pLVX-DD-AcGFP1-Actin

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pLVX-DD-AcGFP1-Actin载体质粒基本信息

出品公司: Clontech
载体名称: pLVX-DD-AcGFP1-Actin
质粒类型: 慢病毒载体;荧光报告载体
高拷贝/低拷贝: 高拷贝
克隆方法: 限制性内切酶,多克隆位点
启动子: CMV
载体大小: 10219 bp
5' 测序引物及序列: CMV Forward: CGCAAATGGGCGGTAGGCGTG 
3' 测序引物及序列: --
载体标签: DD-AcGFP (N-端)
载体抗性: 氨苄青霉素
筛选标记: 嘌呤霉素(Puromycin)
克隆菌株: Stbl3
宿主细胞(系): 常规细胞系(293、CV-1、CHO等)
备注: 慢病毒载体pLVX-DD-AcGFP1-Actin组成型表达N端DD-AcGFP1标记的肌动蛋白Actin;
DD标签是一个大小为12KD的FKBP (L106P)去稳定结构域,带有DD标签的目的蛋白将很快被降解;
向培养基中加入Shield1配体后,细胞中目的蛋白的量则迅速提高。
产品目录号: 631077
稳定性: 稳表达
组成型/诱导型: 组成型
病毒/非病毒: 慢病毒

pLVX-DD-AcGFP1-Actin质粒图谱载体图谱和pLVX-DD-AcGFP1-Actin载体序列质粒序列多克隆位点信息

pLVX-DD-AcGFP1-Actin载体图谱



pLVX-DD-AcGFP1-Actin 载体特征

pLVX-DD-AcGFP1-Actin质粒载体简介

pLVX-DD-AcGFP1-Actin is an HIV-1-based, lentiviral vector that expresses the ligand-dependent, destabilized fusion protein DD-AcGFP1-Actin. This protein is composed of human cytoplasmic b-actin fused to AcGFP1, a monomeric green fluorescent protein derived from Aequorea coerulescens (excitation and emission maxima: 475 nm and 505 nm, respectively; 1). The fusion protein also contains an N-terminal ProteoTuner destabilization domain (DD; 2), which—in the absence of the stabilizing ligand Shield1— causes rapid, proteasomal degradation of the fusion protein. When added to the culture medium, Shield1 binds to the destabilization domain, preventing degradation of DD-AcGFP1-Actin. The stabilized fusion protein can then be incorporated into growing actin filaments, allowing actin-containing subcellular structures to be visualized in living and fixed cells (3, 4).
pLVX-DD-AcGFP-Actin contains all of the viral processing elements necessary for the production of replication-incompetent lentivirus, as well as elements to improve viral titer, transgene expression, and overall vector function. The woodchuck hepatitis virus posttranscriptional regulatory element (WPRE) promotes RNA processing events and enhances nuclear export of viral and transgene RNA (5), leading to increased viral titers from packaging cells, and enhanced expression of your gene of interest in target cells. In addition, the vector includes a Rev-response element (RRE), which further increases viral titers by enhancing the transport of unspliced viral RNA out of the nucleus (6). Finally, pLVX-DD-AcGFP-Actin also contains a central polypurine tract/central termination sequence element (cPPT/CTS). During target cell infection, this element creates a central DNA flap that increases nuclear import of the viral genome, resulting in improved vector integration and more efficient transduction (7).
In addition to lentiviral elements, pLVX-DD-AcGFP-Actin contains a puromycin resistance gene (Puror) under the control of the murine phosphoglycerate kinase promoter (PPGK) for the selection of stable transductants. The vector also contains a pUC origin of replication and an E. coli ampicillin resistance gene (Ampr) for propagation and selection in bacteria.

