| 质粒名称: | px458_2A_GFP_sgRNA_TIAL1 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 106096 |
| 存储实验室: | Thomas Tuschl |
|
载体骨架基本信息 |
|
| 载体名称: | px458 |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | 人细胞色素c氧化酶亚基VIII（human cytochrome C oxidase subunit VIII） |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 新霉素（G418） |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | gRNA TIAL1 |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | --- |
| -- | |
| 标签/融合蛋白: | 人细胞色素c氧化酶亚基VIII（N端） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | AgeI(未破坏) |
| 3' 克隆位点: | NotI (未破坏) |
| 5' 测序引物 | CMV-F |
| 3' 测序引物 | SV40pA-R |
|
文献引用方法 |
|
| 材料和方法部分: | px458_2A_GFP_sgRNA_TIAL1 was a gift from Thomas Tuschl (Addgene plasmid # 106096 ; http://n2t.net/addgene:106096 ; RRID:Addgene_106096) |
| 参考文献部分: | The TIA1 RNA-Binding Protein Family Regulates EIF2AK2-Mediated Stress Response and Cell Cycle Progression. Meyer C, Garzia A, Mazzola M, Gerstberger S, Molina H, Tuschl T. Mol Cell. 2018 Feb 15;69(4):622-635.e6. doi: 10.1016/j.molcel.2018.01.011. 10.1016/j.molcel.2018.01.011 PubMed 29429924 |
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 Exported 9291 bp ds-DNA circular SYN 07-AUG-2018
DEFINITION CRISPR knockout. Expresses Cas9, EGFP, and sgRNA targeting TIAL1.
ACCESSION .
VERSION .
KEYWORDS px458_2A_GFP_sgRNA_TIAL1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9291)
TITLE Tuschl lab plasmids
REFERENCE 2 (bases 1 to 9291)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 7, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..9291
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 1..241
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
primer_bind 1..21
/label=hU6-F
/note="Human U6 promoter, forward primer"
primer_bind 172..191
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
misc_RNA 271..346
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
enhancer 443..728
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer;
contains an 18-bp deletion relative to the standard CMV
enhancer"
promoter 730..1007
/label=chicken beta-actin promoter
intron 1008..1235
/label=hybrid intron
/note="hybrid between chicken beta-actin (CBA) and minute
virus of mice (MMV) introns (Gray et al., 2011)"
regulatory 1247..1256
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 1256..1321
/codon_start=1
/product="three tandem FLAG(R) epitope tags, followed by an
enterokinase cleavage site"
/label=3xFLAG
/translation="DYKDHDGDYKDHDIDYKDDDDK"
CDS 1328..1348
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 1373..5473
/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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
ATLIHQSITGLYETRIDLSQLGGD"
CDS 5474..5521
/codon_start=1
/product="bipartite nuclear localization signal from
nucleoplasmin"
/label=nucleoplasmin NLS
/translation="KRPAATKKAGQAKKKK"
CDS 5537..5590
/codon_start=1
/product="2A peptide from Thosea asigna virus capsid
protein"
/label=T2A
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="EGRGSLLTCGDVEENPGP"
CDS 5591..6304
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
FVTAAGITLGMDELYK"
primer_bind complement(5633..5654)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(5894..5913)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 6241..6262
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind complement(6332..6349)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 6338..6545
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
repeat_region 6554..6694
/label=AAV2 ITR
/note="inverted terminal repeat of adeno-associated virus
serotype 2"
repeat_region 6554..6683
/label=AAV2 ITR
rep_origin 6769..7224
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(6856..6875)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 7066..7087
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(7241..7260)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 7360..7382
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(7420..7438)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 7506..7610
/gene="bla"
/label=AmpR promoter
CDS 7611..8471
/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 complement(7829..7848)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 8642..9230
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 9131..9150
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag
61 ataattggaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga
121 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat
181 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga
241 cgaaacaccg aaatcagcat ttgccccctt gttttagagc tagaaatagc aagttaaaat
301 aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt ttgttttaga
361 gctagaaata gcaagttaaa ataaggctag tccgttttta gcgcgtgcgc caattctgca
421 gacaaatggc tctagaggta cccgttacat aacttacggt aaatggcccg cctggctgac
481 cgcccaacga cccccgccca ttgacgtcaa tagtaacgcc aatagggact ttccattgac
