| 质粒名称: | TETv4 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 167983 |
| 存储实验室: | Luke Gilbert |
|
载体骨架基本信息 |
|
| 载体名称: | CAG expression plasmid |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 5574 bp |
| 完整质粒大小: | 11874 bp |
| 载体类型: | 哺乳动物表达，CRISPR，合成生物学 |
| 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: | TETv4 (TET1CD-XTEN80-dCas9) |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 6581 bp |
| 启动子 | CAG |
| 标签/融合蛋白: |
TET1 catalytic domain（插入件上的N端）
3xNLS（插入件上的C端子）
tagBFP（插入件上的C端子）
|
| 克隆方法: | Gibson克隆 |
| 5' 测序引物 | ggcaaagaattctgcagtcg |
| 3' 测序引物 | ccaccaccttctgataggc |
|
文献引用方法 |
|
| 材料和方法部分: | TETv4 was a gift from Luke Gilbert (Addgene plasmid # 167983 ; http://n2t.net/addgene:167983 ; RRID:Addgene_167983) |
| 参考文献部分: | Genome-wide programmable transcriptional memory by CRISPR-based epigenome editing. Nunez JK, Chen J, Pommier GC, Cogan JZ, Replogle JM, Adriaens C, Ramadoss GN, Shi Q, Hung KL, Samelson AJ, Pogson AN, Kim JYS, Chung A, Leonetti MD, Chang HY, Kampmann M, Bernstein BE, Hovestadt V, Gilbert LA, Weissman JS. Cell. 2021 Apr 7. pii: S0092-8674(21)00353-6. doi: 10.1016/j.cell.2021.03.025. 10.1016/j.cell.2021.03.025 PubMed 33838111 |
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 TETv4 11679 bp ds-DNA circular SYN 13-MAY-2021
DEFINITION Expresses TETv4 (TET1CD-XTEN80-dCas9-3xNLS-P2A-BFP) downstream of
the CAG promoter.
ACCESSION .
VERSION .
KEYWORDS TETv4
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 11679)
AUTHORS Nunez JK, Chen J, Pommier GC, Cogan JZ, Replogle JM, Adriaens C,
Ramadoss GN, Shi Q, Hung KL, Samelson AJ, Pogson AN, Kim JYS, Chung
A, Leonetti MD, Chang HY, Kampmann M, Bernstein BE, Hovestadt V,
Gilbert LA, Weissman JS
TITLE Genome-wide programmable transcriptional memory by CRISPR-based
epigenome editing.
JOURNAL Cell. 2021 Apr 7. pii: S0092-8674(21)00353-6. doi:
10.1016/j.cell.2021.03.025.
PUBMED 33838111
REFERENCE 2 (bases 1 to 11679)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..11679
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 5..384
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 386..661
/label=chicken beta-actin promoter
primer_bind 1116..1138
/label=pCAGGS-5
/note="Chimeric intron in CAG promoter, forward primer"
primer_bind 1200..1219
/label=pCAG-F
/note="Rabbit beta-globin intron, for pCAG plasmids,
forward primer"
regulatory 1250..1259
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 3656..7759
/codon_start=1
/product="catalytically dead mutant of the Cas9
endonuclease from the Streptococcus pyogenes Type II
CRISPR/Cas system"
/label=dCas9
/note="RNA-guided DNA-binding protein that lacks
endonuclease activity due to the D10A mutation in the RuvC
catalytic domain and the H840A mutation in the HNH
catalytic domain"
/translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
DATLIHQSITGLYETRIDLSQLGGD"
CDS 7775..7795
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 7799..7819
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 7823..7843
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 7856..7912
/codon_start=1
/product="2A peptide from porcine teschovirus-1
polyprotein"
/label=P2A
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="ATNFSLLKQAGDVEENPGP"
CDS 7913..8611
/codon_start=1
/product="monomeric blue fluorescent protein"
/label=TagBFP
/note="mammalian codon-optimized"
/translation="SELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKVV
EGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTAT
QDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALKL
VGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYCD
