| 出品公司: | Clontech |
|---|---|
| 载体名称: | pRetroX-Tight-Hyg |
| 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: | 7419 bp |
| 5' 测序引物及序列: | 5’-ATCTGAGGCCCTTTCGTCTTCACT-3’ |
| 3' 测序引物及序列: | -- |
| 载体标签: | 无 |
| 载体抗性: | 氨苄青霉素 |
| 筛选标记: | 潮霉素（Hygromycin） |
| 克隆菌株: | DH5α |
| 宿主细胞（系）: | 常规细胞系（293、CV-1、CHO等） |
| 备注: | 自我失活型逆病毒载体pRetroX-Tight-Hyg是四环素诱导载体 |
| 产品目录号: | 631034 |
| 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 |
买家导航
pRetroX-Tight-Hyg载体质粒基本信息
pRetroX-Tight-Hyg质粒图谱载体图谱和pRetroX-Tight-Hyg载体序列质粒序列多克隆位点信息
pRetroX-Tight-Hyg质粒载体简介
pRetroX-Tight-Hyg载体描述
pRetroX-Tight-Hyg is an inducible, self-inactivating, retroviral expression vector designed to express a gene of interest under the control of PTight, a modifi ed 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 modifi ed minimal CMV promoter (1). This vector is designed for use 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-Hyg is optimized to eliminate promoter interference through LTR selfinactivation. 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 effi cient expression. In this way, pRetroX-Tight-Hyg supports high viral titers, yet eliminates potential downstream interference between the inducible PTight promoter and its adjacent viral LTR.
pRetroX-Tight-Hyg 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-Hyg contains a hygromycin resistance gene (Hygr) 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-Hyg 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 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 attached sequence fi le has been 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 sequence.
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.
pRetroX-Tight-Hyg质粒序列载体序列
LOCUS pRetroX-Tight-Hyg 7419 bp DNA SYN
DEFINITION pRetroX-Tight-Hyg
ACCESSION
KEYWORDS
SOURCE
ORGANISM other sequences; artificial sequences; vectors.
FEATURES Location/Qualifiers
source 1..7419
/organism="pRetroX-Tight-Hyg"
/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..3515
/label="ORF frame 3"
/translation="MEAGRPLGQRPIAALLLRFLGSEAGKGWVRGRAQGRAQGRGGRP
KVLRRPGILHASKAHVCRAVLLFLISGPFDLHPATMKKPELTATSVEKFLIEKFDSVS
