| 出品公司: | Clontech |
|---|---|
| 载体名称: | pRetroX-Tight-Pur |
| 质粒类型: | 逆病毒载体；四环素调控载体 |
| 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: | Tight |
| 载体大小: | 6568 bp |
| 5 测序引物及序列: | -- |
| 3 测序引物及序列: | -- |
| 载体标签: | -- |
| 载体抗性: | 氨苄青霉素 |
| 筛选标记: | 嘌呤霉素（Puromycin） |
| 克隆菌株: | DH5α |
| 宿主细胞（系）: | 常规细胞系（293、CV-1、CHO等） |
| 备注: | 逆病毒载体pRetroX-Tight-Pur是四环素调控系统载体 |
| 产品目录号: | 632104 |
| 稳定性: | 稳表达 |
| 组成型/诱导型: | 诱导型 |
| 病毒/非病毒: | 逆转录病毒 |
pRetroX-Tight-Pur逆病毒载体；四环素调控载体
产品名称： pRetroX-Tight-Pur逆病毒载体；四环素调控载体
英文名称： pRetroX-Tight-Pur逆病毒载体；四环素调控载体
产品编号： BioVector-pRetroX-Ti
产品价格： 0
产品产地： USA
品牌商标： BioVector NTCC Inc.
更新时间： null
使用范围： null
来福生物技术有限公司
- 联系人 :
- 地址 : 北京海淀区学院路
- 邮编 : 100860
- 所在区域 : 北京
- 电话 : 400-800-2947 点击查看
- 传真 : 点击查看
- 邮箱 : ntccbio@163.com
载体基本信息
载体质粒图谱和多克隆位点信息
载体简介
pRetroX-Tight-Pur载体描述
pRetroX-Tight-Pur is an inducible, self-inactivating, retroviral expression vector designed to express a gene of interest under the control of PTight, a modified Tet-responsive promoter. PTight consists of seven direct repeats of a 36 bp regulatory sequence that contains the 19 bp tet operator sequence (tetO), and a modified minimal CMV promoter (1). This vector is supplied with our Retro-XTM Tet-On Advanced and Tet-Off Advanced Inducible Expression Systems (Cat. Nos. 632104 and 632105). These systems provide ready access to the inducible gene expression strategy of Gossen & Bujard, and incorporate major improvements described by Urlinger, et al. (2-6).
pRetroX-Tight-Pur is optimized to eliminate promoter interference through LTR self-inactivation. The hybrid 5’ LTR consists of the cytomegalovirus (CMV) type I enhancer and the mouse sarcoma virus (MSV) promoter. This promoter drives high levels of viral genome transcription in HEK 293-based packaging cell lines due, in part, to the presence of adenoviral E1A (7-8) in these cells. The self-inactivating feature of the vector is provided by a deletion in the 3’ LTR enhancer region (U3). During reverse transcription of the retroviral RNA, a copy of the inactivated 3’ LTR U3 region replaces the corresponding region of the 5’ LTR, resulting in the inactivation of the 5’ LTR CMV enhancer sequence. This mechanism can reduce the phenomenon known as promoter interference (9-10) and allow more efficient expression. In this way, pRetroX-Tight-Pur supports high viral titers, yet eliminates potential downstream interference between the inducible PTight promoter and its adjacent viral LTR.
pRetroX-Tight-Pur contains all of the necessary viral RNA processing elements; these include the 5’ and 3’ LTRs, the packaging signal (Ψ+), and the tRNA primer binding site. For safety reasons, however, the vector lacks the structural genes (gag, pol, and env) necessary for retroviral particle formation and replication. pRetroX-Tight-Pur contains a puromycin resistance gene (Purr) under the control of the murine phosphoglycerate kinase (PGK) promoter (PPGK) for the selection of stable transfectants. In addition, the vector contains a ColE1 origin of replication and an E. coli Ampr gene for propagation and selection in bacteria.
pRetroX-Tight-Pur can be used as either a plasmid or retroviral expression vector. When used as a retroviral expression vector, it must be transfected into a packaging cell line, such as GP2-293; we recommend the Retro-X Universal Packaging System (Cat. No. 631530). Packaging cell lines allow you to produce infectious, replication-incompetent retroviral particles. These retroviral particles can infect a wide range of target cells and transmit your gene of interest, but they cannot replicate within these cells due to the absence of viral structural genes. The separate introduction and integration of the structural genes into the packaging cell line minimizes the chance of producing replication-competent virus due to recombination events during cell proliferation.
