pHR-OptoCAR-mScarlet

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: pHR-OptoCAR-mScarlet
出品公司: Addgene
目录编号: 170463
存储实验室: John James

载体骨架基本信息
载体名称: pHR-SIN
载体出品公司: Clontech
完整质粒大小: 11966bp
载体类型: 哺乳细胞表达, Lentiviral
筛选标记: 新霉素G418
载体抗性: 氨苄霉素
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: OptoCAR
物种来源: Synthetic
插入大小: 3000bp
启动子: SFFV
标签/融合蛋白: mScarlet (C terminal on insert)
克隆方法: 限制性内切酶
5’克隆位点: AsiSI (not destroyed)
3’克隆位点: NotI (not destroyed)
5’测序引物 cttctgttcgcgcgcttctgcttc
3’测序引物 CCAGAGGTTGATTATCGATAAGC

文献引用方法
材料和方法部分:
pHR-OptoCAR-mScarlet was a gift from John James (Addgene plasmid # 170463 ; http://n2t.net/addgene:170463 ; RRID:Addgene_170463)
 
参考文献部分: Quantifying persistence in the T-cell signaling network using an optically controllable antigen receptor. Harris MJ, Fuyal M, James JR. Mol Syst Biol. 2021 May;17(5):e10091. doi: 10.15252/msb.202010091. 10.15252/msb.202010091 PubMed 33988299

