pLVCT-tTR-KRAB

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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: pLVCT-tTR-KRAB 
出品公司: 爱迪基因
目录编号: 11643
存储实验室: Patrick Aebischer, Didier Trono

载体骨架基本信息
载体名称: --
载体出品公司: --
载体抗性: 氨苄青霉素
空载体大小: 12877 bp
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: --
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: CAG promoters, GFP, tTR-KRAB, Tet-on
物种来源: --
插入大小: --
基因突变: --
GenBank编号 --
启动子 --
标签/融合蛋白: --
克隆方法: 限制性内切酶
5' 克隆位点: --
3' 克隆位点: --
5' 测序引物 See map
3' 测序引物 --

文献引用方法
材料和方法部分: pLVCT-tTR-KRAB was a gift from Patrick Aebischer & Didier Trono (Addgene plasmid # 11643 ; http://n2t.net/addgene:11643 ; RRID:Addgene_11643)
参考文献部分: A versatile tool for conditional gene expression and knockdown. Szulc J, Wiznerowicz M, Sauvain MO, Trono D, Aebischer P. Nat Methods. 2006 Feb . 3(2):109-16. 10.1038/nmeth846 PubMed 16432520

载体图谱质粒图谱和多克隆位点信息

pLVCT-tTR-KRAB质粒图谱

载体简介

载体序列

LOCUS       pLVCT-tTR-KRAB               12884 bp ds-DNA     circular SYN 23-MAR-2020
DEFINITION  Tet-regulated (Tet-on) lentiviral vector for transgene (CAG 
            promoter) - OR - shRNA (H1 promoter when subcloned from pLVTHM 
            (Addgene#12247)) - 2nd generation.
ACCESSION   .
VERSION     .
KEYWORDS    pLVCT-tTR-KRAB
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12884)
  AUTHORS   Szulc J, Wiznerowicz M, Sauvain MO, Trono D, Aebischer P
  TITLE     A versatile tool for conditional gene expression and knockdown.
  JOURNAL   Nat Methods. 2006 Feb . 3(2):109-16.
  PUBMED    16432520
REFERENCE   2  (bases 1 to 12884)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Mar 23, 2020 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..12884
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     LTR             6..639
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    686..811
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1304..1537
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     enhancer        2061..2440
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2442..2719
                     /label=chicken beta-actin promoter
     intron          2720..3736
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     3660..3682
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     3744..3763
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     misc_feature    3822..3939
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     regulatory      4016..4025
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             4022..4738
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4067..4088)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4328..4347)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4675..4696
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    4834..5385
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4978..4995)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5205..5224
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5388..6005
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MARLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESG"
     primer_bind     complement(5461..5480)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     CDS             6009..6029
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6132..6326
                     /codon_start=1
                     /gene="ZNF10"
                     /gene_synonym="KOX1"
                     /product="Kruppel-associated box (KRAB) transcriptional 
                     repression domain from the human zinc finger protein ZNF10 
                     (Margolin et al., 1994)"
                     /label=KRAB
                     /translation="RTLVTFKDVFVDFTREEWKLLDTAQQIVYRNVMLENYKNLVSLGY
                     QLTKPDVILRLEKGEEPWLV"
     misc_feature    6469..7057
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6522..6542)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6940..6951)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    7195..7465
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     LTR             7504..7684
