出品公司: | Clontech |
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载体名称: | pRetroQ-DsRed-Monomer-C1 |
质粒类型: | 逆病毒载体;荧光报告载体 |
高拷贝/低拷贝: | 低拷贝 |
克隆方法: | 限制性内切酶,多克隆位点 |
启动子: | CMV IE |
载体大小: | 7573 bp |
5' 测序引物及序列: | -- |
3' 测序引物及序列: | -- |
载体标签: | DsRed-Monomer(N-端) |
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),原代细胞 |
备注: |
pRetroQ-DsRed-Monomer-C1载体是自我失活型逆病毒载体,表达N端DsRed-Monomer融合蛋白; pRetroQ-DsRed-Monomer-C1载体能够高效递送目的基因到靶细胞中,尤其适用于原代细胞和其 它难转染的细胞系; DsRed-Monomer是DsRed的单体突变体,与DsRed相比,编码序列经过了人工优化,适用于在哺乳动物细胞中的高水平表达。 |
产品目录号: | 632508 |
稳定性: | 稳表达 |
组成型/诱导型: | 组成型 |
病毒/非病毒: | 逆转录病毒 |
买家导航
pRetroQ-DsRed-Monomer-C1载体质粒基本信息
pRetroQ-DsRed-Monomer-C1质粒图谱载体图谱和pRetroQ-DsRed-Monomer-C1载体序列质粒序列多克隆位点信息
pRetroQ-DsRed-Monomer-C1质粒载体简介
pRetroQ-DsRed-Monomer-C1载体描述
pRetroQ-DsRed-Monomer-C1 is a high-titer, self-inactivating retroviral vector that facilitates efficient delivery and expression of DsRed-monomer (DsRed.M1) as well as C terminal fusions of DsRed monomer to target cells. DsRed.M1 is a monomeric mutant derived from the tetrameric Discosoma sp. red fluorescent protein DsRed (1).
DsRed-Monomer contains forty-five amino acid substitutions. When DsRed-Monomer is expressed in mammalian cell cultures, red fluorescent cells can be detected by either fluorescence microscopy or flow cytometry 12–16 hours after transfection or 24–48 hours after infection (DsRed-Monomer excitation and emission maxima = 557 nm and 592 nm, respectively). The DsRed-Monomer coding sequence is human codon-optimized for high expression in mammalian cells (2) The MCS in pRetroQ-DsRed Monomer-C1 follows the DsRed-Monomer coding sequences. Genes cloned into the MCS are expressed as fusions to the C-terminus of DsRed Monomer when they are in the same reading frame as DsRed Monomer and there are no intervening stop codons.
The RetroQ retroviral vector backbone incorporates several unique features. The hybrid 5’ long terminal repeat (LTR) consists of the CMV type I enhancer and the murine sarcoma virus (MSV) promoter. This vector demonstrates high levels of transcription in HEK 293-based packaging cell lines due, in part, to the presence of adenoviral E1A (3–6) 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, the inactivated 3’ LTR is copied and replaces the 5’ LTR CMV enhancer sequences. This can reduce the phenomenon known as promoter interference (7) and allow more efficient expression. This vector also contains a puromycin resistance cassette (Puror) driven by the PGK promoter for selection of positively infected cells (2).
Additionally, the viral genomic transcript contains the necessary viral RNA processing elements including the LTRs, packaging signal (ψ+), and tRNA primer-binding site. pRetroQ-DsRed Monomer-C1 contains a bacterial origin of replication, an E. coli Ampr gene for propagation and selection in bacteria, and an SV40 origin for replication in mammalian cells expressing the SV40 T antigen.
pRetro-DsRed Monomer-C1 is designed to efficiently deliver and express fusions to the C terminus of DsRed-Monomer into primary cells or cells that are difficult to transfect. Fusions to the C terminus of DsRed-Monomer retain the fluorescent properties of the native protein, allowing the in vivo localization of the fusion protein. The target gene should be cloned into pDsRed Monomer-C1 so that it is in frame with the DsRed-Monomer coding sequences, with no intervening in-frame stop codons. The inserted gene should include the initiating ATG codon. The recombinant DsRed Monomer vector can be infected or transfected into mammalian cells.
