pDsRed-Express-DR

pDsRed-Express-DR

载体基本信息

出品公司:	Ybscience
载体名称:	pDsRed-Express-DR
质粒类型:	无启动子载体；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:	无启动子
载体大小:	4240 bp
5' 测序引物及序列:	5'd[GTACTGGAACTGGGGGGACAG]
3' 测序引物及序列:	--
载体标签:	红色荧光蛋白DsRed-Express-DR
载体抗性:	卡那霉素
筛选标记:	新霉素（Neomycin）
克隆菌株:	TOP10, DH5α, HB101
宿主细胞（系）:	真核细胞
备注:	pDsRed-Express-DR载体不含启动子，将待研究的目的启动子插入MCS区，用于研究启动子及顺式转录元件在细胞内的作用水平； DsRed-Express-DR与DsRed-Express相比，半衰期短，适用于需要报告基因逆转速度快的研究领域。
稳定性:	瞬表达 或 稳表达
组成型/诱导型:	--
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-Express-DR载体描述

pDsRed-Express-DR is a promoterless vector that encodes DsRed-Express-DR, a destabilized variant of Discosoma sp. red fluorescent protein (DsRed). In contrast to the original protein, which is extremely stable, DsRed-Express-DR has a short half-life, making it well suited for studies that require rapid reporter turnover. Though its geneology can be traced to wild-type DsRed (1), DsRed-Express-DR is directly derived from DsRed-Express, a DsRed variant engineered for rapid fluorescence development (2, 3). DsRed-Express contains nine amino acid substitutions (listed on page 2), which improve the protein’s solubility and decrease the time from transfection to detection of red fluorescence (2). The substitutions also reduce the level of residual green emission (2). The destabilized variant, DsRed-Express-DR, was constructed by fusing the C-terminus of the protein to amino acid residues 422–461 of mouse ornithine decarboxylase (MODC), one of the most shortlived proteins in mammalian cells (4). This region of MODC contains a PEST sequence that targets the protein for degradation, resulting in rapid protein turnover (4, 5). The DsRed-Express-DR gene is human codon-optimized for high expression in mammalian cells (6), and sequences upstream of the gene have been converted to a Kozak consensus translation initiation site (7) to enhance translation efficiency in eukaryotic cells.
pDsRed-Express-DR can be used to monitor transcription from different promoters and promoter/ enhancer combinations inserted into the multiple cloning site (MCS), located upstream of the DsRed-Express-DR coding sequence. SV40 polyadenylation signals downstream of the DsRed- Express-DR gene direct proper processing of the 3' end of the DsRed-Express-DR mRNA. The vector backbone 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. A neomycin-resistance cassette (Neor) allows stably transfected eukaryotic cells to be selected using G418. This cassette consists of 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 expresses kanamycin resistance in E. coli.


DsRed-Express-DR can be used as an in vivo reporter of gene expression. Because of its rapid turnover rate, its expression from a promoter of interest provides a more accurate assessment of the promoter's activity over time than does the more stable DsRed-Express. Promoter/enhancer elements should be inserted into the MCS upstream of the DsRed-Express-DR coding sequence. Without the addition of a functional promoter, this vector will not express DsRed-Express-DR. The recombinant pDsRed-Express-DR vector can be transfected into mammalian cells using any standard transfection method. If required, stable transformants can be selected using G418 (8).