载体图谱质粒图谱和多克隆位点信息

pHR-OptoCAR-mScarlet载体图谱

载体简介

载体序列

LOCUS       pHR-OptoCAR-mScarlet               11966 bp ds-DNA     circular SYN 04-AUG-2021
DEFINITION  Expresses an optically-controllable chimeric antigen receptor that 
            is dependent on dimerizer drug, fused to mScarlet fluorophore.
ACCESSION   .
VERSION     .
KEYWORDS    pHR-OptoCAR-mScarlet
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11966)
  AUTHORS   Harris MJ, Fuyal M, James JR
  TITLE     Quantifying persistence in the T-cell signaling network using an 
            optically controllable antigen receptor.
  JOURNAL   Mol Syst Biol. 2021 May;17(5):e10091. doi: 10.15252/msb.202010091.
  PUBMED    33988299
REFERENCE   2  (bases 1 to 11966)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 4, 2021 from SnapGene Server 1.1.58
            http://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..11966
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        1..408
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     regulatory      446..455
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             515..691
                     /codon_start=1
                     /product="""Zdark"" protein, derived from the Z subunit of 
                     protein A, that binds to the dark state of LOV2 and is 
                     released upon irradiation (Wang et al., 2016)"
                     /label=Zdk1
                     /translation="MVDNKFNKEKTRAGAEIHSLPNLNVEQKFAFIVSLFDDPSQSANL
                     LAEAKKLNDAQAPK"
     regulatory      1028..1037
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1034..1729
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(1167..1185)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1435..1454
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1751..1807
                     /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             1868..1894
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1895..2215
                     /codon_start=1
                     /product="F36V mutant of FK506-binding protein FKBP12"
                     /label=DmrB
                     /note="binds synthetic ligands that do not bind wild-type 
                     FKBP"
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKVDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     misc_feature    3511..4099
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3564..3584)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3982..3993)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4102..4118)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4103..4119)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             4209..4442
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     promoter        complement(4467..4485)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4468..4485)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(4775..4794)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(5008..5027)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5127..5149
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5187..5205)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5273..5377
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5378..6238
                     /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(5596..5615)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6409..6997
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6898..6917
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7151..7168
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(7238..7258)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7243..7572
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7423..7558
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7485..7504
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     intron          8706..8771
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             8901..8921
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    9346..9480
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9383..9402)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9437..9456
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             9648..10281
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    10328..10453
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    10950..11183
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             11368..11412
                     /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    11697..11814
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 gtaacgccat tttgcaaggc atggaaaaat accaaaccaa gaatagagaa gttcagatca
       61 agggcgggta catgaaaata gctaacgttg ggccaaacag gatatctgcg gtgagcagtt