DLMQLSEGEESRAFSFDVGGRGYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIG
EFSESLTYCISRRAQGVTLQDLPETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQ
GIGQYTTWRDFICAIADPHVYHWQTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHA
DFGSNNVLTDNGRITAVIDWSEAMFGDSQYEVANIFFWRPWLACMEQQTRYFERRHPE
LAGSPRLRAYMLRIGLDQLYQSLVDGNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRS
AAVWTDGCVEVLADSGNRRPSTRPRAKE*"
misc_feature 2405..2424
/label="mPGK_F_primer"
/translation="MEAGRPLGQRPIAALLLRFLGSEAGKGWVRGRAQGRAQGRGGRP
KVLRRPGILHASKAHVCRAVLLFLISGPFDLHPATMKKPELTATSVEKFLIEKFDSVS
DLMQLSEGEESRAFSFDVGGRGYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIG
EFSESLTYCISRRAQGVTLQDLPETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQ
GIGQYTTWRDFICAIADPHVYHWQTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHA
DFGSNNVLTDNGRITAVIDWSEAMFGDSQYEVANIFFWRPWLACMEQQTRYFERRHPE
LAGSPRLRAYMLRIGLDQLYQSLVDGNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRS
AAVWTDGCVEVLADSGNRRPSTRPRAKE*"
CDS complement(2418..3248)
/label="ORF frame 2"
/translation="MPPLEVARLLLHTSQPRPPEEDVGDLVLGIPEHRLAPVNDRCYA
AIVRQDIVGAEIRVHEVPDFGAVLGPKHQLIESLRDGRTDGVVHHSLPVIHMGISNRA
YEITPCSVLTDSLRSEWAEPARLAKIGRSDRIHGLRDRLQNSGQFGFRQVLQRDTLCT
AGDAIGQALAELPNVKHFRNRERGRCKVPINITIFVETIGAAIYPQDISTPSYIEAES
TRFFALRELHQVGDAVELFDQKLLDRRRGEFRLFHGGGMQVERPGDEEEENSAADVRF
*"
gene 2505..3512
/label="hygroB"
/gene="hygroB"
/translation="MPPLEVARLLLHTSQPRPPEEDVGDLVLGIPEHRLAPVNDRCYA
AIVRQDIVGAEIRVHEVPDFGAVLGPKHQLIESLRDGRTDGVVHHSLPVIHMGISNRA
YEITPCSVLTDSLRSEWAEPARLAKIGRSDRIHGLRDRLQNSGQFGFRQVLQRDTLCT
AGDAIGQALAELPNVKHFRNRERGRCKVPINITIFVETIGAAIYPQDISTPSYIEAES
TRFFALRELHQVGDAVELFDQKLLDRRRGEFRLFHGGGMQVERPGDEEEENSAADVRF
*"
misc_feature complement(4653..4675)
/label="pGEX_3_primer"
/translation="MPPLEVARLLLHTSQPRPPEEDVGDLVLGIPEHRLAPVNDRCYA
AIVRQDIVGAEIRVHEVPDFGAVLGPKHQLIESLRDGRTDGVVHHSLPVIHMGISNRA
YEITPCSVLTDSLRSEWAEPARLAKIGRSDRIHGLRDRLQNSGQFGFRQVLQRDTLCT
AGDAIGQALAELPNVKHFRNRERGRCKVPINITIFVETIGAAIYPQDISTPSYIEAES
TRFFALRELHQVGDAVELFDQKLLDRRRGEFRLFHGGGMQVERPGDEEEENSAADVRF
*"
misc_feature complement(4811..4831)
/label="pBABE_3_primer"
/translation="MPPLEVARLLLHTSQPRPPEEDVGDLVLGIPEHRLAPVNDRCYA
