| 质粒名称: | pMJ920 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 42234 |
| 存储实验室: | Jennifer Doudna |
|
载体骨架基本信息 |
|
| 载体名称: | MacroLab 6D |
| 载体出品公司: | QB3 MacroLab |
| 空载体大小: | 6145 bp |
| 完整质粒大小: | 10357 bp |
| 载体类型: | 哺乳细胞表达, CRISPR |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | Mach1 |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | Cas9、Csn1 |
| 物种来源: | Synthetic; Streptococcus pyogenes |
| 插入大小: | 4209 bp |
| 基因突变: |
codon-optimized synthetic DNA sequence
GenBank IDNP_269215.1 NC_002737.1 (854757..858863)
|
| 启动子: | CMV |
| 标签/融合蛋白: |
HA epitope (C terminal on insert)
NLS (nuclear localization signal) (C terminal on insert)
GFP (C terminal on backbone)
|
| 克隆方法: | Ligation Independent Cloning |
| 5’克隆位点: | T7 promoter |
| 3’克隆位点: | BGH polyA |
|
文献引用方法 |
|
| 材料和方法部分: | pMJ920 was a gift from Jennifer Doudna (Addgene plasmid # 42234 ; http://n2t.net/addgene:42234 ; RRID:Addgene_42234) |
| 参考文献部分: |
RNA-programmed genome editing in human cells. Jinek M, East A, Cheng A, Lin S, Ma E, Doudna J. elife. 2013;2:e00471. doi: 10.7554/eLife.00471. Epub 2013 Jan 29. 10.7554/eLife.00471 PubMed 23386978
|
买家导航
载体基本信息
载体图谱质粒图谱和多克隆位点信息
载体简介
载体序列
LOCUS pMJ920 10357 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pMJ920
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10357)
AUTHORS Jinek M, East A, Cheng A, Lin S, Ma E, Doudna J
TITLE RNA-programmed genome editing in human cells.
JOURNAL elife. 2013;2:e00471. doi: 10.7554/eLife.00471. Epub 2013 Jan 29.
PUBMED 23386978
REFERENCE 2 (bases 1 to 10357)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..10357
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(44..63)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 235..614
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 615..818
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 769..789
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 863..882
/label=T7
/note="T7 promoter, forward primer"
promoter 863..881
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
regulatory 929..938
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 935..5038
/codon_start=1
/product="Cas9 (Csn1) endonuclease from the Streptococcus
pyogenes Type II CRISPR/Cas system"
/label=Cas9
/note="generates RNA-guided double strand breaks in DNA"
/translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
DATLIHQSITGLYETRIDLSQLGGD"
CDS 5042..5068
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
CDS 5087..5107
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 5114..5134
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 5174..5890
/codon_start=1
/product="enhanced GFP (EGFP) derivative that exhibits
robust folding and enhanced photostability (Zhong et al.,
2019)"
/label=SiriusGFP
/note="mammalian codon-optimized"
/translation="VSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
FVTAAGITLGMDELYK"
primer_bind complement(5216..5237)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind 5824..5845
/label=EGFP-C
/note="EGFP, forward primer"
promoter complement(5909..5927)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(5910..5927)
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind complement(5947..5964)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 5953..6177
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 6223..6651
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(6310..6329)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 6520..6541
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(6660..6680)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 6665..6995
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 6846..6981
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 6908..6927
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 7062..7856
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(7116..7135)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 7726..7745
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 8030..8151
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(8067..8086)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 8121..8140
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(8200..8216)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(8200..8216)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(8213..8235)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 8224..8240
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(8248..8278)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 8293..8314
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(8431..8448)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(8602..9190)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(8682..8701)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(9361..10221)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 9984..10003
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(10222..10326)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
901 gagctcggat ccctacttcc aatccaatgc caccatggac aagaagtaca gcatcggcct
961 ggacatcggc accaactctg tgggctgggc cgtgatcacc gacgagtaca aggtgcccag
1021 caagaaattc aaggtgctgg gcaacaccga ccggcacagc atcaagaaga acctgatcgg