      121 tcggccccgg cccggggcca agaacagatg gtcaccgcag tttcggcccc ggcccgaggc
      181 caagaacaga tggtccccag atatggccca accctcagca gtttcttaag acccatcaga
      241 tgtttccagg ctcccccaag gacctgaaat gaccctgcgc cttatttgaa ttaaccaatc
      301 agcctgcttc tcgcttctgt tcgcgcgctt ctgcttcccg agctctataa aagagctcac
      361 aacccctcac tcggcgcgcc agtcctccga cagactgagt cgcccggggg ggatctggag
      421 ctctcgagaa ttctcacgcg cgatcgccac catgggctgt ggctgcagct cacacccgga
      481 agatgacggc ggatctggag gctccggcgg atctatggtg gataacaaat tcaataaaga
      541 aaagacacgt gccggtgcgg aaatccattc tctgccaaac ctgaatgttg agcagaagtt
      601 tgccttcatc gtgagcctgt ttgatgatcc atctcagagc gccaatctgc tggccgaagc
      661 caaaaaactg aacgatgccc aggccccaaa aactagcgca gacgcccccg cgtaccagca
      721 gggccagaac cagctctata acgagctcaa tctaggacga agagaggagt acgatgtttt
      781 ggacaagaga cgtggccggg accctgagat ggggggaaag ccgcagagaa ggaagaaccc
      841 tcaggaaggc ctgtacaatg aactgcagaa agataagatg gcggaggcct acagtgagat
      901 tgggatgaaa ggcgagcgcc ggaggggcaa ggggcacgat ggcctttacc agggtctcag
      961 tacagccacc aaggacacct acgacgccct tcacatgcag gccctgcccc ctcgggatcc
     1021 accggtcgcc accatggtga gcaagggcga ggcagtgatc aaggagttca tgcggttcaa
     1081 ggtgcacatg gagggctcca tgaacggcca cgagttcgag atcgagggcg agggcgaggg
     1141 ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggtg gccccctgcc
     1201 cttctcctgg gacatcctgt cccctcagtt catgtacggc tccagggcct tcaccaagca
     1261 ccccgccgac atccccgact actataagca gtccttcccc gagggcttca agtgggagcg
     1321 cgtgatgaac ttcgaggacg gcggcgccgt gaccgtgacc caggacacct ccctggagga
     1381 cggcaccctg atctacaagg tgaagctccg cggcaccaac ttccctcctg acggccccgt
     1441 aatgcagaag aagacaatgg gctgggaagc gtccaccgag cggttgtacc ccgaggacgg
     1501 cgtgctgaag ggcgacatta agatggccct gcgcctgaag gacggaggcc gctacctggc
     1561 ggacttcaag accacctaca aggccaagaa gcccgtgcag atgcccggcg cctacaacgt
     1621 cgaccgcaag ttggacatca cctcccacaa cgaggactac accgtggtgg aacagtacga
     1681 acgctccgag ggccgccact ccaccggcgg catggacgag ctgtacaaga ccagttacca
     1741 gggccctggc gccacgaact tctctctgtt aaagcaagca ggagacgtgg aagaaaaccc
     1801 cggtcccacg cgtatgggag tcaaagttct gtttgccctg atctgcatcg ctgtggccga
     1861 ggccaaatat ccgtacgacg taccagacta cgcaggcgtc caagtcgaaa ccattagtcc
     1921 cggcgatggc agaacatttc ctaaaagggg acaaacatgt gtcgtccatt atacaggcat
     1981 gttggaggac ggcaaaaagg tggacagtag tagagatcgc aataaacctt tcaaattcat
     2041 gttgggaaaa caagaagtca ttaggggatg ggaggagggc gtggctcaaa tgtccgtcgg
     2101 ccaacgcgct aagctcacca tcagccccga ctacgcatac ggcgctaccg gacatcccgg
     2161 aattattccc cctcacgcta ccttggtgtt tgacgtcgaa ctgttgaagc tcgagggcgg
     2221 aagcggaggt agtggtggaa gcctgaagat tcaagcttat ttcaatgaga ctgcagacct
     2281 gccatgccaa tttgcaaact ctcaaaacca aagcctgagt gagctagtag tattttggca
     2341 ggaccaggaa aacttggttc tgaatgaggt atacttaggc aaagagaaat ttgacagtgt
     2401 tcattccaag tatatgggcc gcacaagttt tgattcggac agttggaccc tgagacttca
     2461 caatcttcag atcaaggaca agggcttgta tcaatgtatc atccatcaca aaaagcccac
     2521 aggaatgatt cgcatccacc agatgaattc tgaactgtca gtgcttgcta acttcagtca
     2581 acctgaaata gtaccaattt ctaatataac agaaaatgtg tacataaatt tgacctgctc
     2641 atctatacac ggttacccag aacctaagaa gatgagtgtt ttgctaagaa ccaagaattc
     2701 aactatcgag tatgatggta ttatgcagaa atctcaagat aatgtcacag aactgtacga
     2761 cgtttccatc agcttgtctg tttcattccc tgatgttacg agcaatatga ccatcttctg
     2821 tattctggaa actgacaaga cgcggctttt atcttcacct ttctctatag agcttgagga
     2881 ccctcagcct cccccagacc acattccttg gattacagct gtacttccaa cagttattat
     2941 atgtgtgatg gttttctgtc taattctatg gaaatggaag aagaagaagc ggcctcggac
     3001 tagtggcgga agcggaagtt tggctactac acttgaacgt attgagaaga actttgtcat
     3061 tactgaccca agattgccag ataatcccat tatattcgcg tccgatagtt tcttgcagtt
     3121 gacagaatat agccgtgaag aaattttggg aagaaactgc aggtttctac aaggtcctga
     3181 aactgatcgc gcgacagtga gaaaaattag agatgccata gataaccaaa cagaggtcac
     3241 tgttcagctg attaattata caaagagtgg taaaaagttc tggaacctct ttcacttgca
     3301 gcctatgcga gatcagaagg gagatgtcca gtactttatt ggggttcagt tggatggaac
     3361 tgagcatgtc cgagatgctg ccgagagaga gggagtcatg ctgattaaga aaactgcaga
     3421 aaatattgat gaggcggcaa aagaactttg agcggccgcg actctagagt cgacctgcag
     3481 gcatgcaagc ttgatatcaa gcttatcgat aatcaacctc tggattacaa aatttgtgaa
     3541 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     3601 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     3661 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
     3721 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
     3781 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
     3841 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
     3901 gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg
