| 质粒名称: | pspCas9-POLI-ST2-com vector |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 176234 |
| 存储实验室: | Baisong Lu |
|
载体骨架基本信息 |
|
| 载体名称: | Addgene# 136269 |
| 完整质粒大小: | 11588bp |
| 载体类型: | 细菌细胞表达 |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | NEB Stable |
| 拷贝数: | 未知 |
|
文献引用方法 |
|
| 材料和方法部分: |
pspCas9-POLI-ST2-com vector was a gift from Baisong Lu (Addgene plasmid # 176234 ; http://n2t.net/addgene:176234 ; RRID:Addgene_176234)
|
| 参考文献部分: | Targeting DNA polymerase to DNA double-strand breaks reduces DNA deletion size and increases templated insertions generated by CRISPR/Cas9. Yoo KW, Yadav MK, Song Q, Atala A, Lu B. Nucleic Acids Res. 2022 Mar 22. pii: 6552087. doi: 10.1093/nar/gkac186. 10.1093/nar/gkac186 PubMed 35323942 |
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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 pspCas9-POLI-ST2-com vector 11588 bp ds-DNA circular SYN 29-NOV-2021
DEFINITION Vector plasmid for expressing spCas9-POLI fusion protein and sgRNA.
There are two copies of HBB 3' UTR in the 3'UTR of spCas9-POLI to
enhance expression. The ST2 loop of the sgRNA scaffold was replace.
ACCESSION .
VERSION .
KEYWORDS pspCas9-POLI-ST2-com vector
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 11588)
TITLE Targeting DNA Polymerase to DNA double strand breaks reduced the
sizes of CRISPR/Cas9-induced DNA deletions in human cells
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 11588)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Nov 29, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..11588
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 6..291
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer;
contains an 18-bp deletion relative to the standard CMV
enhancer"
intron 571..798
/label=hybrid intron
/note="hybrid between chicken beta-actin (CBA) and minute
virus of mice (MMV) introns (Gray et al., 2011)"
regulatory 810..819
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 819..884
/codon_start=1
/product="three tandem FLAG(R) epitope tags, followed by an
enterokinase cleavage site"
/label=3xFLAG
/translation="DYKDHDGDYKDHDIDYKDDDDK"
CDS 891..911
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 936..5036
/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 5037..5084
/codon_start=1
/product="bipartite nuclear localization signal from
nucleoplasmin"
/label=nucleoplasmin NLS
/translation="KRPAATKKAGQAKKKK"
RBS 5867..5875
/label=Shine-Dalgarno sequence
/note="full consensus sequence for ribosome-binding sites
upstream of start codons in E. coli; complementary to a
region in the 3' end of the 16S rRNA (Chen et al., 1994)"
primer_bind complement(8247..8264)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 8253..8460
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
repeat_region 8472..8612
/label=AAV2 ITR
/note="inverted terminal repeat of adeno-associated virus
serotype 2"
repeat_region 8472..8601
/label=AAV2 ITR
rep_origin 8687..9142
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(8774..8793)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 8984..9005
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(9159..9178)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 9278..9300
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(9338..9356)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 9424..9528
/gene="bla"
/label=AmpR promoter
CDS 9529..10389