If required, stable transformants can be selected using Puromycin. pRetroQ DsRed Monomer-C1 can also be used simply to express DsRed Monomer in a cell line of interest (e.g., as an infection marker).Once pRetroQ-DsRed Monomer-C1 is transfected into a packaging cell line (such as the RetroPack PT67 Cell line , AmphoPack-293, EcoPack2-293, Pantropic Expression System, or Retro-X Universal Packaging System, RNA from the vector is packaged into non-infectious, replication-incompetent retroviral particles, since pRetroQ-DsRed Monomer-C1 lacks the structural genes (gag, pol, and env) necessary for particle formation and replication. These genes, however, are stably integrated as part of the packaging cell genome. Once a high-titer supernatant is produced, these retroviral particles can infect target cells and transmit the gene of interest but 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 chances of producing replication-competent virus due to recombination events during cell proliferation.
Propagation in E. coli
Suitable host strains: DH5α, Fusion Blue, 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: low
pRetroQ-DsRed-Monomer-C1质粒序列载体序列
LOCUS pRetroQ-DsRed-Monomer-C1 7573 bp DNA SYN DEFINITION pRetroQ-DsRed-Monomer-C1 ACCESSION KEYWORDS SOURCE ORGANISM other sequences; artificial sequences; vectors. FEATURES Location/Qualifiers source 1..7573 /organism="pRetroQ-DsRed-Monomer-C1" /mol_type="other DNA" promoter 2..507 /label="CMV_immearly_promoter" misc_feature 6..689 /label="5_LTR" misc_feature 9..297 /label="CAG_enhancer" misc_feature 464..484 /label="CMV_fwd_primer" misc_feature 759..1568 /label="psi_plus_pack" misc_feature 1155..1562 /label="gag" misc_feature 1498..1520 /label="MSCV_primer" misc_feature 1498..1520 /label="pLXSN_5_primer" misc_feature 1536..1552 /label="pBABE_5_primer" promoter 1592..2144 /label="CMV_immearly_promoter" misc_feature 1647..1934 /label="CAG_enhancer" misc_feature 2101..2121 /label="CMV_fwd_primer" promoter 2102..2171 /label="CMV_promoter" CDS 2195..2950 /label="ORF frame 2" /translation="MDNTEDVIKEFMQFKVRMEGSVNGHYFEIEGEGEGKPYEGTQTA KLQVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYMKLSFPEGFTWERSMNFEDG GVVEVQQDSSLQDGTFIYKVKFKGVNFPADGPVMQKKTAGWEPSTEKLYPQDGVLKGE ISHALKLKDGGHYTCDFKTVYKAKKPVQLPGNHYVDSKLDITNHNEDYTVVEQYEHAE ARHSGSQSGLRSRAQASNSAVDGTAGPGSTGSR*" gene 