pDsRed-Monomer-N In-Fusion Rea

pDsRed-Monomer-N In-Fusion Ready

载体基本信息

出品公司:	Ybscience
载体名称:	pDsRed-Monomer-N In-Fusion Ready
质粒类型:	哺乳动物细胞表达载体；荧光报告载体
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:	CMV
载体大小:	4741 bp
5' 测序引物及序列:	5’-GTACTGGAACTGGGGGGACAG-3’
3' 测序引物及序列:	--
载体标签:	DsRed-Monomer(C-端）
载体抗性:	卡那霉素
筛选标记:	新霉素（Neomycin）
克隆菌株:	DH5α, HB101
宿主细胞（系）:	常规细胞系，293、CV-1、CHO等
备注:	pDsRed-Monomer-N In-Fusion Ready载体是荧光报告载体，表达C端DsRed-Monomer融合蛋白； DsRed-Monomer是DsRed的单体突变体，与DsRed相比，编码序列经过了优化，适用于在哺乳动物细胞中的高水平表达。
稳定性:	瞬表达 或 稳表达
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

载体简介

 

pDsRed-Monomer-N In-Fusion Ready载体描述

pDsRed-Monomer-N In-Fusion Ready vector is a linearized mammalian expression vector that encodes DsRed-Monomer (DsRed. M1), a monomeric mutant derived from the tetrameric Discosoma sp. red fluorescent protein, DsRed (1). DsRed-Monomer contains a total of 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 hr after transfection. DsRed-Monomer has an excitation maximum at 557 nm and an emission maximum at 592 nm. The DsRed-Monomer coding sequence is human codon-optimized for high expression levels in mammalian cells (2).The linearized vector allows direct cloning of PCR products without any need for restriction digestion when using the In-Fusion HD Cloning Plus (638909). This is accomplished by the use of a specific 15 nucleotide long sequence within the 5’ ends of the sense and antisense primers that overlap with the cut ends of the linearized vector.

The primers that will be used to amplify In-Fusion Ready PCR products require the following 15 nucleotides on their 5’ends:
Sense primer: 5’ AAGGCCTCTGTCGAC followed by sequence of amplification target 3’
Antisense primer: 5’ AGAATTCGCAAGCTT followed by sequence of amplification target 3’
If the sequence of the gene of interest is added in-frame immediately after the 15 nucleotides mentioned above, this sequence will automatically be in-frame with the DsRed-Monomer sequence downstream and therefore be expressed as a fusion protein to the N-terminus of DsRed-Monomer.
A Kozak consensus translation initiation site upstream of DsRed-Monomer increases the translation efficiency in eukaryotic cells (3). SV40 polyadenylation signals downstream of the DsRed-Monomer gene direct proper processing of the 3' end of the mRNA. transcript. The vector backbone contains an SV40 origin of replication in mammalian cells expressing the SV40 T antigen, a pUC origin of replication for propagation in E. coli, and an f1 origin for single-stranded DNA production.
A neomycin-resistance cassette (Neor) allows selection of stably transfected eukaryotic cells using G418. This cassette contains the SV40 early promoter, the neomycin/kanamycin resistance gene of Tn5, and polyadenylation signals from the Herpes simplex virus thymidine kinase (HSV TK) gene. A bacterial promoter upstream of the cassette allows the plasmid to confer kanamycin resistance in E. coli.
Fusions to the N-terminus of DsRed-Monomer retain the fluorescent properties of the native protein and allow monitoring of the fusion protein localization in vivo. The PCR-amplified gene of interest is directly cloned into the pDsRed-Monomer-N In-Fusion Ready Vector so that it is in-frame with the DsRed-Monomer coding sequences with no intervening in-frame stop codons. This can be accomplished by using the suggested primers. The inserted gene must include the initiating ATG codon. This recombinant vector can be transfected into mammalian cells using any standard transfection method. Stable transformants can be selected using G418 (2).
The DsRed1-N Sequencing Primer (Cat. No. 632387) can be used to sequence genes cloned adjacent to the 5’ end of the DsRed-Monomer coding region.
For Western blotting, the Living Colors DsRed Polyclonal Antibody (Cat. No. 632496) can be used to recognize the DsRed-Monomer protein. For optimal results for immunoprecipitation, we suggest you titrate the amount of antibody needed for quantitative recovery of the antigen. As a starting point, however, we recommend using the antibody at a 1:1,000 dilution.

