| 质粒名称: | pBLO1811_Cas9_noNLS_human |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 74490 |
| 存储实验室: | David Savage |
|
载体骨架基本信息 |
|
| 载体名称: | px330 |
| 载体抗性: | 氨苄青霉素 |
| 载体类型: | 哺乳细胞表达 |
| 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: | Cas9,allosterically regulated Cas9 |
| 物种来源: | H. sapiens (human) |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | bsaI（克隆过程中销毁） |
| 3' 克隆位点: | bsaI（克隆过程中销毁） |
| 5' 测序引物 | GAGGGCCTATTTCCCATGATTCCT |
| 3' 测序引物 | TTCCTCATTTTATTAGGAAAGGACAGTG |
|
文献引用方法 |
|
| 材料和方法部分: | pBLO1811_Cas9_noNLS_human was a gift from David Savage (Addgene plasmid # 74490 ; http://n2t.net/addgene:74490 ; RRID:Addgene_74490) |
| 参考文献部分: | Profiling of engineering hotspots identifies an allosteric CRISPR-Cas9 switch. Oakes BL, Nadler DC, Flamholz A, Fellmann C, Staahl BT, Doudna JA, Savage DF. Nat Biotechnol. 2016 May 2. doi: 10.1038/nbt.3528. 10.1038/nbt.3528 PubMed 27136077 |
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 pBLO1811_Cas9_noNLS_human 9152 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Human codon optimized plasmid to express Cas9 t2a mCherry w/o NLS
and a sgRNA.
ACCESSION .
VERSION .
KEYWORDS pBLO1811_Cas9_noNLS_human
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9152)
AUTHORS Oakes BL, Nadler DC, Flamholz A, Fellmann C, Staahl BT, Doudna JA,
Savage DF
TITLE Profiling of engineering hotspots identifies an allosteric
CRISPR-Cas9 switch.
JOURNAL Nat Biotechnol. 2016 May 2. doi: 10.1038/nbt.3528.
PUBMED 27136077
REFERENCE 2 (bases 1 to 9152)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..9152
/organism="synthetic DNA construct"
/mol_type="other DNA"
rep_origin 1..589
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 490..509
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
promoter 651..891
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
primer_bind 651..671
/label=hU6-F
/note="Human U6 promoter, forward primer"
primer_bind 822..841
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
enhancer 1100..1385
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer;
contains an 18-bp deletion relative to the standard CMV
enhancer"
promoter 1387..1664
/label=chicken beta-actin promoter
intron 1665..1893
/label=hybrid intron
/note="hybrid between chicken beta-actin (CBA) and minute
virus of mice (MMV) introns (Gray et al., 2011)"
CDS 1908..1931
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
CDS 1932..6032
/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 6054..6107
/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 6108..6818
/codon_start=1
/product="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
/label=mCherry
/note="mammalian codon-optimized"
/translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
EGRHSTGGMDELYK"
primer_bind complement(6253..6271)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind complement(6843..6860)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 6849..7056
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
repeat_region 7065..7205
/label=AAV2 ITR
/note="inverted terminal repeat of adeno-associated virus
serotype 2"
repeat_region 7065..7194
/label=AAV2 ITR
rep_origin 7280..7735
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(7367..7386)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 7577..7598
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(7752..7771)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 7871..7893
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(7931..7949)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 8017..8121
/gene="bla"
/label=AmpR promoter
CDS 8122..8982
/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(8340..8359)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
ORIGIN
1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
121 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt gagggcctat
661 ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag ataattggaa
721 ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga aagtaataat
781 ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat atgcttaccg
841 taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga cgaaacaccg
901 ggtcttcgag aagacctgtt taagagctat gctggaaaca gcatagcaag tttaaataag
961 gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgcttttttg ttttagagct
1021 agaaatagca agttaaaata aggctagtcc gtttttagcg cgtgcgccaa ttctgcagac
1081 aaatggctct agaggtaccc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
1141 ccaacgaccc ccgcccattg acgtcaatag taacgccaat agggactttc cattgacgtc
