| 质粒名称: | pLVX-EF1alpha-SARS-CoV-2-N-2xStrep-IRES-Puro |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 141391 |
| 存储实验室: | Nevan Krogan |
|
载体骨架基本信息 |
|
| 载体名称: | pLVX-EF1alpha-IRES-Puro |
| 载体出品公司: | Takara Bio |
| 空载体大小: | 8825 bp |
| 完整质粒大小: | 8825 bp |
| 载体类型: | 哺乳细胞表达, Lentiviral |
| 筛选标记: | Puromycin |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 30 ℃ |
| 宿主菌: | NEB Stable |
| 拷贝数: | 未知 |
|
插入基因信息 |
|
| 插入基因名称: | SARS-CoV-2-N |
| 物种来源: | SARS-CoV-2 |
| 基因突变: | human codon optimized |
| 启动子: | EF1alpha |
| 标签/融合蛋白: | 2xStrep (C terminal on insert) |
| 克隆方法: | 未知 |
|
文献引用方法 |
|
| 材料和方法部分: |
pLVX-EF1alpha-SARS-CoV-2-N-2xStrep-IRES-Puro was a gift from Nevan Krogan (Addgene plasmid # 141391 ; http://n2t.net/addgene:141391 ; RRID:Addgene_141391)
For viral preps, please replace (Addgene plasmid # 141391) in the above sentence with: (Addgene viral prep # 141391-LV)
|
| 参考文献部分: | A SARS-CoV-2 protein interaction map reveals targets for drug repurposing. Gordon DE, Jang GM, Bouhaddou M, Xu J, Obernier K, White KM, O'Meara MJ, Rezelj VV, Guo JZ, Swaney DL, Tummino TA, Huttenhain R, Kaake RM, Richards AL, Tutuncuoglu B, Foussard H, Batra J, Haas K, Modak M, Kim M, Haas P, Polacco BJ, Braberg H, Fabius JM, Eckhardt M, Soucheray M, Bennett MJ, Cakir M, McGregor MJ, Li Q, Meyer B, Roesch F, Vallet T, Mac Kain A, Miorin L, Moreno E, Naing ZZC, Zhou Y, Peng S, Shi Y, Zhang Z, Shen W, Kirby IT, Melnyk JE, Chorba JS, Lou K, Dai SA, Barrio-Hernandez I, Memon D, Hernandez-Armenta C, Lyu J, Mathy CJP, Perica T, Pilla KB, Ganesan SJ, Saltzberg DJ, Rakesh R, Liu X, Rosenthal SB, Calviello L, Venkataramanan S, Liboy-Lugo J, Lin Y, Huang XP, Liu Y, Wankowicz SA, Bohn M, Safari M, Ugur FS, Koh C, Savar NS, Tran QD, Shengjuler D, Fletcher SJ, O'Neal MC, Cai Y, Chang JCJ, Broadhurst DJ, Klippsten S, Sharp PP, Wenzell NA, Kuzuoglu-Ozturk D, Wang HY, Trenker R, Young JM, Cavero DA, Hiatt J, Roth TL, Rathore U, Subramanian A, Noack J, Hubert M, Stroud RM, Frankel AD, Rosenberg OS, Verba KA, Agard DA, Ott M, Emerman M, Jura N, von Zastrow M, Verdin E, Ashworth A, Schwartz O, d'Enfert C, Mukherjee S, Jacobson M, Malik HS, Fujimori DG, Ideker T, Craik CS, Floor SN, Fraser JS, Gross JD, Sali A, Roth BL, Ruggero D, Taunton J, Kortemme T, Beltrao P, Vignuzzi M, Garcia-Sastre A, Shokat KM, Shoichet BK, Krogan NJ. Nature. 2020 Jul;583(7816):459-468. doi: 10.1038/s41586-020-2286-9. Epub 2020 Apr 30. 10.1038/s41586-020-2286-9 PubMed 32353859 |
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 pLVX-EF1alpha-SARS-CoV-2-N-2xStrep-IRES-Puro 10170 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Lentiviral expression of SARS-CoV-2 nucleocapsid (N) protein;
transient expression and generate lentivirus.
ACCESSION .
VERSION .
KEYWORDS pLVX-EF1alpha-SARS-CoV-2-N-2xStrep-IRES-Puro
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10170)
AUTHORS Gordon DE, et al.