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(7709..7727)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(7710..7727)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(8017..8036)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(8250..8269)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8369..8391
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(8429..8447)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8515..8619
                     /gene="bla"
                     /label=AmpR promoter
     CDS             8620..9480
                     /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(8838..8857)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9651..10239
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10140..10159
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10393..10410
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(10480..10500)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        10485..10814
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      10665..10800
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     10727..10746
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     intron          11948..12013
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             12143..12163
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    12588..12722
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(12625..12644)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     12679..12698
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 gtagttggaa gggctaattc actcccaaag aagacaagat atccttgatc tgtggatcta
       61 ccacacacaa ggctacttcc ctgattagca gaactacaca ccagggccag gggtcagata
      121 tccactgacc tttggatggt gctacaagct agtaccagtt gagccagata aggtagaaga
      181 ggccaataaa ggagagaaca ccagcttgtt acaccctgtg agcctgcatg ggatggatga
      241 cccggagaga gaagtgttag agtggaggtt tgacagccgc ctagcatttc atcacgtggc
      301 ccgagagctg catccggagt acttcaagaa ctgctgatat cgagcttgct acaagggact
      361 ttccgctggg gactttccag ggaggcgtgg cctgggcggg actggggagt ggcgagccct
      421 cagatcctgc atataagcag ctgctttttg cctgtactgg gtctctctgg ttagaccaga
      481 tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct
      541 tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat
      601 ccctcagacc cttttagtca gtgtggaaaa tctctagcag tggcgcccga acagggactt
      661 gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg actcggcttg ctgaagcgcg
      721 cacggcaaga ggcgaggggc ggcgactggt gagtacgcca aaaattttga ctagcggagg
      781 ctagaaggag agagatgggt gcgagagcgt cagtattaag cgggggagaa ttagatcgcg
      841 atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa tataaattaa aacatatagt
      901 atgggcaagc agggagctag aacgattcgc agttaatcct ggcctgttag aaacatcaga
      961 aggctgtaga caaatactgg gacagctaca accatccctt cagacaggat cagaagaact
     1021 tagatcatta tataatacag tagcaaccct ctattgtgtg catcaaagga tagagataaa
     1081 agacaccaag gaagctttag acaagataga ggaagagcaa aacaaaagta agaccaccgc
     1141 acagcaagcg gccgctgatc ttcagacctg gaggaggaga tatgagggac aattggagaa
     1201 gtgaattata taaatataaa gtagtaaaaa ttgaaccatt aggagtagca cccaccaagg
     1261 caaagagaag agtggtgcag agagaaaaaa gagcagtggg aataggagct ttgttccttg
     1321 ggttcttggg agcagcagga agcactatgg gcgcagcgtc aatgacgctg acggtacagg
     1381 ccagacaatt attgtctggt atagtgcagc agcagaacaa tttgctgagg gctattgagg
     1441 cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa gcagctccag gcaagaatcc
     1501 tggctgtgga aagataccta aaggatcaac agctcctggg gatttggggt tgctctggaa
     1561 aactcatttg caccactgct gtgccttgga atgctagttg gagtaataaa tctctggaac
     1621 agatttggaa tcacacgacc tggatggagt gggacagaga aattaacaat tacacaagct
     1681 taatacactc cttaattgaa gaatcgcaaa accagcaaga aaagaatgaa caagaattat
     1741 tggaattaga taaatgggca agtttgtgga attggtttaa cataacaaat tggctgtggt
     1801 atataaaatt attcataatg atagtaggag gcttggtagg tttaagaata gtttttgctg
     1861 tactttctat agtgaataga gttaggcagg gatattcacc attatcgttt cagacccacc
     1921 tcccaacccc gaggggaccc gacaggcccg aaggaataga agaagaaggt ggagagagag
     1981 acagagacag atccattcga ttagtgaacg gatctcgacg gtatcgatca cgagactagc
     2041 ctcgaccatc gatgtcgatc gacattgatt attgactagt tattaatagt aatcaattac
     2101 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
     2161 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