Propagation in E. coli
 Suitable host strains: Stellar Competent Cells.
 Selectable marker: plasmid confers resistance to kanamycin (50 μg/ml) in E. coli hosts.
 E. coli replication origin: pUC
 Copy number: ~ 500
 Plasmid incompatibility group: pMB1/ColE1

Excitation and emission maxima of DsRed-Monomer
 Excitation maximum = 557 nm
 Emission maximum = 592 nm

 

载体序列

LOCUS       pDsRed-Monomer-N In-Fusion Ready	4655 bp 	DNA	SYN
DEFINITION  pDsRed-Monomer-N In-Fusion Ready
ACCESSION   
KEYWORDS    
SOURCE      
  ORGANISM  other sequences; artificial sequences; vectors.
FEATURES             Location/Qualifiers
     source          1..4655
                     /organism="pDsRed-Monomer-N In-Fusion Ready"
                     /mol_type="other DNA"
     gene            30..701
                     /label="DsRed_Monomer"
                     /gene="DsRed_Monomer"
     CDS             60..704
                     /label="ORF frame 3"
                     /translation="MQFKVRMEGSVNGHYFEIEGEGEGKPYEGTQTAKLQVTKGGPLP
                     FAWDILSPQFQYGSKAYVKHPADIPDYMKLSFPEGFTWERSMNFEDGGVVEVQQDSSL
                     QDGTFIYKVKFKGVNFPADGPVMQKKTAGWEPSTEKLYPQDGVLKGEISHALKLKDGG
                     HYTCDFKTVYKAKKPVQLPGNHYVDSKLDITNHNEDYTVVEQYEHAEARHSGSQ*"
     misc_feature    complement(207..227)
                     /label="dsRed1_N_primer"
                     /translation="MQFKVRMEGSVNGHYFEIEGEGEGKPYEGTQTAKLQVTKGGPLP
                     FAWDILSPQFQYGSKAYVKHPADIPDYMKLSFPEGFTWERSMNFEDGGVVEVQQDSSL
                     QDGTFIYKVKFKGVNFPADGPVMQKKTAGWEPSTEKLYPQDGVLKGEISHALKLKDGG
                     HYTCDFKTVYKAKKPVQLPGNHYVDSKLDITNHNEDYTVVEQYEHAEARHSGSQ*"
     misc_feature    923..942
                     /label="EBV_rev_primer"
                     /translation="MQFKVRMEGSVNGHYFEIEGEGEGKPYEGTQTAKLQVTKGGPLP
                     FAWDILSPQFQYGSKAYVKHPADIPDYMKLSFPEGFTWERSMNFEDGGVVEVQQDSSL
                     QDGTFIYKVKFKGVNFPADGPVMQKKTAGWEPSTEKLYPQDGVLKGEISHALKLKDGG
                     HYTCDFKTVYKAKKPVQLPGNHYVDSKLDITNHNEDYTVVEQYEHAEARHSGSQ*"
     rep_origin      complement(1092..1398)
                     /label="f1_origin"
                     /translation="MQFKVRMEGSVNGHYFEIEGEGEGKPYEGTQTAKLQVTKGGPLP
                     FAWDILSPQFQYGSKAYVKHPADIPDYMKLSFPEGFTWERSMNFEDGGVVEVQQDSSL