LPSKLGHKLN"
primer_bind complement(8701..8720)
/label=Bglob-pA-R
/note="Rabbit beta-globin polyA region, reverse primer"
polyA_signal 8766..8821
/label=-beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal (Gil and
Proudfoot, 1987)"
primer_bind complement(8820..8839)
/label=rbglobpA-R
/note="Rabbit beta-globin polyA, reverse primer. Also
called rb-glob-pA-term-R"
primer_bind complement(9182..9198)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(9182..9198)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(9195..9217)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 9206..9222
/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(9230..9260)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 9275..9296
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 9355..9550
/label=SV40 promoter
/note="SV40 early promoter"
rep_origin 9401..9536
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 9463..9482
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
polyA_signal 9556..9690
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(9593..9612)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 9647..9666
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(9758..9775)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(9929..10517)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(10009..10028)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(10688..11548)
/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 11311..11330
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(11549..11653)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gsggggsggg gsgrggggsg
541 gggsggggsg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
661 gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc
721 gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc
781 tccgggctgt aattagcgct tggtttaatg acggcttgtt tcttttctgt ggctgcgtga
841 aagccttgag gggctccggg agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg
901 cgtgtgtgtg tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg
961 ctgcgggcgc ggggcgaagc ggtgcggcgc cggcaggaag gaaatgggcg gggagggcct
1021 tcgtgcgtcg ccgcgccgcc gtccccttct ccctctccag cctcggggct gtccgcgggg
1081 ggacggctgc cttcgggggg gacggggcag ggcggggttc ggcttctggc gtgtgaccgg
1141 cggctctaga gcctctgcta accatgttca tgccttcttc tttttcctac agctcctggg
1201 caacgtgctg gttattgtgc tgtctcatca ttttggcaaa gaattctgcg ccaccatggc
1261 cctgcccacc tgcagctgtc ttgatcgagt tatacaaaaa gacaaaggcc catattatac
1321 acaccttggg gcaggaccaa gtgttgctgc tgtcagggaa atcatggaga ataggtatgg
1381 tcaaaaagga aacgcaataa ggatagaaat agtagtgtac accggtaaag aagggaaaag
1441 ctctcatggg tgtccaattg ctaagtgggt tttaagaaga agcagtgatg aagaaaaagt
1501 tctttgtttg gtccggcagc gtacaggcca ccactgtcca actgctgtga tggtggtgct
1561 catcatggtg tgggatggca tccctcttcc aatggccgac cggctataca cagagctcac
1621 agagaatcta aagtcataca atgggcaccc taccgacaga agatgcaccc tcaatgaaaa
1681 tcgtacctgt acatgtcaag gaattgatcc agagacttgt ggagcttcat tctcttttgg
1741 ctgttcatgg agtatgtact ttaatggctg taagtttggt agaagcccaa gccccagaag
1801 atttagaatt gatccaagct ctcccttaca tgaaaaaaac cttgaagata acttacagag
1861 tttggctaca cgattagctc caatttataa gcagtatgct ccagtagctt accaaaatca
1921 ggtggaatat gaaaatgttg cccgagaatg tcggcttggc agcaaggaag gtcgaccctt
1981 ctctggggtc actgcttgcc tggacttctg tgctcatccc cacagggaca ttcacaacat
2041 gaataatgga agcactgtgg tttgtacctt aactcgagaa gataaccgct ctttgggtgt
2101 tattcctcaa gatgagcagc tccatgtgct acctctttat aagctttcag acacagatga
2161 gtttggctcc aaggaaggaa tggaagccaa gatcaaatct ggggccatcg aggtcctggc
2221 accccgccgc aaaaaaagaa cgtgtttcac tcagcctgtt ccccgttctg gaaagaagag
