| 质粒名称: | pREDIT_Cas9-MS2-BB_BbsI |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 164802 |
| 存储实验室: | Le Cong |
|
载体骨架基本信息 |
|
| 载体名称: | pX330 |
| 载体出品公司: | -- |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | -- |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | -- |
|
插入基因信息 |
|
| 插入基因名称: | -- |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子: | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | -- |
| 5’克隆位点: | -- |
| 3’克隆位点: | -- |
| 5’测序引物 | -- |
| 3’测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pREDIT_Cas9-MS2-BB_BbsI was a gift from Le Cong (Addgene plasmid # 164802 ; http://n2t.net/addgene:164802 ; RRID:Addgene_164802) |
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 pREDIT_Cas9-MS2-BB_BbsI 9262 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION REDIT backbone for wildtype Cas9,
pU6-MS2-gRNA-backbone(BbsI)-CBH-SpCas9-T2A-EBFP.
ACCESSION .
VERSION .
KEYWORDS pREDIT_Cas9-MS2-BB_BbsI
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9262)
TITLE Microbial Single-strand Annealing Proteins Enable CRISPR
Gene-editing Tools with Improved Knock-in Efficiencies and Reduced
Off-target Effects
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 9262)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..9262
/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 285..303
/label=MS2 stem loop
/note="stem loop that binds the bacteriophage MS2 coat
protein"
misc_RNA 378..396
/label=MS2 stem loop
/note="stem loop that binds the bacteriophage MS2 coat
protein"
enhancer 417..702
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer;
contains an 18-bp deletion relative to the standard CMV
enhancer"
intron 982..1209
/label=hybrid intron
/note="hybrid between chicken beta-actin (CBA) and minute
virus of mice (MMV) introns (Gray et al., 2011)"
regulatory 1221..1230
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 1230..1295
/codon_start=1
/product="three tandem FLAG(R) epitope tags, followed by an
enterokinase cleavage site"
/label=3xFLAG
/translation="DYKDHDGDYKDHDIDYKDDDDK"
CDS 1302..1322
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 1347..5447
/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 5448..5495
/codon_start=1
/product="bipartite nuclear localization signal from
nucleoplasmin"
/label=nucleoplasmin NLS
/translation="KRPAATKKAGQAKKKK"
CDS 5499..5552
/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 5559..6278
/codon_start=1
/product="enhanced blue variant of GFP (Ai et al., 2007)"
/label=EBFP2
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
KFICTTGKLPVPWPTLVTTLSHGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GTYKTRAEVKFEGDTLVNRIELKGVDFKEDGNILGHKLEYNFNSHNIYIMAVKQKNGIK
VNFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDSHYLSTQSVLSKDPNEKRDHMVLL
EFRTAAGITLGMDELYK"
primer_bind complement(5604..5625)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(6303..6320)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 6309..6516
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
repeat_region 6525..6665
/label=AAV2 ITR
/note="inverted terminal repeat of adeno-associated virus
serotype 2"
repeat_region 6525..6654
/label=AAV2 ITR
rep_origin 6740..7195
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(6827..6846)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 7037..7058
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(7212..7231)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 7331..7353
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(7391..7409)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 7477..7581
/gene="bla"
/label=AmpR promoter
CDS 7582..8442
/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(7800..7819)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 8613..9201
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 9102..9121
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag
61 ataattggaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtggcgtaga
121 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat
181 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga
241 cgaaacaccg ggtcttcgag aagacctgtt taagagctag gccaacatga ggatcaccca
301 tgtctgcagg gcctagcaag tttaaataag gctagtccgt tatcaacttg aaaaagtggc
361 accgagtcgg tgcgcgcaca tgaggatcac ccatgtgctt tttttctaga ggtacccgtt
421 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
481 tcaatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
541 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
601 aatgacggta aatggcccgc ctggcattgt gcccagtaca tgaccttatg ggactttcct
661 acttggcagt acatctacgt attagtcatc gctattacca tggtcgaggt gagccccacg
721 ttctgcttca ctctccccat ctcccccccc tccccacccc caattttgta tttatttatt
781 ttttaattat tttgtgcagc gatgggggcg gggggggggg gggggcgcgc gccggggggg
