| 质粒名称: | pVEGFR1_TEV(C)_TCS(Q'G)_NLS-HA-dCas9(C)-VP64_T2A_MCP-P65-HSF1 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 101103 |
| 存储实验室: | Tudor Fulga |
|
载体骨架基本信息 |
|
| 载体名称: | pTMt_TCS(Q’G)_NLS-HA-dCas9(C)-VP64_T2A_MCP-P65-HSF1 |
| 载体出品公司: | -- |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | 从质粒pDONR223-FLT1（来自William Hahn和David Root的gift，Addgene质粒#23912）PCR扩增包含VEGFR1（FLT1）先导肽、胞外结构域和跨膜结构域的序列，并用于替换pTMt_TCS（Q'G）_NLS-HA-dCas9（C）-VP64_T2A_MCP-P65-HSF1中的TMt，如前所述（Wehr等人，2006），从全长TEV蛋白酶中扩增C端TEV片段，并在VEGFR1跨膜结构域和TEV切割位点之间克隆。 |
| 载体类型: | 哺乳细胞表达,CRISPR，合成生物学 |
| 筛选标记: | -- |
| 载体抗性: | 氨苄霉素100 μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | VEGFR1-dCas9(C) |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子: | -- |
| 标签/融合蛋白: | HA |
| 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: | pVEGFR1_TEV(C)_TCS(Q'G)_NLS-HA-dCas9(C)-VP64_T2A_MCP-P65-HSF1 was a gift from Tudor Fulga (Addgene plasmid # 101103 ; http://n2t.net/addgene:101103 ; RRID:Addgene_101103) |
| 参考文献部分: | Engineering Synthetic Signaling Pathways with Programmable dCas9-Based Chimeric Receptors. Baeumler TA, Ahmed AA, Fulga TA. Cell Rep. 2017 Sep 12;20(11):2639-2653. doi: 10.1016/j.celrep.2017.08.044. 10.1016/j.celrep.2017.08.044 PubMed 28903044 |
pVEGFR1_TEV(C)_TCS(Q'G)_NLS-HA-dCas9(C)-VP64_T2A_MCP-P65-HSF
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
_TCS(Q'G)_NLS-HA-dCas9(C)-VP64_T2A_MCP-P65-HSF1载体图谱.png)
载体简介
载体序列
LOCUS pVEGFR1_TEV(C)_TCS(Q'G)_NLS-HA-dCas9(C)-VP64_T2A_MCP-P65-HSF1 10869 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Encodes dCas9(C)-VP64 with SAM components fused to VEGFR1.
ACCESSION .
VERSION .
KEYWORDS pVEGFR1_TEV(C)_TCS(Q'G)_NLS-HA-dCas9(C)-VP64_T2A_MCP-P65-HSF1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10869)
AUTHORS Baeumler TA, Ahmed AA, Fulga TA
TITLE Engineering Synthetic Signaling Pathways with Programmable
dCas9-Based Chimeric Receptors.
JOURNAL Cell Rep. 2017 Sep 12;20(11):2639-2653. doi:
10.1016/j.celrep.2017.08.044.
PUBMED 28903044
REFERENCE 2 (bases 1 to 10869)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..10869
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 12..297
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer;
contains an 18-bp deletion relative to the standard CMV
enhancer"
promoter 299..576
/label=chicken beta-actin promoter
intron 577..804
/label=hybrid intron
/note="hybrid between chicken beta-actin (CBA) and minute
virus of mice (MMV) introns (Gray et al., 2011)"
regulatory 816..825
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 3567..3587
/codon_start=1
/product="tobacco etch virus (TEV) protease recognition and
cleavage site"
/label=TEV site
/translation="ENLYFQG"
CDS 3588..3608
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 3627..3653
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
CDS 3681..6179
/codon_start=1
/product="C-terminal portion of catalytically inactive
Streptococcus pyogenes dCas9 carrying the N863A mutation
(Zetsche et al., 2015)"
/label=dCas9(C)
/translation="KPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEI
SGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTY
AHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIH
DDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPE
NIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYL
QNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKARGKSDNVPSEEV
VKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQI
LDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVV
GTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITL
ANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESIL
PKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIME
RSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALP
SKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLD
KVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATL
IHQSITGLYETRIDLSQLGGD"
CDS 6183..6203
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 6222..6371
/codon_start=1
/product="tetrameric repeat of the minimal activation
domain of herpes simplex virus VP16 (Beerli et al., 1998)"
/label=VP64
/translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
DLDML"
CDS 6399..6452
/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 6468..6854
/codon_start=1
/product="bacteriophage MS2 coat protein"
/label=MS2-N55K
/note="binds with high affinity to a specific stem-loop
structure in the viral RNA (Lim et al., 1994)"
/translation="ASNFTQFVLVDNGGTGDVTVAPSNFANGVAEWISSNSRSQAYKVT
CSVRQSSAQKRKYTIKVEVPKVATQTVGGVELPVAAWRSYLNMELTIPIFATNSDCELI
VKAMQGLLKDGNPIPSAIAANSGIY"
CDS 6909..6929
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 6945..7487
/codon_start=1
/product="C-terminal portion of the p65 subunit of mouse
NF-kappa-B"
/label=p65
/translation="PSGQISNQALALAPSSAPVLAQTMVPSSAMVPLAQPPAPAPVLTP
GPPQSLSAPVPKSTQAGEGTLSEALLHLQFDADEDLGALLGNSTDPGVFTDLASVDNSE
FQQLLNQGVSMSHSTAEPMLMEYPEAITRLVTGSQRPPDPAPTPLGTSGLPNGLSGDED
FSSIADMDFSALLSQISS"
CDS 7512..7883
/codon_start=1
/product="C-terminal activation domain from the human heat
shock transcription factor HSF1"
/label=HSF1
/translation="GFSVDTSALLDLFSPSVTVPDMSLPDLDSSLASIQELLSPQEPPR
PPEAENSSPDSGKQLVHYTAQPLFLLDPGSVDTGSNDLPVLFELGEGSYFSEGDGFAED
PTISLLTGSEPPKAKDPTVS"
primer_bind complement(7911..7928)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 7917..8124
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
repeat_region 8133..8273
/label=AAV2 ITR
/note="inverted terminal repeat of adeno-associated virus
serotype 2"
repeat_region 8133..8262
/label=AAV2 ITR
rep_origin 8348..8803
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(8435..8454)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 8645..8666
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(8820..8839)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 8939..8961
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(8999..9017)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 9085..9189
/gene="bla"
/label=AmpR promoter
CDS 9190..10050
/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(9408..9427)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 10221..10809
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 10710..10729
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 ctagaggtac ccgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
61 ccccgcccat tgacgtcaat agtaacgcca atagggactt tccattgacg tcaatgggtg
121 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
181 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attgtgccca gtacatgacc
241 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtc
301 gaggtgagcc ccacgttctg cttcactctc cccatctccc ccccctcccc acccccaatt
361 ttgtatttat ttatttttta attattttgt gcagcgatgg gggcgggggg gggggggggg
421 cgcgcgccag gcggggcggg gcggggcgag gggcggggcg gggcgaggcg gagaggtgcg
481 gcggcagcca atcagagcgg cgcgctccga aagtttcctt ttatggcgag gcggcggcgg
541 cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag tcgctgcgcg ctgccttcgc
601 cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc ggctctgact gaccgcgtta
661 ctcccacagg tgagcgggcg ggacggccct tctcctccgg gctgtaatta gctgagcaag
721 aggtaagggt ttaagggatg gttggttggt ggggtattaa tgtttaatta cctggagcac
781 ctgcctgaaa tcactttttt tcaggttgga ccggtgccac catggtcagc tactgggaca
841 ccggggtcct gctgtgcgcg ctgctcagct gtctgcttct cacaggatct agttcaggtt
901 caaaattaaa agatcctgaa ctgagtttaa aaggcaccca gcacatcatg caagcaggcc
961 agacactgca tctccaatgc aggggggaag cagcccataa atggtctttg cctgaaatgg
