pEF1α-IRES-AcGFP1

价格:1800元

联系方式:I47-825O-882O

买家导航

载体基本信息

出品公司: Clontech
载体名称: pEF1α-IRES-AcGFP1
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: 6064 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、AcGFP1
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-AcGFP1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

载体质粒图谱和多克隆位点信息

pEF1α-IRES-AcGFP1载体图谱



pEF1α-IRES-AcGFP1 多克隆位点

载体简介

pEF1α-IRES-AcGFP1 is a bicistronic mammalian expression vector that allows the simultaneous, constitutive expression of a protein of interest and the green fluorescent protein AcGFP1. Expression of the bicistronic transcript is driven by the human elongation factor 1 alpha (EF1α) promoter, which continues to be constitutively active even after stable integration of the vector into the host cell genome. Stable expression of the bicistronic transcript allows the monitoring of a variety of cellular processes (such as differentiation in primary or stem cells), without the transgene silencing associated with CMV promoters. 

In addition, the vector allows efficient flow cytometric detection of stably or transiently transfected mammalian cells expressing AcGFP1 and a protein of interest, without time-consuming drug and clonal selection. 

pEF1α-IRES-AcGFP1 is designed to simultaneously and constitutively express a protein of interest and AcGFP1 in mammalian cells. AcGFP1 is a human-codon-optimized, monomeric green fluorescent protein derived from Aequorea coerulescens (the excitation and emission maxima of native AcGFP1 are 475 nm and 505 nm, respectively). Simultaneous expression of a protein of interest and AcGFP1 is made possible by the presence of an encephalomyocarditis virus (EMCV) internal ribosome entry site (IRES; 1, 2) positioned between the multiple cloning site (MCS) and the AcGFP1 gene. The IRES allows a protein of interest and AcGFP1 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 (3).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 (4). 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.

Location of Features
PEF1α (human elongation factor 1 alpha promoter): 12–1346
MCS (multiple cloning site): 1348–1422
IRES2 (internal ribosome entry site): 1423–2007
AcGFP1 (human-codon-optimized): 2011–2727
SV40 early polyA signal: 2883–2933
f1 origin of replication: 2980–3435 (complementary)
SV40 origin of replication: 3776–3911
PSV40e (SV40 early promoter and enhancer sequences): 3609–3877
Kanr/Neor(kanamycin/neomycin resistance gene): 3960–4754
HSV TK polyA signals: 4990–5008
pUC origin of replication: 5339–5982

Additional Information
pEF1α-IRES2-AcGFP1 can be used to quickly identify cells expressing a gene of interest by screening for AcGFP1 fluorescence. Genes inserted into the MCS must contain a start codon (ATG) and a stop codon. pEF1α-IRES2-AcGFP1 can be introduced into mammalian cells using any standard transfection method. Cells expressing AcGFP1can 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 of green-emitting cells. If required, stable transformants can be selected using G418 (4).

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 AcGFP1
Excitation: 475 nm
Emission: 505 nm

