pXR002: EF1a-dCasRx-2A-EGFP

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pXR002: EF1a-dCasRx-2A-EGFP载体质粒基本信息

质粒名称: pXR002: EF1a-dCasRx-2A-EGFP 
出品公司: 爱迪基因
目录编号: 109050
存储实验室: Patrick Hsu

载体骨架基本信息
载体名称: lenti(AMP)
载体出品公司: --
载体抗性: 氨苄青霉素
空载体大小: --
完整质粒大小: 12990 bp
载体修饰: --
载体类型: 哺乳动物表达,慢病毒,CRISPR
筛选标记: GFP
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: dCasRx,NLS-dRfxCas13d-NLS
物种来源: H. sapiens (human); Ruminococcus flavefaciens XPD3002
插入大小: 3000 bp
基因突变: R239A/H244A/R858A/H863A
GenBank编号 --
启动子 --
标签/融合蛋白:
NLS(插入件上的N端子)
NLS(插接件上的C端子)
HA(插入件上的C端子)
2A eGFP(主干上的C端)
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: pXR002: EF1a-dCasRx-2A-EGFP was a gift from Patrick Hsu (Addgene plasmid # 109050 ; http://n2t.net/addgene:109050 ; RRID:Addgene_109050)
参考文献部分: Transcriptome Engineering with RNA-Targeting Type VI-D CRISPR Effectors. Konermann S, Lotfy P, Brideau NJ, Oki J, Shokhirev MN, Hsu PD. Cell. 2018 Mar 8. pii: S0092-8674(18)30207-1. doi: 10.1016/j.cell.2018.02.033. 10.1016/j.cell.2018.02.033 PubMed 29551272

