| 质粒名称: | pVEGFR2_TEV(N)_NES_TCS(Q'L)_HA-dCas9(N)_P2A_Puro-WPRE |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 101104 |
| 存储实验室: | Tudor Fulga |
|
载体骨架基本信息 |
|
| 载体名称: | pTMt_NES_TCS(Q’G)_HA-dCas9(N)_P2A-Puro-WPRE |
| 载体出品公司: | -- |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | 含有VEGFR2（KDR）先导肽、胞外结构域和跨膜结构域的序列从质粒pDONR223 KDR（来自William Hahn和David Root的gift，Addgene质粒#23925）中PCR扩增，并用于替换pTMt_NES_TCS（Q'G）_HA-dCas9（N）_P2A-Puro-WPRE中的TMt。然后，如前所述（Wehr等人，2006），从全长TEV蛋白酶中扩增N端TEV片段，并与VEGFR2跨膜结构域的C端融合。此外，插入弱TEV切割位点（ENLYFQL）代替TCS（Q’G）。 |
| 载体类型: | 哺乳细胞表达,CRISPR,合成生物学 |
| 筛选标记: | 嘌呤霉素 |
| 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: | VEGFR2-dCas9(N) |
| 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: | pVEGFR2_TEV(N)_NES_TCS(Q'L)_HA-dCas9(N)_P2A_Puro-WPRE was a gift from Tudor Fulga (Addgene plasmid # 101104 ; http://n2t.net/addgene:101104 ; RRID:Addgene_101104) |
| 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 |
pVEGFR2_TEV(N)_NES_TCS(Q'L)_HA-dCas9(N)_P2A_Puro-WPRE
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
_NES_TCS(Q'L)_HA-dCas9(N)_P2A_Puro-WPRE载体图谱.png)
载体简介
载体序列
LOCUS pVEGFR2_TEV(N)_NES_TCS(Q'L)_HA-dCas9(N)_P2A_Puro-WPRE 9530 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Encodes dCas9(N) fused to VEGFR2.
ACCESSION .
VERSION .
KEYWORDS pVEGFR2_TEV(N)_NES_TCS(Q'L)_HA-dCas9(N)_P2A_Puro-WPRE
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9530)
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 9530)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..9530
/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..805
/label=hybrid intron
/note="hybrid between chicken beta-actin (CBA) and minute
virus of mice (MMV) introns (Gray et al., 2011)"
CDS 3643..3669
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
CDS 3685..5286
/codon_start=1
/product="N-terminal portion of catalytically inactive
Streptococcus pyogenes dCas9 carrying the D10A mutation
(Zetsche et al., 2015)"
/label=dCas9(N)
/translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
TVYNELTKVKYVTEGMR"
CDS 5317..5373
/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"
primer_bind complement(5380..5399)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 5876..5896
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
primer_bind complement(6013..6033)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(6431..6442)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
primer_bind complement(6573..6590)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 6579..6786
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
repeat_region 6795..6935
/label=AAV2 ITR
/note="inverted terminal repeat of adeno-associated virus
serotype 2"
repeat_region 6795..6924
/label=AAV2 ITR
rep_origin 7010..7465
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(7097..7116)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 7307..7328
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(7482..7501)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 7601..7623
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(7661..7679)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 7747..7851
/gene="bla"
/label=AmpR promoter
CDS 7852..8712
/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(8070..8089)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 8883..9471
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 9372..9391
/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 tcgctgcgac gctgccttcg
601 ccccgtgccc cgctccgccg ccgcctcgcg ccgcccgccc cggctctgac tgaccgcgtt
661 actcccacag gtgagcgggc gggacggccc ttctcctccg ggctgtaatt agctgagcaa
721 gaggtaaggg tttaagggat ggttggttgg tggggtatta atgtttaatt acctggagca
781 cctgcctgaa atcacttttt ttcaggttgg accggtgcca ccatgcagag caaggtgctg
841 ctggccgtcg ccctgtggct ctgcgtggag acccgggccg cctctgtggg tttgcctagt
901 gtttctcttg atctgcccag gctcagcata caaaaagaca tacttacaat taaggctaat
961 acaactcttc aaattacttg caggggacag agggacttgg actggctttg gcccaataat
1021 cagagtggca gtgagcaaag ggtggaggtg actgagtgca gcgatggcct cttctgtaag
1081 acactcacaa ttccaaaagt gatcggaaat gacactggag cctacaagtg cttctaccgg
1141 gaaactgact tggcctcggt catttatgtc tatgttcaag attacagatc tccatttatt
1201 gcttctgtta gtgaccaaca tggagtcgtg tacattactg agaacaaaaa caaaactgtg
1261 gtgattccat gtctcgggtc catttcaaat ctcaacgtgt cactttgtgc aagataccca
1321 gaaaagagat ttgttcctga tggtaacaga atttcctggg acagcaagaa gggctttact
1381 attcccagct acatgatcag ctatgctggc atggtcttct gtgaagcaaa aattaatgat