pDsRed-Express-DR载体含有以下元件：
 MCS: 12–89
 Destabilized Discosoma sp. Red Fluorescent Protein (DsRed-Express-DR) gene
Kozak consensus translation initiation site: 90–100
Start codon (ATG): 97–99; Stop codon: 904–906
CGC→GCC (Arg-2 to Ala) mutation: 100–102
AAG→GAG (Lys-5 to Glu) mutation: 109–111
AAC→GAC (Asn-6 to Asp) mutation: 112–114
ACC→TCC (Thr-21 to Ser) mutation: 157–159
CAC→ACC (His-41 to Thr) mutation: 217–219
AAC→CAG (Asn-42 to Gln) mutation: 220–222
GTG→GCC (Val-44 to Ala) mutation: 226–228
TGC→TCC (Cys-117 to Ser) mutation: 445–447
ACC→GCC (Thr-217 to Ala) mutation: 745–747
Mouse ornithine decarboxylase PEST sequence: 784–906
 SV40 early mRNA polyadenylation signal
Polyadenylation signals: 1059–1064 & 1088–1093
mRNA 3' ends: 1097 & 1109
 f1 single-strand DNA origin: 1156–1611
(Packages noncoding strand of DsRed-Express-DR.)
 Ampicillin resistance (β-lactamase) promoter
–35 region: 1673–1678; –10 region: 1696–1701
Transcription start point: 1708
 SV40 origin of replication: 1952–2087
 SV40 early promoter
Enhancer (72-bp tandem repeats): 1783–1856 & 1857–1928
21-bp repeats: 1932–1952, 1953–1973 & 1975–1995
Early promoter element: 2008–2014
Major transcription start points: 2004, 2042, 2048 & 2053
 Kanamycin/neomycin resistance gene
Neomycin phosphotransferase coding sequences:
Start codon (ATG): 2136–2138; stop codon: 2928–2930
G→A mutation to remove Pst I site: 2318

C→A (Arg→Ser) mutation to remove BssH II site: 2664
 Herpes simplex virus (HSV) thymidine kinase (TK) polyadenylation signal
Polyadenylation signals: 3166–3171 & 3179–3184
 pUC plasmid replication origin: 3515–4158

Propagation in E. Coli
 Suitable host strains: DH5α, HB101 and other general purpose strains. Single-stranded DNA production requires
a host containing an F plasmid such as JM109 or XL1-Blue.
 Selectable marker: plasmid confers resistance to kanamycin (50 μg/ml) to E. coli hosts.
 E. coli replication origin: pUC
 Copy number: ~500
 Plasmid incompatibility group: pMB1/Col E1

Excitation and emission maxima of DsRed-Express
 Excitation maximum = 557 nm
 Emission maximum = 579 nm

 