2281 ggctgcgatg atgacagagg ttcttgcaca taagataagg gcagtggaaa agaaacctat
2341 tccccgaatc aagcggaaga ataactcaac aacaacaaac aacagtaagc cttcgtcact
2401 gccaacctta gggagtaaca ctgagaccgt gcaacctgaa gtaaaaagtg aaaccgaacc
2461 ccattttatc ttaaaaagtt cagacaacac taaaacttat tcgctgatgc catccgctcc
2521 tcacccagtg aaagaggcat ctccaggctt ctcctggtcc ccgaagactg cttcagccac
2581 accagctcca ctgaagaatg acgcaacagc ctcatgcggg ttttcagaaa gaagcagcac
2641 tccccactgt acgatgcctt cgggaagact cagtggtgcc aatgctgcag ctgctgatgg
2701 ccctggcatt tcacagcttg gcgaagtggc tcctctcccc accctgtctg ctcctgtgat
2761 ggagcccctc attaattctg agccttccac tggtgtgact gagccgctaa cgcctcatca
2821 gccaaaccac cagccctcct tcctcacctc tcctcaagac cttgcctctt ctccaatgga
2881 agaagatgag cagcattctg aagcagatga gcctccatca gacgaacccc tatctgatga
2941 ccccctgtca cctgctgagg agaaattgcc ccacattgat gagtattggt cagacagtga
3001 gcacatcttt ttggatgcaa atattggtgg ggtggccatc gcacctgctc acggctcggt
3061 tttgattgag tgtgcccggc gagagctgca cgctaccact cctgttgagc accccaaccg
3121 taatcatcca acccgcctct cccttgtctt ttaccagcac aaaaacctaa ataagcccca
3181 acatggtttt gaactaaaca agattaagtt tgaggctaaa gaagctaaga ataagaaaat
3241 gaaggcctca gagcaaaaag accaggcagc taatgaaggt ccagaacagt cctctgaagt
3301 aaatgaattg aaccaaattc cttctcataa agcattaaca ttaacccatg acaatgttgt
3361 caccgtgtcc ccttatgctc tcacacacgt tgcggggccc tataaccatt gggtcggagg
3421 gccgagctct ggcgcacccc caccaagtgg agggtctcct gccgggtccc caacatctac
3481 tgaagaaggc accagcgaat ccgcaacgcc cgagtcaggc cctggtacct ccacagaacc
3541 atctgaaggt agtgcgcctg gttccccagc tggaagccct acttccaccg aagaaggcac
3601 gtcaaccgaa ccaagtgaag gatctgcccc tgggaccagc actgaaccat ctgagatgga
3661 caagaagtat tctatcggac tggccatcgg gactaatagc gtcgggtggg ccgtgatcac
3721 tgacgagtac aaggtgccct ctaagaagtt caaggtgctc gggaacaccg accggcattc
3781 catcaagaaa aatctgatcg gagctctcct ctttgattca ggggagaccg ctgaagcaac
3841 ccgcctcaag cggactgcta gacggcggta caccaggagg aagaaccgga tttgttacct
3901 tcaagagata ttctccaacg aaatggcaaa ggtcgacgac agcttcttcc ataggctgga
3961 agaatcattc ctcgtggaag aggataagaa gcatgaacgg catcccatct tcggtaatat
4021 cgtcgacgag gtggcctatc acgagaaata cccaaccatc taccatcttc gcaaaaagct
4081 ggtggactca accgacaagg cagacctccg gcttatctac ctggccctgg cccacatgat
4141 caagttcaga ggccacttcc tgatcgaggg cgacctcaat cctgacaata gcgatgtgga
4201 taaactgttc atccagctgg tgcagactta caaccagctc tttgaagaga accccatcaa
4261 tgcaagcgga gtcgatgcca aggccattct gtcagcccgg ctgtcaaaga gccgcagact
4321 tgagaatctt atcgctcagc tgccgggtga aaagaaaaat ggactgttcg ggaacctgat
4381 tgctctttca cttgggctga ctcccaattt caagtctaat ttcgacctgg cagaggatgc
4441 caagctgcaa ctgtccaagg acacctatga tgacgatctc gacaacctcc tggcccagat
4501 cggtgaccaa tacgccgacc ttttccttgc tgctaagaat ctttctgacg ccatcctgct
4561 gtctgacatt ctccgcgtga acactgaaat caccaaggcc cctctttcag cttcaatgat
4621 taagcggtat gatgagcacc accaggacct gaccctgctt aaggcactcg tccggcagca
4681 gcttccggag aagtacaagg aaatcttctt tgaccagtca aagaatggat acgccggcta
4741 catcgacgga ggtgcctccc aagaggaatt ttataagttt atcaaaccta tccttgagaa
4801 gatggacggc accgaagagc tcctcgtgaa actgaatcgg gaggatctgc tgcggaagca
4861 gcgcactttc gacaatggga gcattcccca ccagatccat cttggggagc ttcacgccat
4921 ccttcggcgc caagaggact tctacccctt tcttaaggac aacagggaga agattgagaa
4981 aattctcact ttccgcatcc cctactacgt gggacccctc gccagaggaa atagccggtt
5041 tgcttggatg accagaaagt cagaagaaac tatcactccc tggaacttcg aagaggtggt
5101 ggacaaggga gccagcgctc agtcattcat cgaacggatg actaacttcg ataagaacct
5161 ccccaatgag aaggtcctgc cgaaacattc cctgctctac gagtacttta ccgtgtacaa
5221 cgagctgacc aaggtgaaat atgtcaccga agggatgagg aagcccgcat tcctgtcagg
