pJFRC7-RFPnls-PQRv2-mCD8:GFP

价格:1800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: pJFRC7-RFPnls-PQRv2-mCD8:GFP
出品公司: Addgene
目录编号: 73974
存储实验室: Brian Chen

载体骨架基本信息
载体名称: pJFRC7
载体出品公司: Janelia Farms
空载体大小: 8142bp
完整质粒大小: 10413 bp
载体类型: Insect Expression
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1: sfGFP / folder GFP
插入大小: 1202 bp
克隆方法: 20XUAS-HS
标签/融合蛋白: mCD8 (N terminal on insert)
克隆方法: Gibson Cloning
5’测序引物 GGCCACCTGATCTGCAAC
3’测序引物 CCATTCATCAGTTCCATAGG
插入基因名称2: RFP / TagRFP-T
物种来源: D. melanogaster (fly)
插入大小: 816bp
克隆方法: 20XUAS-HS
标签/融合蛋白: nls (C terminal on insert)
克隆方法: Gibson Cloning
5’测序引物 GAGCGCCGGAGTATAAATAGAG
3’测序引物 TCTGGAAGAGCCAAGAGCAT

文献引用方法
材料和方法部分: pJFRC7-RFPnls-PQRv2-mCD8:GFP was a gift from Brian Chen (Addgene plasmid # 73974 ; http://n2t.net/addgene:73974 ; RRID:Addgene_73974)
参考文献部分: Quantification of Protein Levels in Single Living Cells. Lo CA, Kays I, Emran F, Lin TJ, Cvetkovska V, Chen BE. Cell Rep. 2015 Dec 22;13(11):2634-44. doi: 10.1016/j.celrep.2015.11.048. Epub 2015 Dec 10. 10.1016/j.celrep.2015.11.048 PubMed 26686644

载体图谱质粒图谱和多克隆位点信息

pJFRC7-RFPnls-PQRv2-mCD8:GFP载体图谱

载体简介

载体序列

LOCUS       pJFRC7-RFPnls-PQRv2-mCD8:GFP               10413 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Protein Quantitation Reporter (PQR) to quantify a protein of 
            interest in insect cells..
ACCESSION   .
VERSION     .
KEYWORDS    pJFRC7-RFPnls-PQRv2-mCD8:GFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10413)
  AUTHORS   Lo CA, Kays I, Emran F, Lin TJ, Cvetkovska V, Chen BE
  TITLE     Quantification of Protein Levels in Single Living Cells.
  JOURNAL   Cell Rep. 2015 Dec 22;13(11):2634-44. doi: 
            10.1016/j.celrep.2015.11.048. Epub 2015 Dec 10.
  PUBMED    26686644
REFERENCE   2  (bases 1 to 10413)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..10413
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        31..135
                     /gene="bla"
                     /label=AmpR promoter
     CDS             136..996
                     /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     complement(354..373)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1167..1755
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1656..1675
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    1842..1911
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     primer_bind     complement(2136..2154)
                     /label=pCasper-F
                     /note="Drosophila mini-white gene, reverse primer"
     CDS             join(2606..2677,3079..3352,3427..4081,4143..4458,4662..4793,
                     4864..5478)
                     /codon_start=1
                     /gene="white"
                     /product="Drosophila white gene eye color pigment"
                     /label=mini-white
                     /note="This is a modified version of the white gene lacking
                     part of the first intron."