载体序列

LOCUS       pDsREd-Express-DR	4240 bp 	DNA	SYN
DEFINITION  pDsREd-Express-DR
ACCESSION   
KEYWORDS    
SOURCE      
  ORGANISM  other sequences; artificial sequences; vectors.
FEATURES             Location/Qualifiers
     source          1..4240
                     /organism="pDsREd-Express-DR"
                     /mol_type="other DNA"
     CDS             complement(25..789)
                     /label="ORF frame 2"
                     /translation="MADITSQEQVVAALGALVLLHDGVVLVVGGDVQLGVHVVVAGQL
                     HGLLGHVDGLELHQVVAAVLQLQGLVDLALQHAVAGVQALGGGLPAHSLLLHYGAVGG
                     EVHADELHLVDEGAVLQGGVLGHGHHAAVLEVHHALPLEALGEGQLLVVGDVGGVLHV
                     HLGAVLELGGQDVPGEGQGAALGHLQLGGLGALVGAALALALDLELVAVHGALHAHLE
                     AHELLDDVLGGGHGGDRWIPGPRYRRLQNSKLELEI*"
     CDS             97..906
                     /label="ORF frame 1"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDG
                     GVVTVTQDSSLQDGSFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGE
                     IHKALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERAE
                     GRHHLFLTSDISHGFPPAVAAQDDGTLPMSCAQESGMDRHPAACASARINV*"
     gene            100..771
                     /label="DsRed_Express"
                     /gene="DsRed_Express"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDG
                     GVVTVTQDSSLQDGSFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGE
                     IHKALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERAE
                     GRHHLFLTSDISHGFPPAVAAQDDGTLPMSCAQESGMDRHPAACASARINV*"
     misc_feature    complement(277..297)
                     /label="dsRed1_N_primer"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDG
                     GVVTVTQDSSLQDGSFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGE
                     IHKALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERAE
                     GRHHLFLTSDISHGFPPAVAAQDDGTLPMSCAQESGMDRHPAACASARINV*"
     misc_feature    689..712
                     /label="dsRed1_C_primer"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDG
                     GVVTVTQDSSLQDGSFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGE
                     IHKALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERAE
                     GRHHLFLTSDISHGFPPAVAAQDDGTLPMSCAQESGMDRHPAACASARINV*"
     misc_feature    1119..1138
                     /label="EBV_rev_primer"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDG
                     GVVTVTQDSSLQDGSFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGE
                     IHKALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERAE
                     GRHHLFLTSDISHGFPPAVAAQDDGTLPMSCAQESGMDRHPAACASARINV*"
     rep_origin      complement(1288..1594)
                     /label="f1_origin"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDG
                     GVVTVTQDSSLQDGSFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGE
                     IHKALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERAE
                     GRHHLFLTSDISHGFPPAVAAQDDGTLPMSCAQESGMDRHPAACASARINV*"
     promoter        1673..1701
                     /label="AmpR_promoter"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDG
                     GVVTVTQDSSLQDGSFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGE
                     IHKALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERAE
                     GRHHLFLTSDISHGFPPAVAAQDDGTLPMSCAQESGMDRHPAACASARINV*"
     misc_feature    complement(1767..1787)
                     /label="pBABE_3_primer"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDG
                     GVVTVTQDSSLQDGSFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGE
                     IHKALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERAE
                     GRHHLFLTSDISHGFPPAVAAQDDGTLPMSCAQESGMDRHPAACASARINV*"
     misc_feature    complement(1773..1988)
                     /label="SV40_enhancer"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDG
                     GVVTVTQDSSLQDGSFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGE
                     IHKALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERAE
                     GRHHLFLTSDISHGFPPAVAAQDDGTLPMSCAQESGMDRHPAACASARINV*"
     promoter        1785..2053
                     /label="SV40_promoter"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDG
                     GVVTVTQDSSLQDGSFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGE
                     IHKALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERAE
                     GRHHLFLTSDISHGFPPAVAAQDDGTLPMSCAQESGMDRHPAACASARINV*"
     rep_origin      1952..2029
                     /label="SV40_origin"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDG
                     GVVTVTQDSSLQDGSFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGE
                     IHKALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERAE
                     GRHHLFLTSDISHGFPPAVAAQDDGTLPMSCAQESGMDRHPAACASARINV*"
     misc_feature    2014..2033
                     /label="SV40pro_F_primer"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDG
                     GVVTVTQDSSLQDGSFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGE
                     IHKALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERAE
                     GRHHLFLTSDISHGFPPAVAAQDDGTLPMSCAQESGMDRHPAACASARINV*"
     CDS             2136..2930
                     /label="ORF frame 3"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR
                     PVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL
                     LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRY
                     QDIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
     gene            2139..2927
                     /label="NeoR/KanR"
                     /gene="NeoR/KanR"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR
                     PVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL
                     LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRY
                     QDIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
