcatcaagct gcccaagtac agcctgttcg agctggagaa
    10501 cggccgcaag cggatgctcg cctccgcggg cgagctgcaa aagggcaacg agctggccct
    10561 cccgtcgaag tacgtcaact tcctgtacct cgcgtcccac tacgagaagc tgaagggctc
    10621 gcccgaggac aacgagcaga agcagctctt cgtggagcag cacaagcact acctggacga
    10681 gatcatcgag cagatcagcg agttcagcaa gcgcgtcatc ctggccgacg cgaacctcga
    10741 caaggtgctg tccgcctaca acaagcaccg cgacaagccg atccgggagc aggcggagaa
    10801 catcatccac ctgttcaccc tcaccaacct gggtgccccg gccgccttca agtacttcga
    10861 caccaccatc gaccgcaagc ggtacacctc caccaaggag gtcctcgacg cgaccctgat
    10921 ccaccagagc atcaccggcc tgtacgagac ccgcatcgac ctgagccagc tgggcggcga
    10981 ctcgggcggc tccaccaacc tgtcggacat catcgagaag gagaccggca agcagctggt
    11041 gatccaggag tccatcctga tgctgccgga ggaagtggag gaagtgatcg gcaacaagcc
    11101 ggagtcggac atcctggtgc acaccgccta cgacgagtcg accgacgaga acgtcatgct
    11161 gctgacctcg gacgccccgg agtacaagcc gtgggccctg gtcatccagg actccaacgg
    11221 cgagaacaag atcaagatgc tgtgagaatt cagatctacg cgttccccgc aaaagcggcc
    11281 tttgactccc tgcaagcctc agcgaccgaa tatatcggtt atgcgtgggc gatggttgtt
    11341 gtcattgtcg gcgcaactat cggtatcaag ctgtttaaga aattcacctc gaaagcaagc
    11401 tgataaaccg atacaattaa aggctccttt tggagccttt ttttaagctt gcatgcctgc
    11461 aggtcgacgg atcttttccg ctgcataacc ctgcttcggg gtcattatag cgattttttc
    11521 ggtatatcca tcctttttcg cacgatatac aggattttgc caaagggttc gtgtagactt
    11581 tccttggtgt atccaacggc gtcagccggg caggataggt gaagtaggcc cacccgcgag
    11641 cgggtgttcc ttcttcactg tcccttattc gcacctggcg gtgctcaacg ggaatcctgc
    11701 tctgcgaggc tggccggcta ccgccggcgt aacagatgag ggcaagcgga tggctgatga
    11761 aaccaagcca accaggaagg gcagcccacc tatcaaggtg tactgccttc cagacgaacg
    11821 aagagcgatt gaggaaaagg cggcggcggc cggcatgagc ctgtcggcct acctgctggc
    11881 cgtcggccag ggctacaaaa tcacgggcgt cgtggactat gagcacgtcc gcgagctggc
    11941 ccgcatcaat ggcgacctgg gccgcctggg cggcctgctg aaactctggc tcaccgacga
    12001 cccgcgcacg gcgcggttcg gtgatgccac gatcctcgcc ctgctggcga agatcgaaga
    12061 gaagcaggac gagcttggca aggtcatgat gggcgtggtc cgcccgaggg cagagccatg
    12121 acttttttag ccgctaaaac ggccgggggg tgcgcgtgat tgccaagcac gtccccatgc
    12181 gctccatcaa gaagagcgac ttcgcggagc tggtgaagta catcaccgac gagcaaggca
    12241 agaccgatcc ccggggacct gcaggtcgac tctagagttt tagagctaga aatagcaagt
    12301 taaaataagg ctagtccgtt atcaacttga aaaagtggca ccgagtcggt gctaaaaaaa
    12361 aaccccgccc ctgacagggc ggggtttttt ttgtagcctc aggagttgta gctgttcaca
    12421 ttcgaacggt ctctgctttg acaacatgct gtgcggtgtt gtaaagtcgt ggccgctagc
    12481 tatttacccg caggacatat ccacgccctc ctacatcgaa gctgaaagca cgagattctt
    12541 cgccctccga gagctgcatc aggtcggaga cgctgtcgaa cttttcgatc agaaacttct
    12601 cgacagacgt cgcggtgagt tcaggctttt tcatatctca ttgcccccgg acgagcgtct
    12661 gctccgccat tcgccgtccg ccgtgccaat cggatcagcc gtccaaatgc gggattttcg
    12721 ttagtcggag gccaaacggc attgagcgtc agcatatcat cagcgagctg aagaaagaca
    12781 atccccgatc cgctccacgt gttgccccag caatcagcgc gaccttgccc ctccaacgtc
    12841 atctcgttct ccgctcatga gctcagccaa tcgactggcg agcggcatcg cattcttcgc
    12901 atcccgcctc tggcggatgc aggaagatca acggatctcg gcccagttga cccagggctg
    12961 tcgccacaat gtcgcgggag cggatcaacc gagcaaaggc atgaccgact ggaccttcct
    13021 tctgaaggct cttctccttg agccacctgt ccgccaaggc aaagcgctca cagcagtggt
    13081 cattctcgag ataatcgacg cgtaccaact tgccatcctg aagaatggtg cagtgtctcg
    13141 gcaccccata gggaaccttt gccatcaact cggcaagatg cagcgtcgtg ttggcatcgt
    13201 gtcccacgcc gaggagaagt acctgcccat cgagttcatg gacacgggcg accgggcttg
    13261 caggcgagtg aggtggcagg ggcaatggat cagagatgat ctgctctgcc tgtggccccg
    13321 ctgccgcaaa ggcaaatgga tgggcgctgc gctttacatt tggcaggcgc cagaatgtgt
    13381 cagagacaac tccaaggtcc ggtgtaacgg gcgacgtggc aggatcgaac ggctcgtcgt
    13441 ccagacctga ccacgagggc atgacgagcg tccctcccgg acccagcgca gcacgcaggg
    13501 cctcgatcag tccaagtggc ccatcttcga ggggccggac gctacggaag gagctgtgga
    13561 ccagcagcac accgccgggg gtaaccccaa ggttgagaag ctgaccgatg
//

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