2198..2869 /label="DsRed_Monomer" /gene="DsRed_Monomer" /translation="MDNTEDVIKEFMQFKVRMEGSVNGHYFEIEGEGEGKPYEGTQTA KLQVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYMKLSFPEGFTWERSMNFEDG GVVEVQQDSSLQDGTFIYKVKFKGVNFPADGPVMQKKTAGWEPSTEKLYPQDGVLKGE ISHALKLKDGGHYTCDFKTVYKAKKPVQLPGNHYVDSKLDITNHNEDYTVVEQYEHAE ARHSGSQSGLRSRAQASNSAVDGTAGPGSTGSR*" misc_feature complement(2375..2395) /label="dsRed1_N_primer" /translation="MDNTEDVIKEFMQFKVRMEGSVNGHYFEIEGEGEGKPYEGTQTA KLQVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYMKLSFPEGFTWERSMNFEDG GVVEVQQDSSLQDGTFIYKVKFKGVNFPADGPVMQKKTAGWEPSTEKLYPQDGVLKGE ISHALKLKDGGHYTCDFKTVYKAKKPVQLPGNHYVDSKLDITNHNEDYTVVEQYEHAE ARHSGSQSGLRSRAQASNSAVDGTAGPGSTGSR*" misc_feature complement(3003..3026) /label="MSCV_rev_primer" /translation="MDNTEDVIKEFMQFKVRMEGSVNGHYFEIEGEGEGKPYEGTQTA KLQVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYMKLSFPEGFTWERSMNFEDG GVVEVQQDSSLQDGTFIYKVKFKGVNFPADGPVMQKKTAGWEPSTEKLYPQDGVLKGE ISHALKLKDGGHYTCDFKTVYKAKKPVQLPGNHYVDSKLDITNHNEDYTVVEQYEHAE ARHSGSQSGLRSRAQASNSAVDGTAGPGSTGSR*" CDS 3248..4087 /label="ORF frame 2" /translation="MEAGRPLGQRPIAALLLRFLGSEAGKGWVRGRAQGRAQGRGGRP KVLRRPGILHASKAHVCRAVLLFLISGPFDLQPKLTMTEYKPTVRLATRDDVPRAVRT LAAAFADYPATRHTVDPDRHIERVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPE SVEAGAVFAEIGPRMAELSGSRLAAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKG LGSAVVLPGVEAAERAGVPAFLETSAPRNLPFYERLGFTVTADVEVPEGPRTWCMTRK PGA*" misc_feature 3400..3419 /label="mPGK_F_primer" /translation="MEAGRPLGQRPIAALLLRFLGSEAGKGWVRGRAQGRAQGRGGRP KVLRRPGILHASKAHVCRAVLLFLISGPFDLQPKLTMTEYKPTVRLATRDDVPRAVRT LAAAFADYPATRHTVDPDRHIERVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPE SVEAGAVFAEIGPRMAELSGSRLAAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKG LGSAVVLPGVEAAERAGVPAFLETSAPRNLPFYERLGFTVTADVEVPEGPRTWCMTRK PGA*" CDS complement(3413..4138) /label="ORF frame 1" /translation="MGSCAPFGRALRVVGRASGTGLAGHAPGARSFGHLDVGGDGEAE PLVEGEVAGRGGLQEGGHPGALGRLHSGEHDGAAQTLALVVGRDADGGQEPRGLLGPV RRQEAFHLLLRGQPGTAQLGHARADLGEHRPRFDALRRGPDRHRGAVVRDPHLADVEP DAREEEFLQLGDPLDVAVRIDGVARGGVVGERGGEGAYGPGDVVAGGEAHRGLVLGHG KLGLQVERPGDEEEENSAADVRF*" gene 3488..4087 /label="puro (variant)" /gene="puro (variant)" /translation="MGSCAPFGRALRVVGRASGTGLAGHAPGARSFGHLDVGGDGEAE PLVEGEVAGRGGLQEGGHPGALGRLHSGEHDGAAQTLALVVGRDADGGQEPRGLLGPV