AIVRQDIVGAEIRVHEVPDFGAVLGPKHQLIESLRDGRTDGVVHHSLPVIHMGISNRA
YEITPCSVLTDSLRSEWAEPARLAKIGRSDRIHGLRDRLQNSGQFGFRQVLQRDTLCT
AGDAIGQALAELPNVKHFRNRERGRCKVPINITIFVETIGAAIYPQDISTPSYIEAES
TRFFALRELHQVGDAVELFDQKLLDRRRGEFRLFHGGGMQVERPGDEEEENSAADVRF
*"
misc_feature complement(4817..5032)
/label="SV40_enhancer"
/translation="MPPLEVARLLLHTSQPRPPEEDVGDLVLGIPEHRLAPVNDRCYA
AIVRQDIVGAEIRVHEVPDFGAVLGPKHQLIESLRDGRTDGVVHHSLPVIHMGISNRA
YEITPCSVLTDSLRSEWAEPARLAKIGRSDRIHGLRDRLQNSGQFGFRQVLQRDTLCT
AGDAIGQALAELPNVKHFRNRERGRCKVPINITIFVETIGAAIYPQDISTPSYIEAES
TRFFALRELHQVGDAVELFDQKLLDRRRGEFRLFHGGGMQVERPGDEEEENSAADVRF
*"
promoter 4829..5097
/label="SV40_promoter"
/translation="MPPLEVARLLLHTSQPRPPEEDVGDLVLGIPEHRLAPVNDRCYA
AIVRQDIVGAEIRVHEVPDFGAVLGPKHQLIESLRDGRTDGVVHHSLPVIHMGISNRA
YEITPCSVLTDSLRSEWAEPARLAKIGRSDRIHGLRDRLQNSGQFGFRQVLQRDTLCT
AGDAIGQALAELPNVKHFRNRERGRCKVPINITIFVETIGAAIYPQDISTPSYIEAES
TRFFALRELHQVGDAVELFDQKLLDRRRGEFRLFHGGGMQVERPGDEEEENSAADVRF
*"
rep_origin 4996..5073
/label="SV40_origin"
/translation="MPPLEVARLLLHTSQPRPPEEDVGDLVLGIPEHRLAPVNDRCYA
AIVRQDIVGAEIRVHEVPDFGAVLGPKHQLIESLRDGRTDGVVHHSLPVIHMGISNRA
YEITPCSVLTDSLRSEWAEPARLAKIGRSDRIHGLRDRLQNSGQFGFRQVLQRDTLCT
AGDAIGQALAELPNVKHFRNRERGRCKVPINITIFVETIGAAIYPQDISTPSYIEAES
TRFFALRELHQVGDAVELFDQKLLDRRRGEFRLFHGGGMQVERPGDEEEENSAADVRF
*"
misc_feature 5058..5077
/label="SV40pro_F_primer"
/translation="MPPLEVARLLLHTSQPRPPEEDVGDLVLGIPEHRLAPVNDRCYA
AIVRQDIVGAEIRVHEVPDFGAVLGPKHQLIESLRDGRTDGVVHHSLPVIHMGISNRA
YEITPCSVLTDSLRSEWAEPARLAKIGRSDRIHGLRDRLQNSGQFGFRQVLQRDTLCT
AGDAIGQALAELPNVKHFRNRERGRCKVPINITIFVETIGAAIYPQDISTPSYIEAES
TRFFALRELHQVGDAVELFDQKLLDRRRGEFRLFHGGGMQVERPGDEEEENSAADVRF
*"
gene complement(6197..7057)
/label="Ampicillin"
/gene="Ampicillin"
/translation="MPPLEVARLLLHTSQPRPPEEDVGDLVLGIPEHRLAPVNDRCYA
AIVRQDIVGAEIRVHEVPDFGAVLGPKHQLIESLRDGRTDGVVHHSLPVIHMGISNRA
YEITPCSVLTDSLRSEWAEPARLAKIGRSDRIHGLRDRLQNSGQFGFRQVLQRDTLCT
AGDAIGQALAELPNVKHFRNRERGRCKVPINITIFVETIGAAIYPQDISTPSYIEAES
TRFFALRELHQVGDAVELFDQKLLDRRRGEFRLFHGGGMQVERPGDEEEENSAADVRF
*"
CDS complement(6197..7057)
/label="ORF frame 3"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE
YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
DRWEPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