                     /translation="MGQEDQELLIRGGSKHPSAEHLNNGDSGAASQSCINQGFGQAKNY
                     GTLRPPSPPEDSGSGSGQLAENLTYAWHNMDIFGAVNQPGSGWRQLVNRTRGLFCNERH
                     IPAPRKHLLKNVCGVAYPGELLAVMGSSGAGKTTLLNALAFRSPQGIQVSPSGMRLLNG
                     QPVDAKEMQARCAYVQQDDLFIGSLTAREHLIFQAMVRMPRHLTYRQRVARVDQVIQEL
                     SLSKCQHTIIGVPGRVKGLSGGERKRLAFASEALTDPPLLICDEPTSGLDSFTAHSVVQ
                     VLKKLSQKGKTVILTIHQPSSELFELFDKILLMAEGRVAFLGTPSEAVDFFSYVGAQCP
                     TNYNPADFYVQVLAVVPGREIESRDRIAKICDNFAISKVARDMEQLLATKNLEKPLEQP
                     ENGYTYKATWFMQFRAVLWRSWLSVLKEPLLVKVRLIQTTMVAILIGLIFLGQQLTQVG
                     VMNINGAIFLFLTNMTFQNVFATINVFTSELPVFMREARSRLYRCDTYFLGKTIAELPL
                     FLTVPLVFTAIAYPMIGLRAGVLHFFNCLALVTLVANVSTSFGYLISCASSSTSMALSV
                     GPPVIIPFLLFGGFFLNSGSVPVYLKWLSYLSWFRYANEGLLINQWADVEPGEISCTSS
                     NTTCPSSGKVILETLNFSAADLPLDYVGLAILIVSFRVLAYLALRLRARRKE"
     protein_bind    6508..6602
                     /label=5X UAS
                     /bound_moiety="GAL4"
                     /note="five tandem copies of the ""ScaI site"" 17-mer 
                     CGGAGTACTGTCCTCCG, an upstream activating sequence (UAS) 
                     that efficiently binds yeast Gal4 (Webster et al., 1988; 
                     Pfeiffer et al., 2010)"
     protein_bind    6636..6730
                     /label=5X UAS
                     /bound_moiety="GAL4"
                     /note="five tandem copies of the ""ScaI site"" 17-mer 
                     CGGAGTACTGTCCTCCG, an upstream activating sequence (UAS) 
                     that efficiently binds yeast Gal4 (Webster et al., 1988; 
                     Pfeiffer et al., 2010)"
     protein_bind    6764..6858
                     /label=5X UAS
                     /bound_moiety="GAL4"
                     /note="five tandem copies of the ""ScaI site"" 17-mer 
                     CGGAGTACTGTCCTCCG, an upstream activating sequence (UAS) 
                     that efficiently binds yeast Gal4 (Webster et al., 1988; 
                     Pfeiffer et al., 2010)"
     protein_bind    6892..6986
                     /label=5X UAS
                     /bound_moiety="GAL4"
                     /note="five tandem copies of the ""ScaI site"" 17-mer 
                     CGGAGTACTGTCCTCCG, an upstream activating sequence (UAS) 
                     that efficiently binds yeast Gal4 (Webster et al., 1988; 
                     Pfeiffer et al., 2010)"
     promoter        7011..7249
                     /label=hsp70 promoter
                     /note="Drosophila melanogaster hsp70Bb promoter"
     primer_bind     7196..7217
                     /label=pCasper-hs
                     /note="Drosophila Hsp70 promoter, forward primer"
     intron          7264..7330
                     /label=Mhc intron
                     /note="modified Drosophila myosin heavy chain intron 16 
                     (Pfeiffer et al., 2010)"
     CDS             7339..8070
                     /codon_start=1
                     /product="photostable variant of TagRFP (Shaner et al., 
                     2008)"
                     /label=TagRFP-T
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFPSNCPVMQKKTLGWEANTEMLYPADGGLEGRTD
                     MALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAV
                     ARYCDLPSKLGHKLNGMDELYK"
     CDS             8080..8100
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8104..8124
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8128..8148