pXR002: EF1a-dCasRx-2A-EGFP质粒图谱载体图谱和pXR002: EF1a-dCasRx-2A-EGFP载体序列质粒序列多克隆位点信息

pXR002EF1a-dCasRx-2A-EGFP质粒图谱

pXR002: EF1a-dCasRx-2A-EGFP质粒载体简介

pXR002: EF1a-dCasRx-2A-EGFP质粒序列载体序列

LOCUS       pXR002: EF1a-dCasRx-2A-EGFP               12991 bp ds-DNA     circular SYN 20-JUN-2019
DEFINITION  Catalytically inactive dCasRx with 2A-EGFP.
ACCESSION   .
VERSION     .
KEYWORDS    pXR002: EF1a-dCasRx-2A-EGFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12991)
  AUTHORS   Konermann S, Lotfy P, Brideau NJ, Oki J, Shokhirev MN, Hsu PD
  TITLE     Transcriptome Engineering with RNA-Targeting Type VI-D CRISPR 
            Effectors.
  JOURNAL   Cell. 2018 Mar 8. pii: S0092-8674(18)30207-1. doi: 
            10.1016/j.cell.2018.02.033.
  PUBMED    29551272
REFERENCE   2  (bases 1 to 12991)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 20, 2019 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..12991
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(42..61)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        233..612
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        614..812
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     767..787
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             830..1010
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1057..1182
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1675..1908
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    2435..2552
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2677..3855
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2908..3846
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3803..3823
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             3874..3894
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6811..6831
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6841..6867
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             6883..6936
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             6937..7653
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(6979..7000)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(7240..7259)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7587..7608
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    7678..8266
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7731..7751)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8149..8160)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(8269..8285)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8270..8286)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             8791..8971
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(8997..9014)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9003..9227
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      9273..9701
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9360..9379)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9570..9591
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9710..9730)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        9715..10044
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      9895..10030
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     9957..9976
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        10092..10139
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             10158..10532
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    10662..10783
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10699..10718)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10753..10772
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(10832..10848)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10832..10848)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10845..10867)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10856..10872
                     /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(10880..10910)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10925..10946
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11063..11080)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11234..11822)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11314..11333)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(11993..12853)
                     /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     12616..12635
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12854..12958)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc
       61 gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg
      121 agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt
      181 agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg ttgacattga
      241 ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
      301 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
      361 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
      421 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
      481 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
      541 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
      601 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
      661 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
      721 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
      781 aggcgtgtac ggtgggaggt ctatataagc agcgcgtttt gcctgtactg ggtctctctg
      841 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
      901 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
      961 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtggcgcccg
     1021 aacagggact tgaaagcgaa agggaaacca gaggagctct ctcgacgcag gactcggctt
     1081 gctgaagcgc gcacggcaag aggcgagggg cggcgactgg tgagtacgcc aaaaattttg
     1141 actagcggag gctagaagga gagagatggg tgcgagagcg tcagtattaa gcgggggaga
     1201 attagatcgc gatgggaaaa aattcggtta aggccagggg gaaagaaaaa atataaatta
     1261 aaacatatag tatgggcaag cagggagcta gaacgattcg cagttaatcc tggcctgtta
     1321 gaaacatcag aaggctgtag acaaatactg ggacagctac aaccatccct tcagacagga
     1381 tcagaagaac ttagatcatt atataataca gtagcaaccc tctattgtgt gcatcaaagg
     1441 atagagataa aagacaccaa ggaagcttta gacaagatag aggaagagca aaacaaaagt
     1501 aagaccaccg cacagcaagc ggccgctgat cttcagacct ggaggaggag atatgaggga
     1561 caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat taggagtagc
     1621 acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg gaataggagc
     1681 tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt caatgacgct
     1741 gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca atttgctgag
     1801 ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca agcagctcca
