pINDUCER21 (ORF-EG)

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: pINDUCER21 (ORF-EG) 
出品公司: 爱迪基因
目录编号: 46948
存储实验室: Stephen Elledge, Thomas Westbrook

载体骨架基本信息
载体名称: pHAGE-CAGS-W
载体出品公司: Dr. Richard Mulligan
载体抗性: 氯霉素和氨苄青霉素
空载体大小: 4874 bp
完整质粒大小: --
载体修饰: pINDUCER21(ORF-UG)是用NdeI和SphI消化pINDUCER20制成的。eGFP分别在5′端和3′端用AgeI和SphI进行PCR扩增,然后用AgeI和SphI进行酶切。将载体与eGFP连接,并进行序列验证。pINDUCER20(ORF-UN)是通过用BamHI和SpeI从pSLIK Neo-TGmiR-Luc载体中消化TRE2,并将钝端克隆到SpeI和XhoI消化的噬菌体-CAGS-W载体(Richard Mulligan博士,哈佛医学院,波士顿)的骨干中来产生噬菌体TRE载体。用SpeI从噬菌体EF-DEST载体(G.H.)中切出含有网关受体盒的片段并插入噬菌体TRE中以产生噬菌体TRE-DEST载体。用PacI和ClaI从pSLIK-Neo-TGmiR-Luc载体中切出Ubc-rtTA3-IRES-Neo片段,并将其克隆到SphI和ClaI消化的噬菌体tredest的骨架中。
载体类型: 慢病毒载体表达
筛选标记: eGFP
高拷贝/低拷贝: --
生长条件: 30 ℃

插入基因信息
插入基因名称: --
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: --
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: pINDUCER21 (ORF-EG) was a gift from Stephen Elledge & Thomas Westbrook (Addgene plasmid # 46948 ; http://n2t.net/addgene:46948 ; RRID:Addgene_46948)
参考文献部分: The pINDUCER lentiviral toolkit for inducible RNA interference in vitro and in vivo. Meerbrey KL, Hu G, Kessler JD, Roarty K, Li MZ, Fang JE, Herschkowitz JI, Burrows AE, Ciccia A, Sun T, Schmitt EM, Bernardi RJ, Fu X, Bland CS, Cooper TA, Schiff R, Rosen JM, Westbrook TF, Elledge SJ. Proc Natl Acad Sci U S A. 2011 Mar 1;108(9):3665-70. doi: 10.1073/pnas.1019736108. Epub 2011 Feb 9. 10.1073/pnas.1019736108 PubMed 21307310