5281 cgaacaaaag aaggcaattg tggaccttct gttcaagacc aatagaaagg tgaccgtgaa
5341 gcagctgaag gaggactatt tcaagaaaat tgaatgcttc gactctgtgg agattagcgg
5401 ggtcgaagat cggttcaacg caagcctggg tacctaccat gatctgctta agatcatcaa
5461 ggacaaggat tttctggaca atgaggagaa cgaggacatc cttgaggaca ttgtcctgac
5521 tctcactctg ttcgaggacc gggaaatgat cgaggagagg cttaagacct acgcccatct
5581 gttcgacgat aaagtgatga agcaacttaa acggagaaga tataccggat ggggacgcct
5641 tagccgcaaa ctcatcaacg gaatccggga caaacagagc ggaaagacca ttcttgattt
5701 ccttaaaagc gacggattcg ctaatcgcaa cttcatgcaa cttatccatg atgattccct
5761 gacctttaag gaggacatcc agaaggccca agtgtctgga caaggtgact cactgcacga
5821 gcatatcgca aatctggctg gttcacccgc tattaagaag ggtattctcc agaccgtgaa
5881 agtcgtggac gagctggtca aggtgatggg tcgccataaa ccagagaaca ttgtcatcga
5941 gatggccagg gaaaaccaga ctacccagaa gggacagaag aacagcaggg agcggatgaa
6001 aagaattgag gaagggatta aggagctcgg gtcacagatc cttaaagagc acccggtgga
6061 aaacacccag cttcagaatg agaagctcta tctgtactac cttcaaaatg gacgcgatat
6121 gtatgtggac caagagcttg atatcaacag gctctcagac tacgacgtgg acgccatcgt
6181 ccctcagagc ttcctcaaag acgactcaat tgacaataag gtgctgactc gctcagacaa
6241 gaaccgggga aagtcagata acgtgccctc agaggaagtc gtgaaaaaga tgaagaacta
6301 ttggcgccag cttctgaacg caaagctgat cactcagcgg aagttcgaca atctcactaa
6361 ggctgagagg ggcggactga gcgaactgga caaagcagga ttcattaaac ggcaacttgt
6421 ggagactcgg cagattacta aacatgtcgc ccaaatcctt gactcacgca tgaataccaa
6481 gtacgacgaa aacgacaaac ttatccgcga ggtgaaggtg attaccctga agtccaagct
6541 ggtcagcgat ttcagaaagg actttcaatt ctacaaagtg cgggagatca ataactatca
6601 tcatgctcat gacgcatatc tgaatgccgt ggtgggaacc gccctgatca agaagtaccc
6661 aaagctggaa agcgagttcg tgtacggaga ctacaaggtc tacgacgtgc gcaagatgat
6721 tgccaaatct gagcaggaga tcggaaaggc caccgcaaag tacttcttct acagcaacat
6781 catgaatttc ttcaagaccg aaatcaccct tgcaaacggt gagatccgga agaggccgct
6841 catcgagact aatggggaga ctggcgaaat cgtgtgggac aagggcagag atttcgctac
6901 cgtgcgcaaa gtgctttcta tgcctcaagt gaacatcgtg aagaaaaccg aggtgcaaac
6961 cggaggcttt tctaaggaat caatcctccc caagcgcaac tccgacaagc tcattgcaag
7021 gaagaaggat tgggacccta agaagtacgg cggattcgat tcaccaactg tggcttattc
7081 tgtcctggtc gtggctaagg tggaaaaagg aaagtctaag aagctcaaga gcgtgaagga
7141 actgctgggt atcaccatta tggagcgcag ctccttcgag aagaacccaa ttgactttct
7201 cgaagccaaa ggttacaagg aagtcaagaa ggaccttatc atcaagctcc caaagtatag
7261 cctgttcgaa ctggagaatg ggcggaagcg gatgctcgcc tccgctggcg aacttcagaa
7321 gggtaatgag ctggctctcc cctccaagta cgtgaatttc ctctaccttg caagccatta
7381 cgagaagctg aaggggagcc ccgaggacaa cgagcaaaag caactgtttg tggagcagca
7441 taagcattat ctggacgaga tcattgagca gatttccgag ttttctaaac gcgtcattct
7501 cgctgatgcc aacctcgata aagtccttag cgcatacaat aagcacagag acaaaccaat
7561 tcgggagcag gctgagaata tcatccacct gttcaccctc accaatcttg gtgcccctgc
7621 cgcattcaag tacttcgaca ccaccatcga ccggaaacgc tatacctcca ccaaagaagt
7681 gctggacgcc accctcatcc accagagcat caccggactt tacgaaactc ggattgacct
7741 ctcacagctc ggaggggatg gtggcggagg ctcgccaaaa aagaagagaa aggtagaccc
7801 aaagaaaaaa cgaaaagtag atccgaaaaa gaagaggaag gtgggaggta gtggggctac
7861 taacttcagc ctgctgaagc aggctggaga cgtggaggag aaccctggac ctagcgagct
7921 gattaaggag aacatgcaca tgaagctgta catggagggc accgtggaca accatcactt
7981 caagtgcaca tccgagggcg aaggcaagcc ctacgagggc acccagacca tgagaatcaa
8041 ggtggtcgag ggcggccctc tccccttcgc cttcgacatc ctggctacta gcttcctcta
8101 cggcagcaag accttcatca accacaccca gggcatcccc gacttcttca agcagtcctt
8161 ccctgagggc ttcacatggg agagagtcac cacatacgaa gacgggggcg tgctgaccgc
8221 tacccaggac accagcctcc aggacggctg cctcatctac aacgtcaaga tcagaggggt
8281 gaacttcaca tccaacggcc ctgtgatgca gaagaaaaca ctcggctggg aggccttcac
