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 |
买家导航
pLVX-DD-AcGFP1-Actin载体质粒基本信息
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 //