RRQEAFHLLLRGQPGTAQLGHARADLGEHRPRFDALRRGPDRHRGAVVRDPHLADVEP DAREEEFLQLGDPLDVAVRIDGVARGGVVGERGGEGAYGPGDVVAGGEAHRGLVLGHG KLGLQVERPGDEEEENSAADVRF*" misc_feature complement(4807..4829) /label="pGEX_3_primer" /translation="MGSCAPFGRALRVVGRASGTGLAGHAPGARSFGHLDVGGDGEAE PLVEGEVAGRGGLQEGGHPGALGRLHSGEHDGAAQTLALVVGRDADGGQEPRGLLGPV RRQEAFHLLLRGQPGTAQLGHARADLGEHRPRFDALRRGPDRHRGAVVRDPHLADVEP DAREEEFLQLGDPLDVAVRIDGVARGGVVGERGGEGAYGPGDVVAGGEAHRGLVLGHG KLGLQVERPGDEEEENSAADVRF*" misc_feature complement(4965..4985) /label="pBABE_3_primer" /translation="MGSCAPFGRALRVVGRASGTGLAGHAPGARSFGHLDVGGDGEAE PLVEGEVAGRGGLQEGGHPGALGRLHSGEHDGAAQTLALVVGRDADGGQEPRGLLGPV RRQEAFHLLLRGQPGTAQLGHARADLGEHRPRFDALRRGPDRHRGAVVRDPHLADVEP DAREEEFLQLGDPLDVAVRIDGVARGGVVGERGGEGAYGPGDVVAGGEAHRGLVLGHG KLGLQVERPGDEEEENSAADVRF*" misc_feature complement(4971..5186) /label="SV40_enhancer" /translation="MGSCAPFGRALRVVGRASGTGLAGHAPGARSFGHLDVGGDGEAE PLVEGEVAGRGGLQEGGHPGALGRLHSGEHDGAAQTLALVVGRDADGGQEPRGLLGPV RRQEAFHLLLRGQPGTAQLGHARADLGEHRPRFDALRRGPDRHRGAVVRDPHLADVEP DAREEEFLQLGDPLDVAVRIDGVARGGVVGERGGEGAYGPGDVVAGGEAHRGLVLGHG KLGLQVERPGDEEEENSAADVRF*" promoter 4983..5251 /label="SV40_promoter" /translation="MGSCAPFGRALRVVGRASGTGLAGHAPGARSFGHLDVGGDGEAE PLVEGEVAGRGGLQEGGHPGALGRLHSGEHDGAAQTLALVVGRDADGGQEPRGLLGPV RRQEAFHLLLRGQPGTAQLGHARADLGEHRPRFDALRRGPDRHRGAVVRDPHLADVEP DAREEEFLQLGDPLDVAVRIDGVARGGVVGERGGEGAYGPGDVVAGGEAHRGLVLGHG KLGLQVERPGDEEEENSAADVRF*" rep_origin 5150..5227 /label="SV40_origin" /translation="MGSCAPFGRALRVVGRASGTGLAGHAPGARSFGHLDVGGDGEAE PLVEGEVAGRGGLQEGGHPGALGRLHSGEHDGAAQTLALVVGRDADGGQEPRGLLGPV RRQEAFHLLLRGQPGTAQLGHARADLGEHRPRFDALRRGPDRHRGAVVRDPHLADVEP DAREEEFLQLGDPLDVAVRIDGVARGGVVGERGGEGAYGPGDVVAGGEAHRGLVLGHG KLGLQVERPGDEEEENSAADVRF*" misc_feature 5212..5231 /label="SV40pro_F_primer" /translation="MGSCAPFGRALRVVGRASGTGLAGHAPGARSFGHLDVGGDGEAE PLVEGEVAGRGGLQEGGHPGALGRLHSGEHDGAAQTLALVVGRDADGGQEPRGLLGPV RRQEAFHLLLRGQPGTAQLGHARADLGEHRPRFDALRRGPDRHRGAVVRDPHLADVEP DAREEEFLQLGDPLDVAVRIDGVARGGVVGERGGEGAYGPGDVVAGGEAHRGLVLGHG KLGLQVERPGDEEEENSAADVRF*" gene complement(6351..7211) /label="Ampicillin" /gene="Ampicillin" /translation="MGSCAPFGRALRVVGRASGTGLAGHAPGARSFGHLDVGGDGEAE PLVEGEVAGRGGLQEGGHPGALGRLHSGEHDGAAQTLALVVGRDADGGQEPRGLLGPV RRQEAFHLLLRGQPGTAQLGHARADLGEHRPRFDALRRGPDRHRGAVVRDPHLADVEP DAREEEFLQLGDPLDVAVRIDGVARGGVVGERGGEGAYGPGDVVAGGEAHRGLVLGHG