/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(9747..9766)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 10560..11148
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 11049..11068
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
promoter 11210..11450
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
primer_bind 11210..11230
/label=hU6-F
/note="Human U6 promoter, forward primer"
primer_bind 11381..11400
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
ORIGIN
1 gtacccgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
61 ccattgacgt caatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
121 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct
181 attgacgtca atgacggtaa atggcccgcc tggcattgtg cccagtacat gaccttatgg
241 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtcgaggtg
301 agccccacgt tctgcttcac tctccccatc tcccccccct ccccaccccc aattttgtat
361 ttatttattt tttaattatt ttgtgcagcg atgggggcgg gggggggggg ggggcgcgcg
421 ccrggsgggg sggggggggg sggggggsgg ggsggggsgr ggsggrgrgg kgcggcggca
481 gccaatcaga gcggcgcgct ccgaaagttt ccttttatgg cgaggcggcg gcggcggcgg
541 ccctataaaa agcgaagcgc gcggcgggcg ggagtcgctg cgcgctgcct tcgccccgtg
601 ccccgctccg ccgccgcctc gcgccgcccg ccccggctct gactgaccgc gttactccca
661 caggtgagcg ggcgggacgg cccttctcct ccgggctgta attagctgag caagaggtaa
721 gggtttaagg gatggttggt tggtggggta ttaatgttta attacctgga gcacctgcct
781 gaaatcactt tttttcaggt tggaccggtg ccaccatgga ctataaggac cacgacggag
841 actacaagga tcatgatatt gattacaaag acgatgacga taagatggcc ccaaagaaga
901 agcggaaggt cggtatccac ggagtcccag cagccgacaa gaagtacagc atcggcctgg
961 acatcggcac caactctgtg ggctgggccg tgatcaccga cgagtacaag gtgcccagca
1021 agaaattcaa ggtgctgggc aacaccgacc ggcacagcat caagaagaac ctgatcggag
1081 ccctgctgtt cgacagcggc gaaacagccg aggccacccg gctgaagaga accgccagaa
1141 gaagatacac cagacggaag aaccggatct gctatctgca agagatcttc agcaacgaga
1201 tggccaaggt ggacgacagc ttcttccaca gactggaaga gtccttcctg gtggaagagg
1261 ataagaagca cgagcggcac cccatcttcg gcaacatcgt ggacgaggtg gcctaccacg
1321 agaagtaccc caccatctac cacctgagaa agaaactggt ggacagcacc gacaaggccg
1381 acctgcggct gatctatctg gccctggccc acatgatcaa gttccggggc cacttcctga
1441 tcgagggcga cctgaacccc gacaacagcg acgtggacaa gctgttcatc cagctggtgc
1501 agacctacaa ccagctgttc gaggaaaacc ccatcaacgc cagcggcgtg gacgccaagg
1561 ccatcctgtc tgccagactg agcaagagca gacggctgga aaatctgatc gcccagctgc
1621 ccggcgagaa gaagaatggc ctgttcggaa acctgattgc cctgagcctg ggcctgaccc
1681 ccaacttcaa gagcaacttc gacctggccg aggatgccaa actgcagctg agcaaggaca
1741 cctacgacga cgacctggac aacctgctgg cccagatcgg cgaccagtac gccgacctgt
1801 ttctggccgc caagaacctg tccgacgcca tcctgctgag cgacatcctg agagtgaaca
1861 ccgagatcac caaggccccc ctgagcgcct ctatgatcaa gagatacgac gagcaccacc
1921 aggacctgac cctgctgaaa gctctcgtgc ggcagcagct gcctgagaag tacaaagaga
1981 ttttcttcga ccagagcaag aacggctacg ccggctacat tgacggcgga gccagccagg
2041 aagagttcta caagttcatc aagcccatcc tggaaaagat ggacggcacc gaggaactgc
2101 tcgtgaagct gaacagagag gacctgctgc ggaagcagcg gaccttcgac aacggcagca
2161 tcccccacca gatccacctg ggagagctgc acgccattct gcggcggcag gaagattttt
2221 acccattcct gaaggacaac cgggaaaaga tcgagaagat cctgaccttc cgcatcccct
2281 actacgtggg ccctctggcc aggggaaaca gcagattcgc ctggatgacc agaaagagcg
2341 aggaaaccat caccccctgg aacttcgagg aagtggtgga caagggcgct tccgcccaga
2401 gcttcatcga gcggatgacc aacttcgata agaacctgcc caacgagaag gtgctgccca
2461 agcacagcct gctgtacgag tacttcaccg tgtataacga gctgaccaaa gtgaaatacg