                     QDGTFIYKVKFKGVNFPADGPVMQKKTAGWEPSTEKLYPQDGVLKGEISHALKLKDGG
                     HYTCDFKTVYKAKKPVQLPGNHYVDSKLDITNHNEDYTVVEQYEHAEARHSGSQ*"
     promoter        1477..1505
                     /label="AmpR_promoter"
                     /translation="MQFKVRMEGSVNGHYFEIEGEGEGKPYEGTQTAKLQVTKGGPLP
                     FAWDILSPQFQYGSKAYVKHPADIPDYMKLSFPEGFTWERSMNFEDGGVVEVQQDSSL
                     QDGTFIYKVKFKGVNFPADGPVMQKKTAGWEPSTEKLYPQDGVLKGEISHALKLKDGG
                     HYTCDFKTVYKAKKPVQLPGNHYVDSKLDITNHNEDYTVVEQYEHAEARHSGSQ*"
     misc_feature    complement(1571..1591)
                     /label="pBABE_3_primer"
                     /translation="MQFKVRMEGSVNGHYFEIEGEGEGKPYEGTQTAKLQVTKGGPLP
                     FAWDILSPQFQYGSKAYVKHPADIPDYMKLSFPEGFTWERSMNFEDGGVVEVQQDSSL
                     QDGTFIYKVKFKGVNFPADGPVMQKKTAGWEPSTEKLYPQDGVLKGEISHALKLKDGG
                     HYTCDFKTVYKAKKPVQLPGNHYVDSKLDITNHNEDYTVVEQYEHAEARHSGSQ*"
     misc_feature    complement(1577..1792)
                     /label="SV40_enhancer"
                     /translation="MQFKVRMEGSVNGHYFEIEGEGEGKPYEGTQTAKLQVTKGGPLP
                     FAWDILSPQFQYGSKAYVKHPADIPDYMKLSFPEGFTWERSMNFEDGGVVEVQQDSSL
                     QDGTFIYKVKFKGVNFPADGPVMQKKTAGWEPSTEKLYPQDGVLKGEISHALKLKDGG
                     HYTCDFKTVYKAKKPVQLPGNHYVDSKLDITNHNEDYTVVEQYEHAEARHSGSQ*"
     promoter        1589..1857
                     /label="SV40_promoter"
                     /translation="MQFKVRMEGSVNGHYFEIEGEGEGKPYEGTQTAKLQVTKGGPLP
                     FAWDILSPQFQYGSKAYVKHPADIPDYMKLSFPEGFTWERSMNFEDGGVVEVQQDSSL
                     QDGTFIYKVKFKGVNFPADGPVMQKKTAGWEPSTEKLYPQDGVLKGEISHALKLKDGG
                     HYTCDFKTVYKAKKPVQLPGNHYVDSKLDITNHNEDYTVVEQYEHAEARHSGSQ*"
     rep_origin      1756..1833
                     /label="SV40_origin"
                     /translation="MQFKVRMEGSVNGHYFEIEGEGEGKPYEGTQTAKLQVTKGGPLP
                     FAWDILSPQFQYGSKAYVKHPADIPDYMKLSFPEGFTWERSMNFEDGGVVEVQQDSSL
                     QDGTFIYKVKFKGVNFPADGPVMQKKTAGWEPSTEKLYPQDGVLKGEISHALKLKDGG
                     HYTCDFKTVYKAKKPVQLPGNHYVDSKLDITNHNEDYTVVEQYEHAEARHSGSQ*"
     misc_feature    1818..1837
                     /label="SV40pro_F_primer"
                     /translation="MQFKVRMEGSVNGHYFEIEGEGEGKPYEGTQTAKLQVTKGGPLP
