SECURE BE3(R33A/K34A)-P2A-EGFP (pJUL1410)

价格:1200元

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: SECURE BE3(R33A/K34A)-P2A-EGFP (pJUL1410) 
出品公司: 爱迪基因
目录编号: 123615
存储实验室: Keith Joung

载体骨架基本信息
载体名称: CAG
载体出品公司: pSQT817 (Addgene #53373)
载体抗性: 氨苄青霉素
空载体大小: 4843 bp
完整质粒大小: 10768 bp
载体类型: 哺乳细胞表达
高拷贝/低拷贝: 高拷贝
生长条件: 37 ℃

插入基因信息
插入基因名称: BE3(R33A/K34A)-P2A-EGFP,rAPOBEC1(R33A/K34A)-XTEN-hSpCas9n(D10A)-UGI-NLS(SV40)-P2A-EGFP
物种来源: R. norvegicus (rat), Synthetic; Streptococcus pyogenes, Bacillus subtilis bacteriophage PBS1
插入大小: 5925 bp
基因突变: R33A/K34A in rAPOBEC1, D10A in SpCas9
启动子 CAG
克隆方法: Gibson克隆
5' 测序引物 CTCTGCTAACCATGTTCATGCC
3' 测序引物 CAGAGGGAAAAAGATCTCAGTGG

文献引用方法
材料和方法部分: SECURE BE3(R33A/K34A)-P2A-EGFP (pJUL1410) was a gift from Keith Joung (Addgene plasmid # 123615 ; http://n2t.net/addgene:123615 ; RRID:Addgene_123615)
参考文献部分: Transcriptome-wide off-target RNA editing induced by CRISPR-guided DNA base editors. Grunewald J, Zhou R, Garcia SP, Iyer S, Lareau CA, Aryee MJ, Joung JK. Nature. 2019 Apr 17. pii: 10.1038/s41586-019-1161-z. doi: 10.1038/s41586-019-1161-z. 10.1038/s41586-019-1161-z PubMed 30995674

