| 出品公司: | Clontech |
|---|---|
| 载体名称: | pEF1α-IRES-ZsGreen1 |
| 质粒类型: | 哺乳动物表达载体；双顺反子载体；荧光报告载体 |
| 高拷贝/低拷贝: | 高拷贝 |
| 克隆方法: | 多克隆位点，限制性内切酶 |
| 启动子: | EF1α |
| 载体大小: | 6040 bp |
| 5' 测序引物及序列: | EF1-F: TCAAGCCTCAGACAGTGGTTC |
| 3' 测序引物及序列: | IRES-R |
| 载体标签: | -- |
| 载体抗性: | 卡那霉素 |
| 筛选标记: | Neomycin、ZsGreen1 |
| 克隆菌株: | DH5alpha |
| 宿主细胞（系）: | 大部分细胞类型，包括原代细胞、分化细胞、干细胞。 |
| 备注: |
pEF1α-IRES-ZsGreen1载体是哺乳动物表达载体； 目的基因与报告基因在EF1启动子驱动下转录为双顺反子； 二者之间的IRES元件，使得目的基因 与报告基因独立翻译。 |
| 产品目录号: | 632420 |
| 稳定性: | 瞬表达 |
| 组成型/诱导型: | 组成型 |
| 病毒/非病毒: | 非病毒 |
买家导航
pEF1α-IRES-ZsGreen1载体质粒基本信息
pEF1α-IRES-ZsGreen1质粒图谱载体图谱和pEF1α-IRES-ZsGreen1载体序列质粒序列多克隆位点信息
pEF1α-IRES-ZsGreen1质粒载体简介
载体介绍
pEF1α-IRES-ZsGreen1 is designed to simultaneously and constitutively express a protein of interest and ZsGreen1 in mammalian cells. ZsGreen1 is a human-codon-optimized variant of the reef coral Zoanthus sp. green fluorescent protein (ZsGreen) that has been engineered for brighter fluorescence (1–3). The excitation and emission maxima of native ZsGreen1 are 493 nm and 505 nm, respectively. Simultaneous expression of a protein of interest and ZsGreen1 is made possible by the presence of an encephalomyocarditis virus internal ribosome entry site (IRES2; 4, 5) positioned between the multiple cloning site (MCS) and the ZsGreen1 gene. The IRES allows a protein of interest and ZsGreen1 to be translated from a single bicistronic mRNA. Stable, constitutive expression of the bicistronic transcript is driven by the EF1α promoter (PEF1α), which continues to be constitutively active even after vector integration into the host cell genome (6).
The vector backbone also contains an SV40 origin for replication in mammalian cells expressing the SV40 large T antigen, a pUC origin of replication for propagation in E. coli, and an f1 origin for single-stranded DNA production. This vector also has a neomycin-resistance cassette (Neor) that allows G418 selection of stably transfected eukaryotic cells (7).
This cassette consists of the SV40 early promoter (PSV40 e), a Tn5 kanamycin/neomycin resistance gene, and herpes simplex virus thymidine kinase (HSV TK) polyadenylation signals. A bacterial promoter upstream of the cassette drives expression of the kanamycin resistance gene in E. coli.
载体特征
PEF1α (human elongation factor 1 alpha promoter): 12–1346
MCS (multiple cloning site): 1348–1422
IRES2 (internal ribosome entry site): 1423–2007
ZsGreen1 (human-codon-optimized): 1999–2703
SV40 early polyA signal: 2859–2909
f1 origin of replication: 2956–3411 (complementary)
SV40 origin of replication: 3752–3887
PSV40e (SV40 early promoter and enhancer sequences): 3585–3853
Kanr/Neor (kanamycin/neomycin resistance gene): 3936–4730
HSV TK polyA signals: 4966–4984
pUC origin of replication: 5315–5958
补充信息
pEF1α-IRES2-ZsGreen1 can be used to quickly identify cells expressing a gene of interest by screening for ZsGreen1 fluorescence. Genes inserted into the MCS must contain a start codon (ATG) and a stop codon. pEF1α-IRES2-ZsGreen1 can be introduced into mammalian cells using any standard transfection method. Cells expressing ZsGreen1 can be detected by flow cytometry or fluorescence microscopy 24 hours after transfection. However, in some cases, up to 48 hours may be required for detection. If required, stable transformants can be selected using G418 (7).