1081 cgccctgctg ttcgacagcg gagaaacagc cgaggccacc cggctgaaga gaaccgccag
1141 aagaagatac accagacgga agaaccggat ctgctatctg caagagatct tcagcaacga
1201 gatggccaag gtggacgaca gcttcttcca cagactggaa gagtccttcc tggtggaaga
1261 ggataagaag cacgagcggc accccatctt cggcaacatc gtggacgagg tggcctacca
1321 cgagaagtac cccaccatct accacctgag aaagaaactg gtggacagca ccgacaaggc
1381 cgacctgcgg ctgatctatc tggccctggc ccacatgatc aagttccggg gccacttcct
1441 gatcgagggc gacctgaacc ccgacaacag cgacgtggac aagctgttca tccagctggt
1501 gcagacctac aaccagctgt tcgaggaaaa ccccatcaac gccagcggcg tggacgccaa
1561 ggccatcctg tctgccagac tgagcaagag cagacggctg gaaaatctga tcgcccagct
1621 gcccggcgag aagaagaatg gcctgttcgg caacctgatt gccctgagcc tgggcctgac
1681 ccccaacttc aagagcaact tcgacctggc cgaggatgcc aaactgcagc tgagcaagga
1741 cacctacgac gacgacctgg acaacctgct ggcccagatc ggcgaccagt acgccgacct
1801 gtttctggcc gccaagaacc tgtccgacgc catcctgctg agcgacatcc tgagagtgaa
1861 caccgagatc accaaggccc ccctgagcgc ctctatgatc aagagatacg acgagcacca
1921 ccaggacctg accctgctga aagctctcgt gcggcagcag ctgcctgaga agtacaaaga
1981 gattttcttc gaccagagca agaacggcta cgccggctac atcgatggcg gagccagcca
2041 ggaagagttc tacaagttca tcaagcccat cctggaaaag atggacggca ccgaggaact
2101 gctcgtgaag ctgaacagag aggacctgct gcggaagcag cggaccttcg acaacggcag
2161 catcccccac cagatccacc tgggagagct gcacgccatt ctgcggcggc aggaagattt
2221 ttacccattc ctgaaggaca accgggaaaa gatcgagaag atcctgacct tccgcatccc
2281 ctactacgtg ggccctctgg ccaggggaaa cagcagattc gcctggatga ccagaaagag
2341 cgaggaaacc atcaccccct ggaacttcga ggaagtggtg gacaagggcg ccagcgccca
2401 gagcttcatc gagcggatga ccaacttcga taagaacctg cccaacgaga aggtgctgcc
2461 caagcacagc ctgctgtacg agtacttcac cgtgtacaac gagctgacca aagtgaaata
2521 cgtgaccgag ggaatgagaa agcccgcctt cctgagcggc gagcagaaaa aagccatcgt
2581 ggacctgctg ttcaagacca accggaaagt gaccgtgaag cagctgaaag aggactactt
2641 caagaaaatc gagtgcttcg actccgtgga aatctccggc gtggaagatc ggttcaacgc
2701 ctccctgggc acataccacg atctgctgaa aattatcaag gacaaggact tcctggacaa
2761 tgaggaaaac gaggacattc tggaagatat cgtgctgacc ctgacactgt ttgaggacag
2821 agagatgatc gaggaacggc tgaaaaccta tgcccacctg ttcgacgaca aagtgatgaa
2881 gcagctgaag cggcggagat acaccggctg gggcaggctg agccggaagc tgatcaacgg
2941 catccgggac aagcagtccg gcaagacaat cctggatttc ctgaagtccg acggcttcgc
3001 caacagaaac ttcatgcagc tgatccacga cgacagcctg acctttaaag aggacatcca
3061 gaaagcccag gtgtccggcc agggcgatag cctgcacgag cacattgcca atctggccgg
3121 cagccccgcc attaagaagg gcatcctgca gacagtgaag gtggtggacg agctcgtgaa
3181 agtgatgggc cggcacaagc ccgagaacat cgtgatcgaa atggccagag agaaccagac
3241 cacccagaag ggacagaaga acagccgcga gagaatgaag cggatcgaag agggcatcaa
3301 agagctgggc agccagatcc tgaaagaaca ccccgtggaa aacacccagc tgcagaacga
3361 gaagctgtac ctgtactacc tgcagaatgg gcgggatatg tacgtggacc aggaactgga
3421 catcaaccgg ctgtccgact acgatgtgga ccatatcgtg cctcagagct ttctgaagga
3481 cgactccatc gataacaaag tgctgactcg gagcgacaag aaccggggca agagcgacaa
3541 cgtgccctcc gaagaggtcg tgaagaagat gaagaactac tggcgccagc tgctgaatgc
3601 caagctgatt acccagagga agttcgacaa tctgaccaag gccgagagag gcggcctgag
3661 cgaactggat aaggccggct tcatcaagag acagctggtg gaaacccggc agatcacaaa
3721 gcacgtggca cagatcctgg actcccggat gaacactaag tacgacgaga acgacaaact
3781 gatccgggaa gtgaaagtga tcaccctgaa gtccaagctg gtgtccgatt tccggaagga
3841 tttccagttt tacaaagtgc gcgagatcaa caactaccac cacgcccacg acgcctacct
3901 gaacgccgtc gtgggaaccg ccctgatcaa aaagtaccct aagctggaaa gcgagttcgt
3961 gtacggcgac tacaaggtgt acgacgtgcg gaagatgatc gccaagagcg agcaggaaat
4021 cggcaaggct accgccaagt acttcttcta cagcaacatc atgaactttt tcaagaccga
4081 gattaccctg gccaacggcg agatccggaa gcggcctctg atcgagacaa acggcgaaac
4141 aggcgagatc gtgtgggata agggccggga ctttgccacc gtgcggaaag tgctgtctat
4201 gccccaagtg aatatcgtga aaaagaccga ggtgcagaca ggcggcttca gcaaagagtc
4261 tatcctgccc aagaggaaca gcgacaagct gatcgccaga aagaaggact gggaccctaa
4321 gaagtacggc ggcttcgaca gccccaccgt ggcctattct gtgctggtgg tggccaaagt
4381 ggaaaagggc aagtccaaga aactgaagag tgtgaaagag ctgctgggga tcaccatcat
4441 ggaaagaagc agcttcgaga agaatcccat cgactttctg gaagccaagg gctacaaaga
4501 agtgaaaaag gacctgatca tcaagctgcc taagtactcc ctgttcgagc tggaaaacgg
4561 ccggaagaga atgctggcct ctgccggcga actgcagaag ggaaacgaac tggccctgcc
4621 ctccaaatat gtgaacttcc tgtacctggc cagccactat gagaagctga agggctcccc
4681 cgaggataat gagcagaaac agctgtttgt ggaacagcac aaacactacc tggacgagat
4741 catcgagcag atcagcgagt tctccaagag agtgatcctg gccgacgcta atctggacaa
4801 ggtgctgagc gcctacaaca agcacagaga caagcctatc agagagcagg ccgagaatat
4861 catccacctg tttaccctga ccaatctggg agcccctgcc gccttcaagt actttgacac
4921 caccatcgac cggaagaggt acaccagcac caaagaggtg ctggacgcca ccctgatcca
4981 ccagagcatc accggcctgt acgagacacg gatcgacctg tctcagctgg gaggcgacgc
5041 ctatccctat gacgtgcccg attatgccag cctgggcagc ggctccccca agaaaaaacg
5101 caaggtggaa gatcctaaga aaaagcggaa agtggacggc attggtagtg ggagcaacgg
5161 cagcagcgga tccgtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
5221 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg cgcggcgagg gcgagggcga
5281 tgccaccaac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