KLGLQVERPGDEEEENSAADVRF*" CDS complement(6351..7211) /label="ORF frame 3" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL DRWEPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA EIGASLIKHW*" promoter complement(7253..7281) /label="AmpR_promoter" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL DRWEPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA EIGASLIKHW*" promoter 7503..507 /label="CMV_immearly_promoter" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL DRWEPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA EIGASLIKHW*" misc_feature 7567..689 /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 GCCCGCTCTG GCATTATGCC 241 CAGTACATGA CCTTATGGGA CTTTCCTACT TGGCAGTACA TCTACGTATT AGTCATCGCT 301 ATTACCATGG TGATGCGGTT TTGGCAGTAC ATCAATGGGC GTGGATAGCG GTTTGACTCA 361 CGGGGATTTC CAAGTCTCCA CCCCATTGAC GTCAATGGGA GTTTGTTTTG GCACCAAAAT 421 CAACGGGACT TTCCAAAATG TCGTAACAAC TCCGCCCCAT TGACGCAAAT GGGCGGTAGG 481 CGTGTACGGT GGGAGGTCTA TATAAGCAGA GCTCAATAAA AGAGCCCACA ACCCCTCACT 541 CGGCGCGCCA GTCTTCCGAT AGACTGCGTC GCCCGGGTAC CCGTATTCCC AATAAAGCCT 601 CTTGCTGTTT GCATCCGAAT CGTGGTCTCG CTGTTCCTTG GGAGGGTCTC CTCTGAGTGA 661 TTGACTACCC ACGACGGGGG TCTTTCATTT GGGGGCTCGT CCGGGATTTG GAGACCCCTG 721 CCCAGGGACC ACCGACCCAC CACCGGGAGG TAAGCTGGCC AGCAACTTAT CTGTGTCTGT 781 CCGATTGTCT AGTGTCTATG TTTGATGTTA TGCGCCTGCG TCTGTACTAG TTAGCTAACT 841 AGCTCTGTAT CTGGCGGACC CGTGGTGGAA CTGACGAGTT CTGAACACCC GGCCGCAACC 901 CTGGGAGACG TCCCAGGGAC TTTGGGGGCC GTTTTTGTGG CCCGACCTGA GGAAGGGAGT 961 CGATGTGGAA TCCGACCCCG TCAGGATATG TGGTTCTGGT AGGAGACGAG AACCTAAAAC 1021 AGTTCCCGCC TCCGTCTGAA TTTTTGCTTT CGGTTTGGAA CCGAAGCCGC GCGTCTTGTC 1081 TGCTGCAGCG CTGCAGCATC GTTCTGTGTT GTCTCTGTCT GACTGTGTTT CTGTATTTGT 1141 CTGAAAATTA GGGCCAGACT GTTACCACTC CCTTAAGTTT GACCTTAGGT CACTGGAAAG 1201 ATGTCGAGCG GATCGCTCAC AACCAGTCGG TAGATGTCAA GAAGAGACGT TGGGTTACCT 1261 TCTGCTCTGC AGAATGGCCA ACCTTTAACG TCGGATGGCC GCGAGACGGC ACCTTTAACC 1321 GAGACCTCAT CACCCAGGTT AAGATCAAGG TCTTTTCACC TGGCCCGCAT GGACACCCAG 1381 ACCAGGTCCC CTACATCGTG ACCTGGGAAG CCTTGGCTTT TGACCCCCCT CCCTGGGTCA 1441 AGCCCTTTGT ACACCCTAAG CCTCCGCCTC CTCTTCCTCC ATCCGCCCCG TCTCTCCCCC 1501 TTGAACCTCC TCGTTCGACC CCGCCTCGAT CCTCCCTTTA TCCAGCCCTC ACTCCTTCTC 1561 TAGGCGCCGG AATTGAAGAT CATAGTTATT AATAGTAATC AATTACGGGG TCATTAGTTC 1621 ATAGCCCATA TATGGAGTTC CGCGTTACAT AACTTACGGT