1441 gaaagttacc agtctattat gtacatagtt gtcgttgtag ggtataggat ttatgatgtg
1501 gttctgagtc cgtctcatgg aattgaacta tctgttggag aaaagcttgt cttaaattgt
1561 acagcaagaa ctgaactaaa tgtggggatt gacttcaact gggaataccc ttcttcgaag
1621 catcagcata agaaacttgt aaaccgagac ctaaaaaccc agtctgggag tgagatgaag
1681 aaatttttga gcaccttaac tatagatggt ataacccgga gtgaccaagg attgtacacc
1741 tgtgcagcat ccagtgggct gatgaccaag aagaacagca catttgtcag ggtccatgaa
1801 aaaccttttg ttgcttttgg aagtggcatg gaatctctgg tggaagccac ggtgggggag
1861 cgtgtcagaa tccctgcgaa gtaccttggt tacccacccc cagaaataaa atggtataaa
1921 aatggaatac cccttgagtc caatcacaca attaaagcgg ggcatgtact gacgattatg
1981 gaagtgagtg aaagagacac aggaaattac actgtcatcc ttaccaatcc catttcaaag
2041 gagaagcaga gccatgtggt ctctctggtt gtgtatgtcc caccccagat tggtgagaaa
2101 tctctaatct ctcctgtgga ttcctaccag tacggcacca ctcaaacgct gacatgtacg
2161 gtctatgcca ttcctccccc gcatcacatc cactggtatt ggcagttgga ggaagagtgc
2221 gccaacgagc ccagccaagc tgtctcagtg acaaacccat acccttgtga agaatggaga
2281 agtgtggagg acttccaggg aggaaataaa attgaagtta ataaaaatca atttgctcta
2341 attgaaggaa aaaacaaaac tgtaagtacc cttgttatcc aagcggcaaa tgtgtcagct
2401 ttgtacaaat gtgaagcggt caacaaagtc gggagaggag agagggtgat ctccttccac
2461 gtgaccaggg gtcctgaaat tactttgcaa cctgacatgc agcccactga gcaggagagc
2521 gtgtctttgt ggtgcactgc agacagatct acgtttgaga acctcacatg gtacaagctt
2581 ggcccacagc ctctgccaat ccatgtggga gagttgccca cacctgtttg caagaacttg
2641 gatactcttt ggaaattgaa tgccaccatg ttctctaata gcacaaatga cattttgatc
2701 atggagctta agaatgcatc cttgcaggac caaggagact atgtctgcct tgctcaagac
2761 aggaagacca agaaaagaca ttgcgtggtc aggcagctca cagtcctaga gcgtgtggca
2821 cccacgatca caggaaacct ggagaatcag acgacaagta ttggggaaag catcgaagtc
2881 tcatgcacgg catctgggaa tccccctcca cagatcatgt ggtttaaaga taatgagacc
2941 cttgtagaag actcaggcat tgtattgaag gatgggaacc ggaacctcac tatccgcaga
3001 gtgaggaagg aggacgaagg cctctacacc tgccaggcat gcagtgttct tggctgtgca
3061 aaagtggagg catttttcat aatagaaggt gcccaggaaa agacgaactt ggaaatcatt
3121 attctagtag gcacggcggt gattgccatg ttcttctggc tacttcttgt catcatcggc
3181 ggtggctcgg gtggtggctc gggagaaagc ttgtttaagg ggccgcgtga ttacaacccg
3241 atatcgagca ccatttgtca tttgacgaat gaatctgatg ggcacacaac atcgttgtat
3301 ggtattggat ttggtccctt catcattaca aacaagcact tgtttagaag aaataatgga
3361 acactgttgg tccaatcact acatggtgta ttcaaggtca agaacaccac gactttgcaa
3421 caacacctca ttgatgggag ggacatgata attattcgca tgcctaagga tttcccacca
3481 tttcctcaaa agctgaaatt tagagagcca caaagggaag agcgcatatg tcttgtgaca
3541 accaacttcc aaactggcgg tggctcgcta gatttagcta gcttaatcct cggcaagctt
3601 ggcgaaaacc tgtattttca gctaggcgga ggcagcacta gttacccata cgacgtacca
3661 gattacgctg gaggttctgg atccgacaag aagtacagca tcggcctggc catcggcacc
3721 aactctgtgg gctgggccgt gatcaccgac gagtacaagg tgcccagcaa gaaattcaag
3781 gtgctgggca acaccgaccg gcacagcatc aagaagaacc tgatcggagc cctgctgttc
3841 gacagcggcg aaacagccga ggccacccgg ctgaagagaa ccgccagaag aagatacacc
3901 agacggaaga accggatctg ctatctgcaa gagatcttca gcaacgagat ggccaaggtg
3961 gacgacagct tcttccacag actggaagag tccttcctgg tggaagagga taagaagcac
4021 gagcggcacc ccatcttcgg caacatcgtg gacgaggtgg cctaccacga gaagtacccc
4081 accatctacc acctgagaaa gaaactggtg gacagcaccg acaaggccga cctgcggctg
4141 atctatctgg ccctggccca catgatcaag ttccggggcc acttcctgat cgagggcgac
4201 ctgaaccccg acaacagcga cgtggacaag ctgttcatcc agctggtgca gacctacaac
4261 cagctgttcg aggaaaaccc catcaacgcc agcggcgtgg acgccaaggc catcctgtct
4321 gccagactga gcaagagcag acggctggaa aatctgatcg cccagctgcc cggcgagaag
4381 aagaatggcc tgttcggcaa cctgattgcc ctgagcctgg gcctgacccc caacttcaag
4441 agcaacttcg acctggccga ggatgccaaa ctgcagctga gcaaggacac ctacgacgac
4501 gacctggaca acctgctggc ccagatcggc gaccagtacg ccgacctgtt tctggccgcc
4561 aagaacctgt ccgacgccat cctgctgagc gacatcctga gagtgaacac cgagatcacc
4621 aaggcccccc tgagcgcctc tatgatcaag agatacgacg agcaccacca ggacctgacc
4681 ctgctgaaag ctctcgtgcg gcagcagctg cctgagaagt acaaagagat tttcttcgac
4741 cagagcaaga acggctacgc cggctacatt gacggcggag ccagccagga agagttctac
4801 aagttcatca agcccatcct ggaaaagatg gacggcaccg aggaactgct cgtgaagctg
4861 aacagagagg acctgctgcg gaagcagcgg accttcgaca acggcagcat cccccaccag
4921 atccacctgg gagagctgca cgccattctg cggcggcagg aagattttta cccattcctg
4981 aaggacaacc gggaaaagat cgagaagatc ctgaccttcc gcatccccta ctacgtgggc
5041 cctctggcca ggggaaacag cagattcgcc tggatgacca gaaagagcga ggaaaccatc
5101 accccctgga acttcgagga agtggtggac aagggcgctt ccgcccagag cttcatcgag
5161 cggatgacca acttcgataa gaacctgccc aacgagaagg tgctgcccaa gcacagcctg