载体序列

LOCUS       pEF1α-IRES-AcGFP1	6064 bp 	DNA	SYN
DEFINITION  pEF1α-IRES-AcGFP1
ACCESSION   
KEYWORDS    
SOURCE      
  ORGANISM  other sequences; artificial sequences; vectors.
FEATURES             Location/Qualifiers
     source          1..6064
                     /organism="pEF1α-IRES-AcGFP1"
                     /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..2730
                     /label="ORF frame 1"
                     /translation="MATTMVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATY
                     GKLTLKFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERT
                     IFFEDDGNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTD
                     KAKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNE
                     KRDHMIYFGFVTAAAITHGMDELYK*"
     gene            2014..2727
                     /label="AcGFP1"
                     /gene="AcGFP1"
                     /translation="MATTMVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATY
                     GKLTLKFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERT
                     IFFEDDGNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTD
                     KAKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNE
                     KRDHMIYFGFVTAAAITHGMDELYK*"
     misc_feature    2943..2962
                     /label="EBV_rev_primer"
                     /translation="MATTMVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATY
                     GKLTLKFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERT
                     IFFEDDGNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTD
                     KAKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNE
                     KRDHMIYFGFVTAAAITHGMDELYK*"
     rep_origin      complement(3112..3418)
                     /label="f1_origin"
                     /translation="MATTMVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATY
                     GKLTLKFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERT
                     IFFEDDGNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTD
                     KAKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNE
                     KRDHMIYFGFVTAAAITHGMDELYK*"
     promoter        3497..3525
                     /label="AmpR_promoter"
                     /translation="MATTMVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATY
                     GKLTLKFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERT
                     IFFEDDGNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTD
                     KAKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNE
                     KRDHMIYFGFVTAAAITHGMDELYK*"
     misc_feature    complement(3591..3611)
                     /label="pBABE_3_primer"
                     /translation="MATTMVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATY
                     GKLTLKFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERT
                     IFFEDDGNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTD
                     KAKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNE
                     KRDHMIYFGFVTAAAITHGMDELYK*"
     misc_feature    complement(3597..3812)
                     /label="SV40_enhancer"
                     /translation="MATTMVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATY
                     GKLTLKFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERT
                     IFFEDDGNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTD
                     KAKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNE
                     KRDHMIYFGFVTAAAITHGMDELYK*"
     promoter        3609..3877
                     /label="SV40_promoter"
                     /translation="MATTMVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATY
                     GKLTLKFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERT
                     IFFEDDGNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTD
                     KAKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNE
                     KRDHMIYFGFVTAAAITHGMDELYK*"
     rep_origin      3776..3853
                     /label="SV40_origin"
                     /translation="MATTMVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATY
                     GKLTLKFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERT
                     IFFEDDGNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTD
                     KAKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNE
                     KRDHMIYFGFVTAAAITHGMDELYK*"
     misc_feature    3838..3857
                     /label="SV40pro_F_primer"
                     /translation="MATTMVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATY
                     GKLTLKFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERT
                     IFFEDDGNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTD
                     KAKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNE
                     KRDHMIYFGFVTAAAITHGMDELYK*"
     CDS             3960..4754
                     /label="ORF frame 3"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR
                     PVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL
                     LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRY
                     QDIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
     gene            3963..4751
                     /label="NeoR/KanR"
                     /gene="NeoR/KanR"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR
                     PVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL
                     LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRY
                     QDIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
     CDS             complement(4269..4805)
                     /label="ORF frame 3"
                     /translation="MAGWASLGRSFRTPESRSEELVKKAIEGDALRIGSGDTVKHEEA
                     VSPFAAKLFSNITGSQRYVLIAVRHTQPATVDESRKAAIFHHDIRQAGIAMGHDEILA
                     VGHARLEPGEQFGWREPLMLFVQIILIDKTGFHPSTCSLDAMFRLVVEWAGSRIKRMQ
                     PPHCISHDGYFLGRSKVR*"
     terminator      4929..5198
                     /label="TK_PA_terminator"
                     /translation="MAGWASLGRSFRTPESRSEELVKKAIEGDALRIGSGDTVKHEEA
                     VSPFAAKLFSNITGSQRYVLIAVRHTQPATVDESRKAAIFHHDIRQAGIAMGHDEILA
                     VGHARLEPGEQFGWREPLMLFVQIILIDKTGFHPSTCSLDAMFRLVVEWAGSRIKRMQ
                     PPHCISHDGYFLGRSKVR*"
     rep_origin      5346..5965
                     /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 ATGGTGAGCA AGGGCGCCGA GCTGTTCACC
 2041 GGCATCGTGC CCATCCTGAT CGAGCTGAAT GGCGATGTGA ATGGCCACAA GTTCAGCGTG
 2101 AGCGGCGAGG GCGAGGGCGA TGCCACCTAC GGCAAGCTGA CCCTGAAGTT CATCTGCACC
 2161 ACCGGCAAGC TGCCTGTGCC CTGGCCCACC CTGGTGACCA CCCTGAGCTA CGGCGTGCAG
 2221 TGCTTCTCAC GCTACCCCGA TCACATGAAG CAGCACGACT TCTTCAAGAG CGCCATGCCT
 2281 GAGGGCTACA TCCAGGAGCG CACCATCTTC TTCGAGGATG ACGGCAACTA CAAGTCGCGC
 2341 GCCGAGGTGA AGTTCGAGGG CGATACCCTG GTGAATCGCA TCGAGCTGAC CGGCACCGAT
 2401 TTCAAGGAGG ATGGCAACAT CCTGGGCAAT AAGATGGAGT ACAACTACAA CGCCCACAAT
 2461 GTGTACATCA TGACCGACAA GGCCAAGAAT GGCATCAAGG TGAACTTCAA GATCCGCCAC
 2521 AACATCGAGG ATGGCAGCGT GCAGCTGGCC GACCACTACC AGCAGAATAC CCCCATCGGC
 2581 GATGGCCCTG TGCTGCTGCC CGATAACCAC TACCTGTCCA CCCAGAGCGC CCTGTCCAAG
 2641 GACCCCAACG AGAAGCGCGA TCACATGATC TACTTCGGCT TCGTGACCGC CGCCGCCATC
 2701 ACCCACGGCA TGGATGAGCT GTACAAGTGA GCGGCCGCGA CTCTAGATCA TAATCAGCCA
 2761 TACCACATTT GTAGAGGTTT TACTTGCTTT AAAAAACCTC CCACACCTCC CCCTGAACCT
 2821 GAAACATAAA ATGAATGCAA TTGTTGTTGT TAACTTGTTT ATTGCAGCTT ATAATGGTTA