     3961 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
     4021 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
     4081 ctttgggccg cctccccgca tcgataccgt cgacctcgag ggaattaatt cgagctcggt
     4141 acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa
     4201 ggggggactg gaagggctaa ttcactccca acgaagacaa gatctgcttt ttgcttgtac
     4261 tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc
     4321 actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt
     4381 gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag
     4441 cagcatctag aattaattcc gtgtattcta tagtgtcacc taaatcgtat gtgtatgata
     4501 cataaggtta tgtattaatt gtagccgcgt tctaacgaca atatgtacaa gcctaattgt
     4561 gtagcatctg gcttactgaa gcagacccta tcatctctct cgtaaactgc cgtcagagtc
     4621 ggtttggttg gacgaacctt ctgagtttct ggtaacgccg tcccgcaccc ggaaatggtc
     4681 agcgaaccaa tcagcagggt catcgctagc cagatcctct acgccggacg catcgtggcc
     4741 ggcatcaccg gcgccacagg tgcggttgct ggcgcctata tcgccgacat caccgatggg
     4801 gaagatcggg ctcgccactt cgggctcatg agcgcttgtt tcggcgtggg tatggtggca
     4861 ggccccgtgg ccgggggact gttgggcgcc atctccttgc atgcaccatt ccttgcggcg
     4921 gcggtgctca acggcctcaa cctactactg ggctgcttcc taatgcagga gtcgcataag
     4981 ggagagcgtc gaatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     5041 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     5101 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     5161 atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     5221 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     5281 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     5341 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     5401 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     5461 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     5521 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     5581 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     5641 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     5701 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     5761 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     5821 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     5881 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     5941 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     6001 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     6061 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     6121 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     6181 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     6241 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     6301 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     6361 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     6421 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     6481 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     6541 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     6601 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     6661 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     6721 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     6781 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     6841 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     6901 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     6961 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     7021 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     7081 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     7141 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
     7201 tctccccgcg cgttggccga ttcattaatg cagctgtgga atgtgtgtca gttagggtgt
     7261 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     7321 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     7381 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     7441 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     7501 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     7561 ggcttttgca aaaagcttgg acacaagaca ggcttgcgag atatgtttga gaataccact
     7621 ttatcccgcg tcagggagag gcagtgcgta aaaagacgcg gactcatgtg aaatactggt
     7681 ttttagtgcg ccagatctct ataatctcgc gcaacctatt ttcccctcga acacttttta
     7741 agccgtagat aaacaggctg ggacacttca catgagcgaa aaatacatcg tcacctggga
     7801 catgttgcag atccatgcac gtaaactcgc aagccgactg atgccttctg aacaatggaa
     7861 aggcattatt gccgtaagcc gtggcggtct gtaccgggtg cgttactggc gcgtgaactg
     7921 ggtattcgtc atgtcgatac cgtttgtatt tccagctacg atcacgacaa ccagcgcgag
     7981 cttaaagtgc tgaaacgcgc agaaggcgat ggcgaaggct tcatcgttat tgatgacctg
     8041 gtggataccg gtggtactgc ggttgcgatt cgtgaaatgt atccaaaagc gcactttgtc
     8101 accatcttcg caaaaccggc tggtcgtccg ctggttgatg actatgttgt tgatatcccg
     8161 caagatacct ggattgaaca gccgtgggat atgggcgtcg tattcgtccc gccaatctcc