Propagation in E. coli
Suitable host strains: DH5α and other general purpose strains. Single-stranded DNA production requires a host containing an F plasmid such as JM101 or XL1-Blue.
Selectable marker: plasmid confers resistance to kanamycin (50 μg/ml) in E. coli hosts.
E. coli replication origin: pUC
Copy number: high
Excitation and Emission Maxima of ZsGreen1
Excitation: 493 nm
Emission: 505 nm
pEF1α-IRES-ZsGreen1质粒序列载体序列
LOCUS pEF1α-IRES-ZsGreen1 6040 bp DNA SYN
DEFINITION pEF1α-IRES-ZsGreen1
ACCESSION
KEYWORDS
SOURCE
ORGANISM other sequences; artificial sequences; vectors.
FEATURES Location/Qualifiers
source 1..6040
/organism="pEF1α-IRES-ZsGreen1"
/mol_type="other DNA"
promoter 159..1346
/label="EF1a_promoter"
CDS complement(439..1086)
/label="ORF frame 2"
/translation="MKRRLRTERPFSFVWVTHPPALPSAASSILSSLQQGREAAIFPL
TQLVPTGPALPPRAGRYTAARGQAPEQAGQLETTPVRFSVAALAGPASPNMCAGTHGP
RRRPRPQKPKYQCADLGPHLQDYLARKKASQQVIKNFKWLETYRKQRDRREGATRFAR
GGPSAQARPQLKHEAKGLLKRKASNSPTHFQPEARDQESRTAARGVEVIQGTQGP*"
misc_feature 1294..1314
/label="EF1a_fwd_primer"
/translation="MKRRLRTERPFSFVWVTHPPALPSAASSILSSLQQGREAAIFPL
TQLVPTGPALPPRAGRYTAARGQAPEQAGQLETTPVRFSVAALAGPASPNMCAGTHGP
RRRPRPQKPKYQCADLGPHLQDYLARKKASQQVIKNFKWLETYRKQRDRREGATRFAR
GGPSAQARPQLKHEAKGLLKRKASNSPTHFQPEARDQESRTAARGVEVIQGTQGP*"
misc_feature 1423..2007
/label="IRES"
/translation="MKRRLRTERPFSFVWVTHPPALPSAASSILSSLQQGREAAIFPL
TQLVPTGPALPPRAGRYTAARGQAPEQAGQLETTPVRFSVAALAGPASPNMCAGTHGP
RRRPRPQKPKYQCADLGPHLQDYLARKKASQQVIKNFKWLETYRKQRDRREGATRFAR
GGPSAQARPQLKHEAKGLLKRKASNSPTHFQPEARDQESRTAARGVEVIQGTQGP*"
CDS 1999..2706
/label="ORF frame 1"
/translation="MATTMAQSKHGLTKEMTMKYRMEGCVDGHKFVITGEGIGYPFKG
KQAINLCVVEGGPLPFAEDILSAAFMYGNRVFTEYPQDIVDYFKNSCPAGYTWDRSFL
FEDGAVCICNADITVSVEENCMYHESKFYGVNFPADGPVMKKMTDNWEPSCEKIIPVP
KQGILKGDVSMYLLLKDGGRLRCQFDTVYKAKSVPRKMPDWHFIQHKLTREDRSDAKN
QKWHLTEHAIASGSALP*"
gene 2014..2703
/label="Zs_Green"
/gene="Zs_Green"
/translation="MATTMAQSKHGLTKEMTMKYRMEGCVDGHKFVITGEGIGYPFKG
KQAINLCVVEGGPLPFAEDILSAAFMYGNRVFTEYPQDIVDYFKNSCPAGYTWDRSFL
FEDGAVCICNADITVSVEENCMYHESKFYGVNFPADGPVMKKMTDNWEPSCEKIIPVP
KQGILKGDVSMYLLLKDGGRLRCQFDTVYKAKSVPRKMPDWHFIQHKLTREDRSDAKN
QKWHLTEHAIASGSALP*"
misc_feature 2919..2938