1201 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
1261 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tgtgcccagt
1321 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
1381 ccatggtcga ggtgagcccc acgttctgct tcactctccc catctccccc ccctccccac
1441 ccccaatttt gtatttattt attttttaat tattttgtgc agcgatgggg gcgggggggg
1501 ggggggggcg cgcgccaggc ggggcggggc ggggcgaggg gcggggcggg gcgaggcgga
1561 gaggtgcggc ggcagccaat cagagcggcg cgctccgaaa gtttcctttt atggcgaggc
1621 ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg ggcgggagtc gctgcgacgc
1681 tgccttcgcc ccgtgccccg ctccgccgcc gcctcgcgcc gcccgccccg gctctgactg
1741 accgcgttac tcccacaggt gagcgggcgg gacggccctt ctcctccggg ctgtaattag
1801 ctgagcaaga ggtaagggtt taagggatgg ttggttggtg gggtattaat gtttaattac
1861 ctggagcacc tgcctgaaat cacttttttt caggttggac cggtatggat tacaaagacg
1921 atgacgataa ggacaagaag tacagcatcg gcctggacat cggcaccaac tctgtgggct
1981 gggccgtgat caccgacgag tacaaggtgc ccagcaagaa attcaaggtg ctgggcaaca
2041 ccgaccggca cagcatcaag aagaacctga tcggagccct gctgttcgac agcggcgaaa
2101 cagccgaggc cacccggctg aagagaaccg ccagaagaag atacaccaga cggaagaacc
2161 ggatctgcta tctgcaagag atcttcagca acgagatggc caaggtggac gacagcttct
2221 tccacagact ggaagagtcc ttcctggtgg aagaggataa gaagcacgag cggcacccca
2281 tcttcggcaa catcgtggac gaggtggcct accacgagaa gtaccccacc atctaccacc
2341 tgagaaagaa actggtggac agcaccgaca aggccgacct gcggctgatc tatctggccc
2401 tggcccacat gatcaagttc cggggccact tcctgatcga gggcgacctg aaccccgaca
2461 acagcgacgt ggacaagctg ttcatccagc tggtgcagac ctacaaccag ctgttcgagg
2521 aaaaccccat caacgccagc ggcgtggacg ccaaggccat cctgtctgcc agactgagca
2581 agagcagacg gctggaaaat ctgatcgccc agctgcccgg cgagaagaag aatggcctgt
2641 tcggcaacct gattgccctg agcctgggcc tgacccccaa cttcaagagc aacttcgacc
2701 tggccgagga tgccaaactg cagctgagca aggacaccta cgacgacgac ctggacaacc
2761 tgctggccca gatcggcgac cagtacgccg acctgtttct ggccgccaag aacctgtccg
2821 acgccatcct gctgagcgac atcctgagag tgaacaccga gatcaccaag gcccccctga
2881 gcgcctctat gatcaagaga tacgacgagc accaccagga cctgaccctg ctgaaagctc
2941 tcgtgcggca gcagctgcct gagaagtaca aagagatttt cttcgaccag agcaagaacg
3001 gctacgccgg ctacattgac ggcggagcca gccaggaaga gttctacaag ttcatcaagc
3061 ccatcctgga aaagatggac ggcaccgagg aactgctcgt gaagctgaac agagaggacc
3121 tgctgcggaa gcagcggacc ttcgacaacg gcagcatccc ccaccagatc cacctgggag
3181 agctgcacgc cattctgcgg cggcaggaag atttttaccc attcctgaag gacaaccggg
3241 aaaagatcga gaagatcctg accttccgca tcccctacta cgtgggccct ctggccaggg
3301 gaaacagcag attcgcctgg atgaccagaa agagcgagga aaccatcacc ccctggaact
3361 tcgaggaagt ggtggacaag ggcgcttccg cccagagctt catcgagcgg atgaccaact
3421 tcgataagaa cctgcccaac gagaaggtgc tgcccaagca cagcctgctg tacgagtact
3481 tcaccgtgta taacgagctg accaaagtga aatacgtgac cgagggaatg agaaagcccg
3541 ccttcctgag cggcgagcag aaaaaggcca tcgtggacct gctgttcaag accaaccgga
3601 aagtgaccgt gaagcagctg aaagaggact acttcaagaa aatcgagtgc ttcgactccg
3661 tggaaatctc cggcgtggaa gatcggttca acgcctccct gggcacatac cacgatctgc
3721 tgaaaattat caaggacaag gacttcctgg acaatgagga aaacgaggac attctggaag
3781 atatcgtgct gaccctgaca ctgtttgagg acagagagat gatcgaggaa cggctgaaaa
3841 cctatgccca cctgttcgac gacaaagtga tgaagcagct gaagcggcgg agatacaccg
3901 gctggggcag gctgagccgg aagctgatca acggcatccg ggacaagcag tccggcaaga
3961 caatcctgga tttcctgaag tccgacggct tcgccaacag aaacttcatg cagctgatcc
4021 acgacgacag cctgaccttt aaagaggaca tccagaaagc ccaggtgtcc ggccagggcg
4081 atagcctgca cgagcacatt gccaatctgg ccggcagccc cgccattaag aagggcatcc
4141 tgcagacagt gaaggtggtg gacgagctcg tgaaagtgat gggccggcac aagcccgaga
4201 acatcgtgat cgaaatggcc agagagaacc agaccaccca gaagggacag aagaacagcc
4261 gcgagagaat gaagcggatc gaagagggca tcaaagagct gggcagccag atcctgaaag
4321 aacaccccgt ggaaaacacc cagctgcaga acgagaagct gtacctgtac tacctgcaga
4381 atgggcggga tatgtacgtg gaccaggaac tggacatcaa ccggctgtcc gactacgatg
4441 tggaccatat cgtgcctcag agctttctga aggacgactc catcgacaac aaggtgctga
4501 ccagaagcga caagaaccgg ggcaagagcg acaacgtgcc ctccgaagag gtcgtgaaga
4561 agatgaagaa ctactggcgg cagctgctga acgccaagct gattacccag agaaagttcg
4621 acaatctgac caaggccgag agaggcggcc tgagcgaact ggataaggcc ggcttcatca
4681 agagacagct ggtggaaacc cggcagatca caaagcacgt ggcacagatc ctggactccc
4741 ggatgaacac taagtacgac gagaatgaca agctgatccg ggaagtgaaa gtgatcaccc
4801 tgaagtccaa gctggtgtcc gatttccgga aggatttcca gttttacaaa gtgcgcgaga
4861 tcaacaacta ccaccacgcc cacgacgcct acctgaacgc cgtcgtggga accgccctga
4921 tcaaaaagta ccctaagctg gaaagcgagt tcgtgtacgg cgactacaag gtgtacgacg
4981 tgcggaagat gatcgccaag agcgagcagg aaatcggcaa ggctaccgcc aagtacttct
