pTYB2

价格： ¥800 - 1200

品牌：Ybscience

货号：YB-0146

产品详情

载体基本信息

出品公司:	Ybscience
载体名称:	pTYB2
质粒类型:	大肠杆菌表达载体；蛋白纯化
高拷贝/低拷贝:	--
启动子:	--
克隆方法:	多克隆位点，限制性内切酶
载体大小:	7474 bp
5' 测序引物及序列:	T7: TAATACGACTCACTATAGGG
3' 测序引物及序列:	--
载体标签:	Intein
载体抗性:	Ampicillin
筛选标记:	--
备注:	--
稳定性:	--
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

pTYB2载体图谱
pTYB2多克隆位点

载体简介

pTYB2 is an E. coli cloning and expression vector (7474 bp) used in the IMPACT™ Kit (NEB #E6901) which allows the overexpression of a target protein as a fusion to a self-cleavable affinity tag (1,2). This C-terminal fusion vector is designed for the in-frame insertion of a target gene into the polylinker upstream of an intein tag (the Sce VMA intein/chitin binding domain, 55 kDa) (1,2). This results in the fusion of the C-terminus of the target protein to the N-terminus of the intein tag. Thiol-induced self-cleavage of the intein releases the target protein from the chitin-bound intein tag, resulting in a single column purification of the target protein. For fusion of the N-terminus of the target protein to the intein tag, use pTYB21 (NEB #N6709) or pTYB22 (NEB #N6710). This vector can be used in conjuction with pTYB22 (NEB #N6710) to test which fusion constructin (N-terminal or C-terminal) maximizes the expression and yield of a target protein (3).FeaturesThe NdeI site in the polylinker contains an ATG sequence for translation initiation.The SmaI site is used for cloning the 3´ end of the target gene and will yield a target protein with a single glycine residue added to its C-terminus after cleavage of the intein.A pBR322 derivative with a ColE1 replication origin.Expression of the fusion gene is under the control of an IPTG-inducible T7 promoter (4).Expression requires an E. coli host that carries the T7 RNA Polymerase gene [e.g., T7 Express Competent E. coli (High Efficiency) NEB #C2566), BL21(DE3) Competent E. coli (NEB #C2527) and derivatives].Ampicillin resistance.M13 allows for the production of single-stranded DNA by helper phage superinfection of cells bearing the plasmid. M13K07 helper phage is available from (NEB #N0315).Other IMPACT vectors are available which allow for fusion of a target gene to N- or C- terminus of an intein, the cleavage reaction may be induced by thiol reagent or temperature/pH shift.The sites in the polylinker region are identical to or compatible with (i.e., NheI of pTYB2 and SpeI of pTYB12) those of pTYB12 (NEB #N6902). This allows the same amplified target gene to be cloned into either vector for optimizing protein expression. Vector derived residues may be present at the N- and/or C-termini of the target proteins.