Propagation in E. coli
Suitable host strains: DH5α, DH10B and other general purpose strains.
Selectable marker: plasmid confers resistance to ampicillin (100 μg/ml) in E. coli hosts.
E. coli replication origin: ColE1
Copy number: high
Notes:
The vector sequence was compiled from information in the sequence databases, published literature, and other sources, together with partial sequences obtained by Clontech. This vector has not been completely sequenced.
The viral supernatants produced by this retroviral vector could contain potentially hazardous recombinant virus. Due caution must be exercised in the production and handling of recombinant retrovirus. Appropriate NIH, regional, and institutional guidelines apply.
载体序列
LOCUS pRetroX-Tight-Pur 6567 bp DNA SYN
DEFINITION pRetroX-Tight-Pur
ACCESSION
KEYWORDS
SOURCE
ORGANISM other sequences; artificial sequences; vectors.
FEATURES Location/Qualifiers
source 1..6567
/organism="pRetroX-Tight-Pur"
/mol_type="other DNA"
promoter 2..506
/label="CMV_immearly_promoter"
misc_feature 6..688
/label="5_LTR"
misc_feature 9..296
/label="CAG_enhancer"
misc_feature 463..483
/label="CMV_fwd_primer"
misc_feature 758..1567
/label="psi_plus_pack"
misc_feature 1154..1561
/label="gag"
misc_feature 1497..1519
/label="MSCV_primer"
misc_feature 1497..1519
/label="pLXSN_5_primer"
misc_feature 1535..1551
/label="pBABE_5_primer"
CDS 1669..2196
/label="ORF frame 1"
/translation="MSSLLPISDRERMSSLLPISDRERMSSLLPISDRERMSSLSLSV
IENVCRVYSLSVIENVCRGRRVRWEAYISRARLVNRQIAWRRIRGRAGSRSRTREFYR
VGEALFPRQSGACALAAPLGTWRYTSGLWPRTHSTSTGRRQPAPFFGGPFAPPSTPPL
VRKFPPAPQLASCRT*"
misc_feature 1888..1907
/label="pCEP_fwd_primer"
/translation="MSSLLPISDRERMSSLLPISDRERMSSLLPISDRERMSSLSLSV
IENVCRVYSLSVIENVCRGRRVRWEAYISRARLVNRQIAWRRIRGRAGSRSRTREFYR
VGEALFPRQSGACALAAPLGTWRYTSGLWPRTHSTSTGRRQPAPFFGGPFAPPSTPPL
VRKFPPAPQLASCRT*"
misc_feature 1890..1914
/label="LNCX_primer"
/translation="MSSLLPISDRERMSSLLPISDRERMSSLLPISDRERMSSLSLSV
IENVCRVYSLSVIENVCRGRRVRWEAYISRARLVNRQIAWRRIRGRAGSRSRTREFYR
VGEALFPRQSGACALAAPLGTWRYTSGLWPRTHSTSTGRRQPAPFFGGPFAPPSTPPL
VRKFPPAPQLASCRT*"
misc_feature complement(2008..2031)
/label="MSCV_rev_primer"
/translation="MSSLLPISDRERMSSLLPISDRERMSSLLPISDRERMSSLSLSV
IENVCRVYSLSVIENVCRGRRVRWEAYISRARLVNRQIAWRRIRGRAGSRSRTREFYR
VGEALFPRQSGACALAAPLGTWRYTSGLWPRTHSTSTGRRQPAPFFGGPFAPPSTPPL
VRKFPPAPQLASCRT*"
CDS 2253..3092
/label="ORF frame 3"
/translation="MEAGRPLGQRPIAALLLRFLGSEAGKGWVRGRAQGRAQGRGGRP
KVLRRPGILHASKAHVCRAVLLFLISGPFDLQPKLTMTEYKPTVRLATRDDVPRAVRT
LAAAFADYPATRHTVDPDRHIERVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPE
SVEAGAVFAEIGPRMAELSGSRLAAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKG
LGSAVVLPGVEAAERAGVPAFLETSAPRNLPFYERLGFTVTADVEVPEGPRTWCMTRK
PGA*"
misc_feature 2405..2424
/label="mPGK_F_primer"
/translation="MEAGRPLGQRPIAALLLRFLGSEAGKGWVRGRAQGRAQGRGGRP
KVLRRPGILHASKAHVCRAVLLFLISGPFDLQPKLTMTEYKPTVRLATRDDVPRAVRT
LAAAFADYPATRHTVDPDRHIERVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPE
SVEAGAVFAEIGPRMAELSGSRLAAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKG
LGSAVVLPGVEAAERAGVPAFLETSAPRNLPFYERLGFTVTADVEVPEGPRTWCMTRK
PGA*"
CDS complement(2418..3143)
/label="ORF frame 2"
/translation="MGSCAPFGRALRVVGRASGTGLAGHAPGARSFGHLDVGGDGEAE
PLVEGEVAGRGGLQEGGHPGALGRLHSGEHDGAAQTLALVVGRDADGGQEPRGLLGPV
RRQEAFHLLLRGQPGTAQLGHARADLGEHRPRFDALRRGPDRHRGAVVRDPHLADVEP
DAREEEFLQLGDPLDVAVRIDGVARGGVVGERGGEGAYGPGDVVAGGEAHRGLVLGHG
KLGLQVERPGDEEEENSAADVRF*"
gene 2493..3092
/label="puro (variant)"
/gene="puro (variant)"