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8170..8223
                     /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             8224..8889
                     /codon_start=1
                     /product="mouse lymphocyte antigen CD8 alpha chain"
                     /label=mCD8
                     /translation="MASPLTRFLSLNLLLLGESIILGSGEAKPQAPELRIFPKKMDAEL
                     GQKVDLVCEVLGSVSQGCSWLFQNSSSKLPQPTFVVYMASSHNKITWDEKLNSSKLFSA
                     MRDTNNKYVLTLNKFSKENEGYYFCSVISNSVMYFSSVVPVLQKVNSTTTKPVLRTPSP
                     VHPTGTSQPQRPEDCRPRGSVKGTGLDFACDIYIWAPLAGICVALLLSLIITLICYHSR
                     "
     CDS             8896..9609
                     /codon_start=1
                     /product="green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(8926..8954)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     9549..9569
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     intron          9696..9761
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             9891..9911
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    10183..10317
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10220..10239)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10274..10293
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(10359..10377)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gactcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
       61 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
      121 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
      181 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
      241 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
      301 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
      361 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
      421 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
      481 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
      541 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
      601 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
      661 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
      721 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
      781 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
      841 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
      901 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
      961 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     1021 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     1081 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     1141 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     1201 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     1261 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     1321 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     1381 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     1441 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     1501 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     1561 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     1621 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     1681 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     1741 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     1801 ccttttgctc acatgttacc gtcgacgatg taggtcacgg tctcgaagcc gcggtgcggg
     1861 tgccagggcg tgcccttggg ctccccgggc gcgtactcca cctcacccat ctggtccatc
     1921 atgatgaacg ggtcgaggtg gcggtagttg atcccggcga acgcgcggcg caccgggaag