载体图谱质粒图谱和多克隆位点信息

pINDUCER21 (ORF-EG)质粒图谱

载体简介

载体序列

LOCUS       pINDUCER21 (ORF-EG)            12152 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Inducible lentiviral gene expression vector. Chloramphenicol cm + 
            ccdB cassette may be removed to clone in gene of interest.
ACCESSION   .
VERSION     .
KEYWORDS    pINDUCER21 (ORF-EG)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12152)
  AUTHORS   Meerbrey KL, Hu G, Kessler JD, Roarty K, Li MZ, Fang JE, 
            Herschkowitz JI, Burrows AE, Ciccia A, Sun T, Schmitt EM, Bernardi 
            RJ, Fu X, Bland CS, Cooper TA, Schiff R, Rosen JM, Westbrook TF, 
            Elledge SJ
  TITLE     The pINDUCER lentiviral toolkit for inducible RNA interference in 
            vitro and in vivo.
  JOURNAL   Proc Natl Acad Sci U S A. 2011 Mar 1;108(9):3665-70. doi: 
            10.1073/pnas.1019736108. Epub 2011 Feb 9.
  PUBMED    21307310
REFERENCE   2  (bases 1 to 12152)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..12152
                     /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"
     protein_bind    2230..2500
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        2533..2571
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2568..2592
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    2701..2825
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        2850..2880
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             2934..3593
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3000..3019)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             3935..4240
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     4119..4138
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(4281..4405)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             4458..4484
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        4704..5882
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          4935..5873
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     5830..5850
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    complement(5952..5985)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     CDS             5992..6738
                     /codon_start=1
                     /product="improved tetracycline-controlled transactivator"
                     /label=rtTA-Advanced
                     /note="In the Tet-On(R) system, rtTA-Advanced binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the presence of tetracycline or doxycycline."
                     /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     misc_feature    6791..7364
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(6958..6975)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     7185..7204
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             7380..8099
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             7380..8096
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(7425..7446)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(7686..7705)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     8033..8054
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     8235..8254
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     misc_feature    8542..9130
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8595..8615)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9013..9024)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9205..9438
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(9557..9575)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        9643..9747
                     /gene="bla"
                     /label=AmpR promoter
     CDS             9748..10608
                     /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(9966..9985)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10779..11367
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11268..11287
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     11521..11538
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     11630..11649
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     protein_bind    11778..11799
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        11814..11844
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11852..11868
                     /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     11857..11879
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     11876..11892
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     11876..11892
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     11942..11961
                     /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 gactaggatc
     2221 ctttaccact ccctatcagt gatagagaaa agtgaaagtc gagtttacca ctccctatca
     2281 gtgatagaga aaagtgaaag tcgagtttac cactccctat cagtgataga gaaaagtgaa
     2341 agtcgagttt accactccct atcagtgata gagaaaagtg aaagtcgagt ttaccactcc
     2401 ctatcagtga tagagaaaag tgaaagtcga gtttaccact ccctatcagt gatagagaaa
     2461 agtgaaagtc gagtttacca ctccctatca gtgatagaga aaagtgaaag tcgagctcgg
     2521 tacccgggtc gaggtaggcg tgtacggtgg gaggcctata taagcagagc tcgtttagtg
     2581 aaccgtcaga tcgcctggag acgccatcca cgctgttttg acctccatag aagacaccgg
     2641 gaccgatcca gcctccgcgg ccccgaacta gtccagtgtg gtggaattct gcagatatca
     2701 acaagtttgt acaaaaaagc tgaacgagaa acgtaaaatg atataaatat caatatatta
     2761 aattagattt tgcataaaaa acagactaca taatactgta aaacacaaca tatccagtca
     2821 ctatggcggc cgcattaggc accccaggct ttacacttta tgcttccggc tcgtataatg