     8221 ggtcgctaat cttttcaacg cctggcactg ccgggcgttg ttctttttaa cttcaggcgg
     8281 gttacaatag tttccagtaa gtattctgga ggctgcatcc atgacacagg caaacctgag
     8341 cgaaaccctg ttcaaacccc gctttaaaca tcctgaaacc tcgacgctag tccgccgctt
     8401 taatcacggc gcacaaccgc ctgtgcagtc ggcccttgat ggtaaaacca tccctcactg
     8461 gtatcgcatg attaaccgtc tgatgtggat ctggcgcggc attgacccac gcgaaatcct
     8521 cgacgtccag gcacgtattg tgatgagcga tgccgaacgt accgacgatg atttatacga
     8581 tacggtgatt ggctaccgtg gcggcaactg gatttatgag tgggccccgg atctttgtga
     8641 aggaacctta cttctgtggt gtgacataat tggacaaact acctacagag atttaaagct
     8701 ctaaggtaaa tataaaattt ttaagtgtat aatgtgttaa actactgatt ctaattgttt
     8761 gtgtatttta gattccaacc tatggaactg atgaatggga gcagtggtgg aatgccttta
     8821 atgaggaaaa cctgttttgc tcagaagaaa tgccatctag tgatgatgag gctactgctg
     8881 actctcaaca ttctactcct ccaaaaaaga agagaaaggt agaagacccc aaggactttc
     8941 cttcagaatt gctaagtttt ttgagtcatg ctgtgtttag taatagaact cttgcttgct
     9001 ttgctattta caccacaaag gaaaaagctg cactgctata caagaaaatt atggaaaaat
     9061 attctgtaac ctttataagt aggcataaca gttataatca taacatactg ttttttctta
     9121 ctccacacag gcatagagtg tctgctatta ataactatgc tcaaaaattg tgtaccttta
     9181 gctttttaat ttgtaaaggg gttaataagg aatatttgat gtatagtgcc ttgactagag
     9241 atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
     9301 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
     9361 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     9421 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc
     9481 aactggataa ctcaagctaa ccaaaatcat cccaaacttc ccaccccata ccctattacc
     9541 actgccaatt acctgtggtt tcatttactc taaacctgtg attcctctga attattttca
     9601 ttttaaagaa attgtatttg ttaaatatgt actacaaact tagtagttgg aagggctaat
     9661 tcactcccaa agaagacaag atatccttga tctgtggatc taccacacac aaggctactt
     9721 ccctgattag cagaactaca caccagggcc aggggtcaga tatccactga cctttggatg
     9781 gtgctacaag ctagtaccag ttgagccaga taaggtagaa gaggccaata aaggagagaa
     9841 caccagcttg ttacaccctg tgagcctgca tgggatggat gacccggaga gagaagtgtt
     9901 agagtggagg tttgacagcc gcctagcatt tcatcacgtg gcccgagagc tgcatccgga
     9961 gtacttcaag aactgctgat atcgagcttg ctacaaggga ctttccgctg gggactttcc
    10021 agggaggcgt ggcctgggcg ggactgggga gtggcgagcc ctcagatcct gcatataagc
    10081 agctgctttt tgcctgtact gggtctctct ggttagacca gatctgagcc tgggagctct
    10141 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag
    10201 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt
    10261 cagtgtggaa aatctctagc agtggcgccc gaacagggac ttgaaagcga aagggaaacc
    10321 agaggagctc tctcgacgca ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg
    10381 gcggcgactg gtgagtacgc caaaaatttt gactagcgga ggctagaagg agagagatgg
    10441 gtgcgagagc gtcagtatta agcgggggag aattagatcg cgatgggaaa aaattcggtt
    10501 aaggccaggg ggaaagaaaa aatataaatt aaaacatata gtatgggcaa gcagggagct
    10561 agaacgattc gcagttaatc ctggcctgtt agaaacatca gaaggctgta gacaaatact
    10621 gggacagcta caaccatccc ttcagacagg atcagaagaa cttagatcat tatataatac
    10681 agtagcaacc ctctattgtg tgcatcaaag gatagagata aaagacacca aggaagcttt
    10741 agacaagata gaggaagagc aaaacaaaag taagaccacc gcacagcaag cggccggccg
    10801 ctgatcttca gacctggagg aggagatatg agggacaatt ggagaagtga attatataaa
    10861 tataaagtag taaaaattga accattagga gtagcaccca ccaaggcaaa gagaagagtg
    10921 gtgcagagag aaaaaagagc agtgggaata ggagctttgt tccttgggtt cttgggagca
    10981 gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg tacaggccag acaattattg
    11041 tctggtatag tgcagcagca gaacaatttg ctgagggcta ttgaggcgca acagcatctg
    11101 ttgcaactca cagtctgggg catcaagcag ctccaggcaa gaatcctggc tgtggaaaga
    11161 tacctaaagg atcaacagct cctggggatt tggggttgct ctggaaaact catttgcacc
    11221 actgctgtgc cttggaatgc tagttggagt aataaatctc tggaacagat ttggaatcac
    11281 acgacctgga tggagtggga cagagaaatt aacaattaca caagcttaat acactcctta
    11341 attgaagaat cgcaaaacca gcaagaaaag aatgaacaag aattattgga attagataaa
    11401 tgggcaagtt tgtggaattg gtttaacata acaaattggc tgtggtatat aaaattattc
    11461 ataatgatag taggaggctt ggtaggttta agaatagttt ttgctgtact ttctatagtg
    11521 aatagagtta ggcagggata ttcaccatta tcgtttcaga cccacctccc aaccccgagg
    11581 ggacccgaca ggcccgaagg aatagaagaa gaaggtggag agagagacag agacagatcc
    11641 attcgattag tgaacggatc tcgacggtat cgccaaatgg cagtattcat ccacaatttt
    11701 aaaagaaaag gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca
    11761 acagacatac aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt
    11821 tattacaggg acagcagaga tccagtttgg atcgataagc ttgatatcga attcctgcag
    11881 ccccgataaa ataaaagatt ttatttagtc tccagaaaaa ggggggaatg aaagacccca
    11941 cctgtaggtt tggcaagcta gctgca
//

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