/label="EBV_rev_primer"
/translation="MATTMAQSKHGLTKEMTMKYRMEGCVDGHKFVITGEGIGYPFKG
KQAINLCVVEGGPLPFAEDILSAAFMYGNRVFTEYPQDIVDYFKNSCPAGYTWDRSFL
FEDGAVCICNADITVSVEENCMYHESKFYGVNFPADGPVMKKMTDNWEPSCEKIIPVP
KQGILKGDVSMYLLLKDGGRLRCQFDTVYKAKSVPRKMPDWHFIQHKLTREDRSDAKN
QKWHLTEHAIASGSALP*"
rep_origin complement(3088..3394)
/label="f1_origin"
/translation="MATTMAQSKHGLTKEMTMKYRMEGCVDGHKFVITGEGIGYPFKG
KQAINLCVVEGGPLPFAEDILSAAFMYGNRVFTEYPQDIVDYFKNSCPAGYTWDRSFL
FEDGAVCICNADITVSVEENCMYHESKFYGVNFPADGPVMKKMTDNWEPSCEKIIPVP
KQGILKGDVSMYLLLKDGGRLRCQFDTVYKAKSVPRKMPDWHFIQHKLTREDRSDAKN
QKWHLTEHAIASGSALP*"
promoter 3473..3501
/label="AmpR_promoter"
/translation="MATTMAQSKHGLTKEMTMKYRMEGCVDGHKFVITGEGIGYPFKG
KQAINLCVVEGGPLPFAEDILSAAFMYGNRVFTEYPQDIVDYFKNSCPAGYTWDRSFL
FEDGAVCICNADITVSVEENCMYHESKFYGVNFPADGPVMKKMTDNWEPSCEKIIPVP
KQGILKGDVSMYLLLKDGGRLRCQFDTVYKAKSVPRKMPDWHFIQHKLTREDRSDAKN
QKWHLTEHAIASGSALP*"
misc_feature complement(3567..3587)
/label="pBABE_3_primer"
/translation="MATTMAQSKHGLTKEMTMKYRMEGCVDGHKFVITGEGIGYPFKG
KQAINLCVVEGGPLPFAEDILSAAFMYGNRVFTEYPQDIVDYFKNSCPAGYTWDRSFL
FEDGAVCICNADITVSVEENCMYHESKFYGVNFPADGPVMKKMTDNWEPSCEKIIPVP
KQGILKGDVSMYLLLKDGGRLRCQFDTVYKAKSVPRKMPDWHFIQHKLTREDRSDAKN
QKWHLTEHAIASGSALP*"
misc_feature complement(3573..3788)
/label="SV40_enhancer"
/translation="MATTMAQSKHGLTKEMTMKYRMEGCVDGHKFVITGEGIGYPFKG
KQAINLCVVEGGPLPFAEDILSAAFMYGNRVFTEYPQDIVDYFKNSCPAGYTWDRSFL
FEDGAVCICNADITVSVEENCMYHESKFYGVNFPADGPVMKKMTDNWEPSCEKIIPVP
KQGILKGDVSMYLLLKDGGRLRCQFDTVYKAKSVPRKMPDWHFIQHKLTREDRSDAKN
QKWHLTEHAIASGSALP*"
promoter 3585..3853
/label="SV40_promoter"
/translation="MATTMAQSKHGLTKEMTMKYRMEGCVDGHKFVITGEGIGYPFKG
KQAINLCVVEGGPLPFAEDILSAAFMYGNRVFTEYPQDIVDYFKNSCPAGYTWDRSFL
FEDGAVCICNADITVSVEENCMYHESKFYGVNFPADGPVMKKMTDNWEPSCEKIIPVP
KQGILKGDVSMYLLLKDGGRLRCQFDTVYKAKSVPRKMPDWHFIQHKLTREDRSDAKN
QKWHLTEHAIASGSALP*"
rep_origin 3752..3829
/label="SV40_origin"
/translation="MATTMAQSKHGLTKEMTMKYRMEGCVDGHKFVITGEGIGYPFKG
KQAINLCVVEGGPLPFAEDILSAAFMYGNRVFTEYPQDIVDYFKNSCPAGYTWDRSFL
FEDGAVCICNADITVSVEENCMYHESKFYGVNFPADGPVMKKMTDNWEPSCEKIIPVP
KQGILKGDVSMYLLLKDGGRLRCQFDTVYKAKSVPRKMPDWHFIQHKLTREDRSDAKN
QKWHLTEHAIASGSALP*"
misc_feature 3814..3833
/label="SV40pro_F_primer"