841 gggggggggg gggggggggg ggggggggsg rggsggrgrg gkgcggcggc agccaatcag
901 agcggcgcgc tccgaaagtt tccttttatg gcgaggcggc ggcggcggcg gccctataaa
961 aagcgaagcg cgcggcgggc gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc
1021 gccgccgcct cgcgccgccc gccccggctc tgactgaccg cgttactccc acaggtgagc
1081 gggcgggacg gcccttctcc tccgggctgt aattagctga gcaagaggta agggtttaag
1141 ggatggttgg ttggtggggt attaatgttt aattacctgg agcacctgcc tgaaatcact
1201 ttttttcagg ttggaccggt gccaccatgg actataagga ccacgacgga gactacaagg
1261 atcatgatat tgattacaaa gacgatgacg ataagatggc cccaaagaag aagcggaagg
1321 tcggtatcca cggagtccca gcagccgaca agaagtacag catcggcctg gacatcggca
1381 ccaactctgt gggctgggcc gtgatcaccg acgagtacaa ggtgcccagc aagaaattca
1441 aggtgctggg caacaccgac cggcacagca tcaagaagaa cctgatcgga gccctgctgt
1501 tcgacagcgg cgaaacagcc gaggccaccc ggctgaagag aaccgccaga agaagataca
1561 ccagacggaa gaaccggatc tgctatctgc aagagatctt cagcaacgag atggccaagg
1621 tggacgacag cttcttccac agactggaag agtccttcct ggtggaagag gataagaagc
1681 acgagcggca ccccatcttc ggcaacatcg tggacgaggt ggcctaccac gagaagtacc
1741 ccaccatcta ccacctgaga aagaaactgg tggacagcac cgacaaggcc gacctgcggc
1801 tgatctatct ggccctggcc cacatgatca agttccgggg ccacttcctg atcgagggcg
1861 acctgaaccc cgacaacagc gacgtggaca agctgttcat ccagctggtg cagacctaca
1921 accagctgtt cgaggaaaac cccatcaacg ccagcggcgt ggacgccaag gccatcctgt
1981 ctgccagact gagcaagagc agacggctgg aaaatctgat cgcccagctg cccggcgaga
2041 agaagaatgg cctgttcgga aacctgattg ccctgagcct gggcctgacc cccaacttca
2101 agagcaactt cgacctggcc gaggatgcca aactgcagct gagcaaggac acctacgacg
2161 acgacctgga caacctgctg gcccagatcg gcgaccagta cgccgacctg tttctggccg
2221 ccaagaacct gtccgacgcc atcctgctga gcgacatcct gagagtgaac accgagatca
2281 ccaaggcccc cctgagcgcc tctatgatca agagatacga cgagcaccac caggacctga
2341 ccctgctgaa agctctcgtg cggcagcagc tgcctgagaa gtacaaagag attttcttcg
2401 accagagcaa gaacggctac gccggctaca ttgacggcgg agccagccag gaagagttct
2461 acaagttcat caagcccatc ctggaaaaga tggacggcac cgaggaactg ctcgtgaagc
2521 tgaacagaga ggacctgctg cggaagcagc ggaccttcga caacggcagc atcccccacc
2581 agatccacct gggagagctg cacgccattc tgcggcggca ggaagatttt tacccattcc
2641 tgaaggacaa ccgggaaaag atcgagaaga tcctgacctt ccgcatcccc tactacgtgg
2701 gccctctggc caggggaaac agcagattcg cctggatgac cagaaagagc gaggaaacca
2761 tcaccccctg gaacttcgag gaagtggtgg acaagggcgc ttccgcccag agcttcatcg
2821 agcggatgac caacttcgat aagaacctgc ccaacgagaa ggtgctgccc aagcacagcc
2881 tgctgtacga gtacttcacc gtgtataacg agctgaccaa agtgaaatac gtgaccgagg
2941 gaatgagaaa gcccgccttc ctgagcggcg agcagaaaaa ggccatcgtg gacctgctgt
3001 tcaagaccaa ccggaaagtg accgtgaagc agctgaaaga ggactacttc aagaaaatcg
3061 agtgcttcga ctccgtggaa atctccggcg tggaagatcg gttcaacgcc tccctgggca
3121 cataccacga tctgctgaaa attatcaagg acaaggactt cctggacaat gaggaaaacg
3181 aggacattct ggaagatatc gtgctgaccc tgacactgtt tgaggacaga gagatgatcg
3241 aggaacggct gaaaacctat gcccacctgt tcgacgacaa agtgatgaag cagctgaagc
3301 ggcggagata caccggctgg ggcaggctga gccggaagct gatcaacggc atccgggaca
3361 agcagtccgg caagacaatc ctggatttcc tgaagtccga cggcttcgcc aacagaaact
3421 tcatgcagct gatccacgac gacagcctga cctttaaaga ggacatccag aaagcccagg
3481 tgtccggcca gggcgatagc ctgcacgagc acattgccaa tctggccggc agccccgcca
3541 ttaagaaggg catcctgcag acagtgaagg tggtggacga gctcgtgaaa gtgatgggcc
3601 ggcacaagcc cgagaacatc gtgatcgaaa tggccagaga gaaccagacc acccagaagg
3661 gacagaagaa cagccgcgag agaatgaagc ggatcgaaga gggcatcaaa gagctgggca
3721 gccagatcct gaaagaacac cccgtggaaa acacccagct gcagaacgag aagctgtacc
3781 tgtactacct gcagaatggg cgggatatgt acgtggacca ggaactggac atcaaccggc
3841 tgtccgacta cgatgtggac catatcgtgc ctcagagctt tctgaaggac gactccatcg
3901 acaacaaggt gctgaccaga agcgacaaga accggggcaa gagcgacaac gtgccctccg
3961 aagaggtcgt gaagaagatg aagaactact ggcggcagct gctgaacgcc aagctgatta
4021 cccagagaaa gttcgacaat ctgaccaagg ccgagagagg cggcctgagc gaactggata
4081 aggccggctt catcaagaga cagctggtgg aaacccggca gatcacaaag cacgtggcac
4141 agatcctgga ctcccggatg aacactaagt acgacgagaa tgacaagctg atccgggaag
4201 tgaaagtgat caccctgaag tccaagctgg tgtccgattt ccggaaggat ttccagtttt
4261 acaaagtgcg cgagatcaac aactaccacc acgcccacga cgcctacctg aacgccgtcg
4321 tgggaaccgc cctgatcaaa aagtacccta agctggaaag cgagttcgtg tacggcgact
4381 acaaggtgta cgacgtgcgg aagatgatcg ccaagagcga gcaggaaatc ggcaaggcta
4441 ccgccaagta cttcttctac agcaacatca tgaacttttt caagaccgag attaccctgg
4501 ccaacggcga gatccggaag cggcctctga tcgagacaaa cggcgaaacc ggggagatcg
4561 tgtgggataa gggccgggat tttgccaccg tgcggaaagt gctgagcatg ccccaagtga
4621 atatcgtgaa aaagaccgag gtgcagacag gcggcttcag caaagagtct atcctgccca
4681 agaggaacag cgataagctg atcgccagaa agaaggactg ggaccctaag aagtacggcg
4741 gcttcgacag ccccaccgtg gcctattctg tgctggtggt ggccaaagtg gaaaagggca
4801 agtccaagaa actgaagagt gtgaaagagc tgctggggat caccatcatg gaaagaagca