AAATGGCCCG CCTGGCTGAC 1681 CGCCCAACGA CCCCCGCCCA TTGACGTCAA TAATGACGTA TGTTCCCATA GTAACGCCAA 1741 TAGGGACTTT CCATTGACGT CAATGGGTGG AGTATTTACG GTAAACTGCC CACTTGGCAG 1801 TACATCAAGT GTATCATATG CCAAGTACGC CCCCTATTGA CGTCAATGAC GGTAAATGGC 1861 CCGCCTGGCA TTATGCCCAG TACATGACCT TATGGGACTT TCCTACTTGG CAGTACATCT 1921 ACGTATTAGT CATCGCTATT ACCATGGTGA TGCGGTTTTG GCAGTACATC AATGGGCGTG 1981 GATAGCGGTT TGACTCACGG GGATTTCCAA GTCTCCACCC CATTGACGTC AATGGGAGTT 2041 TGTTTTGGCA CCAAAATCAA CGGGACTTTC CAAAATGTCG TAACAACTCC GCCCCATTGA 2101 CGCAAATGGG CGGTAGGCGT GTACGGTGGG AGGTCTATAT AAGCAGAGCT GGTTTAGTGA 2161 ACCGTCAGAT CCGCTAGCGC TACCGGTCGC CACCATGGAC AACACCGAGG ACGTCATCAA 2221 GGAGTTCATG CAGTTCAAGG TGCGCATGGA GGGCTCCGTG AACGGCCACT ACTTCGAGAT 2281 CGAGGGCGAG GGCGAGGGCA AGCCCTACGA GGGCACCCAG ACCGCCAAGC TGCAGGTGAC 2341 CAAGGGCGGC CCCCTGCCCT TCGCCTGGGA CATCCTGTCC CCCCAGTTCC AGTACGGCTC 2401 CAAGGCCTAC GTGAAGCACC CCGCCGACAT CCCCGACTAC ATGAAGCTGT CCTTCCCCGA 2461 GGGCTTCACC TGGGAGCGCT CCATGAACTT CGAGGACGGC GGCGTGGTGG AGGTGCAGCA 2521 GGACTCCTCC CTGCAGGACG GCACCTTCAT CTACAAGGTG AAGTTCAAGG GCGTGAACTT 2581 CCCCGCCGAC GGCCCCGTAA TGCAGAAGAA GACTGCCGGC TGGGAGCCCT CCACCGAGAA 2641 GCTGTACCCC CAGGACGGCG TGCTGAAGGG CGAGATCTCC CACGCCCTGA AGCTGAAGGA 2701 CGGCGGCCAC TACACCTGCG ACTTCAAGAC CGTGTACAAG GCCAAGAAGC CCGTGCAGCT 2761 GCCCGGCAAC CACTACGTGG ACTCCAAGCT GGACATCACC AACCACAACG AGGACTACAC 2821 CGTGGTGGAG CAGTACGAGC ACGCCGAGGC CCGCCACTCC GGCTCCCAGT CCGGACTCAG 2881 ATCTCGAGCT CAAGCTTCGA ATTCTGCAGT CGACGGTACC GCGGGCCCGG GATCCACCGG 2941 ATCTAGATAA CTGATCATAA TTCTACCGGG TAGGGGAGGC GCTTTTCCCA AGGCAGTCTG 3001 GAGCATGCGC TTTAGCAGCC CCGCTGGGCA CTTGGCGCTA CACAAGTGGC CTCTGGCCTC 3061 GCACACATTC CACATCCACC GGTAGGCGCC AACCGGCTCC GTTCTTTGGT GGCCCCTTCG 3121 CGCCACCTTC TACTCCTCCC CTAGTCAGGA AGTTCCCCCC CGCCCCGCAG CTCGCGTCGT 3181 GCAGGACGTG ACAAATGGAA GTAGCACGTC TCACTAGTCT CGTGCAGATG GACAGCACCG 3241 CTGAGCAATG GAAGCGGGTA GGCCTTTGGG GCAGCGGCCA ATAGCAGCTT TGCTCCTTCG 3301 CTTTCTGGGC TCAGAGGCTG GGAAGGGGTG GGTCCGGGGG CGGGCTCAGG GGCGGGCTCA 3361 GGGGCGGGGC GGGCGCCCGA AGGTCCTCCG GAGGCCCGGC ATTCTGCACG CTTCAAAAGC 3421 GCACGTCTGC CGCGCTGTTC TCCTCTTCCT CATCTCCGGG CCTTTCGACC TGCAGCCCAA 3481 GCTTACCATG ACCGAGTACA AGCCCACGGT GCGCCTCGCC ACCCGCGACG ACGTCCCCAG 3541 GGCCGTACGC ACCCTCGCCG CCGCGTTCGC CGACTACCCC GCCACGCGCC ACACCGTCGA 3601 TCCGGACCGC CACATCGAGC GGGTCACCGA