     2221 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
     2281 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
     2341 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
     2401 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtcgaggtga gccccacgtt
     2461 ctgcttcact ctccccatct cccccccctc cccaccccca attttgtatt tatttatttt
     2521 ttaattattt tgtgcagcga tgggggcggg gggggggggg gggcgcgcgc naggsggggc
     2581 ggggcggggc gaggggcggg gcggggcgag gcggagaggt gcggcggcag ccaatcagag
     2641 cggcgcgctc cgaaagtttc cttttatggc gaggcggcgg cggcggcggc cctataaaaa
     2701 gcgaagcgcg cggcgggcgg gagtcgctgc gcgctgcctt cgccccgtgc cccgctccgc
     2761 cgccgcctcg cgccgcccgc cccggctctg actgaccgcg ttactcccac aggtgagcgg
     2821 gcgggacggc ccttctcctc cgggctgtaa ttagcgcttg gtttaatgac ggcttgtttc
     2881 ttttctgtgg ctgcgtgaaa gccttgaggg gctccgggag ggccctttgt gcggggggag
     2941 cggctcgggg ggtgcgtgcg tgtgtgtgtg cgtggggagc gccgcgtgcg gctccgcgct
     3001 gcccggcggc tgtgagcgct gcgggcgcgg cgcggggctt tgtgcgctcc gcagtgtgcg
     3061 cgaggggagc gcggccgggg gcggtgcccc gcggtgcggg gggggctgcg aggggaacaa
     3121 aggctgcgtg cggggtgtgt gcgtgggggg gtgagcaggg ggtgtgggcg cgtcggtcgg
     3181 gctgcaaccc cccctgcacc cccctccccg agttgctgag cacggcccgg cttcgggtgc
     3241 ggggctccgt acggggcgtg gcgcggggct cgccgtgccg ggcggggggt ggcggcaggt
     3301 gggggtgccg ggcggggcgg ggccgcctcg ggccggggag ggctcggggg aggggcgcgg
     3361 cggcccccgg agcgccggcg gctgtcgagg cgcggcgagc cgcagccatt gccttttatg
     3421 gtaatcgtgc gagagggcgc agggacttcc tttgtcccaa atctgtgcgg agccgaaatc
     3481 tgggaggcgc cgccgcaccc cctctagcgg gcgcggggcg aagcggtgcg gcgccggcag
     3541 gaaggaaatg ggcggggagg gccttcgtgc gtcgccgcgc cgccgtcccc ttctccctct
     3601 ccagcctcgg ggctgtccgc ggggggacgg ctgccttcgg gggggacggg gcagggcggg
     3661 gttcggcttc tggcgtgtga ccggcggctc tagagcctct gctaaccatg ttcatgcctt
     3721 cttctttttc ctacagctcc tgggcaacgt gctggttatt gtgctgtctc atcattttgg
     3781 caaagaatta aatttaatta atctcgacgg tatcggttaa cttttaaaag aaaagggggg
     3841 attggggggt acagtgcagg ggaaagaata gtagacataa tagcaacaga catacaaact
     3901 aaagaattac aaaaacaaat tacaaaaatt caaaatttta tcgatcacga gactagcctc
     3961 gaggtttaaa ctacgggatc caggcctaag cttacgcgtc ctagcgctac cggtcgccac
     4021 catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
     4081 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta
     4141 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
     4201 cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa
     4261 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
     4321 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
     4381 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     4441 caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa
     4501 cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc
     4561 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     4621 ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt
     4681 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagtc
     4741 cggactcaga tctcgactag ctagtagcta gctagctagt cgagctcaag cttcgaattc
     4801 ccgggctgca ggaattccgc cccccccccc ctaacgttac tggccgaagc cgcttggaat
     4861 aaggccggtg tgcgtttgtc tatatgttat tttccaccat attgccgtct tttggcaatg
     4921 tgagggcccg gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc
     4981 tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct ctggaagctt
     5041 cttgaagaca aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg
     5101 acaggtgcct ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac
     5161 cccagtgcca cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg
     5221 tattcaacaa ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg
     5281 ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt aaaaaacgtc taggcccccc
     5341 gaaccacggg gacgtggttt tcctttgaaa aacacgatga taataccatg gctagattag
     5401 ataaaagtaa agtgattaac agcgcattag agctgcttaa tgaggtcgga atcgaaggtt
     5461 taacaacccg taaactcgcc cagaagctag gtgtagagca gcctacattg tattggcatg
     5521 taaaaaataa gcgggctttg ctcgacgcct tagccattga gatgttagat aggcaccata
     5581 ctcacttttg ccctttagaa ggggaaagct ggcaagattt tttacgtaat aacgctaaaa
     5641 gttttagatg tgctttacta agtcatcgcg atggagcaaa agtacattta ggtacacggc