/translation="MGSCAPFGRALRVVGRASGTGLAGHAPGARSFGHLDVGGDGEAE
PLVEGEVAGRGGLQEGGHPGALGRLHSGEHDGAAQTLALVVGRDADGGQEPRGLLGPV
RRQEAFHLLLRGQPGTAQLGHARADLGEHRPRFDALRRGPDRHRGAVVRDPHLADVEP
DAREEEFLQLGDPLDVAVRIDGVARGGVVGERGGEGAYGPGDVVAGGEAHRGLVLGHG
KLGLQVERPGDEEEENSAADVRF*"
misc_feature complement(3801..3823)
/label="pGEX_3_primer"
/translation="MGSCAPFGRALRVVGRASGTGLAGHAPGARSFGHLDVGGDGEAE
PLVEGEVAGRGGLQEGGHPGALGRLHSGEHDGAAQTLALVVGRDADGGQEPRGLLGPV
RRQEAFHLLLRGQPGTAQLGHARADLGEHRPRFDALRRGPDRHRGAVVRDPHLADVEP
DAREEEFLQLGDPLDVAVRIDGVARGGVVGERGGEGAYGPGDVVAGGEAHRGLVLGHG
KLGLQVERPGDEEEENSAADVRF*"
misc_feature complement(3959..3979)
/label="pBABE_3_primer"
/translation="MGSCAPFGRALRVVGRASGTGLAGHAPGARSFGHLDVGGDGEAE
PLVEGEVAGRGGLQEGGHPGALGRLHSGEHDGAAQTLALVVGRDADGGQEPRGLLGPV
RRQEAFHLLLRGQPGTAQLGHARADLGEHRPRFDALRRGPDRHRGAVVRDPHLADVEP
DAREEEFLQLGDPLDVAVRIDGVARGGVVGERGGEGAYGPGDVVAGGEAHRGLVLGHG
KLGLQVERPGDEEEENSAADVRF*"
misc_feature complement(3965..4180)
/label="SV40_enhancer"
/translation="MGSCAPFGRALRVVGRASGTGLAGHAPGARSFGHLDVGGDGEAE
PLVEGEVAGRGGLQEGGHPGALGRLHSGEHDGAAQTLALVVGRDADGGQEPRGLLGPV
RRQEAFHLLLRGQPGTAQLGHARADLGEHRPRFDALRRGPDRHRGAVVRDPHLADVEP
DAREEEFLQLGDPLDVAVRIDGVARGGVVGERGGEGAYGPGDVVAGGEAHRGLVLGHG
KLGLQVERPGDEEEENSAADVRF*"
promoter 3977..4245
/label="SV40_promoter"
/translation="MGSCAPFGRALRVVGRASGTGLAGHAPGARSFGHLDVGGDGEAE
PLVEGEVAGRGGLQEGGHPGALGRLHSGEHDGAAQTLALVVGRDADGGQEPRGLLGPV
RRQEAFHLLLRGQPGTAQLGHARADLGEHRPRFDALRRGPDRHRGAVVRDPHLADVEP
DAREEEFLQLGDPLDVAVRIDGVARGGVVGERGGEGAYGPGDVVAGGEAHRGLVLGHG
KLGLQVERPGDEEEENSAADVRF*"
rep_origin 4144..4221
/label="SV40_origin"
/translation="MGSCAPFGRALRVVGRASGTGLAGHAPGARSFGHLDVGGDGEAE
PLVEGEVAGRGGLQEGGHPGALGRLHSGEHDGAAQTLALVVGRDADGGQEPRGLLGPV
RRQEAFHLLLRGQPGTAQLGHARADLGEHRPRFDALRRGPDRHRGAVVRDPHLADVEP
DAREEEFLQLGDPLDVAVRIDGVARGGVVGERGGEGAYGPGDVVAGGEAHRGLVLGHG
KLGLQVERPGDEEEENSAADVRF*"
misc_feature 4206..4225
/label="SV40pro_F_primer"
/translation="MGSCAPFGRALRVVGRASGTGLAGHAPGARSFGHLDVGGDGEAE
PLVEGEVAGRGGLQEGGHPGALGRLHSGEHDGAAQTLALVVGRDADGGQEPRGLLGPV
RRQEAFHLLLRGQPGTAQLGHARADLGEHRPRFDALRRGPDRHRGAVVRDPHLADVEP
DAREEEFLQLGDPLDVAVRIDGVARGGVVGERGGEGAYGPGDVVAGGEAHRGLVLGHG
KLGLQVERPGDEEEENSAADVRF*"
gene complement(5345..6205)
/label="Ampicillin"
/gene="Ampicillin"
/translation="MGSCAPFGRALRVVGRASGTGLAGHAPGARSFGHLDVGGDGEAE
PLVEGEVAGRGGLQEGGHPGALGRLHSGEHDGAAQTLALVVGRDADGGQEPRGLLGPV
RRQEAFHLLLRGQPGTAQLGHARADLGEHRPRFDALRRGPDRHRGAVVRDPHLADVEP
DAREEEFLQLGDPLDVAVRIDGVARGGVVGERGGEGAYGPGDVVAGGEAHRGLVLGHG
KLGLQVERPGDEEEENSAADVRF*"
CDS complement(5345..6205)
/label="ORF frame 3"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE
YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
DRWEPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
EIGASLIKHW*"
promoter complement(6247..6275)
/label="AmpR_promoter"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE
YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
DRWEPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
EIGASLIKHW*"
promoter 6497..506
/label="CMV_immearly_promoter"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE
YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
DRWEPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
EIGASLIKHW*"
misc_feature 6561..688
/label="5_LTR"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE
YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
DRWEPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
EIGASLIKHW*"
ORIGIN
1 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
61 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
121 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