5221 ctgtacgagt acttcaccgt gtataacgag ctgaccaaag tgaaatacgt gaccgaggga
5281 atgagaggtg gcggtggctc gggtaccgga tctggagcca ccaatttttc tctgctgaag
5341 caggctggtg atgttgaaga aaaccctgga cctgaattca tgaccgagta caagcccacg
5401 gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc
5461 gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc
5521 gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg
5581 gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg
5641 ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa
5701 cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc
5761 gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga
5821 gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac
5881 ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga
5941 ccgcgcacct ggtgcatgac ccgcaagggg taactaagta aggatcgatc caagatatcg
6001 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
6061 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
6121 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
6181 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
6241 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
6301 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac
6361 gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg
6421 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
6481 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
6541 ctccccgcca attcctagag ctcgctgatc agcctcgact gtgccttcta gttgccagcc
6601 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
6661 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
6721 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagagaata gcaggcatgc
6781 tggggagcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg
6841 ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc
6901 ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt attttctcct
6961 tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt
7021 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
7081 agcgccttag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
7141 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
7201 cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga
7261 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
7321 caaactggaa caacactcaa ctctatctcg ggctattctt ttgatttata agggattttg
7381 ccgatttcgg tctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
7441 aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc
7501 gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
7561 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
7621 aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt
7681 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
7741 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
7801 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
7861 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
7921 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
7981 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
8041 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
8101 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
8161 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
8221 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
8281 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
8341 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
8401 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
8461 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
8521 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtggaag ccgcggtatc
8581 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
8641 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
8701 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
8761 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
8821 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
8881 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
8941 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
9001 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
9061 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
9121 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
9181 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
9241 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
9301 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
9361 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
9421 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
9481 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
//
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