     1981 ccctcgccct cgaaaccgct gggcgcggtg gtcacggtga gcacgggacg tgcgacggcg
     2041 tcggcgggtg cggatacgcg gggcagcgtc agcgggttct cgacggtcac ggcgggcatg
     2101 tcgacaagcc gaacatatgg gcgcgcctag tatgtatgta agttaataaa acccattttt
     2161 gcggaaagta gataaaaaaa acattttttt tttttactgc actggatatc attgaactta
     2221 tctgatcagt tttaaattta cttcgatcca agggtatttg atgtaccagg ttctttcgat
     2281 tacctctcac tcaaaatgac attccactca aagtcagcgc tgtttgcctc cttctctgtc
     2341 cacagaaata tcgccgtctc tttcgccgct gcgtccgcta tctctttcgc caccgtttgt
     2401 agcgttacgt agcgtcaatg tccgccttca gttgcatttt gtcagcggtt tcgtgacgaa
     2461 gctccaagcg gtttacgcca tcaattaaac acaaagtgct gtgccaaaac tcctctcgct
     2521 tcttattttt gtttgttttt tgagtgattg gggtggtgat tggttttggg tgggtaagca
     2581 ggggaaagtg tgaaaaatcc cggcaatggg ccaagaggat caggagctat taattcgcgg
     2641 aggcagcaaa cacccatctg ccgagcatct gaacaatgtg agtagtacat gtgcatacat
     2701 cttaagttca cttgatctat aggaactgcg attgcaacat caaattgtat gcggcgtgag
     2761 aactgcgacc cacaaaaatc ccaaaccgca attgcacaaa caaatagtga cacgaaacag
     2821 attattctgg tagctgttct cgctatataa gacaattttt gagatcatat catgatcaag
     2881 acatctaaag gcattcattt tcgactatat tcttttttac aaaaaatata acaaccagat
     2941 attttaagct gatcctagat gcacaaaaaa taaataaaag tataaaccta cttcgtagga
     3001 tacttcgggg tactttttgt tcggggttag atgagcataa cgctagtagt tgatatttga
     3061 gatcccctat cattgcaggg tgacagcgga gcggcttcgc agagctgcat taaccagggc
     3121 ttcgggcagg ccaaaaacta cggcacgctc cggccaccca gtccgccgga ggactccggt
     3181 tcagggagcg gccaactagc cgagaacctc acctatgcct ggcacaatat ggacatcttt
     3241 ggggcggtca atcagccggg ctccggatgg cggcagctgg tcaaccggac acgcggacta
     3301 ttctgcaacg agcgacacat accggcgccc aggaaacatt tgctcaagaa cggtgagttt
     3361 ctattcgcag tcggctgatc tgtgtgaaat cttaataaag ggtccaatta ccaatttgaa
     3421 actcagtttg cggcgtggcc tatccgggcg aacttttggc cgtgatgggc agttccggtg
     3481 ccggaaagac gaccctgctg aatgcccttg cctttcgatc gccgcagggc atccaagtat
     3541 cgccatccgg gatgcgactg ctcaatggcc aacctgtgga cgccaaggag atgcaggcca
     3601 ggtgcgccta tgtccagcag gatgacctct ttatcggctc cctaacggcc agggaacacc
     3661 tgattttcca agccatggtg cggatgccac gacatctgac ctatcggcag cgagtggccc
     3721 gcgtggatca ggtgatccag gagctttcgc tcagcaaatg tcagcacacg atcatcggtg
     3781 tgcccggcag ggtgaaaggt ctgtccggcg gagaaaggaa gcgtctggca ttcgcctccg
     3841 aggctctaac cgatccgccg cttctgatct gcgatgagcc cacctccgga ctggactcct
     3901 ttaccgccca cagcgtcgtc caggtgctga agaagctgtc gcagaagggc aagaccgtca
     3961 tcctgaccat tcatcagccg tcttccgagc tgtttgagct ctttgacaag atccttctga
     4021 tggccgaggg cagggtagct ttcttgggca ctcccagcga agccgtcgac ttcttttcct
     4081 agtgagttcg atgtgtttat taagggtatc tagtattaca taacatctca actcctatcc
     4141 agcgtgggtg cccagtgtcc taccaactac aatccggcgg acttttacgt acaggtgttg
     4201 gccgttgtgc ccggacggga gatcgagtcc cgtgatcgga tcgccaagat atgcgacaat
     4261 tttgccatta gcaaagtagc ccgggatatg gagcagttgt tggccaccaa aaatctggag
     4321 aagccactgg agcagccgga gaatgggtac acctacaagg ccacctggtt catgcagttc
     4381 cgggcggtcc tgtggcgatc ctggctgtcg gtgctcaagg aaccactcct cgtaaaagtg
     4441 cgacttattc agacaacggt gagtggttcc agtggaaaca aatgatataa cgcttacaat
     4501 tcttggaaac aaattcgcta gattttagat agaattgcct gattccacac ccttcttagt
     4561 ttttttcaat gagatgtata gtttatagtt ttgcagaaga taaataaatt tcatttaact
     4621 cgcgaatatt aatgagatgc gagtaacatt ttaatttgca gatggttgcc atcttgattg
     4681 gcctcatctt tttgggccaa caactcacgc aagtgggtgt gatgaatatc aacggagcca
     4741 tcttcctctt cctgaccaac atgacctttc aaaacgtctt tgccacgata aatgtaagtc
