pHAGE-IRES-puro-NLS-dPspCas13b-EGFP-NLS-3xFlag

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: pHAGE-IRES-puro-NLS-dPspCas13b-EGFP-NLS-3xFlag
出品公司: Addgene
目录编号: 132397
存储实验室: Ling-Ling Chen

载体骨架基本信息
载体名称: pHAGE-EF1a-IRES-puro
载体出品公司: --
空载体大小: --
完整质粒大小: --
载体修饰: --
载体类型: 哺乳动物表达,慢病毒
筛选标记: --
载体抗性: 氨苄青霉素
生长温度: 37 ℃
宿主菌: NEB Stable
拷贝数: 高拷贝

插入基因信息
插入基因名称: dPspCas13b
物种来源: Prevotella sp. P5-125
插入大小: --
基因突变: H133A; H1058A
GenBank编号 --
启动子: --
标签/融合蛋白: flag(插入件上的N端子)
克隆方法: Ligation Independent 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: pHAGE-IRES-puro-NLS-dPspCas13b-EGFP-NLS-3xFlag was a gift from Ling-Ling Chen (Addgene plasmid # 132397 ; http://n2t.net/addgene:132397 ; RRID:Addgene_132397)
参考文献部分: dCas13 imaging. Liang-Zhong Yang, Yang Wang, Si-Qi Li, Huang Wu, Gordon G. Carmichael, Ling-Ling Chen. Molecular Cell 10.1016/j.molcel.2019.10.024