     2881 tgtggatttt gagttaggat ccgtcgagat tttcaggagc taaggaagct aaaatggaga
     2941 aaaaaatcac tggatatacc accgttgata tatcccaatg gcatcgtaaa gaacattttg
     3001 aggcatttca gtcagttgct caatgtacct ataaccagac cgttcagctg gatattacgg
     3061 cctttttaaa gaccgtaaag aaaaataagc acaagtttta tccggccttt attcacattc
     3121 ttgcccgcct gatgaatgct catccggaat tccgtatggc aatgaaagac ggtgagctgg
     3181 tgatatggga tagtgttcac ccttgttaca ccgttttcca tgagcaaact gaaacgtttt
     3241 catcgctctg gagtgaatac cacgacgatt tccggcagtt tctacacata tattcgcaag
     3301 atgtggcgtg ttacggtgaa aacctggcct atttccctaa agggtttatt gagaatatgt
     3361 ttttcgtctc agccaatccc tgggtgagtt tcaccagttt tgatttaaac gtggccaata
     3421 tggacaactt cttcgccccc gttttcacca tgggcaaata ttatacgcaa ggcgacaagg
     3481 tgctgatgcc gctggcgatt caggttcatc atgccgtttg tgatggcttc catgtcggca
     3541 gaatgcttaa tgaattacaa cagtactgcg atgagtggca gggcggggcg taaagatctg
     3601 gatccggctt actaaaagcc agataacagt atgcgtattt gcgcgctgat ttttgcggta
     3661 taagaatata tactgatatg tatacccgaa gtatgtcaaa aagaggtatg ctatgaagca
     3721 gcgtattaca gtgacagttg acagcgacag ctatcagttg ctcaaggcat atatgatgtc
     3781 aatatctccg gtctggtaag cacaaccatg cagaatgaag cccgtcgtct gcgtgccgaa
     3841 cgctggaaag cggaaaatca ggaagggatg gctgaggtcg cccggtttat tgaaatgaac
     3901 ggctcttttg ctgacgagaa caggggctgg tgaaatgcag tttaaggttt acacctataa
     3961 aagagagagc cgttatcgtc tgtttgtgga tgtacagagt gatattattg acacgcccgg
     4021 gcgacggatg gtgatccccc tggccagtgc acgtctgctg tcagataaag tctcccgtga
     4081 actttacccg gtggtgcata tcggggatga aagctggcgc atgatgacca ccgatatggc
     4141 cagtgtgccg gtctccgtta tcggggaaga agtggctgat ctcagccacc gcgaaaatga
     4201 catcaaaaac gccattaacc tgatgttctg gggaatataa atgtcaggct cccttataca
     4261 cagccagtct gcaggtcgac catagtgact ggatatgttg tgttttacag tattatgtag
     4321 tctgtttttt atgcaaaatc taatttaata tattgatatt tatatcattt tacgtttctc
     4381 gttcagcttt cttgtacaaa gtggttgata tccagcacag tggcggccgc tcgagtctag
     4441 agggcccgcg gttcgaatac ccatacgacg tcccagacta cgcttagtaa tgattaatta
     4501 aactagaaat tctaccgggt aggggaggcg cttttcccaa ggcagtctgg agtaacccac
     4561 ccaagatctg gcctccgcgc cgggttttgg cgcctcccgc gggcgccccc ctcctcacgg
     4621 cgagccgcgt aagatctggc ctccgcgccg ggttttggcg cctcccgcgg gcgcccccct
     4681 cctcacggcg agcctagtcg tgaggctccg gtgcccgtca gtgggcagag cgcacatcgc
     4741 ccacagtccc cgagaagttg gggggagggg tcggcaattg aaccggtgcc tagagaaggt
     4801 ggcgcggggt aaactgggaa agtgatgtcg tgtactggct ccgccttttt cccgagggtg
     4861 ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg
     4921 ccgccagaac acaggtaagt gccgtgtgtg gttcccgcgg gcctggcctc tttacgggtt
     4981 atggcccttg cgtgccttga attacttcca cctggctgca gtacgtgatt cttgatcccg
     5041 agcttcgggt tggaagtggg tgggagagtt cgaggccttg cgcttaagga gccccttcgc
     5101 ctcgtgcttg agttgaggcc tggcctgggc gctggggccg ccgcgtgcga atctggtggc
     5161 accttcgcgc ctgtctcgct gctttcgata agtctctagc catttaaaat ttttgatgac
     5221 ctgctgcgac gctttttttc tggcaagata gtcttgtaaa tgcgggccaa gatctgcaca
     5281 ctggtatttc ggtttttggg gccgcgggcg gcgacggggc ccgtgcgtcc cagcgcacat
     5341 gttcggcgag gcggggcctg cgagcgcggc caccgagaat cggacggggg tagtctcaag
     5401 ctggccggcc tgctctggtg cctggcctcg cgccgccgtg tatcgccccg ccctgggcgg
     5461 caaggctggc ccggtcggca ccagttgcgt gagcggaaag atggccgctt cccggccctg
     5521 ctgcagggag ctcaaaatgg aggacgcggc gctcgggaga gcgggcgggt gagtcaccca
     5581 cacaaaggaa aagggccttt ccgtcctcag ccgtcgcttc atgtgactcc acggagtacc
     5641 gggcgccgtc caggcacctc gattagttct cgagcttttg gagtacgtcg tctttaggtt
     5701 ggggggaggg gttttatgcg atggagtttc cccacactga gtgggtggag actgaagtta
     5761 ggccagcttg gcacttgatg taattctcct tggaatttgc cctttttgag tttggatctt
     5821 ggttcattct caagcctcag acagtggttc aaagtttttt tcttccattt caggtgtcgt
     5881 gaagcggccg ccatgctagt aaattgtccg ctaaattctg gccgtttttg gcttttttgt
     5941 tagacggatc cataacttcg tataatgtat gctatacgaa gttatgccac catgtctaga
     6001 ctggacaaga gcaaagtcat aaacggagct ctggaattac tcaatggtgt cggtatcgaa
     6061 ggcctgacga caaggaaact cgctcaaaag ctgggagttg agcagcctac cctgtactgg
     6121 cacgtgaaga acaagcgggc cctgctcgat gccctgccaa tcgagatgct ggacaggcat
     6181 catacccact tctgccccct ggaaggcgag tcatggcaag actttctgcg gaacaacgcc
     6241 aagtcatacc gctgtgctct cctctcacat cgcgacgggg ctaaagtgca tctcggcacc
     6301 cgcccaacag agaaacagta cgaaaccctg gaaaatcagc tcgcgttcct gtgtcagcaa
     6361 ggcttctccc tggagaacgc actgtacgct ctgtccgccg tgggccactt tacactgggc
     6421 tgcgtattgg aggaacagga gcatcaagta gcaaaagagg aaagagagac acctaccacc
     6481 gattctatgc ccccacttct gagacaagca attgagctgt tcgaccggca gggagccgaa
     6541 cctgccttcc ttttcggcct ggaactaatc atatgtggcc tggagaaaca gctaaagtgc
     6601 gaaagcggcg ggccgaccga cgcccttgac gattttgact tagacatgct cccagccgat
     6661 gcccttgacg actttgacct tgatatgctg cctgctgacg ctcttgacga ttttgacctt
     6721 gacatgctcc ccgggtaact aagtaaggat ccgcggccgc actagaggaa ttccgcccct
     6781 ctcccccccc cccctctccc tccccccccc ctaacgttac tggccgaagc cgcttggaat
     6841 aaggccggtg tgcgtttgtc tatatgttat tttccaccat attgccgtct tttggcaatg
     6901 tgagggcccg gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc
     6961 tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct ctggaagctt
     7021 cttgaagaca aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg
     7081 acaggtgcct ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac
     7141 cccagtgcca cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg
     7201 tattcaacaa ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg
     7261 ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc
     7321 cgaaccacgg ggacgtggtt ttcctttgaa aaacacgatg ataagcttac cggtccacca
     7381 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
     7441 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg
     7501 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc
     7561 tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc
     7621 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
     7681 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
     7741 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
     7801 agctggagta caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg
     7861 gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg
     7921 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
     7981 acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc
     8041 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaag
     8101 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     8161 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     8221 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     8281 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     8341 tcgccttctt gacgagttct tctgatgtac aagtaaagcg gccgcgactc tagatcataa
     8401 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
     8461 tgaacctgaa acataaaatg aatgcaattg ttgttgttta gtccctccca attcgatatc
     8521 aagcttatcg cgatagatcc taatcaacct ctggattaca aaatttgtga aagattgact
     8581 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     8641 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     8701 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     8761 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     8821 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     8881 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca
     8941 tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc
     9001 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     9061 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
     9121 gcctccccgc ctgagatcct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc
     9181 actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga agacaagatc
     9241 tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg
     9301 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
     9361 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
     9421 tgtggaaaat ctctagcagt agtagttcat gtcatcttat tattcagtat ttataacttg
     9481 caaagaaatg aatatcagag agtgagaggc ccgggttaat taaggaaagg gctagatcat
     9541 tcttgaagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     9601 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     9661 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     9721 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     9781 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     9841 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     9901 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     9961 gttctgctat gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc
    10021 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
    10081 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
    10141 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
    10201 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
    10261 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
    10321 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
    10381 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
    10441 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
    10501 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
    10561 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
    10621 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
    10681 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
    10741 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
    10801 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
    10861 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
    10921 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
    10981 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
    11041 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
    11101 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
    11161 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
    11221 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
    11281 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
    11341 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
    11401 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
    11461 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
    11521 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
    11581 cgttggccga ttcattaatg cagcaagctc atggctgact aatttttttt atttatgcag
    11641 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
    11701 gcctaggctt ttgcaaaaag ctccccgtgg cacgacaggt ttcccgactg gaaagcgggc
    11761 agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac
    11821 tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga
    11881 aacagctatg acatgattac gaatttcaca aataaagcat ttttttcact gcattctagt
    11941 tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatcaactg gataactcaa
    12001 gctaaccaaa atcatcccaa acttcccacc ccatacccta ttaccactgc caattacctg
    12061 tggtttcatt tactctaaac ctgtgattcc tctgaattat tttcatttta aagaaattgt
    12121 atttgttaaa tatgtactac aaacttagta gt
//

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