1021 tgagtaagga aagcgaaagg ctgagcataa ctaaatctgc ctgtggaaga aatggcaaac
1081 aattctgcag tactttaacc ttgaacacag ctcaagcaaa ccacactggc ttctacagct
1141 gcaaatatct agctgtacct acttcaaaga agaaggaaac agaatctgca atctatatat
1201 ttattagtga tacaggtaga cctttcgtag agatgtacag tgaaatcccc gaaattatac
1261 acatgactga aggaagggag ctcgtcattc cctgccgggt tacgtcacct aacatcactg
1321 ttactttaaa aaagtttcca cttgacactt tgatccctga tggaaaacgc ataatctggg
1381 acagtagaaa gggcttcatc atatcaaatg caacgtacaa agaaataggg cttctgacct
1441 gtgaagcaac agtcaatggg catttgtata agacaaacta tctcacacat cgacaaacca
1501 atacaatcat agatgtccaa ataagcacac cacgcccagt caaattactt agaggccata
1561 ctcttgttct caattgtact gctaccactc ccttgaacac gagagttcaa atgacctgga
1621 gttaccctga tgaaaaaaat aagagagctt ccgtaaggcg acgaattgac caaagcaatt
1681 cccatgccaa catattctac agtgttctta ctattgacaa aatgcagaac aaagacaaag
1741 gactttatac ttgtcgtgta aggagtggac catcattcaa atctgttaac acctcagtgc
1801 atatatatga taaagcattc atcactgtga aacatcgaaa acagcaggtg cttgaaaccg
1861 tagctggcaa gcggtcttac cggctctcta tgaaagtgaa ggcatttccc tcgccggaag
1921 ttgtatggtt aaaagatggg ttacctgcga ctgagaaatc tgctcgctat ttgactcgtg
1981 gctactcgtt aattatcaag gacgtaactg aagaggatgc agggaattat acaatcttgc
2041 tgagcataaa acagtcaaat gtgtttaaaa acctcactgc cactctaatt gtcaatgtga
2101 aaccccagat ttacgaaaag gctgtgtcat cgtttccaga cccggctctc tacccactgg
2161 gcagcagaca aatcctgact tgtaccgcat atggtatccc tcaacctaca atcaagtggt
2221 tctggcaccc ctgtaaccat aatcattccg aagcaaggtg tgacttttgt tccaataatg
2281 aagagtcctt tatcctggat gctgacagca acatgggaaa cagaattgag agcatcactc
2341 agcgcatggc aataatagaa ggaaagaata agatggctag caccttggtt gtggctgact
2401 ctagaatttc tggaatctac atttgcatag cttccaataa agttgggact gtgggaagaa
2461 acataagctt ttatatcaca gatgtgccaa atgggtttca tgttaacttg gaaaaaatgc
2521 cgacggaagg agaggacctg aaactgtctt gcacagttaa caagttctta tacagagacg
2581 ttacttggat tttactgcgg acagttaata acagaacaat gcactacagt attagcaagc
2641 aaaaaatggc catcactaag gagcactcca tcactcttaa tcttaccatc atgaatgttt
2701 ccctgcaaga ttcaggcacc tatgcctgca gagccaggaa tgtatacaca ggggaagaaa
2761 tcctccagaa gaaagaaatt acaatcagag atcaggaagc accatacctc ctgcgaaacc
2821 tcagtgatca cacagtggcc atcagcagtt ccaccacttt agactgtcat gctaatggtg
2881 tccccgagcc tcagatcact tggtttaaaa acaaccacaa aatacaacaa gagcctggaa
2941 ttattttagg accaggaagc agcacgctgt ttattgaaag agtcacagaa gaggatgaag
3001 gtgtctatca ctgcaaagcc accaaccaga agggctctgt ggaaagttca gcatacctca
3061 ctgttcaagg aacctcggac aagcctaatc tggagctgat cactctaaca tgcacctgtg
3121 tggctgcgac tctcttctgg ctcctattaa ccctctttat cggcggtggc tcgggtggtg
3181 gctcgaagag catgtctagc atggtgtcag acactagttg cacattccct tcatctgatg
3241 gcatattctg gaagcattgg attcaaacca aggatgggca gtgtggcagt ccattagtat
3301 caactagaga tgggttcatt gttggtatac actcagcatc gaatttcacc aacacaaaca
3361 attatttcac aagcgtgccg aaaaacttca tggaattgtt gacaaatcag gaggcgcagc
3421 agtgggttag tggttggcga ttaaatgctg actcagtatt gtgggggggc cataaagttt
3481 tcatggtgaa acctgaagag ccttttcagc cagttaagga agcgactcaa ctcatgaatc
3541 gtcgtcgccg tcccggcggt ggctcggaaa acctgtattt tcaggggccc aagaagaaga
3601 ggaaggtggg gggaggcagc actagttacc catacgacgt accagattac gctggaggtt
3661 ctggatccgg cggtggctcg aagcccgcct tcctgagcgg cgagcagaaa aaggccatcg
3721 tggacctgct gttcaagacc aaccggaaag tgaccgtgaa gcagctgaaa gaggactact
3781 tcaagaaaat cgagtgcttc gactccgtgg aaatctccgg cgtggaagat cggttcaacg
3841 cctccctggg cacataccac gatctgctga aaattatcaa ggacaaggac ttcctggaca
3901 atgaggaaaa cgaggacatt ctggaagata tcgtgctgac cctgacactg tttgaggaca
3961 gagagatgat cgaggaacgg ctgaaaacct atgcccacct gttcgacgac aaagtgatga
4021 agcagctgaa gcggcggaga tacaccggct ggggcaggct gagccggaag ctgatcaacg
4081 gcatccggga caagcagtcc ggcaagacaa tcctggattt cctgaagtcc gacggcttcg
4141 ccaacagaaa cttcatgcag ctgatccacg acgacagcct gacctttaaa gaggacatcc