载体序列

LOCUS       pTYB2	7474 bp 	DNA	SYNDEFINITION  pTYB2ACCESSION   KEYWORDS    SOURCE        ORGANISM  other sequences; artificial sequences; vectors.FEATURES             Location/Qualifiers     source          1..7474                     /organism="pTYB2"                     /mol_type="other DNA"     promoter        70..98                     /label="AmpR_promoter"     gene            140..1000                     /label="Ampicillin"                     /gene="Ampicillin"     CDS             140..1000                     /label="ORF frame 2"                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY                     IELDLNSGKILESFRPEERSPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA                     EIGASLIKHW*"     rep_origin      complement(1232..1538)                     /label="f1_origin"                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY                     IELDLNSGKILESFRPEERSPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA                     EIGASLIKHW*"     rep_origin      1650..2269                     /label="pBR322_origin"                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY                     IELDLNSGKILESFRPEERSPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA                     EIGASLIKHW*"     misc_feature    2607..2629                     /label="pGEX_3_primer"                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY                     IELDLNSGKILESFRPEERSPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA                     EIGASLIKHW*"     misc_feature    complement(2623..2814)                     /label="ROP"                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY                     IELDLNSGKILESFRPEERSPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA                     EIGASLIKHW*"     CDS             2953..3588                     /label="ORF frame 1"                     /translation="MDAAGPEKNHSGSMPAERQQDVAQRVGRHAGDNGLLLAETFGGG                     TSDEGLSEGVQDSEYRKRQADHRRAPAKAVLAENDPERCRHLSYELHDKEDSHKCGDD                     SHAPRPPEGADWVEGSQGHRSRSRCLMSELTYINCVALTARFPVGKPVVPAALMNRPT                     RGERRFAYWAPGWFFFSPVRRATADCPSPPGPERVAASGPRWFAPAGENPV*"     gene            complement(3010..3309)                     /label="tet (611 - 312)"                     /gene="tet (611 - 312)"                     /translation="MDAAGPEKNHSGSMPAERQQDVAQRVGRHAGDNGLLLAETFGGG                     TSDEGLSEGVQDSEYRKRQADHRRAPAKAVLAENDPERCRHLSYELHDKEDSHKCGDD                     SHAPRPPEGADWVEGSQGHRSRSRCLMSELTYINCVALTARFPVGKPVVPAALMNRPT                     RGERRFAYWAPGWFFFSPVRRATADCPSPPGPERVAASGPRWFAPAGENPV*"     misc_feature    complement(3371..4462)                     /label="lacI"                     /translation="MDAAGPEKNHSGSMPAERQQDVAQRVGRHAGDNGLLLAETFGGG                     TSDEGLSEGVQDSEYRKRQADHRRAPAKAVLAENDPERCRHLSYELHDKEDSHKCGDD                     SHAPRPPEGADWVEGSQGHRSRSRCLMSELTYINCVALTARFPVGKPVVPAALMNRPT                     RGERRFAYWAPGWFFFSPVRRATADCPSPPGPERVAASGPRWFAPAGENPV*"     CDS             complement(3371..4330)                     /label="ORF frame 1"                     /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS                     RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV                     PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW                     HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE                     SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ                     LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"     CDS             complement(4654..5415)                     /label="ORF frame 2"                     /translation="MPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQESVHRQT                     TDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQESVH                     RQTTDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQE                     SVHRQTTDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICP                     TQESVHRQTTDKTKGPVFRLSLSFYLMPGELGINS*"     terminator      4687..4730                     /label="rrnB_T1_terminator"                     /translation="MPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQESVHRQT                     TDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQESVH                     RQTTDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQE                     SVHRQTTDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICP                     TQESVHRQTTDKTKGPVFRLSLSFYLMPGELGINS*"     terminator      4870..4913                     /label="rrnB_T1_terminator"                     /translation="MPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQESVHRQT                     TDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQESVH                     RQTTDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQE                     SVHRQTTDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICP                     TQESVHRQTTDKTKGPVFRLSLSFYLMPGELGINS*"     terminator      5053..5096                     /label="rrnB_T1_terminator"                     /translation="MPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQESVHRQT                     TDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQESVH                     RQTTDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQE                     SVHRQTTDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICP                     