     4801 atgtttagaa tacatttgca tttcaataat ttactaactt tctaatgaat cgattcgatt
     4861 taggtgttca cctcagagct gccagttttt atgagggagg cccgaagtcg actttatcgc
     4921 tgtgacacat actttctggg caaaacgatt gccgaattgc cgctttttct cacagtgcca
     4981 ctggtcttca cggcgattgc ctatccgatg atcggactgc gggccggagt gctgcacttc
     5041 ttcaactgcc tggcgctggt cactctggtg gccaatgtgt caacgtcctt cggatatcta
     5101 atatcctgcg ccagctcctc gacctcgatg gcgctgtctg tgggtccgcc ggttatcata
     5161 ccattcctgc tctttggcgg cttcttcttg aactcgggct cggtgccagt atacctcaaa
     5221 tggttgtcgt acctctcatg gttccgttac gccaacgagg gtctgctgat taaccaatgg
     5281 gcggacgtgg agccgggcga aattagctgc acatcgtcga acaccacgtg ccccagttcg
     5341 ggcaaggtca tcctggagac gcttaacttc tccgccgccg atctgccgct ggactacgtg
     5401 ggtctggcca ttctcatcgt gagcttccgg gtgctcgcat atctggctct aagacttcgg
     5461 gcccgacgca aggagtagcc gacatatatc cgaaataact gcttgttttt tttttttacc
     5521 attattacca tcgtgtttac tgtttattgc cccctcaaaa agctaatgta attatatttg
     5581 tgccaataaa aacaagatat gacctataga atacaagtat ttccccttcg aacatcccca
     5641 caagtagact ttggatttgt cttctaacca aaagacttac acacctgcat accttacatc
     5701 aaaaactcgt ttatcgctac ataaaacacc gggatatatt ttttatatac atacttttca
     5761 aatcgcgcgc cctcttcata attcacctcc accacaccac gtttcgtagt tgctctttcg
     5821 ctgtctccca cccgctctcc gcaacacatt caccttttgt tcgacgacct tggagcgact
     5881 gtcgttagtt ccgcgcgatt cggttcgctc aaatggttcc gagtggttca tttcgtctca
     5941 atagaaatta gtaataaata tttgtatgta caatttattt gctccaatat atttgtatat
     6001 atttccctca cagctatatt tattctaatt taatattatg actttttaag gtaatttttt
     6061 gtgacctgtt cggagtgatt agcgttacaa tttgaactga aagtgacatc cagtgtttgt
     6121 tccttgtgta gatgcatctc aaaaaaatgg tgggcataat agtgttgttt atatatatca
     6181 aaaataacaa ctataataat aagaatacat ttaatttaga aaatgcttgg atttcactgg
     6241 aactagggcg cgcctccgga acataatggt gcagggcgct gacttccgcg tttccagact
     6301 ttacgaaaca cggaaaccga agaccattca tgttgttgct caggtcgcag acgttttgca
     6361 gcagcagtcg cttcacgttc gctcgcgtat cggtgattca ttctgctaac cagtaaggca
     6421 accccgccag cctagccggg tcctcaacga caggagcacg atcatgcgca cccgtggcca
     6481 gggccgcaag cttgcatgcc tgcaggtcgg agtactgtcc tccgagcgga gtactgtcct
     6541 ccgagcggag tactgtcctc cgagcggagt actgtcctcc gagcggagta ctgtcctccg
     6601 agcggagact ctagccctag ggcatgcctg caggtcggag tactgtcctc cgagcggagt
     6661 actgtcctcc gagcggagta ctgtcctccg agcggagtac tgtcctccga gcggagtact
     6721 gtcctccgag cggagactct agcgctagcg catgcctgca ggtcggagta ctgtcctccg
     6781 agcggagtac tgtcctccga gcggagtact gtcctccgag cggagtactg tcctccgagc
     6841 ggagtactgt cctccgagcg gagactctag cactagtgca tgcctgcagg tcggagtact
     6901 gtcctccgag cggagtactg tcctccgagc ggagtactgt cctccgagcg gagtactgtc
     6961 ctccgagcgg agtactgtcc tccgagcgga gactctagcg acgtcgagcg ccggagtata
     7021 aatagaggcg cttcgtctac ggagcgacaa ttcaattcaa acaagcaaag tgaacacgtc
     7081 gctaagcgaa agctaagcaa ataaacaagc gcagctgaac aagctaaaca atctgcagta
     7141 aagtgcaagt taaagtgaat caattaaaag taaccagcaa ccaagtaaat caactgcaac
     7201 tactgaaatc tgccaagaag taattattga atacaagaag agaactctga atagatctaa
     7261 aaggtaggtt caaccactga tgcctaggca caccgaaacg actaacccta attcttatcc
     7321 tttacttcag gcggccgcat ggtgtctaag ggcgaagagc tgattaagga gaacatgcac
     7381 atgaagctgt acatggaggg caccgtgaac aaccaccact tcaagtgcac atccgagggc
     7441 gaaggcaagc cctacgaggg cacccagacc atgagaatca aggtggtcga gggcggccct
     7501 ctccccttcg ccttcgacat cctggctacc agcttcatgt acggcagcag aaccttcatc
     7561 aaccacaccc agggcatccc cgatttcttt aagcagtcct tccctgaggg cttcacatgg