2521 tgaccgaggg aatgagaaag cccgccttcc tgagcggcga gcagaaaaag gccatcgtgg
2581 acctgctgtt caagaccaac cggaaagtga ccgtgaagca gctgaaagag gactacttca
2641 agaaaatcga gtgcttcgac tccgtggaaa tctccggcgt ggaagatcgg ttcaacgcct
2701 ccctgggcac ataccacgat ctgctgaaaa ttatcaagga caaggacttc ctggacaatg
2761 aggaaaacga ggacattctg gaagatatcg tgctgaccct gacactgttt gaggacagag
2821 agatgatcga ggaacggctg aaaacctatg cccacctgtt cgacgacaaa gtgatgaagc
2881 agctgaagcg gcggagatac accggctggg gcaggctgag ccggaagctg atcaacggca
2941 tccgggacaa gcagtccggc aagacaatcc tggatttcct gaagtccgac ggcttcgcca
3001 acagaaactt catgcagctg atccacgacg acagcctgac ctttaaagag gacatccaga
3061 aagcccaggt gtccggccag ggcgatagcc tgcacgagca cattgccaat ctggccggca
3121 gccccgccat taagaagggc atcctgcaga cagtgaaggt ggtggacgag ctcgtgaaag
3181 tgatgggccg gcacaagccc gagaacatcg tgatcgaaat ggccagagag aaccagacca
3241 cccagaaggg acagaagaac agccgcgaga gaatgaagcg gatcgaagag ggcatcaaag
3301 agctgggcag ccagatcctg aaagaacacc ccgtggaaaa cacccagctg cagaacgaga
3361 agctgtacct gtactacctg cagaatgggc gggatatgta cgtggaccag gaactggaca
3421 tcaaccggct gtccgactac gatgtggacc atatcgtgcc tcagagcttt ctgaaggacg
3481 actccatcga caacaaggtg ctgaccagaa gcgacaagaa ccggggcaag agcgacaacg
3541 tgccctccga agaggtcgtg aagaagatga agaactactg gcggcagctg ctgaacgcca
3601 agctgattac ccagagaaag ttcgacaatc tgaccaaggc cgagagaggc ggcctgagcg
3661 aactggataa ggccggcttc atcaagagac agctggtgga aacccggcag atcacaaagc
3721 acgtggcaca gatcctggac tcccggatga acactaagta cgacgagaat gacaagctga
3781 tccgggaagt gaaagtgatc accctgaagt ccaagctggt gtccgatttc cggaaggatt
3841 tccagtttta caaagtgcgc gagatcaaca actaccacca cgcccacgac gcctacctga
3901 acgccgtcgt gggaaccgcc ctgatcaaaa agtaccctaa gctggaaagc gagttcgtgt
3961 acggcgacta caaggtgtac gacgtgcgga agatgatcgc caagagcgag caggaaatcg
4021 gcaaggctac cgccaagtac ttcttctaca gcaacatcat gaactttttc aagaccgaga
4081 ttaccctggc caacggcgag atccggaagc ggcctctgat cgagacaaac ggcgaaaccg
4141 gggagatcgt gtgggataag ggccgggatt ttgccaccgt gcggaaagtg ctgagcatgc
4201 cccaagtgaa tatcgtgaaa aagaccgagg tgcagacagg cggcttcagc aaagagtcta
4261 tcctgcccaa gaggaacagc gataagctga tcgccagaaa gaaggactgg gaccctaaga
4321 agtacggcgg cttcgacagc cccaccgtgg cctattctgt gctggtggtg gccaaagtgg
4381 aaaagggcaa gtccaagaaa ctgaagagtg tgaaagagct gctggggatc accatcatgg
4441 aaagaagcag cttcgagaag aatcccatcg actttctgga agccaagggc tacaaagaag
4501 tgaaaaagga cctgatcatc aagctgccta agtactccct gttcgagctg gaaaacggcc
4561 ggaagagaat gctggcctct gccggcgaac tgcagaaggg aaacgaactg gccctgccct
4621 ccaaatatgt gaacttcctg tacctggcca gccactatga gaagctgaag ggctcccccg
4681 aggataatga gcagaaacag ctgtttgtgg aacagcacaa gcactacctg gacgagatca
4741 tcgagcagat cagcgagttc tccaagagag tgatcctggc cgacgctaat ctggacaaag
4801 tgctgtccgc ctacaacaag caccgggata agcccatcag agagcaggcc gagaatatca
4861 tccacctgtt taccctgacc aatctgggag cccctgccgc cttcaagtac tttgacacca
4921 ccatcgaccg gaagaggtac accagcacca aagaggtgct ggacgccacc ctgatccacc
4981 agagcatcac cggcctgtac gagacacgga tcgacctgtc tcagctggga ggcgacaaaa
5041 ggccggcggc cacgaaaaag gccggccagg caaaaaagaa aaagggatcc tctgagacac
5101 caggcacctc cgagtctgcc acccctgagg gaggaagcgg aggaagcggc tccgtgcaga
5161 tcccacagaa ccccctgatc ctggtggacg gcagctccta cctgtatcgg gcctaccacg
5221 ccttcccacc tctgacaaac tccgccggag agccaaccgg agccatgtat ggcgtgctga
5281 atatgctgag gagcctgatc atgcagtaca agcctacaca cgccgccgtg gtgtttgatg
5341 ccaagggcaa gaccttccgc gacgagctgt ttgagcacta caagagccac aggccaccaa
5401 tgcctgacga tctgagggca cagatcgagc cactgcacgc aatggtgaag gccatgggcc