TITLE A SARS-CoV-2-Human Protein-Protein Interaction Map Reveals Drug
Targets and Potential Drug Repurposing
JOURNAL BioRxiv March 2020
REFERENCE 2 (bases 1 to 10170)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..10170
/organism="synthetic DNA construct"
/mol_type="other DNA"
LTR 1..634
/label=3' LTR
/note="3' long terminal repeat (LTR) from HIV-1"
misc_feature 681..806
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1303..1536
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 1721..1765
/codon_start=1
/product="antigenic peptide corresponding to amino acids
655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
et al., 2013)"
/label=gp41 peptide
/note="recognized by the 2H10 single-chain llama nanobody"
/translation="KNEQELLELDKWASL"
misc_feature 2028..2143
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1 (lacking the first T)"
promoter 2338..3519
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
intron 2568..3510
/label=EF-1-alpha intron A
/note="intron upstream of the start codon of human
EF-1-alpha"
primer_bind 3467..3487
/label=EF1a-F
/note="Human elongation factor-1a promoter, forward primer"
CDS 4825..4848
/codon_start=1
/product="peptide that binds Strep-Tactin(R), an engineered
form of streptavidin"
/label=Strep-Tag II
/translation="WSHPQFEK"
CDS 4885..4908
/codon_start=1
/product="peptide that binds Strep-Tactin(R), an engineered
form of streptavidin"
/label=Strep-Tag II
/translation="WSHPQFEK"
misc_feature 4920..5493
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
primer_bind complement(5087..5104)
/label=IRES reverse
/note="IRES internal ribosome entry site, reverse primer.
Also called pCDH-rev"
primer_bind 5314..5333
/label=IRES-F
/note="IRES internal ribosome entry site, forward primer"
CDS 5526..6125
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/label=PuroR
/note="confers resistance to puromycin"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
primer_bind complement(5526..5545)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 6022..6042
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
misc_feature 6139..6727
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(6192..6212)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(6610..6621)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
LTR 6934..7567
/label=3' LTR
/note="3' long terminal repeat (LTR) from HIV-1"
primer_bind complement(7695..7711)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(7695..7711)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(7708..7730)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 7719..7735
/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(7743..7773)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 7788..7809
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(7926..7943)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(8097..8685)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(8177..8196)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(8856..9716)
/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 9479..9498
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(9717..9821)
/gene="bla"
/label=AmpR promoter
polyA_signal 9869..10003
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(9906..9925)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 9960..9979
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
ORIGIN
1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccttt aaaagaaaag
2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
2101 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
2161 acagcagaga tccagtttat cgatgagtaa ttcatacaaa aggactcgcc cctgccttgg
2221 ggaatcccag ggaccgtcgt taaactccca ctaacgtaga acccagagat cgctgcgttc
2281 ccgccccctc acccgcccgc tctcgtcatc actgaggtgg agaagagcat gcgtgaggct
2341 ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag
2401 gggtcggcaa ttgaaccggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg
2461 tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag
2521 tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacaggta agtgccgtgt
2581 gtggttcccg cgggcctggc ctctttacgg gttatggccc ttgcgtgcct tgaattactt
2641 ccacgcccct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
2701 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
2761 cttgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
2821 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
2881 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
2941 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
3001 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
3061 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
3121 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
3181 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
3241 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
3301 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
3361 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
3421 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
3481 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gatctatttc cggtgaattc
3541 gccgccacca tgagcgataa cggcccccaa aaccaacgga atgcccctcg gataacattt
3601 ggtggcccta gtgattccac aggaagtaac caaaacggag aacggtctgg agcccgctct
3661 aaacagaggc gaccacaggg attgccaaat aacacagcgt catggtttac agccctgact
3721 caacatggaa aggaggactt gaaatttcca cgaggacagg gtgtaccaat aaacacaaac
3781 agtagtccag acgatcagat cgggtattat agaagggcca cgagacgcat cagaggtgga
3841 gatggtaaaa tgaaagatct cagtccgcgc tggtactttt attatttggg aaccggccca
3901 gaagcgggcc tcccatatgg agccaataag gacgggatta tatgggtggc tacagaggga
3961 gccctcaata cgccaaaaga tcacatcgga acaagaaacc ctgctaataa tgccgcaatc
4021 gtgctgcagt tgcctcaggg gacgacgctt cctaaaggct tttacgcaga aggatcacgc
4081 ggcggcagcc aagcatcctc aaggtctagt tccagaagtc gaaacagctc cagaaactcc
4141 acaccagggt ccagtagggg cacaagtccg gcgcggatgg cgggcaacgg cggagacgcc
4201 gcactcgctc tgttgctcct ggaccggctc aaccaacttg aatccaagat gagtggtaag
4261 ggacagcagc aacaaggtca aaccgtaacc aagaaaagcg ctgcagaagc ttctaaaaaa
4321 cctcgacaaa aacggaccgc tacgaaggca tataacgtta ctcaagcctt cgggagacga
4381 gggccggagc aaacccaggg gaattttgga gatcaggagc tcatccggca agggacagat
4441 tacaaacatt ggcctcaaat cgctcagttc gcgccgtccg caagtgcttt tttcggaatg