541 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
601 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattgtgccc
661 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
721 ttaccatggt cgaggtgagc cccacgttct gcttcactct ccccatctcc cccccctccc
781 cacccccaat tttgtattta tttatttttt aattattttg tgcagcgatg ggggcggggg
841 gggggggggg gcgcgcgcca ggcggggcgg ggcggggcga ggggcggggc ggggcgaggc
901 ggagaggtgc ggcggcagcc aatcagagcg gcgcgctccg aaagtttcct tttatggcga
961 ggcggcggcg gcggcggccc tataaaaagc gaagcgcgcg gcgggcggga gtcgctgcgc
1021 gctgccttcg ccccgtgccc cgctccgccg ccgcctcgcg ccgcccgccc cggctctgac
1081 tgaccgcgtt actcccacag gtgagcgggc gggacggccc ttctcctccg ggctgtaatt
1141 agctgagcaa gaggtaaggg tttaagggat ggttggttgg tggggtatta atgtttaatt
1201 acctggagca cctgcctgaa atcacttttt ttcaggttgg accggtgcca ccatggacta
1261 taaggaccac gacggagact acaaggatca tgatattgat tacaaagacg atgacgataa
1321 gatggcccca aagaagaagc ggaaggtcgg tatccacgga gtcccagcag ccgacaagaa
1381 gtacagcatc ggcctggaca tcggcaccaa ctctgtgggc tgggccgtga tcaccgacga
1441 gtacaaggtg cccagcaaga aattcaaggt gctgggcaac accgaccggc acagcatcaa
1501 gaagaacctg atcggagccc tgctgttcga cagcggcgaa acagccgagg ccacccggct
1561 gaagagaacc gccagaagaa gatacaccag acggaagaac cggatctgct atctgcaaga
1621 gatcttcagc aacgagatgg ccaaggtgga cgacagcttc ttccacagac tggaagagtc
1681 cttcctggtg gaagaggata agaagcacga gcggcacccc atcttcggca acatcgtgga
1741 cgaggtggcc taccacgaga agtaccccac catctaccac ctgagaaaga aactggtgga
1801 cagcaccgac aaggccgacc tgcggctgat ctatctggcc ctggcccaca tgatcaagtt
1861 ccggggccac ttcctgatcg agggcgacct gaaccccgac aacagcgacg tggacaagct
1921 gttcatccag ctggtgcaga cctacaacca gctgttcgag gaaaacccca tcaacgccag
1981 cggcgtggac gccaaggcca tcctgtctgc cagactgagc aagagcagac ggctggaaaa
2041 tctgatcgcc cagctgcccg gcgagaagaa gaatggcctg ttcggaaacc tgattgccct
2101 gagcctgggc ctgaccccca acttcaagag caacttcgac ctggccgagg atgccaaact
2161 gcagctgagc aaggacacct acgacgacga cctggacaac ctgctggccc agatcggcga
2221 ccagtacgcc gacctgtttc tggccgccaa gaacctgtcc gacgccatcc tgctgagcga
2281 catcctgaga gtgaacaccg agatcaccaa ggcccccctg agcgcctcta tgatcaagag
2341 atacgacgag caccaccagg acctgaccct gctgaaagct ctcgtgcggc agcagctgcc
2401 tgagaagtac aaagagattt tcttcgacca gagcaagaac ggctacgccg gctacattga
2461 cggcggagcc agccaggaag agttctacaa gttcatcaag cccatcctgg aaaagatgga
2521 cggcaccgag gaactgctcg tgaagctgaa cagagaggac ctgctgcgga agcagcggac
2581 cttcgacaac ggcagcatcc cccaccagat ccacctggga gagctgcacg ccattctgcg
2641 gcggcaggaa gatttttacc cattcctgaa ggacaaccgg gaaaagatcg agaagatcct
2701 gaccttccgc atcccctact acgtgggccc tctggccagg ggaaacagca gattcgcctg
2761 gatgaccaga aagagcgagg aaaccatcac cccctggaac ttcgaggaag tggtggacaa
2821 gggcgcttcc gcccagagct tcatcgagcg gatgaccaac ttcgataaga acctgcccaa
2881 cgagaaggtg ctgcccaagc acagcctgct gtacgagtac ttcaccgtgt ataacgagct
2941 gaccaaagtg aaatacgtga ccgagggaat gagaaagccc gccttcctga gcggcgagca
3001 gaaaaaggcc atcgtggacc tgctgttcaa gaccaaccgg aaagtgaccg tgaagcagct
3061 gaaagaggac tacttcaaga aaatcgagtg cttcgactcc gtggaaatct ccggcgtgga
3121 agatcggttc aacgcctccc tgggcacata ccacgatctg ctgaaaatta tcaaggacaa
3181 ggacttcctg gacaatgagg aaaacgagga cattctggaa gatatcgtgc tgaccctgac
3241 actgtttgag gacagagaga tgatcgagga acggctgaaa acctatgccc acctgttcga
3301 cgacaaagtg atgaagcagc tgaagcggcg gagatacacc ggctggggca ggctgagccg
3361 gaagctgatc aacggcatcc gggacaagca gtccggcaag acaatcctgg atttcctgaa
3421 gtccgacggc ttcgccaaca gaaacttcat gcagctgatc cacgacgaca gcctgacctt
3481 taaagaggac atccagaaag cccaggtgtc cggccagggc gatagcctgc acgagcacat
3541 tgccaatctg gccggcagcc ccgccattaa gaagggcatc ctgcagacag tgaaggtggt
3601 ggacgagctc gtgaaagtga tgggccggca caagcccgag aacatcgtga tcgaaatggc
3661 cagagagaac cagaccaccc agaagggaca gaagaacagc cgcgagagaa tgaagcggat
3721 cgaagagggc atcaaagagc tgggcagcca gatcctgaaa gaacaccccg tggaaaacac
3781 ccagctgcag aacgagaagc tgtacctgta ctacctgcag aatgggcggg atatgtacgt
3841 ggaccaggaa ctggacatca accggctgtc cgactacgat gtggaccata tcgtgcctca
3901 gagctttctg aaggacgact ccatcgacaa caaggtgctg accagaagcg acaagaaccg
3961 gggcaagagc gacaacgtgc cctccgaaga ggtcgtgaag aagatgaaga actactggcg
4021 gcagctgctg aacgccaagc tgattaccca gagaaagttc gacaatctga ccaaggccga
4081 gagaggcggc ctgagcgaac tggataaggc cggcttcatc aagagacagc tggtggaaac
4141 ccggcagatc acaaagcacg tggcacagat cctggactcc cggatgaaca ctaagtacga
4201 cgagaatgac aagctgatcc gggaagtgaa agtgatcacc ctgaagtcca agctggtgtc
4261 cgatttccgg aaggatttcc agttttacaa agtgcgcgag atcaacaact accaccacgc
4321 ccacgacgcc tacctgaacg ccgtcgtggg aaccgccctg atcaaaaagt accctaagct
4381 ggaaagcgag ttcgtgtacg gcgactacaa ggtgtacgac gtgcggaaga tgatcgccaa
4441 gagcgagcag gaaatcggca aggctaccgc caagtacttc ttctacagca acatcatgaa
4501 ctttttcaag accgagatta ccctggccaa cggcgagatc cggaagcggc ctctgatcga
4561 gacaaacggc gaaaccgggg agatcgtgtg ggataagggc cgggattttg ccaccgtgcg
4621 gaaagtgctg agcatgcccc aagtgaatat cgtgaaaaag accgaggtgc agacaggcgg