TQESVHRQTTDKTKGPVFRLSLSFYLMPGELGINS*"     terminator      5236..5279                     /label="rrnB_T1_terminator"                     /translation="MPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQESVHRQT                     TDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQESVH                     RQTTDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQE                     SVHRQTTDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICP                     TQESVHRQTTDKTKGPVFRLSLSFYLMPGELGINS*"     terminator      5419..5462                     /label="rrnB_T1_terminator"                     /translation="MPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQESVHRQT                     TDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQESVH                     RQTTDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQE                     SVHRQTTDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICP                     TQESVHRQTTDKTKGPVFRLSLSFYLMPGELGINS*"     promoter        5637..5655                     /label="T7_promoter"                     /translation="MPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQESVHRQT                     TDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQESVH                     RQTTDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQE                     SVHRQTTDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICP                     TQESVHRQTTDKTKGPVFRLSLSFYLMPGELGINS*"     misc_feature    5655..5682                     /label="lacO"                     /translation="MPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQESVHRQT                     TDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQESVH                     RQTTDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQE                     SVHRQTTDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICP                     TQESVHRQTTDKTKGPVFRLSLSFYLMPGELGINS*"     misc_feature    5700..5716                     /label="T7_transl_en_RBS"                     /translation="MPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQESVHRQT                     TDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQESVH                     RQTTDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQE                     SVHRQTTDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICP                     TQESVHRQTTDKTKGPVFRLSLSFYLMPGELGINS*"     CDS             5725..7326                     /label="ORF frame 1"                     /translation="MASSRVDGGREFLEPGCFAKGTNVLMADGSIECIENIEVGNKVM                     GKDGRPREVIKLPRGRETMYSVVQKSQHRAHKSDSSREVPELLKFTCNATHELVVRTP                     RSVRRLSRTIKGVEYFEVITFEMGQKKAPDGRIVELVKEVSKSYPISEGPERANELVE                     