181 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
241 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
301 ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
361 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
421 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
481 gtgtacggtg ggaggtctat ataagcagag ctcaataaaa gagcccacaa cccctcactc
541 ggcgcgccag tcttccgata gactgcgtcg cccgggtacc cgtattccca ataaagcctc
601 ttgctgtttg catccgaatc gtggtctcgc tgttccttgg gagggtctcc tctgagtgat
661 tgactaccca cgacgggggt ctttcatttg ggggctcgtc cgggatttgg agacccctgc
721 ccagggacca ccgacccacc accgggaggt aagctggcca gcaacttatc tgtgtctgtc
781 cgattgtcta gtgtctatgt ttgatgttat gcgcctgcgt ctgtactagt tagctaacta
841 gctctgtatc tggcggaccc gtggtggaac tgacgagttc tgaacacccg gccgcaaccc
901 tgggagacgt cccagggact ttgggggccg tttttgtggc ccgacctgag gaagggagtc
961 gatgtggaat ccgaccccgt caggatatgt ggttctggta ggagacgaga acctaaaaca
1021 gttcccgcct ccgtctgaat ttttgctttc ggtttggaac cgaagccgcg cgtcttgtct
1081 gctgcagcgc tgcagcatcg ttctgtgttg tctctgtctg actgtgtttc tgtatttgtc
1141 tgaaaattag ggccagactg ttaccactcc cttaagtttg accttaggtc actggaaaga
1201 tgtcgagcgg atcgctcaca accagtcggt agatgtcaag aagagacgtt gggttacctt
1261 ctgctctgca gaatggccaa cctttaacgt cggatggccg cgagacggca cctttaaccg
1321 agacctcatc acccaggtta agatcaaggt cttttcacct ggcccgcatg gacacccaga
1381 ccaggtcccc tacatcgtga cctgggaagc cttggctttt gacccccctc cctgggtcaa
1441 gccctttgta caccctaagc ctccgcctcc tcttcctcca tccgccccgt ctctccccct
1501 tgaacctcct cgttcgaccc cgcctcgatc ctccctttat ccagccctca ctccttctct
1561 aggcgccgga attgaagatc tgaggccctt tcgtcttcac tcgagtttac tccctatcag
1621 tgatagagaa cgtatgtcga gtttactccc tatcagtgat agagaacgat gtcgagttta
1681 ctccctatca gtgatagaga acgtatgtcg agtttactcc ctatcagtga tagagaacgt
1741 atgtcgagtt tactccctat cagtgataga gaacgtatgt cgagtttatc cctatcagtg
1801 atagagaacg tatgtcgagt ttactcccta tcagtgatag agaacgtatg tcgaggtagg
1861 cgtgtacggt gggaggccta tataagcaga gctcgtttag tgaaccgtca gatcgcctgg
1921 agaaggatcc gcggccgcgc cggctctaga tcgcgaacgc gtgaattcta ccgggtaggg
1981 gaggcgcttt tcccaaggca gtctggagca tgcgctttag cagccccgct gggcacttgg
2041 cgctacacaa gtggcctctg gcctcgcaca cattccacat ccaccggtag gcgccaaccg
2101 gctccgttct ttggtggccc cttcgcgcca ccttctactc ctcccctagt caggaagttc
2161 ccccccgccc cgcagctcgc gtcgtgcagg acgtgacaaa tggaagtagc acgtctcact
2221 agtctcgtgc agatggacag caccgctgag caatggaagc gggtaggcct ttggggcagc
2281 ggccaatagc agctttgctc cttcgctttc tgggctcaga ggctgggaag gggtgggtcc
2341 gggggcgggc tcaggggcgg gctcaggggc ggggcgggcg cccgaaggtc ctccggaggc
2401 ccggcattct gcacgcttca aaagcgcacg tctgccgcgc tgttctcctc ttcctcatct
2461 ccgggccttt cgacctgcag cccaagctta ccatgaccga gtacaagccc acggtgcgcc
2521 tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg ttcgccgact
2581 accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc accgagctgc
2641 aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg
2701 gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg
2761 agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg
2821 aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg
2881 tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg
2941 cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct
3001 tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca
3061 cctggtgcat gacccgcaag cccggtgcct gacgcccgcc ccacgacccg cagcgcccga
3121 ccgaaaggag cgcacgaccc catgcatcgg cgtctcgaga tatcagtggt ccaggctcta
3181 gttttgactc aacaatatca ccagctgaag cctatagagt acgagccata gataaaataa