载体图谱质粒图谱和多克隆位点信息

SECURE BE3(R33AK34A)-P2A-EGFP (pJUL1410)质粒图谱

载体简介

载体序列

LOCUS       SECURE BE3(R33A/K34A)-P2A-EGFP (pJUL1410)               10768 bp ds-DNA     circular SYN 26-MAR-2019
DEFINITION  CAG promoter expression plasmid for 
            rAPOBEC1(R33A/K34A)-XTEN-hSpCas9n(D10A)-UGI-NLS(SV40)-P2A-EGFP 
            (SECURE variant BE3-R33A/K34A)..
ACCESSION   .
VERSION     .
KEYWORDS    SECURE BE3(R33A/K34A)-P2A-EGFP (pJUL1410)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10768)
  TITLE     Expression plasmids for Base Editors
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 10768)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Mar 26, 2019 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..10768
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(87..106)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_site      152..207
                     /label=-beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal"
     primer_bind     complement(206..225)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(568..584)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(568..584)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(581..603)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    592..608
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(616..646)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    661..682
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        741..936
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      787..922
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     849..868
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    942..1076
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(979..998)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1033..1052
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1144..1161)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1315..1903)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1395..1414)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2074..2934)
                     /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"
     primer_bind     2697..2716
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2935..3039)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        3070..3449
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3451..3728
                     /label=chicken beta-actin promoter
     intron          3729..4745
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     4669..4691
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     4753..4772
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             5579..9679
                     /codon_start=1
                     /product="nickase mutant of the Cas9 endonuclease from the 
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(D10A)
                     /note="generates RNA-guided single strand nicks in DNA due 
                     to the D10A mutation in the RuvC catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             9692..9940
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             9953..9973
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             9992..10048
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             10049..10768
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(10094..10115)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(10355..10374)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     10702..10723
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 agcggccgca ctcctcaggt gcaggctgcc tatcagaagg tggtggctgg tgtggccaat
       61 gccctggctc acaaatacca ctgagatctt tttccctctg ccaaaaatta tggggacatc
      121 atgaagcccc ttgagcatct gacttctggc taataaagga aatttatttt cattgcaata
      181 gtgtgttgga attttttgtg tctctcactc ggaaggacat atgggagggc aaatcattta
      241 aaacatcaga atgagtattt ggtttagagt ttggcaacat atgcccatat gctggctgcc
      301 atgaacaaag gttggctata aagaggtcat cagtatatga aacagccccc tgctgtccat
      361 tccttattcc atagaaaagc cttgacttga ggttagattt tttttatatt ttgttttgtg
      421 ttattttttt ctttaacatc cctaaaattt tccttacatg ttttactagc cagatttttc
      481 ctcctctcct gactactccc agtcatagct gtccctcttc tcttatggag atccctcgac
      541 ctgcagccca agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
      601 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
      661 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
      721 cctgtcgtgc cagcggatcc gcatctcaat tagtcagcaa ccatagtccc gcccctaact
      781 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
      841 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
      901 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc taacttgttt attgcagctt
      961 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     1021 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatccgct
     1081 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     1141 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     1201 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     1261 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     1321 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     1381 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     1441 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     1501 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     1561 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     1621 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     1681 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     1741 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     1801 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     1861 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     1921 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     1981 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     2041 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     2101 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     2161 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     2221 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     2281 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     2341 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     2401 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     2461 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     2521 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     2581 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     2641 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     2701 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     2761 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     2821 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     2881 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     2941 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     3001 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     3061 acctgggtcg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag
     3121 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
     3181 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
     3241 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
     3301 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
     3361 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     3421 tctacgtatt agtcatcgct attaccatgg tcgaggtgag ccccacgttc tgcttcactc
     3481 tccccatctc ccccccctcc ccacccccaa ttttgtattt atttattttt taattatttt
     3541 gtgcagcgat gggggcgggg gggggggggg ggcgcgcgcc rggggggggg gggggggggg
     3601 gggggggggg gggggsgggg cggagaggtg cggcggcagc caatcagagc ggcgcgctcc
     3661 gaaagtttcc ttttatggcg aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc
     3721 ggcgggcggg agtcgctgcg cgctgccttc gccccgtgcc ccgctccgcc gccgcctcgc
     3781 gccgcccgcc ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc
     3841 cttctcctcc gggctgtaat tagcgcttgg tttaatgacg gcttgtttct tttctgtggc
     3901 tgcgtgaaag ccttgagggg ctccgggagg gccctttgtg cggggggagc ggctcggggg
     3961 gtgcgtgcgt gtgtgtgtgc gtggggagcg ccgcgtgcgg ctccgcgctg cccggcggct
     4021 gtgagcgctg cgggcgcggc gcggggcttt gtgcgctccg cagtgtgcgc gaggggagcg
     4081 cggccggggg cggtgccccg cggtgcgggg ggggctgcga ggggaacaaa ggctgcgtgc
     4141 ggggtgtgtg cgtggggggg tgagcagggg gtgtgggcgc gtcggtcggg ctgcaacccc
     4201 ccctgcaccc ccctccccga gttgctgagc acggcccggc ttcgggtgcg gggctccgta
     4261 cggggcgtgg cgcggggctc gccgtgccgg gcggggggtg gcggcaggtg ggggtgccgg
     4321 gcggggcggg gccgcctcgg gccggggagg gctcggggga ggggcgcggc ggcccccgga
     4381 gcgccggcgg ctgtcgaggc gcggcgagcc gcagccattg ccttttatgg taatcgtgcg
     4441 agagggcgca gggacttcct ttgtcccaaa tctgtgcgga gccgaaatct gggaggcgcc
     4501 gccgcacccc ctctagcggg cgcggggcga agcggtgcgg cgccggcagg aaggaaatgg
     4561 gcggggaggg ccttcgtgcg tcgccgcgcc gccgtcccct tctccctctc cagcctcggg
     4621 gctgtccgcg gggggacggc tgccttcggg ggggacgggg cagggcgggg ttcggcttct
     4681 ggcgtgtgac cggcggctct agagcctctg ctaaccatgt tcatgccttc ttctttttcc
     4741 tacagctcct gggcaacgtg ctggttattg tgctgtctca tcattttggc aaagaattct
     4801 gcagtcgacg gtaccgcggg cccgggatcc accggtcgcc accatgagct cagagactgg
     4861 cccagtggct gtggacccca cattgagacg gcggatcgag ccccatgagt ttgaggtatt
     4921 cttcgatccg agagagctcg ctgctgagac ctgcctgctt tacgaaatta attggggggg
     4981 ccggcactcc atttggcgac atacatcaca gaacactaac aagcacgtcg aagtcaactt
     5041 catcgagaag ttcacgacag aaagatattt ctgtccgaac acaaggtgca gcattacctg
     5101 gtttctcagc tggagcccat gcggcgaatg tagtagggcc atcactgaat tcctgtcaag
     5161 gtatccccac gtcactctgt ttatttacat cgcaaggctg taccaccacg ctgacccccg
     5221 caatcgacaa ggcctgcggg atttgatctc ttcaggtgtg actatccaaa ttatgactga
     5281 gcaggagtca ggatactgct ggagaaactt tgtgaattat agcccgagta atgaagccca
     5341 ctggcctagg tatccccatc tgtgggtacg actgtacgtt cttgaactgt actgcatcat
     5401 actgggcctg cctccttgtc tcaacattct gagaaggaag cagccacagc tgacattctt
     5461 taccatcgct cttcagtctt gtcattacca gcgactgccc ccacacattc tctgggccac
     5521 cgggttgaaa agcggcagcg agactcccgg gacctcagag tccgccacac ccgaaagtga
     5581 taaaaagtat tctattggtt tagccatcgg cactaattcc gttggatggg ctgtcataac
     5641 cgatgaatac aaagtacctt caaagaaatt taaggtgttg gggaacacag accgtcattc
     5701 gattaaaaag aatcttatcg gtgccctcct attcgatagt ggcgaaacgg cagaggcgac
     5761 tcgcctgaaa cgaaccgctc ggagaaggta tacacgtcgc aagaaccgaa tatgttactt
     5821 acaagaaatt tttagcaatg agatggccaa agttgacgat tctttctttc accgtttgga
     5881 agagtccttc cttgtcgaag aggacaagaa acatgaacgg caccccatct ttggaaacat
     5941 agtagatgag gtggcatatc atgaaaagta cccaacgatt tatcacctca gaaaaaagct
     6001 agttgactca actgataaag cggacctgag gttaatctac ttggctcttg cccatatgat
     6061 aaagttccgt gggcactttc tcattgaggg tgatctaaat ccggacaact cggatgtcga
     6121 caaactgttc atccagttag tacaaaccta taatcagttg tttgaagaga accctataaa
     6181 tgcaagtggc gtggatgcga aggctattct tagcgcccgc ctctctaaat cccgacggct
     6241 agaaaacctg atcgcacaat tacccggaga gaagaaaaat gggttgttcg gtaaccttat
     6301 agcgctctca ctaggcctga caccaaattt taagtcgaac ttcgacttag ctgaagatgc