/translation="MATTMAQSKHGLTKEMTMKYRMEGCVDGHKFVITGEGIGYPFKG
KQAINLCVVEGGPLPFAEDILSAAFMYGNRVFTEYPQDIVDYFKNSCPAGYTWDRSFL
FEDGAVCICNADITVSVEENCMYHESKFYGVNFPADGPVMKKMTDNWEPSCEKIIPVP
KQGILKGDVSMYLLLKDGGRLRCQFDTVYKAKSVPRKMPDWHFIQHKLTREDRSDAKN
QKWHLTEHAIASGSALP*"
CDS 3936..4730
/label="ORF frame 3"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR
PVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL
LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
EHQGLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRY
QDIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
gene 3939..4727
/label="NeoR/KanR"
/gene="NeoR/KanR"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR
PVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL
LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
EHQGLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRY
QDIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
CDS complement(4245..4781)
/label="ORF frame 3"
/translation="MAGWASLGRSFRTPESRSEELVKKAIEGDALRIGSGDTVKHEEA
VSPFAAKLFSNITGSQRYVLIAVRHTQPATVDESRKAAIFHHDIRQAGIAMGHDEILA
VGHARLEPGEQFGWREPLMLFVQIILIDKTGFHPSTCSLDAMFRLVVEWAGSRIKRMQ
PPHCISHDGYFLGRSKVR*"
terminator 4905..5174
/label="TK_PA_terminator"
/translation="MAGWASLGRSFRTPESRSEELVKKAIEGDALRIGSGDTVKHEEA
VSPFAAKLFSNITGSQRYVLIAVRHTQPATVDESRKAAIFHHDIRQAGIAMGHDEILA
VGHARLEPGEQFGWREPLMLFVQIILIDKTGFHPSTCSLDAMFRLVVEWAGSRIKRMQ
PPHCISHDGYFLGRSKVR*"
rep_origin 5322..5941
/label="pBR322_origin"
/translation="MAGWASLGRSFRTPESRSEELVKKAIEGDALRIGSGDTVKHEEA
VSPFAAKLFSNITGSQRYVLIAVRHTQPATVDESRKAAIFHHDIRQAGIAMGHDEILA
VGHARLEPGEQFGWREPLMLFVQIILIDKTGFHPSTCSLDAMFRLVVEWAGSRIKRMQ
PPHCISHDGYFLGRSKVR*"
ORIGIN
1 TAGTTATTAA TGAGTAATTC ATACAAAAGG ACTCGCCCCT GCCTTGGGGA ATCCCAGGGA
61 CCGTCGTTAA ACTCCCACTA ACGTAGAACC CAGAGATCGC TGCGTTCCCG CCCCCTCACC
121 CGCCCGCTCT CGTCATCACT GAGGTGGAGA AGAGCATGCG TGAGGCTCCG GTGCCCGTCA
181 GTGGGCAGAG CGCACATCGC CCACAGTCCC CGAGAAGTTG GGGGGAGGGG TCGGCAATTG
241 AACCGGTGCC TAGAGAAGGT GGCGCGGGGT AAACTGGGAA AGTGATGTCG TGTACTGGCT
301 CCGCCTTTTT CCCGAGGGTG GGGGAGAACC GTATATAAGT GCAGTAGTCG CCGTGAACGT
361 TCTTTTTCGC AACGGGTTTG CCGCCAGAAC ACAGGTAAGT GCCGTGTGTG GTTCCCGCGG
421 GCCTGGCCTC TTTACGGGTT ATGGCCCTTG CGTGCCTTGA ATTACTTCCA CGCCCCTGGC