8341 cgagacgctg taccccgctg acggcggcct ggaaggcaga aacgacatgg ccctgaagct
8401 cgtgggcggg agccatctga tcgcaaacat caagaccaca tatagatcca agaaacccgc
8461 taagaacctc aagatgcctg gcgtctacta tgtggactac agactggaaa gaatcaagga
8521 ggccaacaac gagacctacg tcgagcagca cgaggtggca gtggccagat actgcgacct
8581 ccctagcaaa ctggggcaca agcttaatta gtaaagcggc cgcactcctc aggtgcaggc
8641 tgcctatcag aaggtggtgg ctggtgtggc caatgccctg gctcacaaat accactgaga
8701 tctttttccc tctgccaaaa attatgggga catcatgaag ccccttgagc atctgacttc
8761 tggctaataa aggaaattta ttttcattgc aatagtgtgt tggaattttt tgtgtctctc
8821 actcggaagg acatatggga gggcaaatca tttaaaacat cagaatgagt atttggttta
8881 gagtttggca acatatgccc atatgctggc tgccatgaac aaaggttggc tataaagagg
8941 tcatcagtat atgaaacagc cccctgctgt ccattcctta ttccatagaa aagccttgac
9001 ttgaggttag atttttttta tattttgttt tgtgttattt ttttctttaa catccctaaa
9061 attttcctta catgttttac tagccagatt tttcctcctc tcctgactac tcccagtcat
9121 agctgtccct cttctcttat ggagatccct cgacctgcag cccaagcttg gcgtaatcat
9181 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
9241 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
9301 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagcgg atccgcatct
9361 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
9421 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
9481 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
9541 cttttgcaaa aagctaactt gtttattgca gcttataatg gttacaaata aagcaatagc
9601 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
9661 ctcatcaatg tatcttatca tgtctggatc cgctgcatta atgaatcggc caacgcgcgg
9721 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
9781 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
9841 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
9901 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
9961 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
10021 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
10081 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
10141 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
10201 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
10261 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
10321 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
10381 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
10441 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
10501 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
10561 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
10621 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
10681 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
10741 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
10801 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
10861 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
10921 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
10981 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
11041 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
11101 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
11161 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
11221 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
11281 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
11341 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
11401 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
11461 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
11521 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
11581 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
11641 taggggttcc gcgcacattt ccccgaaaag tgccacctg
//
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