4681 cttcagcaaa gagtctatcc tgcccaagag gaacagcgat aagctgatcg ccagaaagaa
4741 ggactgggac cctaagaagt acggcggctt cgacagcccc accgtggcct attctgtgct
4801 ggtggtggcc aaagtggaaa agggcaagtc caagaaactg aagagtgtga aagagctgct
4861 ggggatcacc atcatggaaa gaagcagctt cgagaagaat cccatcgact ttctggaagc
4921 caagggctac aaagaagtga aaaaggacct gatcatcaag ctgcctaagt actccctgtt
4981 cgagctggaa aacggccgga agagaatgct ggcctctgcc ggcgaactgc agaagggaaa
5041 cgaactggcc ctgccctcca aatatgtgaa cttcctgtac ctggccagcc actatgagaa
5101 gctgaagggc tcccccgagg ataatgagca gaaacagctg tttgtggaac agcacaagca
5161 ctacctggac gagatcatcg agcagatcag cgagttctcc aagagagtga tcctggccga
5221 cgctaatctg gacaaagtgc tgtccgccta caacaagcac cgggataagc ccatcagaga
5281 gcaggccgag aatatcatcc acctgtttac cctgaccaat ctgggagccc ctgccgcctt
5341 caagtacttt gacaccacca tcgaccggaa gaggtacacc agcaccaaag aggtgctgga
5401 cgccaccctg atccaccaga gcatcaccgg cctgtacgag acacggatcg acctgtctca
5461 gctgggaggc gacaaaaggc cggcggccac gaaaaaggcc ggccaggcaa aaaagaaaaa
5521 ggaattcggc agtggagagg gcagaggaag tctgctaaca tgcggtgacg tcgaggagaa
5581 tcctggccca gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
5641 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc
5701 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
5761 gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca
5821 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
5881 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
5941 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
6001 ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca
6061 gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca
6121 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
6181 caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca
6241 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
6301 caaggaattc taactagagc tcgctgatca gcctcgactg tgccttctag ttgccagcca
6361 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
6421 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
6481 gggggtgggg tggggcagga cagcaagggg gaggattggg aagagaatag caggcatgct
6541 ggggagcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc
6601 tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc
6661 tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt
6721 acgcatctgt gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta
6781 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
6841 gcgccttagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
6901 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
6961 acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat
7021 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
7081 aaactggaac aacactcaac tctatctcgg gctattcttt tgatttataa gggattttgc
7141 cgatttcggt ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
7201 acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg
7261 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
7321 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
7381 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt
7441 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
7501 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
7561 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
7621 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
7681 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
7741 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
7801 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
7861 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
7921 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
7981 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
8041 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
8101 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
8161 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
8221 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
8281 cggctggctg gtttattgct gataaatctg gagccggtga gcgtggaagc cgcggtatca
8341 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
8401 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
8461 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
8521 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
8581 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
8641 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
8701 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
8761 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
8821 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
8881 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
8941 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
9001 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
9061 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
9121 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
9181 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
9241 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg t
//
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