pLVX-DD-AcGFP1-Actin, available as part of the Lenti-X Actin Dynamics Monitoring Kit (Cat. No. 631077), allows you to monitor how newly synthesized, Shield1-stabilized DD-AcGFP1-Actin is integrated into the dynamic actin filament network. pLVX-DD-AcGFP-Actin expresses a DD-AcGFP-Actin fusion protein when transduced into target cells; the fusion protein is degraded until Shield1 is added to the culture medium. The stabilized fusion protein is incorporated into actin filaments, allowing the visualization of actin-containing subcellular structures 15–20 minutes after the addition of Shield1 to the medium.
Before the vector can be transduced, it must be transfected into 293T packaging cells with our Lenti-X HT Packaging System (Cat. Nos. 632160 and 632161). This packaging system allows you to safely produce high titer, infectious, replication-incompetent, VSV-G pseudotyped lentiviral particles that can infect a wide range of cell types, including non-dividing and primary cells (8). If required, stable transfectants can be selected using puromycin.

Propagation in E. coli
 Recommended host strains: DH5α and other general purpose strains.
 Selectable marker: plasmid confers resistance to ampicillin (100 μg/ml) in E. coli hosts.
 E. coli replication origin: pUC
 Copy number: high

Excitation and emission maxima of AcGFP1
 Excitation maximum = 475 nm
 Emission maximum = 505 nm

References
1. Ponte, P. et al. (1984) Nucleic Acids Res. 12(3):1687–1696.
2. Banaszynski, L. et al. (2006) Cell 126(5):995–1004.
3. Westphal, M. et al. (1997) Curr. Biol. 7(3):176–183.
4. de Hostos, E. L., unpublished data.
5. Zufferey, R. et al. (1999) J. Virol. 73(4):2886–2892.
6. Cochrane, A. W. et al. (1990) Proc. Natl. Acad. Sci. USA 87(3):1198–1202.
7. Zennou, V. et al. (2000) Cell 101(2):173–185.
5. Wu, X. et al. (2000) Mol. Ther. 2(1):47–55.