     1861 ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg ggatttgggg
     1921 ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt ggagtaataa
     1981 atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag aaattaacaa
     2041 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
     2101 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
     2161 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
     2221 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
     2281 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
     2341 tggagagaga gacagagaca gatccattcg attagtgaac ggatcggcac tgcgtgcgcc
     2401 aattctgcag acaaatggca gtattcatcc acaattttaa aagaaaaggg gggattgggg
     2461 ggtacagtgc aggggaaaga atagtagaca taatagcaac agacatacaa actaaagaat
     2521 tacaaaaaca aattacaaaa attcaaaatt ttcgggttta ttacagggac agcagagatc
     2581 cagtttggtt aattaatgca aagatggata aagttttaaa cagagaggaa tctttgcagc
     2641 taatggacct tctaggtctt gaaaggagtg ggaattggct ccggtgcccg tcagtgggca
     2701 gagcgcacat cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgaaccggt
     2761 gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt
     2821 tttcccgagg gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt
     2881 cgcaacgggt ttgccgccag aacacaggta agtgccgtgt gtggttcccg cgggcctggc
     2941 ctctttacgg gttatggccc ttgcgtgcct tgaattactt ccacctggct gcagtacgtg
     3001 attcttgatc ccgagcttcg ggttggaagt gggtgggaga gttcgaggcc ttgcgcttaa
     3061 ggagcccctt cgcctcgtgc ttgagttgag gcctggcctg ggcgctgggg ccgccgcgtg
     3121 cgaatctggt ggcaccttcg cgcctgtctc gctgctttcg ataagtctct agccatttaa
     3181 aatttttgat gacctgctgc gacgcttttt ttctggcaag atagtcttgt aaatgcgggc
     3241 caagatctgc acactggtat ttcggttttt ggggccgcgg gcggcgacgg ggcccgtgcg
     3301 tcccagcgca catgttcggc gaggcggggc ctgcgagcgc ggccaccgag aatcggacgg
     3361 gggtagtctc aagctggccg gcctgctctg gtgcctggcc tcgcgccgcc gtgtatcgcc
     3421 ccgccctggg cggcaaggct ggcccggtcg gcaccagttg cgtgagcgga aagatggccg
     3481 cttcccggcc ctgctgcagg gagctcaaaa tggaggacgc ggcgctcggg agagcgggcg
     3541 ggtgagtcac ccacacaaag gaaaagggcc tttccgtcct cagccgtcgc ttcatgtgac
     3601 tccacggagt accgggcgcc gtccaggcac ctcgattagt tctcgagctt ttggagtacg
     3661 tcgtctttag gttgggggga ggggttttat gcgatggagt ttccccacac tgagtgggtg
     3721 gagactgaag ttaggccagc ttggcacttg atgtaattct ccttggaatt tgcccttttt
     3781 gagtttggat cttggttcat tctcaagcct cagacagtgg ttcaaagttt ttttcttcca
     3841 tttcaggtgt cgtgacgtac ggccaccatg agccccaaga agaagagaaa ggtggaggcc
     3901 agcatcgaaa aaaaaaagtc cttcgccaag ggcatgggcg tgaagtccac actcgtgtcc
     3961 ggctccaaag tgtacatgac aaccttcgcc gaaggcagcg acgccaggct ggaaaagatc
     4021 gtggagggcg acagcatcag gagcgtgaat gagggcgagg ccttcagcgc tgaaatggcc
     4081 gataaaaacg ccggctataa gatcggcaac gccaaattca gccatcctaa gggctacgcc
     4141 gtggtggcta acaaccctct gtatacagga cccgtccagc aggatatgct cggcctgaag
     4201 gaaactctgg aaaagaggta cttcggcgag agcgctgatg gcaatgacaa tatttgtatc
     4261 caggtgatcc ataacatcct ggacattgaa aaaatcctcg ccgaatacat taccaacgcc
     4321 gcctacgccg tcaacaatat ctccggcctg gataaggaca ttattggatt cggcaagttc
     4381 tccacagtgt atacctacga cgaattcaaa gaccccgagc accatagggc cgctttcaac
     4441 aataacgata agctcatcaa cgccatcaag gcccagtatg acgagttcga caacttcctc
     4501 gataacccca gactcggcta tttcggccag gcctttttca gcaaggaggg cagaaattac
     4561 atcatcaatt acggcaacga atgctatgac attctggccc tcctgagcgg actggcgcac
     4621 tgggtggtcg ctaacaacga agaagagtcc aggatctcca ggacctggct ctacaacctc
     4681 gataagaacc tcgacaacga atacatctcc accctcaact acctctacga caggatcacc
     4741 aatgagctga ccaactcctt ctccaagaac tccgccgcca acgtgaacta tattgccgaa
     4801 actctgggaa tcaaccctgc cgaattcgcc gaacaatatt tcagattcag cattatgaaa
     4861 gagcagaaaa acctcggatt caatatcacc aagctcaggg aagtgatgct ggacaggaag
     4921 gatatgtccg agatcaggaa aaatcataag gtgttcgact ccatcaggac caaggtctac
     4981 accatgatgg actttgtgat ttataggtat tacatcgaag aggatgccaa ggtggctgcc
     5041 gccaataagt ccctccccga taatgagaag tccctgagcg agaaggatat ctttgtgatt
     5101 aacctgaggg gctccttcaa cgacgaccag aaggatgccc tctactacga tgaagctaat
     5161 agaatttgga gaaagctcga aaatatcatg cacaacatca aggaatttag gggaaacaag
     5221 acaagagagt ataagaagaa ggacgcccct agactgccca gaatcctgcc cgctggccgt
     5281 gatgtttccg ccttcagcaa actcatgtat gccctgacca tgttcctgga tggcaaggag
     5341 atcaacgacc tcctgaccac cctgattaat aaattcgata acatccagag cttcctgaag
     5401 gtgatgcctc tcatcggagt caacgctaag ttcgtggagg aatacgcctt tttcaaagac
     5461 tccgccaaga tcgccgatga gctgaggctg atcaagtcct tcgctagaat gggagaacct
     5521 attgccgatg ccaggagggc catgtatatc gacgccatcc gtattttagg aaccaacctg
     5581 tcctatgatg agctcaaggc cctcgccgac accttttccc tggacgagaa cggaaacaag
     5641 ctcaagaaag gcaagcacgg catgagaaat ttcattatta ataacgtgat cagcaataaa
     5701 aggttccact acctgatcag atacggtgat cctgcccacc tccatgagat cgccaaaaac
     5761 gaggccgtgg tgaagttcgt gctcggcagg atcgctgaca tccagaaaaa acagggccag
     5821 aacggcaaga accagatcga caggtactac gaaacttgta tcggaaagga taagggcaag
     5881 agcgtgagcg aaaaggtgga cgctctcaca aagatcatca ccggaatgaa ctacgaccaa
     5941 ttcgacaaga aaaggagcgt cattgaggac accggcaggg aaaacgccga gagggagaag
     6001 tttaaaaaga tcatcagcct gtacctcacc gtgatctacc acatcctcaa gaatattgtc
     6061 aatatcaacg ccaggtacgt catcggattc cattgcgtcg agcgtgatgc tcaactgtac
     6121 aaggagaaag gctacgacat caatctcaag aaactggaag agaagggatt cagctccgtc
     6181 accaagctct gcgctggcat tgatgaaact gcccccgata agagaaagga cgtggaaaag
     6241 gagatggctg aaagagccaa ggagagcatt gacagcctcg agagcgccaa ccccaagctg
     6301 tatgccaatt acatcaaata cagcgacgag aagaaagccg aggagttcac caggcagatt
     6361 aacagggaga aggccaaaac cgccctgaac gcctacctga ggaacaccaa gtggaatgtg
     6421 atcatcaggg aggacctcct gagaattgac aacaagacat gtaccctgtt cgcaaacaag
     6481 gccgtcgccc tggaagtggc caggtatgtc cacgcctata tcaacgacat tgccgaggtc
     6541 aattcctact tccaactgta ccattacatc atgcagagaa ttatcatgaa tgagaggtac
     6601 gagaaaagca gcggaaaggt gtccgagtac ttcgacgctg tgaatgacga gaagaagtac
     6661 aacgataggc tcctgaaact gctgtgtgtg cctttcggct actgtatccc caggtttaag
     6721 aacctgagca tcgaggccct gttcgatagg aacgaggccg ccaagttcga caaggagaaa
     6781 aagaaggtgt ccggcaattc cggatccgga cctaagaaaa agaggaaggt ggcggccgct
     6841 tacccatacg atgttccaga ttacgctgct agcggcagtg gagagggcag aggaagtctg
     6901 ctaacatgcg gtgacgtcga ggagaatcct ggcccagtga gcaagggcga ggagctgttc
     6961 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
     7021 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