EIGASLIKHW*"
promoter complement(7099..7127)
/label="AmpR_promoter"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE
YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
DRWEPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
EIGASLIKHW*"
promoter 7349..506
/label="CMV_immearly_promoter"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE
YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
DRWEPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
EIGASLIKHW*"
misc_feature 7413..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 CGACCTGCAT CCCGCCACCA TGAAAAAGCC TGAACTCACC GCGACGTCTG
2521 TCGAGAAGTT TCTGATCGAA AAGTTCGACA GCGTCTCCGA CCTGATGCAG CTCTCGGAGG
2581 GCGAAGAATC TCGTGCTTTC AGCTTCGATG TAGGAGGGCG TGGATATGTC CTGCGGGTAA
2641 ATAGCTGCGC CGATGGTTTC TACAAAGATC GTTATGTTTA TCGGCACTTT GCATCGGCCG
2701 CGCTCCCGAT TCCGGAAGTG CTTGACATTG GGGAGTTCAG CGAGAGCCTG ACCTATTGCA
2761 TCTCCCGCCG TGCACAGGGT GTCACGTTGC AAGACCTGCC TGAAACCGAA CTGCCCGCTG
2821 TTCTGCAGCC GGTCGCGGAG GCCATGGATG CGATCGCTGC GGCCGATCTT AGCCAGACGA
2881 GCGGGTTCGG CCCATTCGGA CCGCAAGGAA TCGGTCAATA CACTACATGG CGTGATTTCA
2941 TATGCGCGAT TGCTGATCCC CATGTGTATC ACTGGCAAAC TGTGATGGAC GACACCGTCA
3001 GTGCGTCCGT CGCGCAGGCT CTCGATGAGC TGATGCTTTG GGCCGAGGAC TGCCCCGAAG
3061 TCCGGCACCT CGTGCACGCG GATTTCGGCT CCAACAATGT CCTGACGGAC AATGGCCGCA
3121 TAACAGCGGT CATTGACTGG AGCGAGGCGA TGTTCGGGGA TTCCCAATAC GAGGTCGCCA
3181 ACATCTTCTT CTGGAGGCCG TGGTTGGCTT GTATGGAGCA GCAGACGCGC TACTTCGAGC
3241 GGAGGCATCC GGAGCTTGCA GGATCGCCGC GGCTCCGGGC GTATATGCTC CGCATTGGTC
3301 TTGACCAACT CTATCAGAGC TTGGTTGACG GCAATTTCGA TGATGCAGCT TGGGCGCAGG
3361 GTCGATGCGA CGCAATCGTC CGATCCGGAG CCGGGACTGT CGGGCGTACA CAAATCGCCC
3421 GCAGAAGCGC GGCCGTCTGG ACCGATGGCT GTGTAGAAGT ACTCGCCGAT AGTGGAAACC
3481 GACGCCCCAG CACTCGTCCG AGGGCAAAGG AATAGAGTAG ATGCCGACCG AACAAGAGCT
3541 GATTTCGAGA ACGCCTCAGC CAGCAACTCG CGCGAGCCTA GCAAGGCAAA TGCGAGAGAA
3601 CGGCCTTACG CTTGGTGGCA CAGTTCTCGT CCACAGTTCG CTAAGCTCGC TCGGCTGGGT
3661 CGCGGGAGGG CCGGTCGCAG TGATTCAGGC CCTTCTGGAT TGTGTTGGTC CCCAGGGCAC
3721 GATTGTCATG CCCACGCACT CGGGTGATCT GACTGATCCC GCAGATTGGA GATCGCCGCC
3781 CGTGCCTGCC GATTGGGTGC AGATCCGTCG ACCTGCAGCC AAGCTTATCG ATAAAATAAA