481 TGCAGTACGT GATTCTTGAT CCCGAGCTTC GGGTTGGAAG TGGGTGGGAG AGTTCGAGGC
541 CTTGCGCTTA AGGAGCCCCT TCGCCTCGTG CTTGAGTTGA GGCCTGGCTT GGGCGCTGGG
601 GCCGCCGCGT GCGAATCTGG TGGCACCTTC GCGCCTGTCT CGCTGCTTTC GATAAGTCTC
661 TAGCCATTTA AAATTTTTGA TGACCTGCTG CGACGCTTTT TTTCTGGCAA GATAGTCTTG
721 TAAATGCGGG CCAAGATCTG CACACTGGTA TTTCGGTTTT TGGGGCCGCG GGCGGCGACG
781 GGGCCCGTGC GTCCCAGCGC ACATGTTCGG CGAGGCGGGG CCTGCGAGCG CGGCCACCGA
841 GAATCGGACG GGGGTAGTCT CAAGCTGGCC GGCCTGCTCT GGTGCCTGGC CTCGCGCCGC
901 CGTGTATCGC CCCGCCCTGG GCGGCAAGGC TGGCCCGGTC GGCACCAGTT GCGTGAGCGG
961 AAAGATGGCC GCTTCCCGGC CCTGCTGCAG GGAGCTCAAA ATGGAGGACG CGGCGCTCGG
1021 GAGAGCGGGC GGGTGAGTCA CCCACACAAA GGAAAAGGGC CTTTCCGTCC TCAGCCGTCG
1081 CTTCATGTGA CTCCACGGAG TACCGGGCGC CGTCCAGGCA CCTCGATTAG TTCTCGAGCT
1141 TTTGGAGTAC GTCGTCTTTA GGTTGGGGGG AGGGGTTTTA TGCGATGGAG TTTCCCCACA
1201 CTGAGTGGGT GGAGACTGAA GTTAGGCCAG CTTGGCACTT GATGTAATTC TCCTTGGAAT
1261 TTGCCCTTTT TGAGTTTGGA TCTTGGTTCA TTCTCAAGCC TCAGACAGTG GTTCAAAGTT
1321 TTTTTCTTCC ATTTCAGGTG TCGTGACGCT AGCGCTACCG GACTCAGATC TCGAGCTCAA
1381 GCTTCGAATT CTGCAGTCGA CGGTACCGCG GGCCCGGGAT CCGCCCCTCT CCCTCCCCCC
1441 CCCCTAACGT TACTGGCCGA AGCCGCTTGG AATAAGGCCG GTGTGCGTTT GTCTATATGT
1501 TATTTTCCAC CATATTGCCG TCTTTTGGCA ATGTGAGGGC CCGGAAACCT GGCCCTGTCT
1561 TCTTGACGAG CATTCCTAGG GGTCTTTCCC CTCTCGCCAA AGGAATGCAA GGTCTGTTGA
1621 ATGTCGTGAA GGAAGCAGTT CCTCTGGAAG CTTCTTGAAG ACAAACAACG TCTGTAGCGA
1681 CCCTTTGCAG GCAGCGGAAC CCCCCACCTG GCGACAGGTG CCTCTGCGGC CAAAAGCCAC
1741 GTGTATAAGA TACACCTGCA AAGGCGGCAC AACCCCAGTG CCACGTTGTG AGTTGGATAG
1801 TTGTGGAAAG AGTCAAATGG CTCTCCTCAA GCGTATTCAA CAAGGGGCTG AAGGATGCCC
1861 AGAAGGTACC CCATTGTATG GGATCTGATC TGGGGCCTCG GTACACATGC TTTACATGTG
1921 TTTAGTCGAG GTTAAAAAAA CGTCTAGGCC CCCCGAACCA CGGGGACGTG GTTTTCCTTT
1981 GAAAAACACG ATGATAATAT GGCCACAACC ATGGCCCAGT CCAAGCACGG CCTGACCAAG
2041 GAGATGACCA TGAAGTACCG CATGGAGGGC TGCGTGGACG GCCACAAGTT CGTGATCACC
2101 GGCGAGGGCA TCGGCTACCC CTTCAAGGGC AAGCAGGCCA TCAACCTGTG CGTGGTGGAG
2161 GGCGGCCCCT TGCCCTTCGC CGAGGACATC TTGTCCGCCG CCTTCATGTA CGGCAACCGC
2221 GTGTTCACCG AGTACCCCCA GGACATCGTC GACTACTTCA AGAACTCCTG CCCCGCCGGC
2281 TACACCTGGG ACCGCTCCTT CCTGTTCGAG GACGGCGCCG TGTGCATCTG CAACGCCGAC
