| 质粒名称: | pLVX-EF1alpha-SARS-CoV-2-nsp5-C145A-2xStrep-IRES-Puro |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 141371 |
| 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-nsp5-C145A / 3CLPro |
| 物种来源: | SARS-CoV-2 |
| 基因突变: | C145A, 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-nsp5-C145A-2xStrep-IRES-Puro was a gift from Nevan Krogan (Addgene plasmid # 141371 ; http://n2t.net/addgene:141371 ; RRID:Addgene_141371) |
| 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 |
pLVX-EF1alpha-SARS-CoV-2-nsp5-C145A-2xStrep-IRES-Puro
买家导航
pLVX-EF1alpha-SARS-CoV-2-nsp5-C145A-2xStrep-IRES-Puro载体质粒基本信息
pLVX-EF1alpha-SARS-CoV-2-nsp5-C145A-2xStrep-IRES-Puro质粒图谱载体图谱和pLVX-EF1alpha-SARS-CoV-2-nsp5-C145A-2xStrep-IRES-Puro载体序列质粒序列多克隆位点信息
pLVX-EF1alpha-SARS-CoV-2-nsp5-C145A-2xStrep-IRES-Puro质粒载体简介
pLVX-EF1alpha-SARS-CoV-2-nsp5-C145A-2xStrep-IRES-Puro质粒序列载体序列
LOCUS pLVX-EF1alpha-SARS-CoV-2-nsp5-C145A-2xStrep-IRES-Puro 9834 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Lentiviral expression of SARS-CoV-2 protein; transient expression
and generate lentivirus.
ACCESSION .
VERSION .
KEYWORDS pLVX-EF1alpha-SARS-CoV-2-nsp5-C145A-2xStrep-IRES-Puro
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9834)
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 9834)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..9834
/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 4489..4512
/codon_start=1
/product="peptide that binds Strep-Tactin(R), an engineered
form of streptavidin"
/label=Strep-Tag II
/translation="WSHPQFEK"
CDS 4549..4572
/codon_start=1
/product="peptide that binds Strep-Tactin(R), an engineered
form of streptavidin"
/label=Strep-Tag II
/translation="WSHPQFEK"
misc_feature 4584..5157
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
primer_bind complement(4751..4768)
/label=IRES reverse
/note="IRES internal ribosome entry site, reverse primer.
Also called pCDH-rev"
primer_bind 4978..4997
/label=IRES-F
/note="IRES internal ribosome entry site, forward primer"
CDS 5190..5789
/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(5190..5209)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 5686..5706
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
misc_feature 5803..6391
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(5856..5876)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(6274..6285)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
LTR 6598..7231
/label=3' LTR
/note="3' long terminal repeat (LTR) from HIV-1"
primer_bind complement(7359..7375)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(7359..7375)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(7372..7394)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 7383..7399
/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(7407..7437)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 7452..7473
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(7590..7607)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(7761..8349)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(7841..7860)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(8520..9380)
/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 9143..9162
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(9381..9485)
/gene="bla"
/label=AmpR promoter
polyA_signal 9533..9667
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(9570..9589)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 9624..9643
/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 tgtctggttt taggaaaatg gcgttcccca gcggtaaagt tgaaggatgt
3601 atggtccaag taacctgtgg taccactacc cttaatgggc tttggttgga cgacgtagtc
3661 tactgccccc gacacgtaat ctgcactagt gaggatatgc ttaatcccaa ttacgaagac
3721 cttttgattc ggaaatccaa tcacaacttc ctggtccaag cgggcaacgt ccaactcagg
3781 gttattggac atagtatgca gaattgcgta ctgaagctca aagtcgatac tgcaaacccc
3841 aagacgccca agtataaatt cgtccgaatc caaccaggcc aaacattttc cgtattggct