3841 AGATTTTATT TAGTCTCCAG AAAAAGGGGG GAATGAAAGA CCCCACCTGT AGGTTTGGCA
3901 AGCTAGCTTA AGTAACGCCA TTTTGCAAGG CATGGAAAAT ACATAACTGA GAATAGAGAA
3961 GTTCAGATCA AGGTTAGGAA CAGAGAGACA GCAGAATATG GGCCAAACAG GATATCAGTG
4021 GTCCAGGCTC TAGTTTTGAC TCAACAATAT CACCAGCTGA AGCCTATAGA GTACGAGCCA
4081 TAGATAAAAT AAAAGATTTT ATTTAGTCTC CAGAAAAAGG GGGGAATGAA AGACCCCACC
4141 TGTAGGTTTG GCAAGCTAGC TTAAGTAACG CCATTTTGCA AGGCATGGAA AAATACATAA
4201 CTGAGAATAG AGAAGTTCAG ATCAAGGTCA GGAACAGATG GAACAGGGTC GACCCTAGAG
4261 AACCATCAGA TGTTTCCAGG GTGCCCCAAG GACCTGAAAT GACCCTGTGC CTTATTTGAA
4321 CTAACCAATC AGTTCGCTTC TCGCTTCTGT TCGCGCGCTT CTGCTCCCCG AGCTCAATAA
4381 AAGAGCCCAC AACCCCTCAC TCGGGGCGCC AGTCCTCCGA TTGACTGAGT CGCCCGGGTA
4441 CCCGTGTATC CAATAAACCC TCTTGCAGTT GCATCCGACT TGTGGTCTCG CTGTTCCTTG
4501 GGAGGGTCTC CTCTGAGTGA TTGACTACCC GTCAGCGGGG GTCTTTCATT TGGGGGCTCG
4561 TCCGGGATCG GGAGACCCCT GCCCAGGGAC CACCGACCCA CCACCGGGAG GTAAGCTGGC
4621 TGCCTCGCGC GTTTCGGTGA TGACGGTGAA AACCTCTGAC ACATGCAGCT CCCGGAGACG
4681 GTCACAGCTT GTCTGTAAGC GGATGCCGGG AGCAGACAAG CCCGTCAGGG CGCGTCAGCG
4741 GGTGTTGGCG GGTGTCGGGG CGCAGCCATG ACCCAGTCAC GTAGCGATAG CGGAGTGTAG
4801 ATCCGGCTGT GGAATGTGTG TCAGTTAGGG TGTGGAAAGT CCCCAGGCTC CCCAGCAGGC
4861 AGAAGTATGC AAAGCATGCA TCTCAATTAG TCAGCAACCA GGTGTGGAAA GTCCCCAGGC
4921 TCCCCAGCAG GCAGAAGTAT GCAAAGCATG CATCTCAATT AGTCAGCAAC CATAGTCCCG
4981 CCCCTAACTC CGCCCATCCC GCCCCTAACT CCGCCCAGTT CCGCCCATTC TCCGCCCCAT
5041 GGCTGACTAA TTTTTTTTAT TTATGCAGAG GCCGAGGCCG CCTCGGCCTC TGAGCTATTC
5101 CAGAAGTAGT GAGGAGGCTT TTTTGGAGGC CTAGGCTTTT GCAAAAAGCT TACTGGCTTA
5161 ACTATGCGGC ATCAGAGCAG ATTGTACTGA GAGTGCACCA TATGCGGTGT GAAATACCGC
5221 ACAGATGCGT AAGGAGAAAA TACCGCATCA GGCGCTCTTC CGCTTCCTCG CTCACTGACT
5281 CGCTGCGCTC GGTCGTTCGG CTGCGGCGAG CGGTATCAGC TCACTCAAAG GCGGTAATAC
5341 GGTTATCCAC AGAATCAGGG GATAACGCAG GAAAGAACAT GTGAGCAAAA GGCCAGCAAA
5401 AGGCCAGGAA CCGTAAAAAG GCCGCGTTGC TGGCGTTTTT CCATAGGCTC CGCCCCCCTG
5461 ACGAGCATCA CAAAAATCGA CGCTCAAGTC AGAGGTGGCG AAACCCGACA GGACTATAAA
5521 GATACCAGGC GTTTCCCCCT GGAAGCTCCC TCGTGCGCTC TCCTGTTCCG ACCCTGCCGC
5581 TTACCGGATA CCTGTCCGCC TTTCTCCCTT CGGGAAGCGT GGCGCTTTCT CATAGCTCAC