4861 gcttcgagaa gaatcccatc gactttctgg aagccaaggg ctacaaagaa gtgaaaaagg
4921 acctgatcat caagctgcct aagtactccc tgttcgagct ggaaaacggc cggaagagaa
4981 tgctggcctc tgccggcgaa ctgcagaagg gaaacgaact ggccctgccc tccaaatatg
5041 tgaacttcct gtacctggcc agccactatg agaagctgaa gggctccccc gaggataatg
5101 agcagaaaca gctgtttgtg gaacagcaca agcactacct ggacgagatc atcgagcaga
5161 tcagcgagtt ctccaagaga gtgatcctgg ccgacgctaa tctggacaaa gtgctgtccg
5221 cctacaacaa gcaccgggat aagcccatca gagagcaggc cgagaatatc atccacctgt
5281 ttaccctgac caatctggga gcccctgccg ccttcaagta ctttgacacc accatcgacc
5341 ggaagaggta caccagcacc aaagaggtgc tggacgccac cctgatccac cagagcatca
5401 ccggcctgta cgagacacgg atcgacctgt ctcagctggg aggcgacaaa aggccggcgg
5461 ccacgaaaaa ggccggccag gcaaaaaaga aaaagcttga gggcagagga agtctgctaa
5521 catgcggtga cgtggaggag aatcccggcc ctgctagcat ggtgagcaag ggcgaggagc
5581 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
5641 tcagcgtgag gggcgagggc gagggcgatg ccaccaacgg caagctgacc ctgaagttca
5701 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgagccacg
5761 gcgtgcagtg cttcgcccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
5821 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcacctaca
5881 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
5941 gcgtcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aacttcaaca
6001 gccacaacat ctatatcatg gccgtcaagc agaagaacgg catcaaggtg aacttcaaga
6061 tccgccacaa cgtggaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc
6121 ccatcggcga cggccccgtg ctgctgcccg acagccacta cctgagcacc cagtccgtgc
6181 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc cgcaccgccg
6241 ccgggatcac tctcggcatg gacgagctgt acaagtgaga attcctagag ctcgctgatc
6301 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
6361 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
6421 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
6481 ggaggattgg gaagagaata gcaggcatgc tggggagcgg ccgcaggaac ccctagtgat
6541 ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt
6601 cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct
6661 gcaggggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
6721 acgtcaaagc aaccatagta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
6781 gttacgcgca gcgtgaccgc tacacttgcc agcgccttag cgcccgctcc tttcgctttc
6841 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
6901 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt
6961 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
7021 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ctctatctcg
7081 ggctattctt ttgatttata agggattttg ccgatttcgg tctattggtt aaaaaatgag
7141 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aattttatgg
7201 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca
7261 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
7321 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
7381 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
7441 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
7501 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
7561 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
7621 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
7681 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
7741 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
7801 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
7861 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
7921 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
7981 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
8041 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
8101 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
8161 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
8221 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
8281 ggagccggtg agcgtggaag ccgcggtatc attgcagcac tggggccaga tggtaagccc
8341 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
8401 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
8461 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
8521 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
8581 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
8641 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
8701 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
8761 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
8821 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
8881 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
8941 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
9001 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
9061 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
9121 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
9181 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
9241 tgctggcctt ttgctcacat gt
//
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