2341 ATCACCGTGA GCGTGGAGGA GAACTGCATG TACCACGAGT CCAAGTTCTA CGGCGTGAAC
2401 TTCCCCGCCG ACGGCCCCGT GATGAAGAAG ATGACCGACA ACTGGGAGCC CTCCTGCGAG
2461 AAGATCATCC CCGTGCCCAA GCAGGGCATC TTGAAGGGCG ACGTGAGCAT GTACCTGCTG
2521 CTGAAGGACG GTGGCCGCTT GCGCTGCCAG TTCGACACCG TGTACAAGGC CAAGTCCGTG
2581 CCCCGCAAGA TGCCCGACTG GCACTTCATC CAGCACAAGC TGACCCGCGA GGACCGCAGC
2641 GACGCCAAGA ACCAGAAGTG GCACCTGACC GAGCACGCCA TCGCCTCCGG CTCCGCCTTG
2701 CCCTGAGCGG CCGCGACTCT AGATCATAAT CAGCCATACC ACATTTGTAG AGGTTTTACT
2761 TGCTTTAAAA AACCTCCCAC ACCTCCCCCT GAACCTGAAA CATAAAATGA ATGCAATTGT
2821 TGTTGTTAAC TTGTTTATTG CAGCTTATAA TGGTTACAAA TAAAGCAATA GCATCACAAA
2881 TTTCACAAAT AAAGCATTTT TTTCACTGCA TTCTAGTTGT GGTTTGTCCA AACTCATCAA
2941 TGTATCTTAA GGCGTAAATT GTAAGCGTTA ATATTTTGTT AAAATTCGCG TTAAATTTTT
3001 GTTAAATCAG CTCATTTTTT AACCAATAGG CCGAAATCGG CAAAATCCCT TATAAATCAA
3061 AAGAATAGAC CGAGATAGGG TTGAGTGTTG TTCCAGTTTG GAACAAGAGT CCACTATTAA
3121 AGAACGTGGA CTCCAACGTC AAAGGGCGAA AAACCGTCTA TCAGGGCGAT GGCCCACTAC
3181 GTGAACCATC ACCCTAATCA AGTTTTTTGG GGTCGAGGTG CCGTAAAGCA CTAAATCGGA
3241 ACCCTAAAGG GAGCCCCCGA TTTAGAGCTT GACGGGGAAA GCCGGCGAAC GTGGCGAGAA
3301 AGGAAGGGAA GAAAGCGAAA GGAGCGGGCG CTAGGGCGCT GGCAAGTGTA GCGGTCACGC
3361 TGCGCGTAAC CACCACACCC GCCGCGCTTA ATGCGCCGCT ACAGGGCGCG TCAGGTGGCA
3421 CTTTTCGGGG AAATGTGCGC GGAACCCCTA TTTGTTTATT TTTCTAAATA CATTCAAATA
3481 TGTATCCGCT CATGAGACAA TAACCCTGAT AAATGCTTCA ATAATATTGA AAAAGGAAGA
3541 GTCCTGAGGC GGAAAGAACC AGCTGTGGAA TGTGTGTCAG TTAGGGTGTG GAAAGTCCCC
3601 AGGCTCCCCA GCAGGCAGAA GTATGCAAAG CATGCATCTC AATTAGTCAG CAACCAGGTG
3661 TGGAAAGTCC CCAGGCTCCC CAGCAGGCAG AAGTATGCAA AGCATGCATC TCAATTAGTC
3721 AGCAACCATA GTCCCGCCCC TAACTCCGCC CATCCCGCCC CTAACTCCGC CCAGTTCCGC
3781 CCATTCTCCG CCCCATGGCT GACTAATTTT TTTTATTTAT GCAGAGGCCG AGGCCGCCTC
3841 GGCCTCTGAG CTATTCCAGA AGTAGTGAGG AGGCTTTTTT GGAGGCCTAG GCTTTTGCAA
3901 AGATCGATCA AGAGACAGGA TGAGGATCGT TTCGCATGAT TGAACAAGAT GGATTGCACG
3961 CAGGTTCTCC GGCCGCTTGG GTGGAGAGGC TATTCGGCTA TGACTGGGCA CAACAGACAA
4021 TCGGCTGCTC TGATGCCGCC GTGTTCCGGC TGTCAGCGCA GGGGCGCCCG GTTCTTTTTG
4081 TCAAGACCGA CCTGTCCGGT GCCCTGAATG AACTGCAAGA CGAGGCAGCG CGGCTATCGT
4141 GGCTGGCCAC GACGGGCGTT CCTTGCGCAG CTGTGCTCGA CGTTGTCACT GAAGCGGGAA