5641 GCTGTAGGTA TCTCAGTTCG GTGTAGGTCG TTCGCTCCAA GCTGGGCTGT GTGCACGAAC
5701 CCCCCGTTCA GCCCGACCGC TGCGCCTTAT CCGGTAACTA TCGTCTTGAG TCCAACCCGG
5761 TAAGACACGA CTTATCGCCA CTGGCAGCAG CCACTGGTAA CAGGATTAGC AGAGCGAGGT
5821 ATGTAGGCGG TGCTACAGAG TTCTTGAAGT GGTGGCCTAA CTACGGCTAC ACTAGAAGGA
5881 CAGTATTTGG TATCTGCGCT CTGCTGAAGC CAGTTACCTT CGGAAAAAGA GTTGGTAGCT
5941 CTTGATCCGG CAAACAAACC ACCGCTGGTA GCGGTGGTTT TTTTGTTTGC AAGCAGCAGA
6001 TTACGCGCAG AAAAAAAGGA TCTCAAGAAG ATCCTTTGAT CTTTTCTACG GGGTCTGACG
6061 CTCAGTGGAA CGAAAACTCA CGTTAAGGGA TTTTGGTCAT GAGATTATCA AAAAGGATCT
6121 TCACCTAGAT CCTTTTAAAT TAAAAATGAA GTTTTAAATC AATCTAAAGT ATATATGAGT
6181 AAACTTGGTC TGACAGTTAC CAATGCTTAA TCAGTGAGGC ACCTATCTCA GCGATCTGTC
6241 TATTTCGTTC ATCCATAGTT GCCTGACTCC CCGTCGTGTA GATAACTACG ATACGGGAGG
6301 GCTTACCATC TGGCCCCAGT GCTGCAATGA TACCGCGAGA CCCACGCTCA CCGGCTCCAG
6361 ATTTATCAGC AATAAACCAG CCAGCCGGAA GGGCCGAGCG CAGAAGTGGT CCTGCAACTT
6421 TATCCGCCTC CATCCAGTCT ATTAATTGTT GCCGGGAAGC TAGAGTAAGT AGTTCGCCAG
6481 TTAATAGTTT GCGCAACGTT GTTGCCATTG CTGCAGGCAT CGTGGTGTCA CGCTCGTCGT
6541 TTGGTATGGC TTCATTCAGC TCCGGTTCCC AACGATCAAG GCGAGTTACA TGATCCCCCA
6601 TGTTGTGCAA AAAAGCGGTT AGCTCCTTCG GTCCTCCGAT CGTTGTCAGA AGTAAGTTGG
6661 CCGCAGTGTT ATCACTCATG GTTATGGCAG CACTGCATAA TTCTCTTACT GTCATGCCAT
6721 CCGTAAGATG CTTTTCTGTG ACTGGTGAGT ACTCAACCAA GTCATTCTGA GAATAGTGTA
6781 TGCGGCGACC GAGTTGCTCT TGCCCGGCGT CAACACGGGA TAATACCGCG CCACATAGCA
6841 GAACTTTAAA AGTGCTCATC ATTGGAAAAC GTTCTTCGGG GCGAAAACTC TCAAGGATCT
6901 TACCGCTGTT GAGATCCAGT TCGATGTAAC CCACTCGTGC ACCCAACTGA TCTTCAGCAT
6961 CTTTTACTTT CACCAGCGTT TCTGGGTGAG CAAAAACAGG AAGGCAAAAT GCCGCAAAAA
7021 AGGGAATAAG GGCGACACGG AAATGTTGAA TACTCATACT CTTCCTTTTT CAATATTATT
7081 GAAGCATTTA TCAGGGTTAT TGTCTCATGA GCGGATACAT ATTTGAATGT ATTTAGAAAA
7141 ATAAACAAAT AGGGGTTCCG CGCACATTTC CCCGAAAAGT GCCACCTGAC GTCTAAGAAA
7201 CCATTATTAT CATGACATTA ACCTATAAAA ATAGGCGTAT CACGAGGCCC TTTCGTCTTC
7261 AAGAATTAGC TTGGCCATTG CATACGTTGT ATCCATATCA TAATATGTAC ATTTATATTG
7321 GCTCATGTCC AACATTACCG CCATGTTGAC ATTGATTATT GACTAGTTAT TAATAGTAAT
7381 CAATTACGGG GTCATTAGTT CATAGCCCAT ATATGGAGT
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