                     FAWDILSPQFQYGSKAYVKHPADIPDYMKLSFPEGFTWERSMNFEDGGVVEVQQDSSL
                     QDGTFIYKVKFKGVNFPADGPVMQKKTAGWEPSTEKLYPQDGVLKGEISHALKLKDGG
                     HYTCDFKTVYKAKKPVQLPGNHYVDSKLDITNHNEDYTVVEQYEHAEARHSGSQ*"
     CDS             1940..2734
                     /label="ORF frame 2"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR
                     PVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL
                     LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRY
                     QDIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
     gene            1943..2731
                     /label="NeoR/KanR"
                     /gene="NeoR/KanR"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR
                     PVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL
                     LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRY
                     QDIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
     CDS             complement(2249..2785)
                     /label="ORF frame 2"
                     /translation="MAGWASLGRSFRTPESRSEELVKKAIEGDALRIGSGDTVKHEEA
                     VSPFAAKLFSNITGSQRYVLIAVRHTQPATVDESRKAAIFHHDIRQAGIAMGHDEILA
                     VGHARLEPGEQFGWREPLMLFVQIILIDKTGFHPSTCSLDAMFRLVVEWAGSRIKRMQ
                     PPHCISHDGYFLGRSKVR*"
     terminator      2909..3178
                     /label="TK_PA_terminator"
                     /translation="MAGWASLGRSFRTPESRSEELVKKAIEGDALRIGSGDTVKHEEA
                     VSPFAAKLFSNITGSQRYVLIAVRHTQPATVDESRKAAIFHHDIRQAGIAMGHDEILA
                     VGHARLEPGEQFGWREPLMLFVQIILIDKTGFHPSTCSLDAMFRLVVEWAGSRIKRMQ
                     PPHCISHDGYFLGRSKVR*"
     rep_origin      3326..3945
                     /label="pBR322_origin"
                     /translation="MAGWASLGRSFRTPESRSEELVKKAIEGDALRIGSGDTVKHEEA
                     VSPFAAKLFSNITGSQRYVLIAVRHTQPATVDESRKAAIFHHDIRQAGIAMGHDEILA
                     VGHARLEPGEQFGWREPLMLFVQIILIDKTGFHPSTCSLDAMFRLVVEWAGSRIKRMQ
                     PPHCISHDGYFLGRSKVR*"
     promoter        4054..4606
                     /label="CMV_immearly_promoter"
                     /translation="MAGWASLGRSFRTPESRSEELVKKAIEGDALRIGSGDTVKHEEA