4201 agaaagccca ggtgtccggc cagggcgata gcctgcacga gcacattgcc aatctggccg
4261 gcagccccgc cattaagaag ggcatcctgc agacagtgaa ggtggtggac gagctcgtga
4321 aagtgatggg ccggcacaag cccgagaaca tcgtgatcga aatggccaga gagaaccaga
4381 ccacccagaa gggacagaag aacagccgcg agagaatgaa gcggatcgaa gagggcatca
4441 aagagctggg cagccagatc ctgaaagaac accccgtgga aaacacccag ctgcagaacg
4501 agaagctgta cctgtactac ctgcagaatg ggcgggatat gtacgtggac caggaactgg
4561 acatcaaccg gctgtccgac tacgatgtgg accatatcgt gcctcagagc tttctgaagg
4621 acgactccat cgacaacaag gtgctgacca gaagcgacaa ggcccggggc aagagcgaca
4681 acgtgccctc cgaagaggtc gtgaagaaga tgaagaacta ctggcggcag ctgctgaacg
4741 ccaagctgat tacccagaga aagttcgaca atctgaccaa ggccgagaga ggcggcctga
4801 gcgaactgga taaggccggc ttcatcaaga gacagctggt ggaaacccgg cagatcacaa
4861 agcacgtggc acagatcctg gactcccgga tgaacactaa gtacgacgag aatgacaagc
4921 tgatccggga agtgaaagtg atcaccctga agtccaagct ggtgtccgat ttccggaagg
4981 atttccagtt ttacaaagtg cgcgagatca acaactacca ccacgcccac gacgcctacc
5041 tgaacgccgt cgtgggaacc gccctgatca aaaagtaccc taagctggaa agcgagttcg
5101 tgtacggcga ctacaaggtg tacgacgtgc ggaagatgat cgccaagagc gagcaggaaa
5161 tcggcaaggc taccgccaag tacttcttct acagcaacat catgaacttt ttcaagaccg
5221 agattaccct ggccaacggc gagatccgga agcggcctct gatcgagaca aacggcgaaa
5281 ccggggagat cgtgtgggat aagggccggg attttgccac cgtgcggaaa gtgctgagca
5341 tgccccaagt gaatatcgtg aaaaagaccg aggtgcagac aggcggcttc agcaaagagt
5401 ctatcctgcc caagaggaac agcgataagc tgatcgccag aaagaaggac tgggacccta
5461 agaagtacgg cggcttcgac agccccaccg tggcctattc tgtgctggtg gtggccaaag
5521 tggaaaaggg caagtccaag aaactgaaga gtgtgaaaga gctgctgggg atcaccatca
5581 tggaaagaag cagcttcgag aagaatccca tcgactttct ggaagccaag ggctacaaag
5641 aagtgaaaaa ggacctgatc atcaagctgc ctaagtactc cctgttcgag ctggaaaacg
5701 gccggaagag aatgctggcc tctgccggcg aactgcagaa gggaaacgaa ctggccctgc
5761 cctccaaata tgtgaacttc ctgtacctgg ccagccacta tgagaagctg aagggctccc
5821 ccgaggataa tgagcagaaa cagctgtttg tggaacagca caagcactac ctggacgaga
5881 tcatcgagca gatcagcgag ttctccaaga gagtgatcct ggccgacgct aatctggaca
5941 aagtgctgtc cgcctacaac aagcaccggg ataagcccat cagagagcag gccgagaata
6001 tcatccacct gtttaccctg accaatctgg gagcccctgc cgccttcaag tactttgaca
6061 ccaccatcga ccggaagagg tacaccagca ccaaagaggt gctggacgcc accctgatcc
6121 accagagcat caccggcctg tacgagacac ggatcgacct gtctcagctg ggaggcgaca
6181 gccccaagaa gaagagaaag gtggaggcca gcggacgggc tgacgcattg gacgattttg
6241 atctggatat gctgggaagt gacgccctcg atgattttga ccttgacatg cttggttcgg
6301 atgcccttga tgactttgac ctcgacatgc tcggcagtga cgcccttgat gatttcgacc
6361 tggacatgct gattaacggt accgctagcg gcagtggaga gggcagagga agtctgctaa
6421 catgcggtga cgtcgaggag aatcctggcc cagtgagcaa gcttatggct tcaaacttta
6481 ctcagttcgt gctcgtggac aatggtggga caggggatgt gacagtggct ccttctaatt
6541 tcgctaatgg ggtggcagag tggatcagct ccaactcacg gagccaggcc tacaaggtga
6601 catgcagcgt caggcagtct agtgcccaga agagaaagta taccatcaag gtggaggtcc
6661 ccaaagtggc tacccagaca gtgggcggag tcgaactgcc tgtcgccgct tggaggtcct
6721 acctgaacat ggagctcact atcccaattt tcgctaccaa ttctgactgt gaactcatcg
6781 tgaaggcaat gcaggggctc ctcaaagacg gtaatcctat cccttccgcc atcgccgcta
6841 actcaggtat ctacagcgct ggaggaggtg gaagcggagg aggaggaagc ggaggaggag
6901 gtagcggacc taagaaaaag aggaaggtgg cggccgctgg atccccttca gggcagatca
6961 gcaaccaggc cctggctctg gcccctagct ccgctccagt gctggcccag actatggtgc
7021 cctctagtgc tatggtgcct ctggcccagc cacctgctcc agcccctgtg ctgaccccag
7081 gaccacccca gtcactgagc gctccagtgc ccaagtctac acaggccggc gaggggactc
7141 tgagtgaagc tctgctgcac ctgcagttcg acgctgatga ggacctggga gctctgctgg
7201 ggaacagcac cgatcccgga gtgttcacag atctggcctc cgtggacaac tctgagtttc
7261 agcagctgct gaatcagggc gtgtccatgt ctcatagtac agccgaacca atgctgatgg
7321 agtaccccga agccattacc cggctggtga ccggcagcca gcggcccccc gaccccgctc