5341 ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga
5401 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
5461 caccatctcc ttcaaggacg acggcaccta caagacccgc gccgaggtga agttcgaggg
5521 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
5581 cctggggcac aagctggagt acaacttcaa cagccacaac gtctatatca cggccgacaa
5641 gcagaagaac ggcatcaagg cgaacttcaa gatccgccac aacgtcgagg acggcagcgt
5701 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
5761 cgacaaccac tacctgagca cccagtccaa gctgagcaaa gaccccaacg agaagcgcga
5821 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
5881 gtacaagtag taatctagag ggccctattc tatagtgtca cctaaatgct agagctcgct
5941 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
6001 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
6061 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
6121 agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt
6181 ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg
6241 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
6301 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
6361 gtcaagctct aaatcggggc atccctttag ggttccgatt tagtgcttta cggcacctcg
6421 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
6481 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
6541 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttggggattt
6601 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg
6661 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccaggcaggc agaagtatgc
6721 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
6781 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
6841 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
6901 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
6961 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca
7021 ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
7081 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
7141 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
7201 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc
7261 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
7321 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
7381 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
7441 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
7501 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
7561 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga
7621 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
7681 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
7741 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
7801 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
7861 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
7921 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
7981 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
8041 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
8101 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
8161 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
8221 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
8281 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
8341 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
8401 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
8461 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
8521 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
8581 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
8641 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
8701 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
8761 cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag
8821 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
8881 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
8941 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
9001 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
9061 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
9121 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
9181 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
9241 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
9301 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
9361 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
9421 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
9481 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
9541 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
9601 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
9661 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
9721 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
9781 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
9841 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
9901 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
9961 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
10021 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
10081 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
10141 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
10201 acggaaatgt tgaatactca tactcttcct ttttcattat tattgaagca tttatcaggg
10261 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
10321 tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//
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