                     VSPFAAKLFSNITGSQRYVLIAVRHTQPATVDESRKAAIFHHDIRQAGIAMGHDEILA
                     VGHARLEPGEQFGWREPLMLFVQIILIDKTGFHPSTCSLDAMFRLVVEWAGSRIKRMQ
                     PPHCISHDGYFLGRSKVR*"
     misc_feature    4109..4396
                     /label="CAG_enhancer"
                     /translation="MAGWASLGRSFRTPESRSEELVKKAIEGDALRIGSGDTVKHEEA
                     VSPFAAKLFSNITGSQRYVLIAVRHTQPATVDESRKAAIFHHDIRQAGIAMGHDEILA
                     VGHARLEPGEQFGWREPLMLFVQIILIDKTGFHPSTCSLDAMFRLVVEWAGSRIKRMQ
                     PPHCISHDGYFLGRSKVR*"
     misc_feature    4563..4583
                     /label="CMV_fwd_primer"
                     /translation="MAGWASLGRSFRTPESRSEELVKKAIEGDALRIGSGDTVKHEEA
                     VSPFAAKLFSNITGSQRYVLIAVRHTQPATVDESRKAAIFHHDIRQAGIAMGHDEILA
                     VGHARLEPGEQFGWREPLMLFVQIILIDKTGFHPSTCSLDAMFRLVVEWAGSRIKRMQ
                     PPHCISHDGYFLGRSKVR*"
     promoter        4564..4633
                     /label="CMV_promoter"
                     /translation="MAGWASLGRSFRTPESRSEELVKKAIEGDALRIGSGDTVKHEEA
                     VSPFAAKLFSNITGSQRYVLIAVRHTQPATVDESRKAAIFHHDIRQAGIAMGHDEILA
                     VGHARLEPGEQFGWREPLMLFVQIILIDKTGFHPSTCSLDAMFRLVVEWAGSRIKRMQ
                     PPHCISHDGYFLGRSKVR*"
ORIGIN
    1 AGCTTGCGAA TTCTGGCCTA CCGGTCGCGG ACAACACCGA GGACGTCATC AAGGAGTTCA
   61 TGCAGTTCAA GGTGCGCATG GAGGGCTCCG TGAACGGCCA CTACTTCGAG ATCGAGGGCG
  121 AGGGCGAGGG CAAGCCCTAC GAGGGCACCC AGACCGCCAA GCTGCAGGTG ACCAAGGGCG
  181 GCCCCCTGCC CTTCGCCTGG GACATCCTGT CCCCCCAGTT CCAGTACGGC TCCAAGGCCT
  241 ACGTGAAGCA CCCCGCCGAC ATCCCCGACT ACATGAAGCT GTCCTTCCCC GAGGGCTTCA
  301 CCTGGGAGCG CTCCATGAAC TTCGAGGACG GCGGCGTGGT GGAGGTGCAG CAGGACTCCT
  361 CCCTGCAGGA CGGCACCTTC ATCTACAAGG TGAAGTTCAA GGGCGTGAAC TTCCCCGCCG
  421 ACGGCCCCGT AATGCAGAAG AAGACTGCCG GCTGGGAGCC CTCCACCGAG AAGCTGTACC
  481 CCCAGGACGG CGTGCTGAAG GGCGAGATCT CCCACGCCCT GAAGCTGAAG GACGGCGGCC
  541 ACTACACCTG CGACTTCAAG ACCGTGTACA AGGCCAAGAA GCCCGTGCAG CTGCCCGGCA
  601 ACCACTACGT GGACTCCAAG CTGGACATCA CCAACCACAA CGAGGACTAC ACCGTGGTGG
  661 AGCAGTACGA GCACGCCGAG GCCCGCCACT CCGGCTCCCA GTAGGCGGCC GCGACTGCGA
  721 CTCTAGATCA TAATCAGCCA TACCACATTT GTAGAGGTTT TACTTGCTTT AAAAAACCTC
  781 CCACACCTCC CCCTGAACCT GAAACATAAA ATGAATGCAA TTGTTGTTGT TAACTTGTTT