     CDS             complement(2445..2981)
                     /label="ORF frame 3"
                     /translation="MAGWASLGRSFRTPESRSEELVKKAIEGDALRIGSGDTVKHEEA
                     VSPFAAKLFSNITGSQRYVLIAVRHTQPATVDESRKAAIFHHDIRQAGIAMGHDEILA
                     VGHARLEPGEQFGWREPLMLFVQIILIDKTGFHPSTCSLDAMFRLVVEWAGSRIKRMQ
                     PPHCISHDGYFLGRSKVR*"
     terminator      3105..3374
                     /label="TK_PA_terminator"
                     /translation="MAGWASLGRSFRTPESRSEELVKKAIEGDALRIGSGDTVKHEEA
                     VSPFAAKLFSNITGSQRYVLIAVRHTQPATVDESRKAAIFHHDIRQAGIAMGHDEILA
                     VGHARLEPGEQFGWREPLMLFVQIILIDKTGFHPSTCSLDAMFRLVVEWAGSRIKRMQ
                     PPHCISHDGYFLGRSKVR*"
     rep_origin      3522..4141
                     /label="pBR322_origin"
                     /translation="MAGWASLGRSFRTPESRSEELVKKAIEGDALRIGSGDTVKHEEA
                     VSPFAAKLFSNITGSQRYVLIAVRHTQPATVDESRKAAIFHHDIRQAGIAMGHDEILA
                     VGHARLEPGEQFGWREPLMLFVQIILIDKTGFHPSTCSLDAMFRLVVEWAGSRIKRMQ
                     PPHCISHDGYFLGRSKVR*"
ORIGIN
    1 TAGTTATTAC TAGCGCTACC GGACTCAGAT CTCGAGCTCA AGCTTCGAAT TCTGCAGTCG
   61 ACGGTACCGC GGGCCCGGGA TCCACCGGTC GCCACCATGG CCTCCTCCGA GGACGTCATC
  121 AAGGAGTTCA TGCGCTTCAA GGTGCGCATG GAGGGCTCCG TGAACGGCCA CGAGTTCGAG
  181 ATCGAGGGCG AGGGCGAGGG CCGCCCCTAC GAGGGCACCC AGACCGCCAA GCTGAAGGTG
  241 ACCAAGGGCG GCCCCCTGCC CTTCGCCTGG GACATCCTGT CCCCCCAGTT CCAGTACGGC
  301 TCCAAGGTGT ACGTGAAGCA CCCCGCCGAC ATCCCCGACT ACAAGAAGCT GTCCTTCCCC
  361 GAGGGCTTCA AGTGGGAGCG CGTGATGAAC TTCGAGGACG GCGGCGTGGT GACCGTGACC
  421 CAGGACTCCT CCCTGCAGGA CGGCTCCTTC ATCTACAAGG TGAAGTTCAT CGGCGTGAAC
  481 TTCCCCTCCG ACGGCCCCGT AATGCAGAAG AAGACTATGG GCTGGGAGGC CTCCACCGAG
  541 CGCCTGTACC CCCGCGACGG CGTGCTGAAG GGCGAGATCC ACAAGGCCCT GAAGCTGAAG
  601 GACGGCGGCC ACTACCTGGT GGAGTTCAAG TCCATCTACA TGGCCAAGAA GCCCGTGCAG
  661 CTGCCCGGCT ACTACTACGT GGACTCCAAG CTGGACATCA CCTCCCACAA CGAGGACTAC
  721 ACCATCGTGG AGCAGTACGA GCGCGCCGAG GGCCGCCACC ACCTGTTCCT GACTAGTGAT
  781 ATCAGCCATG GCTTCCCGCC GGCGGTGGCG GCGCAGGATG ATGGCACGCT GCCCATGTCT
  841 TGTGCCCAGG AGAGCGGGAT GGACCGTCAC CCTGCAGCCT GTGCTTCTGC TAGGATCAAT
  901 GTGTAGGCGG CCGCGACTCT AGATCATAAT CAGCCATACC ACATTTGTAG AGGTTTTACT
  961 TGCTTTAAAA AACCTCCCAC ACCTCCCCCT GAACCTGAAA CATAAAATGA ATGCAATTGT
 1021 TGTTGTTAAC TTGTTTATTG CAGCTTATAA TGGTTACAAA TAAAGCAATA GCATCACAAA
 1081 TTTCACAAAT AAAGCATTTT TTTCACTGCA TTCTAGTTGT GGTTTGTCCA AACTCATCAA
 1141 TGTATCTTAA GGCGTAAATT GTAAGCGTTA ATATTTTGTT AAAATTCGCG TTAAATTTTT
 1201 GTTAAATCAG CTCATTTTTT AACCAATAGG CCGAAATCGG CAAAATCCCT TATAAATCAA
 1261 AAGAATAGAC CGAGATAGGG TTGAGTGTTG TTCCAGTTTG GAACAAGAGT CCACTATTAA
 1321 AGAACGTGGA CTCCAACGTC AAAGGGCGAA AAACCGTCTA TCAGGGCGAT GGCCCACTAC
 1381 GTGAACCATC ACCCTAATCA AGTTTTTTGG GGTCGAGGTG CCGTAAAGCA CTAAATCGGA
 1441 ACCCTAAAGG GAGCCCCCGA TTTAGAGCTT GACGGGGAAA GCCGGCGAAC GTGGCGAGAA
 1501 AGGAAGGGAA GAAAGCGAAA GGAGCGGGCG CTAGGGCGCT GGCAAGTGTA GCGGTCACGC
 1561 TGCGCGTAAC CACCACACCC GCCGCGCTTA ATGCGCCGCT ACAGGGCGCG TCAGGTGGCA
 1621 CTTTTCGGGG AAATGTGCGC GGAACCCCTA TTTGTTTATT TTTCTAAATA CATTCAAATA