4201 GGGACTGGCT GCTATTGGGC GAAGTGCCGG GGCAGGATCT CCTGTCATCT CACCTTGCTC
4261 CTGCCGAGAA AGTATCCATC ATGGCTGATG CAATGCGGCG GCTGCATACG CTTGATCCGG
4321 CTACCTGCCC ATTCGACCAC CAAGCGAAAC ATCGCATCGA GCGAGCACGT ACTCGGATGG
4381 AAGCCGGTCT TGTCGATCAG GATGATCTGG ACGAAGAGCA TCAGGGGCTC GCGCCAGCCG
4441 AACTGTTCGC CAGGCTCAAG GCGAGCATGC CCGACGGCGA GGATCTCGTC GTGACCCATG
4501 GCGATGCCTG CTTGCCGAAT ATCATGGTGG AAAATGGCCG CTTTTCTGGA TTCATCGACT
4561 GTGGCCGGCT GGGTGTGGCG GACCGCTATC AGGACATAGC GTTGGCTACC CGTGATATTG
4621 CTGAAGAGCT TGGCGGCGAA TGGGCTGACC GCTTCCTCGT GCTTTACGGT ATCGCCGCTC
4681 CCGATTCGCA GCGCATCGCC TTCTATCGCC TTCTTGACGA GTTCTTCTGA GCGGGACTCT
4741 GGGGTTCGAA ATGACCGACC AAGCGACGCC CAACCTGCCA TCACGAGATT TCGATTCCAC
4801 CGCCGCCTTC TATGAAAGGT TGGGCTTCGG AATCGTTTTC CGGGACGCCG GCTGGATGAT
4861 CCTCCAGCGC GGGGATCTCA TGCTGGAGTT CTTCGCCCAC CCTAGGGGGA GGCTAACTGA
4921 AACACGGAAG GAGACAATAC CGGAAGGAAC CCGCGCTATG ACGGCAATAA AAAGACAGAA
4981 TAAAACGCAC GGTGTTGGGT CGTTTGTTCA TAAACGCGGG GTTCGGTCCC AGGGCTGGCA
5041 CTCTGTCGAT ACCCCACCGA GACCCCATTG GGGCCAATAC GCCCGCGTTT CTTCCTTTTC
5101 CCCACCCCAC CCCCCAAGTT CGGGTGAAGG CCCAGGGCTC GCAGCCAACG TCGGGGCGGC
5161 AGGCCCTGCC ATAGCCTCAG GTTACTCATA TATACTTTAG ATTGATTTAA AACTTCATTT
5221 TTAATTTAAA AGGATCTAGG TGAAGATCCT TTTTGATAAT CTCATGACCA AAATCCCTTA
5281 ACGTGAGTTT TCGTTCCACT GAGCGTCAGA CCCCGTAGAA AAGATCAAAG GATCTTCTTG
5341 AGATCCTTTT TTTCTGCGCG TAATCTGCTG CTTGCAAACA AAAAAACCAC CGCTACCAGC
5401 GGTGGTTTGT TTGCCGGATC AAGAGCTACC AACTCTTTTT CCGAAGGTAA CTGGCTTCAG
5461 CAGAGCGCAG ATACCAAATA CTGTTCTTCT AGTGTAGCCG TAGTTAGGCC ACCACTTCAA
5521 GAACTCTGTA GCACCGCCTA CATACCTCGC TCTGCTAATC CTGTTACCAG TGGCTGCTGC
5581 CAGTGGCGAT AAGTCGTGTC TTACCGGGTT GGACTCAAGA CGATAGTTAC CGGATAAGGC
5641 GCAGCGGTCG GGCTGAACGG GGGGTTCGTG CACACAGCCC AGCTTGGAGC GAACGACCTA
5701 CACCGAACTG AGATACCTAC AGCGTGAGCT ATGAGAAAGC GCCACGCTTC CCGAAGGGAG
5761 AAAGGCGGAC AGGTATCCGG TAAGCGGCAG GGTCGGAACA GGAGAGCGCA CGAGGGAGCT
5821 TCCAGGGGGA AACGCCTGGT ATCTTTATAG TCCTGTCGGG TTTCGCCACC TCTGACTTGA
5881 GCGTCGATTT TTGTGATGCT CGTCAGGGGG GCGGAGCCTA TGGAAAAACG CCAGCAACGC
5941 GGCCTTTTTA CGGTTCCTGG CCTTTTGCTG GCCTTTTGCT CACATGTTCT TTCCTGCGTT
6001 ATCCCCTGAT TCTGTGGATA ACCGTATTAC CGCCATGCAT
//