 2881 CAAATAAAGC AATAGCATCA CAAATTTCAC AAATAAAGCA TTTTTTTCAC TGCATTCTAG
 2941 TTGTGGTTTG TCCAAACTCA TCAATGTATC TTAAGGCGTA AATTGTAAGC GTTAATATTT
 3001 TGTTAAAATT CGCGTTAAAT TTTTGTTAAA TCAGCTCATT TTTTAACCAA TAGGCCGAAA
 3061 TCGGCAAAAT CCCTTATAAA TCAAAAGAAT AGACCGAGAT AGGGTTGAGT GTTGTTCCAG
 3121 TTTGGAACAA GAGTCCACTA TTAAAGAACG TGGACTCCAA CGTCAAAGGG CGAAAAACCG
 3181 TCTATCAGGG CGATGGCCCA CTACGTGAAC CATCACCCTA ATCAAGTTTT TTGGGGTCGA
 3241 GGTGCCGTAA AGCACTAAAT CGGAACCCTA AAGGGAGCCC CCGATTTAGA GCTTGACGGG
 3301 GAAAGCCGGC GAACGTGGCG AGAAAGGAAG GGAAGAAAGC GAAAGGAGCG GGCGCTAGGG
 3361 CGCTGGCAAG TGTAGCGGTC ACGCTGCGCG TAACCACCAC ACCCGCCGCG CTTAATGCGC
 3421 CGCTACAGGG CGCGTCAGGT GGCACTTTTC GGGGAAATGT GCGCGGAACC CCTATTTGTT
 3481 TATTTTTCTA AATACATTCA AATATGTATC CGCTCATGAG ACAATAACCC TGATAAATGC
 3541 TTCAATAATA TTGAAAAAGG AAGAGTCCTG AGGCGGAAAG AACCAGCTGT GGAATGTGTG
 3601 TCAGTTAGGG TGTGGAAAGT CCCCAGGCTC CCCAGCAGGC AGAAGTATGC AAAGCATGCA
 3661 TCTCAATTAG TCAGCAACCA GGTGTGGAAA GTCCCCAGGC TCCCCAGCAG GCAGAAGTAT
 3721 GCAAAGCATG CATCTCAATT AGTCAGCAAC CATAGTCCCG CCCCTAACTC CGCCCATCCC
 3781 GCCCCTAACT CCGCCCAGTT CCGCCCATTC TCCGCCCCAT GGCTGACTAA TTTTTTTTAT
 3841 TTATGCAGAG GCCGAGGCCG CCTCGGCCTC TGAGCTATTC CAGAAGTAGT GAGGAGGCTT
 3901 TTTTGGAGGC CTAGGCTTTT GCAAAGATCG ATCAAGAGAC AGGATGAGGA TCGTTTCGCA
 3961 TGATTGAACA AGATGGATTG CACGCAGGTT CTCCGGCCGC TTGGGTGGAG AGGCTATTCG
 4021 GCTATGACTG GGCACAACAG ACAATCGGCT GCTCTGATGC CGCCGTGTTC CGGCTGTCAG
 4081 CGCAGGGGCG CCCGGTTCTT TTTGTCAAGA CCGACCTGTC CGGTGCCCTG AATGAACTGC
 4141 AAGACGAGGC AGCGCGGCTA TCGTGGCTGG CCACGACGGG CGTTCCTTGC GCAGCTGTGC
 4201 TCGACGTTGT CACTGAAGCG GGAAGGGACT GGCTGCTATT GGGCGAAGTG CCGGGGCAGG
 4261 ATCTCCTGTC ATCTCACCTT GCTCCTGCCG AGAAAGTATC CATCATGGCT GATGCAATGC
 4321 GGCGGCTGCA TACGCTTGAT CCGGCTACCT GCCCATTCGA CCACCAAGCG AAACATCGCA
 4381 TCGAGCGAGC ACGTACTCGG ATGGAAGCCG GTCTTGTCGA TCAGGATGAT CTGGACGAAG
 4441 AGCATCAGGG GCTCGCGCCA GCCGAACTGT TCGCCAGGCT CAAGGCGAGC ATGCCCGACG
 4501 GCGAGGATCT CGTCGTGACC CATGGCGATG CCTGCTTGCC GAATATCATG GTGGAAAATG
 4561 GCCGCTTTTC TGGATTCATC GACTGTGGCC GGCTGGGTGT GGCGGACCGC TATCAGGACA
 4621 TAGCGTTGGC TACCCGTGAT ATTGCTGAAG AGCTTGGCGG CGAATGGGCT GACCGCTTCC
 4681 TCGTGCTTTA CGGTATCGCC GCTCCCGATT CGCAGCGCAT CGCCTTCTAT CGCCTTCTTG
 4741 ACGAGTTCTT CTGAGCGGGA CTCTGGGGTT CGAAATGACC GACCAAGCGA CGCCCAACCT
 4801 GCCATCACGA GATTTCGATT CCACCGCCGC CTTCTATGAA AGGTTGGGCT TCGGAATCGT
 4861 TTTCCGGGAC GCCGGCTGGA TGATCCTCCA GCGCGGGGAT CTCATGCTGG AGTTCTTCGC
 4921 CCACCCTAGG GGGAGGCTAA CTGAAACACG GAAGGAGACA ATACCGGAAG GAACCCGCGC
 4981 TATGACGGCA ATAAAAAGAC AGAATAAAAC GCACGGTGTT GGGTCGTTTG TTCATAAACG
 5041 CGGGGTTCGG TCCCAGGGCT GGCACTCTGT CGATACCCCA CCGAGACCCC ATTGGGGCCA
 5101 ATACGCCCGC GTTTCTTCCT TTTCCCCACC CCACCCCCCA AGTTCGGGTG AAGGCCCAGG
 5161 GCTCGCAGCC AACGTCGGGG CGGCAGGCCC TGCCATAGCC TCAGGTTACT CATATATACT
 5221 TTAGATTGAT TTAAAACTTC ATTTTTAATT TAAAAGGATC TAGGTGAAGA TCCTTTTTGA
 5281 TAATCTCATG ACCAAAATCC CTTAACGTGA GTTTTCGTTC CACTGAGCGT CAGACCCCGT
 5341 AGAAAAGATC AAAGGATCTT CTTGAGATCC TTTTTTTCTG CGCGTAATCT GCTGCTTGCA
 5401 AACAAAAAAA CCACCGCTAC CAGCGGTGGT TTGTTTGCCG GATCAAGAGC TACCAACTCT
 5461 TTTTCCGAAG GTAACTGGCT TCAGCAGAGC GCAGATACCA AATACTGTTC TTCTAGTGTA
 5521 GCCGTAGTTA GGCCACCACT TCAAGAACTC TGTAGCACCG CCTACATACC TCGCTCTGCT
 5581 AATCCTGTTA CCAGTGGCTG CTGCCAGTGG CGATAAGTCG TGTCTTACCG GGTTGGACTC
 5641 AAGACGATAG TTACCGGATA AGGCGCAGCG GTCGGGCTGA ACGGGGGGTT CGTGCACACA
 5701 GCCCAGCTTG GAGCGAACGA CCTACACCGA ACTGAGATAC CTACAGCGTG AGCTATGAGA
 5761 AAGCGCCACG CTTCCCGAAG GGAGAAAGGC GGACAGGTAT CCGGTAAGCG GCAGGGTCGG
 5821 AACAGGAGAG CGCACGAGGG AGCTTCCAGG GGGAAACGCC TGGTATCTTT ATAGTCCTGT
 5881 CGGGTTTCGC CACCTCTGAC TTGAGCGTCG ATTTTTGTGA TGCTCGTCAG GGGGGCGGAG
 5941 CCTATGGAAA AACGCCAGCA ACGCGGCCTT TTTACGGTTC CTGGCCTTTT GCTGGCCTTT
 6001 TGCTCACATG TTCTTTCCTG CGTTATCCCC TGATTCTGTG GATAACCGTA TTACCGCCAT
 6061 GCAT
//

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