     7081 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg
     7141 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     7201 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     7261 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     7321 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     7381 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc
     7441 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     7501 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc
     7561 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     7621 atcactctcg gcatggacga gctgtacaag taagaattcg atatcaagct tatcgataat
     7681 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
     7741 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
     7801 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg
     7861 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt
     7921 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt
     7981 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     8041 ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc
     8101 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat
     8161 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc
     8221 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcatcg ataccgtcga
     8281 cctcgagacc tagaaaaaca tggagcaatc acaagtagca atacagcagc taccaatgct
     8341 gattgtgcct ggctagaagc acaagaggag gaggaggtgg gttttccagt cacacctcag
     8401 gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa
     8461 aaggggggac tggaagggct aattcactcc caacgaagac aagatatcct tgatctgtgg
     8521 atctaccaca cacaaggcta cttccctgat tggcagaact acacaccagg gccagggatc
     8581 agatatccac tgacctttgg atggtgctac aagctagtac cagttgagca agagaaggta
     8641 gaagaagcca atgaaggaga gaacacccgc ttgttacacc ctgtgagcct gcatgggatg
     8701 gatgacccgg agagagaagt attagagtgg aggtttgaca gccgcctagc atttcatcac
     8761 atggcccgag agctgcatcc ggactgtact gggtctctct ggttagacca gatctgagcc
     8821 tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga
     8881 gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga
     8941 cccttttagt cagtgtggaa aatctctagc agggcccgtt taaacccgct gatcagcctc
     9001 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     9061 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     9121 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     9181 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga
     9241 aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
     9301 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     9361 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     9421 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     9481 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     9541 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     9601 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     9661 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt
     9721 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     9781 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     9841 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     9901 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     9961 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
    10021 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct
    10081 gatcagcacg tgttgacaat taatcatcgg catagtatat cggcatagta taatacgaca
    10141 aggtgaggaa ctaaaccatg gccaagttga ccagtgccgt tccggtgctc accgcgcgcg
    10201 acgtcgccgg agcggtcgag ttctggaccg accggctcgg gttctcccgg gacttcgtgg
    10261 aggacgactt cgccggtgtg gtccgggacg acgtgaccct gttcatcagc gcggtccagg
    10321 accaggtggt gccggacaac accctggcct gggtgtgggt gcgcggcctg gacgagctgt
    10381 acgccgagtg gtcggaggtc gtgtccacga acttccggga cgcctccggg ccggccatga
    10441 ccgagatcgg cgagcagccg tgggggcggg agttcgccct gcgcgacccg gccggcaact
    10501 gcgtgcactt cgtggccgag gagcaggact gacacgtgct acgagatttc gattccaccg
    10561 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
    10621 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt
    10681 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
    10741 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt
    10801 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
    10861 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
    10921 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
    10981 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
    11041 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
    11101 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
    11161 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
    11221 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
    11281 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
    11341 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
    11401 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
    11461 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
    11521 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
    11581 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
    11641 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
    11701 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
    11761 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
    11821 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
    11881 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
    11941 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
    12001 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
    12061 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
    12121 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
    12181 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
    12241 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
    12301 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
    12361 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
    12421 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
    12481 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
    12541 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
    12601 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
    12661 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
    12721 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
    12781 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
    12841 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
    12901 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
    12961 catttccccg aaaagtgcca cctgacgtcg a
//

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