3241 aagattttat ttagtctcca gaaaaagggg ggaatgaaag accccacctg taggtttggc
3301 aagctagctt aagtaacgcc attttgcaag gcatggaaaa atacataact gagaatagag
3361 aagttcagat caaggtcagg aacagatgga acagggtcga ccctagagaa ccatcagatg
3421 tttccagggt gccccaagga cctgaaatga ccctgtgcct tatttgaact aaccaatcag
3481 ttcgcttctc gcttctgttc gcgcgcttct gctccccgag ctcaataaaa gagcccacaa
3541 cccctcactc ggggcgccag tcctccgatt gactgagtcg cccgggtacc cgtgtatcca
3601 ataaaccctc ttgcagttgc atccgacttg tggtctcgct gttccttggg agggtctcct
3661 ctgagtgatt gactacccgt cagcgggggt ctttcatttg ggggctcgtc cgggatcggg
3721 agacccctgc ccagggacca ccgacccacc accgggaggt aagctggctg cctcgcgcgt
3781 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt
3841 ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg
3901 tgtcggggcg cagccatgac ccagtcacgt agcgatagcg gagtgtagat ccggctgtgg
3961 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
4021 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
4081 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
4141 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
4201 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
4261 ggaggctttt ttggaggcct aggcttttgc aaaaagctta ctggcttaac tatgcggcat
4321 cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa
4381 ggagaaaata ccgcatcagg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
4441 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
4501 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
4561 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
4621 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
4681 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
4741 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
4801 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
4861 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
4921 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
4981 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
5041 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
5101 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
5161 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
5221 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
5281 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
5341 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
5401 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
5461 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
5521 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
5581 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
5641 gcaacgttgt tgccattgct gcaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
5701 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
5761 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
5821 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
5881 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
5941 gttgctcttg cccggcgtca acacgggata ataccgcgcc acatagcaga actttaaaag
6001 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
6061 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
6121 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
6181 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
6241 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
6301 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
6361 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtcttcaa gaattagctt
6421 ggccattgca tacgttgtat ccatatcata atatgtacat ttatattggc tcatgtccaa
6481 cattaccgcc atgttgacat tgattattga ctagttatta atagtaatca attacggggt
6541 cattagttca tagcccatat atggagt
//