     5701 ctacagaaaa acagtatgaa actctcgaaa atcaattagc ctttttatgc caacaaggtt
     5761 tttcactaga gaatgcatta tatgcactca gcgctgtggg gcattttact ttaggttgcg
     5821 tattggaaga tcaagagcat caagtcgcta aagaagaaag ggaaacacct actactgata
     5881 gtatgccgcc attattacga caagctatcg aattatttga tcaccaaggt gcagagccag
     5941 ccttcttatt cggccttgaa ttgatcatat gcggattaga aaaacaactt aaatgtgaaa
     6001 gtgggtcgcc aaaaaagaag agaaaggtcg acggcggtgg tgctttgtct cctcagcact
     6061 ctgctgtcac tcaaggaagt atcatcaaga acaaggaggg catggatgct aagtcactaa
     6121 ctgcctggtc ccggacactg gtgaccttca aggatgtatt tgtggacttc accagggagg
     6181 agtggaagct gctggacact gctcagcaga tcgtgtacag aaatgtgatg ctggagaact
     6241 ataagaacct ggtttccttg ggttatcagc ttactaagcc agatgtgatc ctccggttgg
     6301 agaagggaga agagccctgg ctggtggaga gagaaattca ccaagagacc catcctgatt
     6361 cagagactgc atttgaaatc aaatcatcag tttaaggatc tcgactagct agtagctagc
     6421 tagctagtcg agctcaagct tcgaattcga tgggactagt catatgataa tcaacctctg
     6481 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     6541 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     6601 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
     6661 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
     6721 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
     6781 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     6841 aattccgtgg tgttgtcggg gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc
     6901 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
     6961 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
     7021 cagacgagtc ggatctccct ttgggccgcc tccccgcatc ggtacgtatg gccaggtacc
     7081 tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaaggg
     7141 gggactggaa gggctaattc actcccaacg aagacaagat ctcgagttta ccactcccta
     7201 tcagtgatag agaaaagtga aagtcgagtt taccactccc tatcagtgat agagaaaagt
     7261 gaaagtcgag tttaccactc cctatcagtg atagagaaaa gtgaaagtcg agtttaccac
     7321 tccctatcag tgatagagaa aagtgaaagt cgagtttacc actccctatc agtgatagag
     7381 aaaagtgaaa gtcgagttta ccactcccta tcagtgatag agaaaagtga aagtcgagtt
     7441 taccactccc tatcagtgat agagaaaagt gaaagtcgga tcgatctgct ttttgcttgt
     7501 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
     7561 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
     7621 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
     7681 agcagcatct agaattaatt ccgtgtattc tatagtgtca cctaaatcgt atgtgtatga
     7741 tacataaggt tatgtattaa ttgtagccgc gttctaacga caatatgtac aagcctaatt
     7801 gtgtagcatc tggcttactg aagcagaccc tatcatctct ctcgtaaact gccgtcagag
     7861 tcggtttggt tggacgaacc ttctgagttt ctggtaacgc cgtcccgcac ccggaaatgg
     7921 tcagcgaacc aatcagcagg gtcatcgcta gccagatcct ctacgccgga cgcatcgtgg
     7981 ccggcatcac cggcgccaca ggtgcggttg ctggcgccta tatcgccgac atcaccgatg
     8041 gggaagatcg ggctcgccac ttcgggctca tgagcgcttg tttcggcgtg ggtatggtgg
     8101 caggccccgt ggccggggga ctgttgggcg ccatctcctt gcatgcacca ttccttgcgg
     8161 cggcggtgct caacggcctc aacctactac tgggctgctt cctaatgcag gagtcgcata
     8221 agggagagcg tcgaatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     8281 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     8341 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     8401 tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     8461 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     8521 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     8581 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     8641 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     8701 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     8761 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     8821 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     8881 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     8941 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     9001 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     9061 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     9121 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     9181 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     9241 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     9301 