GCTGCAAGAA CTCTTCCTCA CGCGCGTCGG 3661 GCTCGACATC GGCAAGGTGT GGGTCGCGGA CGACGGCGCC GCGGTGGCGG TCTGGACCAC 3721 GCCGGAGAGC GTCGAAGCGG GGGCGGTGTT CGCCGAGATC GGCCCGCGCA TGGCCGAGTT 3781 GAGCGGTTCC CGGCTGGCCG CGCAGCAACA GATGGAAGGC CTCCTGGCGC CGCACCGGCC 3841 CAAGGAGCCC GCGTGGTTCC TGGCCACCGT CGGCGTCTCG CCCGACCACC AGGGCAAGGG 3901 TCTGGGCAGC GCCGTCGTGC TCCCCGGAGT GGAGGCGGCC GAGCGCGCCG GGGTGCCCGC 3961 CTTCCTGGAG ACCTCCGCGC CCCGCAACCT CCCCTTCTAC GAGCGGCTCG GCTTCACCGT 4021 CACCGCCGAC GTCGAGGTGC CCGAAGGACC GCGCACCTGG TGCATGACCC GCAAGCCCGG 4081 TGCCTGACGC CCGCCCCACG ACCCGCAGCG CCCGACCGAA AGGAGCGCAC GACCCCATGC 4141 ATCGGCGTCT CGATCGAGAT ATCAGTGGTC CAGGCTCTAG TTTTGACTCA ACAATATCAC 4201 CAGCTGAAGC CTATAGAGTA CGAGCCATAG ATAAAATAAA AGATTTTATT TAGTCTCCAG 4261 AAAAAGGGGG GAATGAAAGA CCCCACCTGT AGGTTTGGCA AGCTACTAGC TTAAGTAACG 4321 CCATTTTGCA AGGCATGGAA AAATACATAA CTGAGAATAG AGAAGTTCAG ATCAAGGTCA 4381 GGAACAGATG GAACAGGGTC GATCGACCCT AGAGAACCAT CAGATGTTTC CAGGGTGCCC 4441 CAAGGACCTG AAATGACCCT GTGCCTTATT TGAACTAACC AATCAGTTCG CTTCTCGCTT 4501 CTGTTCGCGC GCTTCTGCTC CCCGAGCTCA ATAAAAGAGC CCACAACCCC TCACTCGGGG 4561 CGCCAGTCCT CCGATTGACT GAGTCGCCCG GGTACCCGTG TATCCAATAA ACCCTCTTGC 4621 AGTTGCATCC GACTTGTGGT CTCGCTGTTC CTTGGGAGGG TCTCCTCTGA GTGATTGACT 4681 ACCCGTCAGC GGGGGTCTTT CATTTGGGGG CTCGTCCGGG ATCGGGAGAC CCCTGCCCAG 4741 GGACCACCGA CCCACCACCG GGAGGTAAGC TGGCTGCCTC GCGCGTTTCG GTGATGACGG 4801 TGAAAACCTC TGACACATGC AGCTCCCGGA GACGGTCACA GCTTGTCTGT AAGCGGATGC 4861 CGGGAGCAGA CAAGCCCGTC AGGGCGCGTC AGCGGGTGTT GGCGGGTGTC GGGGCGCAGC 4921 CATGACCCAG TCACGTAGCG ATAGCGGAGT GTAGATCCGG CTGTGGAATG TGTGTCAGTT 4981 AGGGTGTGGA AAGTCCCCAG GCTCCCCAGC AGGCAGAAGT ATGCAAAGCA TGCATCTCAA 5041 TTAGTCAGCA ACCAGGTGTG GAAAGTCCCC AGGCTCCCCA GCAGGCAGAA GTATGCAAAG 5101 CATGCATCTC AATTAGTCAG CAACCATAGT CCCGCCCCTA ACTCCGCCCA TCCCGCCCCT 5161 AACTCCGCCC AGTTCCGCCC ATTCTCCGCC CCATGGCTGA CTAATTTTTT TTATTTATGC 5221 AGAGGCCGAG GCCGCCTCGG CCTCTGAGCT ATTCCAGAAG TAGTGAGGAG GCTTTTTTGG 5281 AGGCCTAGGC TTTTGCAAAA AGCTTACTGG CTTAACTATG CGGCATCAGA GCAGATTGTA 5341 CTGAGAGTGC ACCATATGCG GTGTGAAATA CCGCACAGAT GCGTAAGGAG AAAATACCGC 5401 ATCAGGCGCT CTTCCGCTTC CTCGCTCACT GACTCGCTGC GCTCGGTCGT TCGGCTGCGG 5461 CGAGCGGTAT CAGCTCACTC AAAGGCGGTA ATACGGTTAT CCACAGAATC AGGGGATAAC 5521 GCAGGAAAGA ACATGTGAGC AAAAGGCCAG CAAAAGGCCA GGAACCGTAA AAAGGCCGCG 5581 TTGCTGGCGT TTTTCCATAG GCTCCGCCCC