     7621 gagagagtca ccacatacga agacgggggc gtgctgaccg ctacccagga caccagcctc
     7681 caggacggct gcctcatcta caacgtcaag atcagagggg tgaacttccc atccaactgc
     7741 cctgtgatgc agaagaaaac actcggctgg gaggccaaca ccgagatgct gtaccccgct
     7801 gacggcggcc tggaaggcag aaccgacatg gccctgaagc tcgtgggcgg gggccacctg
     7861 atctgcaact tcaagaccac atacagatcc aagaaacccg ctaagaacct caagatgccc
     7921 ggcgtctact atgtggacca cagactggaa agaatcaagg aggccgacaa agagacctac
     7981 gtcgagcagc acgaggtggc tgtggccaga tactgcgacc tccctagcaa actggggcac
     8041 aaacttaatg gcatggacga gctgtacaag ctcgaggacc caaagaagaa gcggaaagtg
     8101 gatccaaaga aaaagcgcaa ggtggaccct aagaagaaac ggaaggtgac cggtatcgat
     8161 gggtccgggg aaggtcgtgg tagtctacta acgtgtggtg atgtagaaga aaatcctgga
     8221 cctatggcct caccgttgac ccgctttctg tcgctgaacc tgctgctgct gggtgagtcg
     8281 attatcctgg ggagtggaga agctaagcca caggcacccg aactccgaat ctttccaaag
     8341 aaaatggacg ccgaacttgg tcagaaggtg gacctggtat gtgaagtgtt ggggtccgtt
     8401 tcgcaaggat gctcttggct cttccagaac tccagctcca aactccccca gcccaccttc
     8461 gttgtctata tggcttcatc ccacaacaag ataacgtggg acgagaagct gaattcgtcg
     8521 aaactgtttt ctgccatgag ggacacgaat aataagtacg ttctcaccct gaacaagttc
     8581 agcaaggaaa acgaaggcta ctatttctgc tcagtcatca gcaactcggt gatgtacttc
     8641 agttctgtcg tgccagtcct tcagaaagtg aactctacta ctaccaagcc agtgctgcga
     8701 actccctcac ctgtgcaccc taccgggaca tctcagcccc agagaccaga agattgtcgg
     8761 ccccgtggct cagtgaaggg gaccggattg gacttcgcct gtgatattta catctgggca
     8821 cccttggccg gaatctgcgt ggcccttctg ctgtccttga tcatcactct catctgctac
     8881 cacagccgcg gatccatgag taaaggagaa gaacttttca ctggagttgt cccaattctt
     8941 gttgaattag atggtgatgt taatgggcac aaattttctg tcagtggaga gggtgaaggt
     9001 gatgcaacat acggaaaact tacccttaaa tttatttgca ctactggaaa actacctgtt
     9061 ccatggccaa cacttgtcac tactttaact tatggtgttc aatgcttttc aagataccca
     9121 gatcatatga aacagcatga ctttttcaag agtgccatgc ccgaaggtta tgtccaggaa
     9181 agaactatat ttttcaaaga tgacgggaac tacaagacac gtgctgaagt caagtttgaa
     9241 ggtgataccc ttgttaatag aatcgagtta aaaggtattg attttaaaga agatggaaac
     9301 attcttggac acaaattgga atacaactat aactcacaca atgtatacat catggcagac
     9361 aaacaaaaga atggaatcaa agttaacttc aaaattagac acaacattga agatggaagc
     9421 gttcaactag cagaccatta tcaacaaaat actccaattg gcgatggccc tgtcctttta
     9481 ccagacaacc attacctgtc cacacaatct gccctttcga aagatcccaa cgaaaagaga
     9541 gaccacatgg tccttcttga gtttgtaaca gctgctggga ttacacatgg catggatgaa
     9601 ctatacaaat aatctagagg atctttgtga aggaacctta cttctgtggt gtgacataat
     9661 tggacaaact acctacagag atttaaagct ctaaggtaaa tataaaattt ttaagtgtat
     9721 aatgtgttaa actactgatt ctaattgttt gtgtatttta gattccaacc tatggaactg
     9781 atgaatggga gcagtggtgg aatgccttta atgaggaaaa cctgttttgc tcagaagaaa
     9841 tgccatctag tgatgatgag gctactgctg actctcaaca ttctactcct ccaaaaaaga
     9901 agagaaaggt agaagacccc aaggactttc cttcagaatt gctaagtttt ttgagtcatg
     9961 ctgtgtttag taatagaact cttgcttgct ttgctattta caccacaaag gaaaaagctg
    10021 cactgctata caagaaaatt atggaaaaat atttgatgta tagtgccttg actagagatc
    10081 ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc
    10141 cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct
    10201 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
    10261 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatcgat
    10321 ctggccggcc gtttaaacga attcttgaag acgaaagggc ctcgtgatac gcctattttt
    10381 ataggttaat gtcatgataa taatggtttc tta
//

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