SYRKASNKAYFEWTIEARDLSLLGSHVRKATYQTYAPILYENDHFFDYMQKSKFHLTI                     EGPKVLAYLLGLWIGDGLSDRATFSVDSRDTSLMERVTEYAEKLNLCAEYKDRKEPQV                     AKTVNLYSKVVRGNGIRNNLNTENPLWDAIVGLGFLKDGVKNIPSFLSTDNIGTRETF                     LAGLIDSDGYVTDEHGIKATIKTIHTSVRDGLVSLARSLGLVVSVNAEPAKVDMNVTK                     HKISYAIYMSGGDVLLNVLSKCAGSKKFRPAPAAAFARECRGFYFELQELKEDDYYGI                     TLSDDSDHQFLLGSQVVVHACGGLTGLNSGLTTNPGVSAWQVNTAYTAGQLVTYNGKT                     YKCLQPHTSLAGWEPSNVPALWQLQ*"     terminator      7346..7474                     /label="T7_terminator"                     /translation="MASSRVDGGREFLEPGCFAKGTNVLMADGSIECIENIEVGNKVM                     GKDGRPREVIKLPRGRETMYSVVQKSQHRAHKSDSSREVPELLKFTCNATHELVVRTP                     RSVRRLSRTIKGVEYFEVITFEMGQKKAPDGRIVELVKEVSKSYPISEGPERANELVE                     SYRKASNKAYFEWTIEARDLSLLGSHVRKATYQTYAPILYENDHFFDYMQKSKFHLTI                     EGPKVLAYLLGLWIGDGLSDRATFSVDSRDTSLMERVTEYAEKLNLCAEYKDRKEPQV                     AKTVNLYSKVVRGNGIRNNLNTENPLWDAIVGLGFLKDGVKNIPSFLSTDNIGTRETF                     LAGLIDSDGYVTDEHGIKATIKTIHTSVRDGLVSLARSLGLVVSVNAEPAKVDMNVTK                     HKISYAIYMSGGDVLLNVLSKCAGSKKFRPAPAAAFARECRGFYFELQELKEDDYYGI                     TLSDDSDHQFLLGSQVVVHACGGLTGLNSGLTTNPGVSAWQVNTAYTAGQLVTYNGKT                     YKCLQPHTSLAGWEPSNVPALWQLQ*"     misc_feature    complement(7388..7406)                     /label="T7_Terminal_primer"                     /translation="MASSRVDGGREFLEPGCFAKGTNVLMADGSIECIENIEVGNKVM                     GKDGRPREVIKLPRGRETMYSVVQKSQHRAHKSDSSREVPELLKFTCNATHELVVRTP                     RSVRRLSRTIKGVEYFEVITFEMGQKKAPDGRIVELVKEVSKSYPISEGPERANELVE                     SYRKASNKAYFEWTIEARDLSLLGSHVRKATYQTYAPILYENDHFFDYMQKSKFHLTI                     EGPKVLAYLLGLWIGDGLSDRATFSVDSRDTSLMERVTEYAEKLNLCAEYKDRKEPQV                     AKTVNLYSKVVRGNGIRNNLNTENPLWDAIVGLGFLKDGVKNIPSFLSTDNIGTRETF                     LAGLIDSDGYVTDEHGIKATIKTIHTSVRDGLVSLARSLGLVVSVNAEPAKVDMNVTK                     HKISYAIYMSGGDVLLNVLSKCAGSKKFRPAPAAAFARECRGFYFELQELKEDDYYGI                     TLSDDSDHQFLLGSQVVVHACGGLTGLNSGLTTNPGVSAWQVNTAYTAGQLVTYNGKT                     YKCLQPHTSLAGWEPSNVPALWQLQ*"ORIGIN    1 AACTACGTCA GGTGGCACTT TTCGGGGAAA TGTGCGCGGA ACCCCTATTT GTTTATTTTT   61 CTAAATACAT TCAAATATGT ATCCGCTCAT GAGACAATAA CCCTGATAAA TGCTTCAATA  121 ATATTGAAAA AGGAAGAGTA TGAGTATTCA ACATTTCCGT GTCGCCCTTA TTCCCTTTTT  181 TGCGGCATTT TGCCTTCCTG TTTTTGCTCA CCCAGAAACG CTGGTGAAAG TAAAAGATGC  241 TGAAGATCAG TTGGGTGCAC GAGTGGGTTA CATCGAACTG GATCTCAACA GCGGTAAGAT  301 CCTTGAGAGT TTTCGCCCCG AAGAACGTTC TCCAATGATG AGCACTTTTA AAGTTCTGCT  361 ATGTGGCGCG GTATTATCCC GTGTTGACGC CGGGCAAGAG CAACTCGGTC GCCGCATACA  421 CTATTCTCAG AATGACTTGG TTGAGTACTC ACCAGTCACA GAAAAGCATC TTACGGATGG  481 CATGACAGTA