载体图谱质粒图谱和多克隆位点信息

pHAGE-IRES-puro-NLS-dPspCas13b-EGFP-NLS-3xFlag质粒图谱

载体简介

载体序列

LOCUS       pHAGE-IRES-puro-NLS-dPspCas13b-EGFP-NLS-3xFlag               12449 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  overexpression in human cells.
ACCESSION   .
VERSION     .
KEYWORDS    pHAGE-IRES-puro-NLS-dPspCas13b-EGFP-NLS-3xFlag
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12449)
  AUTHORS   Liang-Zhong Yang, Yang Wang, Si-Qi Li, Huang Wu, Gordon G. 
            Carmichael, Ling-Ling Chen
  TITLE     dCas13 imaging
  JOURNAL   Molecular Cell
REFERENCE   2  (bases 1 to 12449)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..12449
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     LTR             1..634
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1303..1536
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1721..1765
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2058..2175
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2223..3401
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2454..3392
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3349..3369
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             3431..3451
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      6746..6755
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             6752..7468
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             6752..7468
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(6797..6818)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(7058..7077)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7405..7426
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             7496..7543
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7562..7627
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    7634..8201
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(7795..7812)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     8022..8041
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             8221..8820
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(8221..8240)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     8717..8737
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    8839..9427
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8892..8912)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9310..9321)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9502..9735
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(9854..9872)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        9940..10044
                     /gene="bla"
                     /label=AmpR promoter
     CDS             10045..10905
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(10263..10282)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      11076..11664
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11565..11584
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     11818..11835
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     11927..11946
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     protein_bind    12075..12096
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        12111..12141
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12149..12165
                     /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)."
     primer_bind     12154..12176
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     12173..12189
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     12173..12189
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     12239..12258
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccgaa ttcacaaatg
     2041 gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa
     2101 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca
     2161 aaaattcaaa attttcgggt ttattacagg gacagcagag atccagtttg gactagtcgt
     2221 gaggctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg
     2281 ggggaggggt cggcaattga accggtgcct agagaaggtg gcgcggggta aactgggaaa
     2341 gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg
     2401 cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca caggtaagtg
     2461 ccgtgtgtgg ttcccgcggg cctggcctct ttacgggtta tggcccttgc gtgccttgaa
     2521 ttacttccac ctggctgcag tacgtgattc ttgatcccga gcttcgggtt ggaagtgggt
     2581 gggagagttc gaggccttgc gcttaaggag ccccttcgcc tcgtgcttga gttgaggcct
     2641 ggcctgggcg ctggggccgc cgcgtgcgaa tctggtggca ccttcgcgcc tgtctcgctg
     2701 ctttcgataa gtctctagcc atttaaaatt tttgatgacc tgctgcgacg ctttttttct
     2761 ggcaagatag tcttgtaaat gcgggccaag atctgcacac tggtatttcg gtttttgggg
     2821 ccgcgggcgg cgacggggcc cgtgcgtccc agcgcacatg ttcggcgagg cggggcctgc
     2881 gagcgcggcc accgagaatc ggacgggggt agtctcaagc tggccggcct gctctggtgc
     2941 ctggcctcgc gccgccgtgt atcgccccgc cctgggcggc aaggctggcc cggtcggcac
     3001 cagttgcgtg agcggaaaga tggccgcttc ccggccctgc tgcagggagc tcaaaatgga
     3061 ggacgcggcg ctcgggagag cgggcgggtg agtcacccac acaaaggaaa agggcctttc
     3121 cgtcctcagc cgtcgcttca tgtgactcca cggagtaccg ggcgccgtcc aggcacctcg
     3181 attagttctc gagcttttgg agtacgtcgt ctttaggttg gggggagggg ttttatgcga
     3241 tggagtttcc ccacactgag tgggtggaga ctgaagttag gccagcttgg cacttgatgt
     3301 aattctcctt ggaatttgcc ctttttgagt ttggatcttg gttcattctc aagcctcaga
     3361 cagtggttca aagttttttt cttccatttc aggtgtcgtg aagcggccgc accggtctgc