4501 tctcgcatcg gtatggaagt aacaccatcc gggacatggc tcacttatac cggcgccatc
4561 aagctggacg acaaagatcc aaatttcaaa gatcaggtga ttctcttgaa taaacacata
4621 gacgcgtata agacgtttcc acccaccgaa cccaagaagg acaaaaagaa aaaagctgat
4681 gaaacacaag cgttgcccca gcgacagaag aagcagcaaa ccgtcaccct cctgccagcc
4741 gctgatctcg atgacttctc caagcaactt caacagtcca tgagctcagc cgattctact
4801 caggcgctcg aaggcggcgg gggatggagc catccacaat tcgagaaagg cggtggttca
4861 ggaggaggta gcgggggtgg atcatggtca catccgcagt ttgaaaagta aggatcccgc
4921 ccctctccct cccccccccc taacgttact ggccgaagcc gcttggaata aggccggtgt
4981 gcgtttgtct atatgttatt ttccaccata ttgccgtctt ttggcaatgt gagggcccgg
5041 aaacctggcc ctgtcttctt gacgagcatt cctaggggtc tttcccctct cgccaaagga
5101 atgcaaggtc tgttgaatgt cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa
5161 acaacgtctg tagcgaccct ttgcaggcag cggaaccccc cacctggcga caggtgcctc
5221 tgcggccaaa agccacgtgt ataagataca cctgcaaagg cggcacaacc ccagtgccac
5281 gttgtgagtt ggatagttgt ggaaagagtc aaatggctct cctcaagcgt attcaacaag
5341 gggctgaagg atgcccagaa ggtaccccat tgtatgggat ctgatctggg gcctcggtgc
5401 acatgcttta catgtgttta gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg
5461 gacgtggttt tcctttgaaa aacacgatga taagcttgcc acaacccaca aggagacgac
5521 cttccatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtcccccggg
5581 ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgacc
5641 cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc
5701 tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc
5761 cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga
5821 gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca
5881 aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc
5941 tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct
6001 tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca
6061 ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg
6121 cctagacgcg tctggaacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt
6181 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
6241 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
6301 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
6361 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
6421 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
6481 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaagctgacg
6541 tcctttccat ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc
6601 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
6661 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
6721 tccccgcctg gaattaattc tgcagtcgag acctagaaaa acatggagca atcacaagta
6781 gcaatacagc agctaccaat gctgattgtg cctggctaga agcacaagag gaggaggagg
6841 tgggttttcc agtcacacct caggtacctt taagaccaat gacttacaag gcagctgtag
6901 atcttagcca ctttttaaaa gaaaagaggg gactggaagg gctaattcac tcccaacgaa
6961 gacaagatat ccttgatctg tggatctacc acacacaagg ctacttccct gattagcaga
7021 actacacacc agggccaggg gtcagatatc cactgacctt tggatggtgc tacaagctag
7081 taccagttga gccagataag gtagaagagg ccaataaagg agagaacacc agcttgttac
7141 accctgtgag cctgcatggg atggatgacc cggagagaga agtgttagag tggaggtttg
7201 acagccgcct agcatttcat cacgtggccc gagagctgca tccggagtac ttcaagaact
7261 gctgatatcg agcttgctac aagggacttt ccgctgggga ctttccaggg aggcgtggcc
7321 tgggcgggac tggggagtgg cgagccctca gatcctgcat ataagcagct gctttttgcc
7381 tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg
7441 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt
7501 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc
7561 tctagcagta gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga
7621 atatcagaga gtgagaggcc ttgacattgc tagcgtttac cgtcgacctc tagctagagc
7681 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
7741 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
7801 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
7861 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
7921 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
7981 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
8041 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
8101 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
8161 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
8221 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
8281 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
8341 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
8401 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
8461 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
8521 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
8581 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
8641 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
8701 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
8761 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
8821 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
8881 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
8941 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
9001 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
9061 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
9121 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
9181 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
9241 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
9301 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
9361 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
9421 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
9481 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
9541 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
9601 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
9661 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
9721 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
9781 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
9841 ccacctgacg tcgacggatc gggagatcaa cttgtttatt gcagcttata atggttacaa
9901 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
9961 tggtttgtcc aaactcatca atgtatctta tcatgtctgg atcaactgga taactcaagc
10021 taaccaaaat catcccaaac ttcccacccc ataccctatt accactgcca attacctgtg
10081 gtttcattta ctctaaacct gtgattcctc tgaattattt tcattttaaa gaaattgtat
10141 ttgttaaata tgtactacaa acttagtagt
//
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