AGAGAATTAT GCAGTGCTGC CATAACCATG AGTGATAACA CTGCGGCCAA  541 CTTACTTCTG ACAACGATCG GAGGACCGAA GGAGCTAACC GCTTTTTTGC ACAACATGGG  601 GGATCATGTA ACTCGCCTTG ATCGTTGGGA ACCGGAGCTG AATGAAGCCA TACCAAACGA  661 CGAGCGTGAC ACCACGATGC CTGTAGCAAT GGCAACAACG TTGCGCAAAC TATTAACTGG  721 CGAACTACTT ACTCTAGCTT CCCGGCAACA ATTAATAGAC TGGATGGAGG CGGATAAAGT  781 TGCAGGACCA CTTCTGCGCT CGGCCCTTCC GGCTGGCTGG TTTATTGCTG ATAAATCTGG  841 AGCCGGTGAG CGTGGGTCTC GCGGTATCAT TGCAGCACTG GGGCCAGATG GTAAGCCCTC  901 CCGTATCGTA GTTATCTACA CGACGGGGAG TCAGGCAACT ATGGATGAAC GAAATAGACA  961 GATCGCTGAG ATAGGTGCCT CACTGATTAA GCATTGGTAA CTGTCAGACC AAGTTTACTC 1021 ATATATACTT TAGATTGATT TACCCCGGTT GATAATCAGA AAAGCCCCAA AAACAGGAAG 1081 ATTGTATAAG CAAATATTTA AATTGTAAAC GTTAATATTT TGTTAAAATT CGCGTTAAAT 1141 TTTTGTTAAA TCAGCTCATT TTTTAACCAA TAGGCCGAAA TCGGCAAAAT CCCTTATAAA 1201 TCAAAAGAAT AGCCCGAGAT AGGGTTGAGT GTTGTTCCAG TTTGGAACAA GAGTCCACTA 1261 TTAAAGAACG TGGACTCCAA CGTCAAAGGG CGAAAAACCG TCTATCAGGG CGATGGCCCA 1321 CTACGTGAAC CATCACCCAA ATCAAGTTTT TTGGGGTCGA GGTGCCGTAA AGCACTAAAT 1381 CGGAACCCTA AAGGGAGCCC CCGATTTAGA GCTTGACGGG GAAAGCCGGC GAACGTGGCG 1441 AGAAAGGAAG GGAAGAAAGC GAAAGGAGCG GGCGCTAGGG CGCTGGCAAG TGTAGCGGTC 1501 ACGCTGCGCG TAACCACCAC ACCCGCCGCG CTTAATGCGC CGCTACAGGG CGCGTAAAAG 1561 GATCTAGGTG AAGATCCTTT TTGATAATCT CATGACCAAA ATCCCTTAAC GTGAGTTTTC 1621 GTTCCACTGA GCGTCAGACC CCGTAGAAAA GATCAAAGGA TCTTCTTGAG ATCCTTTTTT 1681 TCTGCGCGTA ATCTGCTGCT TGCAAACAAA AAAACCACCG CTACCAGCGG TGGTTTGTTT 1741 GCCGGATCAA GAGCTACCAA CTCTTTTTCC GAAGGTAACT GGCTTCAGCA GAGCGCAGAT 1801 ACCAAATACT GTCCTTCTAG TGTAGCCGTA GTTAGGCCAC CACTTCAAGA ACTCTGTAGC 1861 ACCGCCTACA TACCTCGCTC TGCTAATCCT GTTACCAGTG GCTGCTGCCA GTGGCGATAA 1921 GTCGTGTCTT ACCGGGTTGG ACTCAAGACG ATAGTTACCG GATAAGGCGC AGCGGTCGGG 1981 CTGAACGGGG GGTTCGTGCA CACAGCCCAG CTTGGAGCGA ACGACCTACA CCGAACTGAG 2041 ATACCTACAG CGTGAGCTAT GAGAAAGCGC CACGCTTCCC GAAGGGAGAA AGGCGGACAG 2101 GTATCCGGTA AGCGGCAGGG TCGGAACAGG AGAGCGCACG AGGGAGCTTC CAGGGGGAAA 2161 CGCCTGGTAT CTTTATAGTC CTGTCGGGTT TCGCCACCTC TGACTTGAGC GTCGATTTTT 2221 GTGATGCTCG TCAGGGGGGC GGAGCCTATG GAAAAACGCC AGCAACGCGG CCTTTTTACG 2281 GTTCCTGGCC TTTTGCTGGC CTTTTGCTCA CATGTTCTTT CCTGCGTTAT CCCCTGATTC 2341 TGTGGATAAC CGTATTACCG CCTTTGAGTG AGCTGATACC GCTCGCCGCA GCCGAACGAC 2401 CGAGCGCAGC GAGTCAGTGA GCGAGGAAGC TATGGTGCAC TCTCAGTACA ATCTGCTCTG 2461 ATGCCGCATA GTTAAGCCAG TATACACTCC GCTATCGCTA CGTGACTGGG TCATGGCTGC 2521 GCCCCGACAC CCGCCAACAC CCGCTGACGC GCCCTGACGG GCTTGTCTGC TCCCGGCATC 2581 CGCTTACAGA CAAGCTGTGA CCGTCTCCGG GAGCTGCATG TGTCAGAGGT TTTCACCGTC 2641 ATCACCGAAA CGCGCGAGGC AGCTGCGGTA AAGCTCATCA GCGTGGTCGT GCAGCGATTC 2701 ACAGATGTCT GCCTGTTCAT CCGCGTCCAG CTCGTTGAGT TTCTCCAGAA GCGTTAATGT 2761 CTGGCTTCTG ATAAAGCGGG CCATGTTAAG GGCGGTTTTT TCCTGTTTGG TCACTGATGC 