  841 ATTGCAGCTT ATAATGGTTA CAAATAAAGC AATAGCATCA CAAATTTCAC AAATAAAGCA
  901 TTTTTTTCAC TGCATTCTAG TTGTGGTTTG TCCAAACTCA TCAATGTATC TTAAGGCGTA
  961 AATTGTAAGC GTTAATATTT TGTTAAAATT CGCGTTAAAT TTTTGTTAAA TCAGCTCATT
 1021 TTTTAACCAA TAGGCCGAAA TCGGCAAAAT CCCTTATAAA TCAAAAGAAT AGACCGAGAT
 1081 AGGGTTGAGT GTTGTTCCAG TTTGGAACAA GAGTCCACTA TTAAAGAACG TGGACTCCAA
 1141 CGTCAAAGGG CGAAAAACCG TCTATCAGGG CGATGGCCCA CTACGTGAAC CATCACCCTA
 1201 ATCAAGTTTT TTGGGGTCGA GGTGCCGTAA AGCACTAAAT CGGAACCCTA AAGGGAGCCC
 1261 CCGATTTAGA GCTTGACGGG GAAAGCCGGC GAACGTGGCG AGAAAGGAAG GGAAGAAAGC
 1321 GAAAGGAGCG GGCGCTAGGG CGCTGGCAAG TGTAGCGGTC ACGCTGCGCG TAACCACCAC
 1381 ACCCGCCGCG CTTAATGCGC CGCTACAGGG CGCGTCAGGT GGCACTTTTC GGGGAAATGT
 1441 GCGCGGAACC CCTATTTGTT TATTTTTCTA AATACATTCA AATATGTATC CGCTCATGAG
 1501 ACAATAACCC TGATAAATGC TTCAATAATA TTGAAAAAGG AAGAGTCCTG AGGCGGAAAG
 1561 AACCAGCTGT GGAATGTGTG TCAGTTAGGG TGTGGAAAGT CCCCAGGCTC CCCAGCAGGC
 1621 AGAAGTATGC AAAGCATGCA TCTCAATTAG TCAGCAACCA GGTGTGGAAA GTCCCCAGGC
 1681 TCCCCAGCAG GCAGAAGTAT GCAAAGCATG CATCTCAATT AGTCAGCAAC CATAGTCCCG
 1741 CCCCTAACTC CGCCCATCCC GCCCCTAACT CCGCCCAGTT CCGCCCATTC TCCGCCCCAT
 1801 GGCTGACTAA TTTTTTTTAT TTATGCAGAG GCCGAGGCCG CCTCGGCCTC TGAGCTATTC
 1861 CAGAAGTAGT GAGGAGGCTT TTTTGGAGGC CTAGGCTTTT GCAAAGATCG ATCAAGAGAC
 1921 AGGATGAGGA TCGTTTCGCA TGATTGAACA AGATGGATTG CACGCAGGTT CTCCGGCCGC
 1981 TTGGGTGGAG AGGCTATTCG GCTATGACTG GGCACAACAG ACAATCGGCT GCTCTGATGC
 2041 CGCCGTGTTC CGGCTGTCAG CGCAGGGGCG CCCGGTTCTT TTTGTCAAGA CCGACCTGTC
 2101 CGGTGCCCTG AATGAACTGC AAGACGAGGC AGCGCGGCTA TCGTGGCTGG CCACGACGGG
 2161 CGTTCCTTGC GCAGCTGTGC TCGACGTTGT CACTGAAGCG GGAAGGGACT GGCTGCTATT
 2221 GGGCGAAGTG CCGGGGCAGG ATCTCCTGTC ATCTCACCTT GCTCCTGCCG AGAAAGTATC
 2281 CATCATGGCT GATGCAATGC GGCGGCTGCA TACGCTTGAT CCGGCTACCT GCCCATTCGA
 2341 CCACCAAGCG AAACATCGCA TCGAGCGAGC ACGTACTCGG ATGGAAGCCG GTCTTGTCGA
 2401 TCAGGATGAT CTGGACGAAG AGCATCAGGG GCTCGCGCCA GCCGAACTGT TCGCCAGGCT
 2461 CAAGGCGAGC ATGCCCGACG GCGAGGATCT CGTCGTGACC CATGGCGATG CCTGCTTGCC
 2521 GAATATCATG GTGGAAAATG GCCGCTTTTC TGGATTCATC GACTGTGGCC GGCTGGGTGT
 2581 GGCGGACCGC TATCAGGACA TAGCGTTGGC TACCCGTGAT ATTGCTGAAG AGCTTGGCGG
 2641 CGAATGGGCT GACCGCTTCC TCGTGCTTTA CGGTATCGCC GCTCCCGATT CGCAGCGCAT
 2701 CGCCTTCTAT CGCCTTCTTG ACGAGTTCTT CTGAGCGGGA CTCTGGGGTT CGAAATGACC