pLVX-DD-AcGFP1-Actin质粒序列载体序列

LOCUS       pLVX-DD-AcGFP1-Actin  10222 bp    DNA   circular            21-DEC-2009
COMMENT     Created by Clontech Laboratories Inc. http://www.clontech.com
COMMENT     Cat. No. 631076,631077
FEATURES             Location/Qualifiers
     LTR             1..635
                     /label=5'\LTR
     misc_feature    685..822
                     /label=packaging\signal
     misc_feature    2028..2151
                     /label=cPPT/CTS\(DNA\flap)
     promoter        2185..2787
                     /label=CMV\IE\Promoter
     misc_feature    6190..6781
                     /label=WPRE
     rep_origin      complement(8090..8763)
                     /label=pUC\origin
     CDS             complement(8908..9904)
                     /label=Amp(R)
     misc_binding    1303..1536
                     /label=RRE
     misc_binding    636..653
                     /label=PBS
     misc_feature    2031..2046
                     /label=cPPT
     LTR             6984..7620
                     /label=LTR
     misc_feature    5048..5556
                     /label=PGK\promoter
     misc_feature    5577..6176
                     /label=Puro(R)
     CDS             2821..3144
                     /label=DD
     CDS             3175..3891
                     /label=AcGFP1
     CDS             3913..5037
                     /label=Actin
BASE COUNT     2504 a      2724 c      2709 g      2285 t 
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 tatcgccttt aaaagaaaag 
     2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac 
     2101 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg 
     2161 acagcagaga tccagtttat cgataagctt gggagttccg cgttacataa cttacggtaa 
     2221 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg 
     2281 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt 
     2341 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg 
     2401 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc 
     2461 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc 
     2521 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca 
     2581 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta 
     2641 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa 
     2701 gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc 
     2761 tccatagaag acaccgactc tactagagga tcgctagcgc taccggactc agatctcgag 
     2821 atgggagtgc aggtggaaac catctcccca ggagacgggc gcaccttccc caagcgcggc 
     2881 cagacctgtg tggtgcacta caccgggatg cttgaagatg gaaagaaagt cgattcctcc 
     2941 cgggacagaa acaagccctt taagtttatg ctaggcaagc aggaggtgat ccgaggctgg 
     3001 gaagaagggg ttgcccagat gagtgtgggt cagagagcca aactgactat atctccagat 
     3061 tatgcctatg gtgccactgg gcacccaggc atcatcccac cacatgccac tctcgtcttc 
     3121 gatgtggagc ttctaaaacc ggaagaattc tgcagtcgac ggtaccgcgg gcccatggtg 
     3181 agcaagggcg ccgagctgtt caccggcatc gtgcccatcc tgatcgagct gaatggcgat 
     3241 gtgaatggcc acaagttcag cgtgagcggc gagggcgagg gcgatgccac ctacggcaag 
     3301 ctgaccctga agttcatctg caccaccggc aagctgcctg tgccctggcc caccctggtg 
     3361 accaccctga gctacggcgt gcagtgcttc tcacgctacc ccgatcacat gaagcagcac 
     3421 gacttcttca agagcgccat gcctgagggc tacatccagg agcgcaccat cttcttcgag 
     3481 gatgacggca actacaagtc gcgcgccgag gtgaagttcg agggcgatac cctggtgaat 
     3541 cgcatcgagc tgaccggcac cgatttcaag gaggatggca acatcctggg caataagatg 
     3601 gagtacaact acaacgccca caatgtgtac atcatgaccg acaaggccaa gaatggcatc 
     3661 aaggtgaact tcaagatccg ccacaacatc gaggatggca gcgtgcagct ggccgaccac 
     3721 taccagcaga atacccccat cggcgatggc cctgtgctgc tgcccgataa ccactacctg 
     3781 tccacccaga gcgccctgtc caaggacccc aacgagaagc gcgatcacat gatctacttc 
     3841 ggcttcgtga ccgccgccgc catcacccac ggcatggatg agctgtacaa gtccggactc 
     3901 agatctcgag ctatggatga tgatatcgcc gcgctcgtcg tcgacaacgg ctccggcatg 
     3961 tgcaaggccg gcttcgcggg cgacgatgcc ccccgggccg tcttcccctc catcgtgggg 
     4021 cgccccaggc accagggcgt gatggtgggc atgggtcaga aggattccta tgtgggcgac 
     4081 gaggcccaga gcaagagagg catcctcacc ctgaagtacc ccatcgagca cggcatcgtc 
     4141 accaactggg acgacatgga gaaaatctgg caccacacct tctacaatga gctgcgtgtg 
     4201 gctcccgagg agcaccccgt gctgctgacc gaggcccccc tgaaccccaa ggccaaccgc 
     4261 gagaagatga cccagatcat gtttgagacc ttcaacaccc cagccatgta cgttgctatc 
     4321 caggctgtgc tatccctgta cgcctctggc cgtaccactg gcatcgtgat ggactccggt 
     4381 gacggggtca cccacactgt gcccatctac gaggggtatg ccctccccca tgccatcctg 
     4441 cgtctggacc tggctggccg ggacctgact gactacctca tgaagatcct caccgagcgc 
     4501 ggctacagct tcaccaccac ggccgagcgg gaaatcgtgc gtgacattaa ggagaagctg 
     4561 tgctacgtcg ccctggactt cgagcaagag atggccacgg ctgcttccag ctcctccctg 
     4621 gagaagagct acgagctgcc tgacggccag gtcatcacca ttggcaatga gcggttccgc 
     4681 tgccctgagg cactcttcca gccttccttc ctgggcatgg agtcctgtgg catccacgaa 
     4741 actaccttca actccatcat gaagtgtgac gtggacatcc gcaaagacct gtacgccaac 
     4801 acagtgctgt ctggcggcac caccatgtac cctggcattg ccgacaggat gcagaaggag 
     4861 atcactgccc tggcacccag cacaatgaag atcaagatca ttgctcctcc tgagcgcaag 
     4921 tactccgtgt ggatcggcgg ctccatcctg gcctcgctgt ccaccttcca gcagatgtgg 
     4981 atcagcaagc aggagtatga cgagtccggc ccctccatcg tccaccgcaa atgcttctag 
     5041 tctagataat tctaccgggt aggggaggcg cttttcccaa ggcagtctgg agcatgcgct 
     5101 ttagcagccc cgctgggcac ttggcgctac acaagtggcc tctggcctcg cacacattcc 
     5161 acatccaccg gtaggcgcca accggctccg ttctttggtg gccccttcgc gccaccttct 
     5221 actcctcccc tagtcaggaa gttccccccc gccccgcagc tcgcgtcgtg caggacgtga 
     5281 caaatggaag tagcacgtct cactagtctc gtgcagatgg acagcaccgc tgagcaatgg 
     5341 aagcgggtag gcctttgggg cagcggccaa tagcagcttt gctccttcgc tttctgggct 