2821 CTCCGTGTAA GGGGGATTTC TGTTCATGGG GGTAATGATA CCGATGAAAC GAGAGAGGAT 2881 GCTCACGATA CGGGTTACTG ATGATGAACA TGCCCGGTTA CTGGAACGTT GTGAGGGTAA 2941 ACAACTGGCG GTATGGATGC GGCGGGACCA GAGAAAAATC ACTCAGGGTC AATGCCAGCC 3001 GAACGCCAGC AAGACGTAGC CCAGCGCGTC GGCCGCCATG CCGGCGATAA TGGCCTGCTT 3061 CTCGCCGAAA CGTTTGGTGG CGGGACCAGT GACGAAGGCT TGAGCGAGGG CGTGCAAGAT 3121 TCCGAATACC GCAAGCGACA GGCCGATCAT CGTCGCGCTC CAGCGAAAGC GGTCCTCGCC 3181 GAAAATGACC CAGAGCGCTG CCGGCACCTG TCCTACGAGT TGCATGATAA AGAAGACAGT 3241 CATAAGTGCG GCGACGATAG TCATGCCCCG CGCCCACCGG AAGGAGCTGA CTGGGTTGAA 3301 GGCTCTCAAG GGCATCGGTC GAGATCCCGG TGCCTAATGA GTGAGCTAAC TTACATTAAT 3361 TGCGTTGCGC TCACTGCCCG CTTTCCAGTC GGGAAACCTG TCGTGCCAGC TGCATTAATG 3421 AATCGGCCAA CGCGCGGGGA GAGGCGGTTT GCGTATTGGG CGCCAGGGTG GTTTTTCTTT 3481 TCACCAGTGA GACGGGCAAC AGCTGATTGC CCTTCACCGC CTGGCCCTGA GAGAGTTGCA 3541 GCAAGCGGTC CACGCTGGTT TGCCCCAGCA GGCGAAAATC CTGTTTGATG GTGGTTAACG 3601 GCGGGATATA ACATGAGCTG TCTTCGGTAT CGTCGTATCC CACTACCGAG ATATCCGCAC 3661 CAACGCGCAG CCCGGACTCG GTAATGGCGC GCATTGCGCC CAGCGCCATC TGATCGTTGG 3721 CAACCAGCAT CGCAGTGGGA ACGATGCCCT CATTCAGCAT TTGCATGGTT TGTTGAAAAC 3781 CGGACATGGC ACTCCAGTCG CCTTCCCGTT CCGCTATCGG CTGAATTTGA TTGCGAGTGA 3841 GATATTTATG CCAGCCAGCC AGACGCAGAC GCGCCGAGAC AGAACTTAAT GGGCCCGCTA 3901 ACAGCGCGAT TTGCTGGTGA CCCAATGCGA CCAGATGCTC CACGCCCAGT CGCGTACCGT 3961 CTTCATGGGA GAAAATAATA CTGTTGATGG GTGTCTGGTC AGAGACATCA AGAAATAACG 4021 CCGGAACATT AGTGCAGGCA GCTTCCACAG CAATGGCATC CTGGTCATCC AGCGGATAGT 4081 TAATGATCAG CCCACTGACG CGTTGCGCGA GAAGATTGTG CACCGCCGCT TTACAGGCTT 4141 CGACGCCGCT TCGTTCTACC ATCGACACCA CCACGCTGGC ACCCAGTTGA TCGGCGCGAG 4201 ATTTAATCGC CGCGACAATT TGCGACGGCG CGTGCAGGGC CAGACTGGAG GTGGCAACGC 4261 CAATCAGCAA CGACTGTTTG CCCGCCAGTT GTTGTGCCAC GCGGTTGGGA ATGTAATTCA 4321 GCTCCGCCAT CGCCGCTTCC ACTTTTTCCC GCGTTTTCGC AGAAACGTGG CTGGCCTGGT 4381 TCACCACGCG GGAAACGGTC TGATAAGAGA CACCGGCATA CTCTGCGACA TCGTATAACG 4441 TTACTGGTTT CACATTCACC ACCCTGAATT GACTCTCTTC CGGGCGCTAT CATGCCATAC 4501 CGCGAAAGGT TTTGCGCCAT TCGATGGTGT CCGGGATCTC GACGCTCTCC CTTATGCGAC 4561 TCCTGCATTA GGAAGCAGCC CAGTAGTAGG TTGAGGCCGT TGAGCACCGC CGCCGCAAGG 4621 AATGGTGCAT GCCGGCATGC CGCCCTTTCG TCTTCAAGAA TTAATTCCCA ATTCCCCAGG 4681 CATCAAATAA AACGAAAGGC TCAGTCGAAA GACTGGGCCT TTCGTTTTAT CTGTTGTTTG 4741 TCGGTGAACG CTCTCCTGAG TAGGACAAAT CCGCCGGGAG CGGATTTGAA CGTTGCGAAG 4801 CAACGGCCCG GAGGGTGGCG GGCAGGACGC CCGCCATAAA CTGCCAGGAA TTAATTCCCC 4861 AGGCATCAAA TAAAACGAAA GGCTCAGTCG AAAGACTGGG CCTTTCGTTT TATCTGTTGT 4921 TTGTCGGTGA ACGCTCTCCT GAGTAGGACA AATCCGCCGG GAGCGGATTT GAACGTTGCG 4981 AAGCAACGGC CCGGAGGGTG GCGGGCAGGA CGCCCGCCAT AAACTGCCAG GAATTAATTC 5041 CCCAGGCATC AAATAAAACG AAAGGCTCAG TCGAAAGACT GGGCCTTTCG TTTTATCTGT 5101 TGTTTGTCGG TGAACGCTCT CCTGAGTAGG ACAAATCCGC CGGGAGCGGA