     3421 agctagcatg cccaaaaaga agaggaaagt gaacatcccc gctctggtgg aaaaccagaa
     3481 gaagtacttt ggcacctaca gcgtgatggc catgctgaac gctcagaccg tgctggacca
     3541 catccagaag gtggccgata ttgagggcga gcagaacgag aacaacgaga atctgtggtt
     3601 tcaccccgtg atgagccacc tgtacaacgc caagaacggc tacgacaagc agcccgagaa
     3661 aaccatgttc atcatcgagc ggctgcagag ctacttccca ttcctgaaga tcatggccga
     3721 gaaccagaga gagtacagca acggcaagta caagcagaac cgcgtggaag tgaacagcaa
     3781 cgacatcttc gaggtgctga agcgcgcctt cggcgtgctg aagatgtaca gggacctgac
     3841 caacgcctac aagacctacg aggaaaagct gaacgacggc tgcgagttcc tgaccagcac
     3901 agagcaacct ctgagcggca tgatcaacaa ctactacaca gtggccctgc ggaacatgaa
     3961 cgagagatac ggctacaaga cagaggacct ggccttcatc caggacaagc ggttcaagtt
     4021 cgtgaaggac gcctacggca agaaaaagtc ccaagtgaat accggattct tcctgagcct
     4081 gcaggactac aacggcgaca cacagaagaa gctgcacctg agcggagtgg gaatcgccct
     4141 gctgatctgc ctgttcctgg acaagcagta catcaacatc tttctgagca ggctgcccat
     4201 cttctccagc tacaatgccc agagcgagga acggcggatc atcatcagat ccttcggcat
     4261 caacagcatc aagctgccca aggaccggat ccacagcgag aagtccaaca agagcgtggc
     4321 catggatatg ctcaacgaag tgaagcggtg ccccgacgag ctgttcacaa cactgtctgc
     4381 cgagaagcag tcccggttca gaatcatcag cgacgaccac aatgaagtgc tgatgaagcg
     4441 gagcagcgac agattcgtgc ctctgctgct gcagtatatc gattacggca agctgttcga
     4501 ccacatcagg ttccacgtga acatgggcaa gctgagatac ctgctgaagg ccgacaagac
     4561 ctgcatcgac ggccagacca gagtcagagt gatcgagcag cccctgaacg gcttcggcag
     4621 actggaagag gccgagacaa tgcggaagca agagaacggc accttcggca acagcggcat
     4681 ccggatcaga gacttcgaga acatgaagcg ggacgacgcc aatcctgcca actatcccta
     4741 catcgtggac acctacacac actacatcct ggaaaacaac aaggtcgaga tgtttatcaa
     4801 cgacaaagag gacagcgccc cactgctgcc cgtgatcgag gatgatagat acgtggtcaa
     4861 gacaatcccc agctgccgga tgagcaccct ggaaattcca gccatggcct tccacatgtt
     4921 tctgttcggc agcaagaaaa ccgagaagct gatcgtggac gtgcacaacc ggtacaagag
     4981 actgttccag gccatgcaga aagaagaagt gaccgccgag aatatcgcca gcttcggaat
     5041 cgccgagagc gacctgcctc agaagatcct ggatctgatc agcggcaatg cccacggcaa
     5101 ggatgtggac gccttcatca gactgaccgt ggacgacatg ctgaccgaca ccgagcggag
     5161 aatcaagaga ttcaaggacg accggaagtc cattcggagc gccgacaaca agatgggaaa
     5221 gagaggcttc aagcagatct ccacaggcaa gctggccgac ttcctggcca aggacatcgt
     5281 gctgtttcag cccagcgtga acgatggcga gaacaagatc accggcctga actaccggat
     5341 catgcagagc gccattgccg tgtacgatag cggcgacgat tacgaggcca agcagcagtt
     5401 caagctgatg ttcgagaagg cccggctgat cggcaagggc acaacagagc ctcatccatt
     5461 tctgtacaag gtgttcgccc gcagcatccc cgccaatgcc gtcgagttct acgagcgcta
     5521 cctgatcgag cggaagttct acctgaccgg cctgtccaac gagatcaaga aaggcaacag
     5581 agtggatgtg cccttcatcc ggcgggacca gaacaagtgg aaaacacccg ccatgaagac
     5641 cctgggcaga atctacagcg aggatctgcc cgtggaactg cccagacaga tgttcgacaa
     5701 tgagatcaag tcccacctga agtccctgcc acagatggaa ggcatcgact tcaacaatgc
     5761 caacgtgacc tatctgatcg ccgagtacat gaagagagtg ctggacgacg acttccagac
     5821 cttctaccag tggaaccgca actaccggta catggacatg cttaagggcg agtacgacag
     5881 aaagggctcc ctgcagcact gcttcaccag cgtggaagag agagaaggcc tctggaaaga
     5941 gcgggcctcc agaacagagc ggtacagaaa gcaggccagc aacaagatcc gcagcaaccg
     6001 gcagatgaga aacgccagca gcgaagagat cgagacaatc ctggataagc ggctgagcaa
     6061 cagccggaac gagtaccaga aaagcgagaa agtgatccgg cgctacagag tgcaggatgc
     6121 cctgctgttt ctgctggcca aaaagaccct gaccgaactg gccgatttcg acggcgagag
     6181 gttcaaactg aaagaaatca tgcccgacgc cgagaaggga atcctgagcg agatcatgcc
     6241 catgagcttc accttcgaga aaggcggcaa gaagtacacc atcaccagcg agggcatgaa
     6301 gctgaagaac tacggcgact tctttgtgct ggctagcgac aagaggatcg gcaacctgct
     6361 ggaactcgtg ggcagcgaca tcgtgtccaa agaggatatc atggaagagt tcaacaaata
     6421 cgaccagtgc aggcccgaga tcagctccat cgtgttcaac ctggaaaagt gggccttcga
     6481 cacatacccc gagctgtctg ccagagtgga ccgggaagag aaggtggact tcaagagcat
     6541 cctgaaaatc ctgctgaaca acaagaacat caacaaagag cagagcgaca tcctgcggaa
     6601 gatccggaac gccttcgatg ccaacaatta ccccgacaaa ggcgtggtgg aaatcaaggc
     6661 cctgcctgag atcgccatga gcatcaagaa ggcctttggg gagtacgcca tcatgaagga
     6721 gggagctcta gaggcgctac cggtcgccac catggtgagc aagggcgagg agctgttcac
     6781 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
     6841 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac
     6901 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca
     6961 gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
     7021 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
     7081 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
     7141 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa
     7201 cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca
     7261 caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
     7321 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa
     7381 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
     7441 cactctcggc atggacgagc tgtacaagtc gaattctaag ccaggattcc atatgaagcg
     7501 acctgccgcc acaaagaagg ctggacaggc taagaagaag aaacaacgcg tgcagctaag
     7561 cgactacaaa gaccatgacg gtgattataa agatcatgac atcgattaca aggatgacga
     7621 tgacaagtaa ggatccctcc ccccccccta acgttactgg ccgaagccgc ttggaataag
     7681 gccggtgtgc gtttgtctat atgttatttt ccaccatatt gccgtctttt ggcaatgtga
     7741 gggcccggaa acctggccct gtcttcttga cgagcattcc taggggtctt tcccctctcg