     5401 cagaggctgg gaaggggtgg gtccgggggc gggctcaggg gcgggctcag gggcggggcg 
     5461 ggcgcccgaa ggtcctccgg aggcccggca ttctgcacgc ttcaaaagcg cacgtctgcc 
     5521 gcgctgttct cctcttcctc atctccgggc ctttcgacct gcagcccaag cttaccatga 
     5581 ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtccccagg gccgtacgca 
     5641 ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat ccggaccgcc 
     5701 acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg 
     5761 gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg 
     5821 tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc 
     5881 ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg 
     5941 cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg 
     6001 ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga 
     6061 cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg 
     6121 tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctgaccgc 
     6181 gtctggaaca atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac 
     6241 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt 
     6301 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat 
     6361 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca 
     6421 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc 
     6481 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg 
     6541 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca 
     6601 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct 
     6661 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt 
     6721 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct 
     6781 ggaattaatt ctgcagtcga gacctagaaa aacatggagc aatcacaagt agcaatacag 
     6841 cagctaccaa tgctgattgt gcctggctag aagcacaaga ggaggaggag gtgggttttc 
     6901 cagtcacacc tcaggtacct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc 
     6961 actttttaaa agaaaagagg ggactggaag ggctaattca ctcccaacga agacaagata 
     7021 tccttgatct gtggatctac cacacacaag gctacttccc tgattagcag aactacacac 
     7081 cagggccagg ggtcagatat ccactgacct ttggatggtg ctacaagcta gtaccagttg 
     7141 agccagataa ggtagaagag gccaataaag gagagaacac cagcttgtta caccctgtga 
     7201 gcctgcatgg gatggatgac ccggagagag aagtgttaga gtggaggttt gacagccgcc 
     7261 tagcatttca tcacgtggcc cgagagctgc atccggagta cttcaagaac tgctgatatc 
     7321 gagcttgcta caagggactt tccgctgggg actttccagg gaggcgtggc ctgggcggga 
     7381 ctggggagtg gcgagccctc agatcctgca tataagcagc tgctttttgc ctgtactggg 
     7441 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg 
     7501 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt 
     7561 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt 
     7621 agtagttcat gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag 
     7681 agtgagaggc cttgacattg ctagcgtttt accgtcgacc tctagctaga gcttggcgta 
     7741 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat 
     7801 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt 
     7861 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta 
     7921 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc 
     7981 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa 
     8041 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa 
     8101 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct 
     8161 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac 
     8221 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc 
     8281 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc 
     8341 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg 
     8401 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga 
     8461 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag 
     8521 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta 
     8581 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag 
     8641 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg 
     8701 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac 
     8761 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc 
     8821 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag 
     8881 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc 
     8941 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac 
     9001 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc 
     9061 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg 
     9121 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag 
     9181 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc 
     9241 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac 
     9301 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag 
     9361 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac 
     9421 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg 
     9481 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc 
     9541 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact 
     9601 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg 
     9661 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa 
     9721 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt 
     9781 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg 
     9841 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga 
     9901 cgtcgacgga tcgggagatc aacttgttta ttgcagctta taatggttac aaataaagca 
     9961 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt 
    10021 ccaaactcat caatgtatct tatcatgtct ggatcaactg gataactcaa gctaaccaaa 
    10081 atcatcccaa acttcccacc ccatacccta ttaccactgc caattacctg tggtttcatt 
    10141 tactctaaac ctgtgattcc tctgaattat tttcatttta aagaaattgt atttgttaaa 
    10201 tatgtactac aaacttagta gt 
//

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