 1681 TGTATCCGCT CATGAGACAA TAACCCTGAT AAATGCTTCA ATAATATTGA AAAAGGAAGA
 1741 GTCCTGAGGC GGAAAGAACC AGCTGTGGAA TGTGTGTCAG TTAGGGTGTG GAAAGTCCCC
 1801 AGGCTCCCCA GCAGGCAGAA GTATGCAAAG CATGCATCTC AATTAGTCAG CAACCAGGTG
 1861 TGGAAAGTCC CCAGGCTCCC CAGCAGGCAG AAGTATGCAA AGCATGCATC TCAATTAGTC
 1921 AGCAACCATA GTCCCGCCCC TAACTCCGCC CATCCCGCCC CTAACTCCGC CCAGTTCCGC
 1981 CCATTCTCCG CCCCATGGCT GACTAATTTT TTTTATTTAT GCAGAGGCCG AGGCCGCCTC
 2041 GGCCTCTGAG CTATTCCAGA AGTAGTGAGG AGGCTTTTTT GGAGGCCTAG GCTTTTGCAA
 2101 AGATCGATCA AGAGACAGGA TGAGGATCGT TTCGCATGAT TGAACAAGAT GGATTGCACG
 2161 CAGGTTCTCC GGCCGCTTGG GTGGAGAGGC TATTCGGCTA TGACTGGGCA CAACAGACAA
 2221 TCGGCTGCTC TGATGCCGCC GTGTTCCGGC TGTCAGCGCA GGGGCGCCCG GTTCTTTTTG
 2281 TCAAGACCGA CCTGTCCGGT GCCCTGAATG AACTGCAAGA CGAGGCAGCG CGGCTATCGT
 2341 GGCTGGCCAC GACGGGCGTT CCTTGCGCAG CTGTGCTCGA CGTTGTCACT GAAGCGGGAA
 2401 GGGACTGGCT GCTATTGGGC GAAGTGCCGG GGCAGGATCT CCTGTCATCT CACCTTGCTC
 2461 CTGCCGAGAA AGTATCCATC ATGGCTGATG CAATGCGGCG GCTGCATACG CTTGATCCGG
 2521 CTACCTGCCC ATTCGACCAC CAAGCGAAAC ATCGCATCGA GCGAGCACGT ACTCGGATGG
 2581 AAGCCGGTCT TGTCGATCAG GATGATCTGG ACGAAGAGCA TCAGGGGCTC GCGCCAGCCG
 2641 AACTGTTCGC CAGGCTCAAG GCGAGCATGC CCGACGGCGA GGATCTCGTC GTGACCCATG
 2701 GCGATGCCTG CTTGCCGAAT ATCATGGTGG AAAATGGCCG CTTTTCTGGA TTCATCGACT
 2761 GTGGCCGGCT GGGTGTGGCG GACCGCTATC AGGACATAGC GTTGGCTACC CGTGATATTG
 2821 CTGAAGAGCT TGGCGGCGAA TGGGCTGACC GCTTCCTCGT GCTTTACGGT ATCGCCGCTC
 2881 CCGATTCGCA GCGCATCGCC TTCTATCGCC TTCTTGACGA GTTCTTCTGA GCGGGACTCT
 2941 GGGGTTCGAA ATGACCGACC AAGCGACGCC CAACCTGCCA TCACGAGATT TCGATTCCAC
 3001 CGCCGCCTTC TATGAAAGGT TGGGCTTCGG AATCGTTTTC CGGGACGCCG GCTGGATGAT
 3061 CCTCCAGCGC GGGGATCTCA TGCTGGAGTT CTTCGCCCAC CCTAGGGGGA GGCTAACTGA
 3121 AACACGGAAG GAGACAATAC CGGAAGGAAC CCGCGCTATG ACGGCAATAA AAAGACAGAA
 3181 TAAAACGCAC GGTGTTGGGT CGTTTGTTCA TAAACGCGGG GTTCGGTCCC AGGGCTGGCA
 3241 CTCTGTCGAT ACCCCACCGA GACCCCATTG GGGCCAATAC GCCCGCGTTT CTTCCTTTTC
 3301 CCCACCCCAC CCCCCAAGTT CGGGTGAAGG CCCAGGGCTC GCAGCCAACG TCGGGGCGGC
 3361 AGGCCCTGCC ATAGCCTCAG GTTACTCATA TATACTTTAG ATTGATTTAA AACTTCATTT
 3421 TTAATTTAAA AGGATCTAGG TGAAGATCCT TTTTGATAAT CTCATGACCA AAATCCCTTA
 3481 ACGTGAGTTT TCGTTCCACT GAGCGTCAGA CCCCGTAGAA AAGATCAAAG GATCTTCTTG
 3541 AGATCCTTTT TTTCTGCGCG TAATCTGCTG CTTGCAAACA AAAAAACCAC CGCTACCAGC
 3601 GGTGGTTTGT TTGCCGGATC AAGAGCTACC AACTCTTTTT CCGAAGGTAA CTGGCTTCAG
 3661 CAGAGCGCAG ATACCAAATA CTGTCCTTCT AGTGTAGCCG TAGTTAGGCC ACCACTTCAA
 3721 GAACTCTGTA GCACCGCCTA CATACCTCGC TCTGCTAATC CTGTTACCAG TGGCTGCTGC
 3781 CAGTGGCGAT AAGTCGTGTC TTACCGGGTT GGACTCAAGA CGATAGTTAC CGGATAAGGC
 3841 GCAGCGGTCG GGCTGAACGG GGGGTTCGTG CACACAGCCC AGCTTGGAGC GAACGACCTA
 3901 CACCGAACTG AGATACCTAC AGCGTGAGCT ATGAGAAAGC GCCACGCTTC CCGAAGGGAG
 3961 AAAGGCGGAC AGGTATCCGG TAAGCGGCAG GGTCGGAACA GGAGAGCGCA CGAGGGAGCT
 4021 TCCAGGGGGA AACGCCTGGT ATCTTTATAG TCCTGTCGGG TTTCGCCACC TCTGACTTGA
 4081 GCGTCGATTT TTGTGATGCT CGTCAGGGGG GCGGAGCCTA TGGAAAAACG CCAGCAACGC
 4141 GGCCTTTTTA CGGTTCCTGG CCTTTTGCTG GCCTTTTGCT CACATGTTCT TTCCTGCGTT
 4201 ATCCCCTGAT TCTGTGGATA ACCGTATTAC CGCCATGCAT