     7801 ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt
     7861 gaagacaaac aacgtctgta gcgacccttt gcaggcagcg gaacccccca cctggcgaca
     7921 ggtgcctctg cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc
     7981 agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa atggctctcc tcaagcgtat
     8041 tcaacaaggg gctgaaggat gcccagaagg taccccattg tatgggatct gatctggggc
     8101 ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga
     8161 accacgggga cgtggttttc ctttgaaaaa cacgatgata aggatccacc ggagcttacc
     8221 atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc cagggccgta
     8281 cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac
     8341 cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac
     8401 atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag
     8461 agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt
     8521 tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag
     8581 cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc
     8641 agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg
     8701 gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc
     8761 gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctga
     8821 cgcccatcga tagatcctaa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     8881 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     8941 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     9001 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     9061 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     9121 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     9181 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg
     9241 tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc
     9301 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     9361 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
     9421 tccccgcctg agatccttta agaccaatga cttacaaggc agctgtagat cttagccact
     9481 ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaaga caagatctgc
     9541 tttttgcttg tactgggtct ctctggttag accagatctg agcctgggag ctctctggct
     9601 aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt
     9661 gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt
     9721 ggaaaatctc tagcagtagt agttcatgtc atcttattat tcagtattta taacttgcaa
     9781 agaaatgaat atcagagagt gagaggcccg ggttaattaa ggaaagggct agatcattct
     9841 tgaagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     9901 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     9961 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
    10021 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
    10081 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
    10141 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
    10201 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
    10261 ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc aagagcaact cggtcgccgc
    10321 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
    10381 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
    10441 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
    10501 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
    10561 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
    10621 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
    10681 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
    10741 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
    10801 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
    10861 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
    10921 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
    10981 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
    11041 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
    11101 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
    11161 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
    11221 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
    11281 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
    11341 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
    11401 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
    11461 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
    11521 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
    11581 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
    11641 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
    11701 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
    11761 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
    11821 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt
    11881 tggccgattc attaatgcag caagctcatg gctgactaat tttttttatt tatgcagagg
    11941 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
    12001 taggcttttg caaaaagctc cccgtggcac gacaggtttc ccgactggaa agcgggcagt
    12061 gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc tttacacttt
    12121 atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac
    12181 agctatgaca tgattacgaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
    12241 ggtttgtcca aactcatcaa tgtatcttat catgtctgga tcaactggat aactcaagct
    12301 aaccaaaatc atcccaaact tcccacccca taccctatta ccactgccaa ttacctgtgg
    12361 tttcatttac tctaaacctg tgattcctct gaattatttt cattttaaag aaattgtatt
    12421 tgttaaatat gtactacaaa cttagtagt
//

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