 2761 GACCAAGCGA CGCCCAACCT GCCATCACGA GATTTCGATT CCACCGCCGC CTTCTATGAA
 2821 AGGTTGGGCT TCGGAATCGT TTTCCGGGAC GCCGGCTGGA TGATCCTCCA GCGCGGGGAT
 2881 CTCATGCTGG AGTTCTTCGC CCACCCTAGG GGGAGGCTAA CTGAAACACG GAAGGAGACA
 2941 ATACCGGAAG GAACCCGCGC TATGACGGCA ATAAAAAGAC AGAATAAAAC GCACGGTGTT
 3001 GGGTCGTTTG TTCATAAACG CGGGGTTCGG TCCCAGGGCT GGCACTCTGT CGATACCCCA
 3061 CCGAGACCCC ATTGGGGCCA ATACGCCCGC GTTTCTTCCT TTTCCCCACC CCACCCCCCA
 3121 AGTTCGGGTG AAGGCCCAGG GCTCGCAGCC AACGTCGGGG CGGCAGGCCC TGCCATAGCC
 3181 TCAGGTTACT CATATATACT TTAGATTGAT TTAAAACTTC ATTTTTAATT TAAAAGGATC
 3241 TAGGTGAAGA TCCTTTTTGA TAATCTCATG ACCAAAATCC CTTAACGTGA GTTTTCGTTC
 3301 CACTGAGCGT CAGACCCCGT AGAAAAGATC AAAGGATCTT CTTGAGATCC TTTTTTTCTG
 3361 CGCGTAATCT GCTGCTTGCA AACAAAAAAA CCACCGCTAC CAGCGGTGGT TTGTTTGCCG
 3421 GATCAAGAGC TACCAACTCT TTTTCCGAAG GTAACTGGCT TCAGCAGAGC GCAGATACCA
 3481 AATACTGTTC TTCTAGTGTA GCCGTAGTTA GGCCACCACT TCAAGAACTC TGTAGCACCG
 3541 CCTACATACC TCGCTCTGCT AATCCTGTTA CCAGTGGCTG CTGCCAGTGG CGATAAGTCG
 3601 TGTCTTACCG GGTTGGACTC AAGACGATAG TTACCGGATA AGGCGCAGCG GTCGGGCTGA
 3661 ACGGGGGGTT CGTGCACACA GCCCAGCTTG GAGCGAACGA CCTACACCGA ACTGAGATAC
 3721 CTACAGCGTG AGCTATGAGA AAGCGCCACG CTTCCCGAAG GGAGAAAGGC GGACAGGTAT
 3781 CCGGTAAGCG GCAGGGTCGG AACAGGAGAG CGCACGAGGG AGCTTCCAGG GGGAAACGCC
 3841 TGGTATCTTT ATAGTCCTGT CGGGTTTCGC CACCTCTGAC TTGAGCGTCG ATTTTTGTGA
 3901 TGCTCGTCAG GGGGGCGGAG CCTATGGAAA AACGCCAGCA ACGCGGCCTT TTTACGGTTC
 3961 CTGGCCTTTT GCTGGCCTTT TGCTCACATG TTCTTTCCTG CGTTATCCCC TGATTCTGTG
 4021 GATAACCGTA TTACCGCCAT GCATTAGTTA TTAATAGTAA TCAATTACGG GGTCATTAGT
 4081 TCATAGCCCA TATATGGAGT TCCGCGTTAC ATAACTTACG GTAAATGGCC CGCCTGGCTG
 4141 ACCGCCCAAC GACCCCCGCC CATTGACGTC AATAATGACG TATGTTCCCA TAGTAACGCC
 4201 AATAGGGACT TTCCATTGAC GTCAATGGGT GGAGTATTTA CGGTAAACTG CCCACTTGGC
 4261 AGTACATCAA GTGTATCATA TGCCAAGTAC GCCCCCTATT GACGTCAATG ACGGTAAATG
 4321 GCCCGCCTGG CATTATGCCC AGTACATGAC CTTATGGGAC TTTCCTACTT GGCAGTACAT
 4381 CTACGTATTA GTCATCGCTA TTACCATGGT GATGCGGTTT TGGCAGTACA TCAATGGGCG
 4441 TGGATAGCGG TTTGACTCAC GGGGATTTCC AAGTCTCCAC CCCATTGACG TCAATGGGAG
 4501 TTTGTTTTGG CACCAAAATC AACGGGACTT TCCAAAATGT CGTAACAACT CCGCCCCATT
 4561 GACGCAAATG GGCGGTAGGC GTGTACGGTG GGAGGTCTAT ATAAGCAGAG CTGGTTTAGT
 4621 GAACCGTCAG ATCCGCTAGC TAAGGCCTCT GTCGA