5041 tctacagcaa catcatgaac tttttcaaga ccgagattac cctggccaac ggcgagatcc
5101 ggaagcggcc tctgatcgag acaaacggcg aaaccgggga gatcgtgtgg gataagggcc
5161 gggattttgc caccgtgcgg aaagtgctga gcatgcccca agtgaatatc gtgaaaaaga
5221 ccgaggtgca gacaggcggc ttcagcaaag agtctatcct gcccaagagg aacagcgata
5281 agctgatcgc cagaaagaag gactgggacc ctaagaagta cggcggcttc gacagcccca
5341 ccgtggccta ttctgtgctg gtggtggcca aagtggaaaa gggcaagtcc aagaaactga
5401 agagtgtgaa agagctgctg gggatcacca tcatggaaag aagcagcttc gagaagaatc
5461 ccatcgactt tctggaagcc aagggctaca aagaagtgaa aaaggacctg atcatcaagc
5521 tgcctaagta ctccctgttc gagctggaaa acggccggaa gagaatgctg gcctctgccg
5581 gcgaactgca gaagggaaac gaactggccc tgccctccaa atatgtgaac ttcctgtacc
5641 tggccagcca ctatgagaag ctgaagggct cccccgagga taatgagcag aaacagctgt
5701 ttgtggaaca gcacaagcac tacctggacg agatcatcga gcagatcagc gagttctcca
5761 agagagtgat cctggccgac gctaatctgg acaaagtgct gtccgcctac aacaagcacc
5821 gggataagcc catcagagag caggccgaga atatcatcca cctgtttacc ctgaccaatc
5881 tgggagcccc tgccgccttc aagtactttg acaccaccat cgaccggaag aggtacacca
5941 gcaccaaaga ggtgctggac gccaccctga tccaccagag catcaccggc ctgtacgaga
6001 cacggatcga cctgtctcag ctgggaggcg acggatccga atccggcagt ggagagggca
6061 gaggaagtct gctaacatgc ggtgacgtcg aggagaatcc tggcccaatg gtgagcaagg
6121 gcgaggagga taacatggcc atcatcaagg agttcatgcg cttcaaggtg cacatggagg
6181 gctccgtgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg
6241 gcacccagac cgccaagctg aaggtgacca agggtggccc cctgcccttc gcctgggaca
6301 tcctgtcccc tcagttcatg tacggctcca aggcctacgt gaagcacccc gccgacatcc
6361 ccgactactt gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg
6421 aggacggcgg cgtggtgacc gtgacccagg actcctccct gcaggacggc gagttcatct
6481 acaaggtgaa gctgcgcggc accaacttcc cctccgacgg ccccgtaatg cagaagaaaa
6541 ccatgggctg ggaggcctcc tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg
6601 agatcaagca gaggctgaag ctgaaggacg gcggccacta cgacgctgag gtcaagacca
6661 cctacaaggc caagaagccc gtgcagctgc ccggcgccta caacgtcaac atcaagttgg
6721 acatcacctc ccacaacgag gactacacca tcgtggaaca gtacgaacgc gccgagggcc
6781 gccactccac cggcggcatg gacgagctgt acaagtaaga attcctagag ctcgctgatc
6841 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
6901 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
6961 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
7021 ggaggattgg gaagagaata gcaggcatgc tggggagcgg ccgcaggaac ccctagtgat
7081 ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt
7141 cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct
7201 gcaggggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
7261 acgtcaaagc aaccatagta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
7321 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
7381 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
7441 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt
7501 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
7561 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
7621 ggctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
7681 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aattttatgg
7741 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca
7801 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
7861 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
7921 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
7981 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
8041 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
8101 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
8161 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
8221 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
8281 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
8341 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
8401 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
8461 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
8521 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
8581 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
8641 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
8701 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
8761 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
8821 ggagccggtg agcgtggaag ccgcggtatc attgcagcac tggggccaga tggtaagccc
8881 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
8941 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
9001 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
9061 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
9121 tcagaccccg tagaaaagat caaaggatct tc
//
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