CCTGACGAGC ATCACAAAAA TCGACGCTCA 5641 AGTCAGAGGT GGCGAAACCC GACAGGACTA TAAAGATACC AGGCGTTTCC CCCTGGAAGC 5701 TCCCTCGTGC GCTCTCCTGT TCCGACCCTG CCGCTTACCG GATACCTGTC CGCCTTTCTC 5761 CCTTCGGGAA GCGTGGCGCT TTCTCATAGC TCACGCTGTA GGTATCTCAG TTCGGTGTAG 5821 GTCGTTCGCT CCAAGCTGGG CTGTGTGCAC GAACCCCCCG TTCAGCCCGA CCGCTGCGCC 5881 TTATCCGGTA ACTATCGTCT TGAGTCCAAC CCGGTAAGAC ACGACTTATC GCCACTGGCA 5941 GCAGCCACTG GTAACAGGAT TAGCAGAGCG AGGTATGTAG GCGGTGCTAC AGAGTTCTTG 6001 AAGTGGTGGC CTAACTACGG CTACACTAGA AGGACAGTAT TTGGTATCTG CGCTCTGCTG 6061 AAGCCAGTTA CCTTCGGAAA AAGAGTTGGT AGCTCTTGAT CCGGCAAACA AACCACCGCT 6121 GGTAGCGGTG GTTTTTTTGT TTGCAAGCAG CAGATTACGC GCAGAAAAAA AGGATCTCAA 6181 GAAGATCCTT TGATCTTTTC TACGGGGTCT GACGCTCAGT GGAACGAAAA CTCACGTTAA 6241 GGGATTTTGG TCATGAGATT ATCAAAAAGG ATCTTCACCT AGATCCTTTT AAATTAAAAA 6301 TGAAGTTTTA AATCAATCTA AAGTATATAT GAGTAAACTT GGTCTGACAG TTACCAATGC 6361 TTAATCAGTG AGGCACCTAT CTCAGCGATC TGTCTATTTC GTTCATCCAT AGTTGCCTGA 6421 CTCCCCGTCG TGTAGATAAC TACGATACGG GAGGGCTTAC CATCTGGCCC CAGTGCTGCA 6481 ATGATACCGC GAGACCCACG CTCACCGGCT CCAGATTTAT CAGCAATAAA CCAGCCAGCC 6541 GGAAGGGCCG AGCGCAGAAG TGGTCCTGCA ACTTTATCCG CCTCCATCCA GTCTATTAAT 6601 TGTTGCCGGG AAGCTAGAGT AAGTAGTTCG CCAGTTAATA GTTTGCGCAA CGTTGTTGCC 6661 ATTGCTGCAG GCATCGTGGT GTCACGCTCG TCGTTTGGTA TGGCTTCATT CAGCTCCGGT 6721 TCCCAACGAT CAAGGCGAGT TACATGATCC CCCATGTTGT GCAAAAAAGC GGTTAGCTCC 6781 TTCGGTCCTC CGATCGTTGT CAGAAGTAAG TTGGCCGCAG TGTTATCACT CATGGTTATG 6841 GCAGCACTGC ATAATTCTCT TACTGTCATG CCATCCGTAA GATGCTTTTC TGTGACTGGT 6901 GAGTACTCAA CCAAGTCATT CTGAGAATAG TGTATGCGGC GACCGAGTTG CTCTTGCCCG 6961 GCGTCAACAC GGGATAATAC CGCGCCACAT AGCAGAACTT TAAAAGTGCT CATCATTGGA 7021 AAACGTTCTT CGGGGCGAAA ACTCTCAAGG ATCTTACCGC TGTTGAGATC CAGTTCGATG 7081 TAACCCACTC GTGCACCCAA CTGATCTTCA GCATCTTTTA CTTTCACCAG CGTTTCTGGG 7141 TGAGCAAAAA CAGGAAGGCA AAATGCCGCA AAAAAGGGAA TAAGGGCGAC ACGGAAATGT 7201 TGAATACTCA TACTCTTCCT TTTTCAATAT TATTGAAGCA TTTATCAGGG TTATTGTCTC 7261 ATGAGCGGAT ACATATTTGA ATGTATTTAG AAAAATAAAC AAATAGGGGT TCCGCGCACA 7321 TTTCCCCGAA AAGTGCCACC TGACGTCTAA GAAACCATTA TTATCATGAC ATTAACCTAT 7381 AAAAATAGGC GTATCACGAG GCCCTTTCGT CTTCAAGAAT TAGCTTGGCC ATTGCATACG 7441 TTGTATCCAT ATCATAATAT GTACATTTAT ATTGGCTCAT GTCCAACATT ACCGCCATGT 7501 TGACATTGAT TATTGACTAG TTATTAATAG TAATCAATTA CGGGGTCATT AGTTCATAGC 7561 CCATATATGG AGT //