7381 caactcccct gggaaccagc ggcctgccta atgggctgtc cggagatgaa gacttctcaa
7441 gcatcgctga tatggacttt agtgccctgc tgtcacagat ttcctctagt gggcagggag
7501 gaggtggaag cggcttcagc gtggacacca gtgccctgct ggacctgttc agcccctcgg
7561 tgaccgtgcc cgacatgagc ctgcctgacc ttgacagcag cctggccagt atccaagagc
7621 tcctgtctcc ccaggagccc cccaggcctc ccgaggcaga gaacagcagc ccggattcag
7681 ggaagcagct ggtgcactac acagcgcagc cgctgttcct gctggacccc ggctccgtgg
7741 acaccgggag caacgacctg ccggtgctgt ttgagctggg agagggctcc tacttctccg
7801 aaggggacgg cttcgccgag gaccccacca tctccctgct gacaggctcg gagcctccca
7861 aagccaagga ccccactgtc tcctaagaat tcctagagct cgctgatcag cctcgactgt
7921 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
7981 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
8041 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
8101 agagaatagc aggcatgctg gggagcggcc gcaggaaccc ctagtgatgg agttggccac
8161 tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc
8221 gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct
8281 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa
8341 ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
8401 gtgaccgcta cacttgccag cgccttagcg cccgctcctt tcgctttctt cccttccttt
8461 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
8521 cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt
8581 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
8641 aatagtggac tcttgttcca aactggaaca acactcaact ctatctcggg ctattctttt
8701 gatttataag ggattttgcc gatttcggtc tattggttaa aaaatgagct gatttaacaa
8761 aaatttaacg cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt
8821 acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac
8881 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
8941 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc
9001 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
9061 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
9121 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
9181 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
9241 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
9301 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
9361 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
9421 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
9481 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
9541 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
9601 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
9661 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
9721 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
9781 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
9841 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
9901 cgtggaagcc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
9961 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
10021 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
10081 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
10141 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
10201 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
10261 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
10321 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
10381 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
10441 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
10501 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
10561 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
10621 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
10681 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
10741 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
10801 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
10861 gctcacatg
//
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