     6361 caaattgcag cttagtaagg acacgtacga tgacgatctc gacaatctac tggcacaaat
     6421 tggagatcag tatgcggact tatttttggc tgccaaaaac cttagcgatg caatcctcct
     6481 atctgacata ctgagagtta atactgagat taccaaggcg ccgttatccg cttcaatgat
     6541 caaaaggtac gatgaacatc accaagactt gacacttctc aaggccctag tccgtcagca
     6601 actgcctgag aaatataagg aaatattctt tgatcagtcg aaaaacgggt acgcaggtta
     6661 tattgacggc ggagcgagtc aagaggaatt ctacaagttt atcaaaccca tattagagaa
     6721 gatggatggg acggaagagt tgcttgtaaa actcaatcgc gaagatctac tgcgaaagca
     6781 gcggactttc gacaacggta gcattccaca tcaaatccac ttaggcgaat tgcatgctat
     6841 acttagaagg caggaggatt tttatccgtt cctcaaagac aatcgtgaaa agattgagaa
     6901 aatcctaacc tttcgcatac cttactatgt gggacccctg gcccgaggga actctcggtt
     6961 cgcatggatg acaagaaagt ccgaagaaac gattactcca tggaattttg aggaagttgt
     7021 cgataaaggt gcgtcagctc aatcgttcat cgagaggatg accaactttg acaagaattt
     7081 accgaacgaa aaagtattgc ctaagcacag tttactttac gagtatttca cagtgtacaa
     7141 tgaactcacg aaagttaagt atgtcactga gggcatgcgt aaacccgcct ttctaagcgg
     7201 agaacagaag aaagcaatag tagatctgtt attcaagacc aaccgcaaag tgacagttaa
     7261 gcaattgaaa gaggactact ttaagaaaat tgaatgcttc gattctgtcg agatctccgg
     7321 ggtagaagat cgatttaatg cgtcacttgg tacgtatcat gacctcctaa agataattaa
     7381 agataaggac ttcctggata acgaagagaa tgaagatatc ttagaagata tagtgttgac
     7441 tcttaccctc tttgaagatc gggaaatgat tgaggaaaga ctaaaaacat acgctcacct
     7501 gttcgacgat aaggttatga aacagttaaa gaggcgtcgc tatacgggct ggggacgatt
     7561 gtcgcggaaa cttatcaacg ggataagaga caagcaaagt ggtaaaacta ttctcgattt
     7621 tctaaagagc gacggcttcg ccaataggaa ctttatgcag ctgatccatg atgactcttt
     7681 aaccttcaaa gaggatatac aaaaggcaca ggtttccgga caaggggact cattgcacga
     7741 acatattgcg aatcttgctg gttcgccagc catcaaaaag ggcatactcc agacagtcaa
     7801 agtagtggat gagctagtta aggtcatggg acgtcacaaa ccggaaaaca ttgtaatcga
     7861 gatggcacgc gaaaatcaaa cgactcagaa ggggcaaaaa aacagtcgag agcggatgaa
     7921 gagaatagaa gagggtatta aagaactggg cagccagatc ttaaaggagc atcctgtgga
     7981 aaatacccaa ttgcagaacg agaaacttta cctctattac ctacaaaatg gaagggacat
     8041 gtatgttgat caggaactgg acataaaccg tttatctgat tacgacgtcg atcacattgt
     8101 accccaatcc tttttgaagg acgattcaat cgacaataaa gtgcttacac gctcggataa
     8161 gaaccgaggg aaaagtgaca atgttccaag cgaggaagtc gtaaagaaaa tgaagaacta
     8221 ttggcggcag ctcctaaatg cgaaactgat aacgcaaaga aagttcgata acttaactaa
     8281 agctgagagg ggtggcttgt ctgaacttga caaggccgga tttattaaac gtcagctcgt
     8341 ggaaacccgc caaatcacaa agcatgttgc acagatacta gattcccgaa tgaatacgaa
     8401 atacgacgag aacgataagc tgattcggga agtcaaagta atcactttaa agtcaaaatt
     8461 ggtgtcggac ttcagaaagg attttcaatt ctataaagtt agggagataa ataactacca
     8521 ccatgcgcac gacgcttatc ttaatgccgt cgtagggacc gcactcatta agaaataccc
     8581 gaagctagaa agtgagtttg tgtatggtga ttacaaagtt tatgacgtcc gtaagatgat
     8641 cgcgaaaagc gaacaggaga taggcaaggc tacagccaaa tacttctttt attctaacat
     8701 tatgaatttc tttaagacgg aaatcactct ggcaaacgga gagatacgca aacgaccttt
     8761 aattgaaacc aatggggaga caggtgaaat cgtatgggat aagggccggg acttcgcgac
     8821 ggtgagaaaa gttttgtcca tgccccaagt caacatagta aagaaaactg aggtgcagac
     8881 cggagggttt tcaaaggaat cgattcttcc aaaaaggaat agtgataagc tcatcgctcg
     8941 taaaaaggac tgggacccga aaaagtacgg tggcttcgat agccctacag ttgcctattc
     9001 tgtcctagta gtggcaaaag ttgagaaggg aaaatccaag aaactgaagt cagtcaaaga
     9061 attattgggg ataacgatta tggagcgctc gtcttttgaa aagaacccca tcgacttcct
     9121 tgaggcgaaa ggttacaagg aagtaaaaaa ggatctcata attaaactac caaagtatag
     9181 tctgtttgag ttagaaaatg gccgaaaacg gatgttggct agcgccggag agcttcaaaa
     9241 ggggaacgaa ctcgcactac cgtctaaata cgtgaatttc ctgtatttag cgtcccatta
     9301 cgagaagttg aaaggttcac ctgaagataa cgaacagaag caactttttg ttgagcagca
     9361 caaacattat ctcgacgaaa tcatagagca aatttcggaa ttcagtaaga gagtcatcct
     9421 agctgatgcc aatctggaca aagtattaag cgcatacaac aagcacaggg ataaacccat
     9481 acgtgagcag gcggaaaata ttatccattt gtttactctt accaacctcg gcgctccagc
     9541 cgcattcaag tattttgaca caacgataga tcgcaaacga tacacttcta ccaaggaggt
     9601 gctagacgcg acactgattc accaatccat cacgggatta tatgaaactc ggatagattt
     9661 gtcacagctt gggggtgact ctggtggttc tactaatctg tcagatatta ttgaaaagga
     9721 gaccggtaag caactggtta tccaggaatc catcctcatg ctcccagagg aggtggaaga
     9781 agtcattggg aacaagccgg aaagcgatat actcgtgcac accgcctacg acgagagcac
     9841 cgacgagaat gtcatgcttc tgactagcga cgcccctgaa tacaagcctt gggctctggt
     9901 catacaggat agcaacggtg agaacaagat taagatgctc tctggtggtt ctcccaagaa
     9961 gaagaggaaa gtcggaggcg gtggaagcgg cgcaacaaac ttctctctgc tgaaacaagc
    10021 cggagatgtc gaagagaatc ctggaccgat ggtgagcaag ggcgaggagc tgttcaccgg
    10081 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc
    10141 cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac
    10201 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg
    10261 cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga
    10321 aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc
    10381 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt
    10441 caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt
    10501 ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa
    10561 catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga
    10621 cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga
    10681 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
    10741 tctcggcatg gacgagctgt acaagtga
//

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