5461 tgcctctgct ggccgtgagc ggagtggagg ccgacgatgt gatcggcaca ctggcaaggg
5521 aggcagagaa ggcaggccgc ccagtgctga tctccaccgg cgacaaggat atggcccagc
5581 tggtgacacc aaacatcacc ctgatcaaca ccatgacaaa taccatcctg ggccccgagg
5641 aggtggtgaa taagtatggc gtgcctccag agctgatcat cgatttcctg gccctgatgg
5701 gcgactctag cgataacatc cctggcgtgc caggagtggg agaaaagacc gcacaggccc
5761 tgctgcaggg cctgggaggc ctggacaccc tgtacgccga gccagagaag atcgccggcc
5821 tgtcctttag gggcgccaag acaatggccg ccaagctgga gcagaataag gaggtggcct
5881 acctgtctta tcagctggcc acaatcaaga ccgacgtgga gctggagctg acctgcgagc
5941 agctggaggt gcagcagcct gcagcagagg agctgctggg cctgttcaag aagtacgagt
6001 ttaagagatg gacagccgat gtggaggccg gcaagtggct gcaggcaaag ggagcaaagc
6061 cagcagcaaa gccacaggag acaagcgtgg cagacgaggc accagaggtg acagccaccg
6121 tgatctccta cgataactat gtgacaatcc tggacgagga gacactgaag gcctggatcg
6181 ccaagctgga gaaggccccc gtgttcgcct ttgatacaga gacagacagc ctggataaca
6241 tctccgccaa tctggtgggc ctgtctttcg ccatcgagcc tggcgtggcc gcctatatcc
6301 cagtggccca cgactacctg gatgcccccg accagatcag cagggagaga gccctggagc
6361 tgctgaagcc tctgctggag gatgagaagg ccctgaaggt cggccagaac ctgaagtatg
6421 acaggggcat cctggccaat tacggcatcg agctgagagg catcgccttt gacaccatgc
6481 tggagtctta tatcctgaat agcgtggcag gccggcacga catggattcc ctggccgaga
6541 ggtggctgaa gcacaagaca atcaccttcg aggagatcgc cggcaagggc aagaaccagc
6601 tgaccttcaa ccagatcgcc ctggaggagg caggcaggta cgcagcagag gacgcagatg
6661 tgaccctgca gctgcacctg aagatgtggc cagatctgca gaagcacaag ggccccctga
6721 acgtctttga gaatatcgag atgcccctgg tgcctgtgct gagccggatc gagcgcaacg
6781 gcgtgaagat cgaccctaag gtgctgcaca atcactccga ggagctgacc ctgagactgg
6841 ccgagctgga gaagaaggcc cacgagatcg ccggcgagga gttcaacctg tcctctacaa
6901 agcagctgca gaccatcctg tttgagaagc agggcatcaa gcccctgaag aaaacccctg
6961 gaggagcacc atctaccagc gaggaggtgc tggaggagct ggccctggat tatcccctgc
7021 ctaaagtgat cctggagtac cggggcctgg ccaagctgaa gtctacatat accgacaagc
7081 tgcccctgat gatcaaccct aagacaggcc gggtgcacac cagctaccac caggcagtga
7141 cagcaaccgg ccgcctgagc tccaccgatc caaacctgca gaatatcccc gtgaggaatg
7201 aggagggcag gagaatcaga caggccttca tcgcccccga ggattatgtg atcgtgtccg
7261 ccgactactc tcagatcgag ctgaggatca tggcccacct gtccagagat aagggcctgc
7321 tgacagcctt cgccgagggc aaggacatcc acagggcaac cgcagcagag gtgtttggcc
7381 tgcctctgga gacagtgacc tccgagcagc ggcgctctgc caaggccatc aacttcggcc
7441 tgatctatgg catgtctgcc tttggcctgg ccaggcagct gaatatccct agaaaggagg
7501 cccagaagta catggacctg tatttcgaga gataccccgg cgtgctggag tacatggaga
7561 ggacacgcgc acaggcaaag gagcagggct atgtggagac actggatggc aggagactgt
7621 acctgccaga catcaagtct agcaacggag caaggagggc agcagcagag agggccgcca
7681 tcaatgcccc catgcagggc acagccgccg atatcatcaa gagagccatg atcgcagtgg
7741 acgcctggct gcaggcagag cagccaaggg tgagaatgat catgcaggtg cacgatgagc
7801 tggtgtttga ggtgcacaag gacgatgtgg acgccgtggc caagcagatc caccagctga
7861 tggagaactg tacccgcctg gatgtgccac tgctggtgga agtgggaagc ggagagaatt
7921 gggaccaggc ccactgagga tcctaagctc gctttcttgc tgtccaattt ctattaaagg
7981 ttcctttgtt ccctaagtcc aactactaaa ctgggggata ttatgaaggg ccttgagcat
8041 ctggattctg cctaataaaa aacatttatt ttcattgcta gctcgctttc ttgctgtcca
8101 atttctatta aaggttcctt tgttccctaa gtccaactac taaactgggg gatattatga
8161 agggccttga gcatctggat tctgcctaat aaaaaacatt tattttcatt gcgaattcta
8221 gaaaattcct agagctcgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
8281 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
8341 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