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     9361 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     9421 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     9481 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     9541 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     9601 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     9661 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     9721 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     9781 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     9841 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     9901 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     9961 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
    10021 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
    10081 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
    10141 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
    10201 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
    10261 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
    10321 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
    10381 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
    10441 cctctccccg cgcgttggcc gattcattaa tgcagctgtg gaatgtgtgt cagttagggt
    10501 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
    10561 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
    10621 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
    10681 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
    10741 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
    10801 taggcttttg caaaaagctt ggacacaaga caggcttgcg agatatgttt gagaatacca
    10861 ctttatcccg cgtcagggag aggcagtgcg taaaaagacg cggactcatg tgaaatactg
    10921 gtttttagtg cgccagatct ctataatctc gcgcaaccta ttttcccctc gaacactttt
    10981 taagccgtag ataaacaggc tgggacactt cacatgagcg aaaaatacat cgtcacctgg
    11041 gacatgttgc agatccatgc acgtaaactc gcaagccgac tgatgccttc tgaacaatgg
    11101 aaaggcatta ttgccgtaag ccgtggcggt ctgtaccggg tgcgttactg gcgcgtgaac
    11161 tgggtattcg tcatgtcgat accgtttgta tttccagcta cgatcacgac aaccagcgcg
    11221 agcttaaagt gctgaaacgc gcagaaggcg atggcgaagg cttcatcgtt attgatgacc
    11281 tggtggatac cggtggtact gcggttgcga ttcgtgaaat gtatccaaaa gcgcactttg
    11341 tcaccatctt cgcaaaaccg gctggtcgtc cgctggttga tgactatgtt gttgatatcc
    11401 cgcaagatac ctggattgaa cagccgtggg atatgggcgt cgtattcgtc ccgccaatct
    11461 ccggtcgcta atcttttcaa cgcctggcac tgccgggcgt tgttcttttt aacttcaggc
    11521 gggttacaat agtttccagt aagtattctg gaggctgcat ccatgacaca ggcaaacctg
    11581 agcgaaaccc tgttcaaacc ccgctttaaa catcctgaaa cctcgacgct agtccgccgc
    11641 tttaatcacg gcgcacaacc gcctgtgcag tcggcccttg atggtaaaac catccctcac
    11701 tggtatcgca tgattaaccg tctgatgtgg atctggcgcg gcattgaccc acgcgaaatc
    11761 ctcgacgtcc aggcacgtat tgtgatgagc gatgccgaac gtaccgacga tgatttatac
    11821 gatacggtga ttggctaccg tggcggcaac tggatttatg agtgggcccc ggatctttgt
    11881 gaaggaacct tacttctgtg gtgtgacata attggacaaa ctacctacag agatttaaag
    11941 ctctaaggta aatataaaat ttttaagtgt ataatgtgtt aaactactga ttctaattgt
    12001 ttgtgtattt tagattccaa cctatggaac tgatgaatgg gagcagtggt ggaatgcctt
    12061 taatgaggaa aacctgtttt gctcagaaga aatgccatct agtgatgatg aggctactgc
    12121 tgactctcaa cattctactc ctccaaaaaa gaagagaaag gtagaagacc ccaaggactt
    12181 tccttcagaa ttgctaagtt ttttgagtca tgctgtgttt agtaatagaa ctcttgcttg
    12241 ctttgctatt tacaccacaa aggaaaaagc tgcactgcta tacaagaaaa ttatggaaaa
    12301 atattctgta acctttataa gtaggcataa cagttataat cataacatac tgttttttct
    12361 tactccacac aggcatagag tgtctgctat taataactat gctcaaaaat tgtgtacctt
    12421 tagcttttta atttgtaaag gggttaataa ggaatatttg atgtatagtg ccttgactag
    12481 agatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
    12541 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
    12601 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
    12661 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga
    12721 tcaactggat aactcaagct aaccaaaatc atcccaaact tcccacccca taccctatta
    12781 ccactgccaa ttacctgtgg tttcatttac tctaaacctg tgattcctct gaattatttt
    12841 cattttaaag aaattgtatt tgttaaatat gtactacaaa ctta
//

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