TTTGAACGTT 5161 GCGAAGCAAC GGCCCGGAGG GTGGCGGGCA GGACGCCCGC CATAAACTGC CAGGAATTAA 5221 TTCCCCAGGC ATCAAATAAA ACGAAAGGCT CAGTCGAAAG ACTGGGCCTT TCGTTTTATC 5281 TGTTGTTTGT CGGTGAACGC TCTCCTGAGT AGGACAAATC CGCCGGGAGC GGATTTGAAC 5341 GTTGCGAAGC AACGGCCCGG AGGGTGGCGG GCAGGACGCC CGCCATAAAC TGCCAGGAAT 5401 TAATTCCCCA GGCATCAAAT AAAACGAAAG GCTCAGTCGA AAGACTGGGC CTTTCGTTTT 5461 ATCTGTTGTT TGTCGGTGAA CGCTCTCCTG AGTAGGACAA ATCCGCCGGG AGCGGATTTG 5521 AACGTTGCGA AGCAACGGCC CGGAGGGTGG CGGGCAGGAC GCCCGCCATA AACTGCCAGG 5581 AATTGGGGAT CGGAATTAAT TCCCGGTTTA AACCGGGGAT CTCGATCCCG CGAAATTAAT 5641 ACGACTCACT ATAGGGGAAT TGTGAGCGGA TAACAATTCC CCTCTAGAAA TAATTTTGTT 5701 TAACTTTAAG AAGGAGATAT ACATATGGCT AGCTCGCGAG TCGACGGCGG CCGCGAATTC 5761 CTCGAGCCCG GGTGCTTTGC CAAGGGTACC AATGTTTTAA TGGCGGATGG GTCTATTGAA 5821 TGTATTGAAA ACATTGAGGT TGGTAATAAG GTCATGGGTA AAGATGGCAG ACCTCGTGAG 5881 GTAATTAAAT TGCCCAGAGG AAGAGAAACT ATGTACAGCG TCGTGCAGAA AAGTCAGCAC 5941 AGAGCCCACA AAAGTGACTC AAGTCGTGAA GTGCCAGAAT TACTCAAGTT TACGTGTAAT 6001 GCGACCCATG AGTTGGTTGT TAGAACACCT CGTAGTGTCC GCCGTTTGTC TCGTACCATT 6061 AAGGGTGTCG AATATTTTGA AGTTATTACT TTTGAGATGG GCCAAAAGAA AGCCCCCGAC 6121 GGTAGAATTG TTGAGCTTGT CAAGGAAGTT TCAAAGAGCT ACCCAATATC TGAGGGGCCT 6181 GAGAGAGCCA ACGAATTAGT AGAATCCTAT AGAAAGGCTT CAAATAAAGC TTATTTTGAG 6241 TGGACTATTG AGGCCAGAGA TCTTTCTCTG TTGGGTTCCC ATGTTCGTAA AGCTACCTAC 6301 CAGACTTACG CTCCAATTCT TTATGAGAAT GACCACTTTT TCGACTACAT GCAAAAAAGT 6361 AAGTTTCATC TCACCATTGA AGGTCCAAAA GTACTTGCTT ATTTACTTGG TTTATGGATT 6421 GGTGATGGAT TGTCTGACAG GGCAACTTTT TCGGTTGATT CCAGAGATAC TTCTTTGATG 6481 GAACGTGTTA CTGAATATGC TGAAAAGTTG AATTTGTGCG CCGAGTATAA GGACAGAAAA 6541 GAACCACAAG TTGCCAAAAC TGTTAATTTG TACTCTAAAG TTGTCAGAGG TAATGGTATT 6601 CGCAATAATC TTAATACTGA GAATCCATTA TGGGACGCTA TTGTTGGCTT AGGATTCTTG 6661 AAGGACGGTG TCAAAAATAT TCCTTCTTTC TTGTCTACGG ACAATATCGG TACTCGTGAA 6721 ACATTTCTTG CTGGTCTAAT TGATTCTGAT GGCTATGTTA CTGATGAGCA TGGTATTAAA 6781 GCAACAATAA AGACAATTCA TACTTCTGTC AGAGATGGTT TGGTTTCCCT TGCTCGTTCT 6841 TTAGGCTTAG TAGTCTCGGT TAACGCAGAA CCTGCTAAGG TTGACATGAA TGTCACCAAA 6901 CATAAAATTA GTTATGCTAT TTATATGTCT GGTGGAGATG TTTTGCTTAA CGTTCTTTCG 6961 AAGTGTGCCG GCTCTAAAAA ATTCAGGCCT GCTCCCGCCG CTGCTTTTGC ACGTGAGTGC 7021 CGCGGATTTT ATTTCGAGTT ACAAGAATTG AAGGAAGACG ATTATTATGG GATTACTTTA 7081 TCTGATGATT CTGATCATCA GTTTTTGCTT GGATCCCAGG TTGTCGTCCA TGCATGCGGT 7141 GGCCTGACCG GTCTGAACTC AGGCCTCACG ACAAATCCTG GTGTATCCGC TTGGCAGGTC 7201 AACACAGCTT ATACTGCGGG ACAATTGGTC ACATATAACG GCAAGACGTA TAAATGTTTG 7261 CAGCCCCACA CCTCCTTGGC AGGATGGGAA CCATCCAACG TTCCTGCCTT GTGGCAGCTT 7321 CAATGACTGC AGGAAGGGGA TCCGGCTGCT AACAAAGCCC GAAAGGAAGC TGAGTTGGCT 7381 GCTGCCACCG CTGAGCAATA ACTAGCATAA CCCCTTGGGG CCTCTAAACG GGTCTTGAGG 7441 GGTTTTTTGC TGAAAGGAGG AACTATATCC GGAT//