8401 tggggtgggg caggacagca agggggagga ttgggaagag aatagcaggc atgctgggga
8461 ggtacggccg caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc
8521 gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc
8581 agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt ttctccttac
8641 gcatctgtgc ggtatttcac accgcatacg tcaaagcaac catagtacgc gccctgtagc
8701 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
8761 gccttagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
8821 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
8881 ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag
8941 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
9001 actggaacaa cactcaactc tatctcgggc tattcttttg atttataagg gattttgccg
9061 atttcggtct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
9121 aaaatattaa cgtttacaat tttatggtgc actctcagta caatctgctc tgatgccgca
9181 tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
9241 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
9301 ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta
9361 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
9421 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
9481 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
9541 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
9601 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
9661 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
9721 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
9781 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
9841 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
9901 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
9961 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
10021 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
10081 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
10141 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
10201 gctggctggt ttattgctga taaatctgga gccggtgagc gtggaagccg cggtatcatt
10261 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
10321 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
10381 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
10441 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
10501 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
10561 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
10621 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
10681 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
10741 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
10801 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
10861 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
10921 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
10981 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
11041 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
11101 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
11161 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtg agggcctatt
11221 tcccatgatt ccttcatatt tgcatatacg atacaaggct gttagagaga taattggaat
11281 taatttgact gtaaacacaa agatattagt acaaaatacg tgacgtagaa agtaataatt
11341 tcttgggtag tttgcagttt taaaattatg ttttaaaatg gactatcata tgcttaccgt
11401 aacttgaaag tatttcgatt tcttggcttt atatatcttg tggaaaggac gaaacaccgg
11461 tgtcttcctc gaggaagacc cgtttgagag ctagaaatag caagttcaaa taaggctagt
11521 ccgttatcaa cttggctgaa tgcctgcgag catcccaccc aagtggcacc gagtcggtgc
11581 tttttttg
//
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