3901 tgctataatg gaagccccag cggtgtctac caatgtgcaa tgagaccaaa ctttacgata
3961 aagggttcat ttctgaacgg ctctgccggt tccgttggtt ttaacatcga ctatgactgt
4021 gtatcctttt gctacatgca ccatatggaa ctccctaccg gtgtccacgc cggtacagat
4081 ctggaaggaa atttctacgg tccgttcgtt gaccggcaaa ccgcgcaagc ggctggaacc
4141 gacacaacga ttacagtgaa tgtgctcgcg tggctgtacg cagcagtcat aaacggagac
4201 aggtggtttc tgaaccgatt tacgacgact ctcaatgact tcaaccttgt tgcgatgaag
4261 tacaattacg agccactcac ccaggaccat gttgatatcc tgggtcccct cagtgcccag
4321 acagggatcg cagttctcga tatgtgcgcg tcactgaagg agcttctcca aaatggaatg
4381 aatgggcgga ccatacttgg ttccgcactc ctcgaagatg aatttactcc atttgacgtg
4441 gtcagacaat gcagtggggt cactttccag ctcgaaggcg gcgggggatg gagccatcca
4501 caattcgaga aaggcggtgg ttcaggagga ggtagcgggg gtggatcatg gtcacatccg
4561 cagtttgaaa agtaaggatc ccgcccctct ccctcccccc cccctaacgt tactggccga
4621 agccgcttgg aataaggccg gtgtgcgttt gtctatatgt tattttccac catattgccg
4681 tcttttggca atgtgagggc ccggaaacct ggccctgtct tcttgacgag cattcctagg
4741 ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt
4801 cctctggaag cttcttgaag acaaacaacg tctgtagcga ccctttgcag gcagcggaac
4861 cccccacctg gcgacaggtg cctctgcggc caaaagccac gtgtataaga tacacctgca
4921 aaggcggcac aaccccagtg ccacgttgtg agttggatag ttgtggaaag agtcaaatgg
4981 ctctcctcaa gcgtattcaa caaggggctg aaggatgccc agaaggtacc ccattgtatg
5041 ggatctgatc tggggcctcg gtgcacatgc tttacatgtg tttagtcgag gttaaaaaaa
5101 cgtctaggcc ccccgaacca cggggacgtg gttttccttt gaaaaacacg atgataagct
5161 tgccacaacc cacaaggaga cgaccttcca tgaccgagta caagcccacg gtgcgcctcg
5221 ccacccgcga cgacgtcccc cgggccgtac gcaccctcgc cgccgcgttc gccgactacc
5281 ccgccacgcg ccacaccgtc gacccggacc gccacatcga gcgggtcacc gagctgcaag
5341 aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg
5401 ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga
5461 tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag
5521 gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct
5581 cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg
5641 ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct
5701 acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct
5761 ggtgcatgac ccgcaagccc ggtgcctaga cgcgtctgga acaatcaacc tctggattac
5821 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga
5881 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc
5941 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa
6001 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc
6061 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc
6121 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc
6181 gtggtgttgt cggggaagct gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg
6241 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct
6301 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg
6361 agtcggatct ccctttgggc cgcctccccg cctggaatta attctgcagt cgagacctag
6421 aaaaacatgg agcaatcaca agtagcaata cagcagctac caatgctgat tgtgcctggc
6481 tagaagcaca agaggaggag gaggtgggtt ttccagtcac acctcaggta cctttaagac
6541 caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag aggggactgg
6601 aagggctaat tcactcccaa cgaagacaag atatccttga tctgtggatc taccacacac
6661 aaggctactt ccctgattag cagaactaca caccagggcc aggggtcaga tatccactga
6721 cctttggatg gtgctacaag ctagtaccag ttgagccaga taaggtagaa gaggccaata
6781 aaggagagaa caccagcttg ttacaccctg tgagcctgca tgggatggat gacccggaga
6841 gagaagtgtt agagtggagg tttgacagcc gcctagcatt tcatcacgtg gcccgagagc
6901 tgcatccgga gtacttcaag aactgctgat atcgagcttg ctacaaggga ctttccgctg
6961 gggactttcc agggaggcgt ggcctgggcg ggactgggga gtggcgagcc ctcagatcct
7021 gcatataagc agctgctttt tgcctgtact gggtctctct ggttagacca gatctgagcc
7081 tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga
7141 gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga
7201 cccttttagt cagtgtggaa aatctctagc agtagtagtt catgtcatct tattattcag
7261 tatttataac ttgcaaagaa atgaatatca gagagtgaga ggccttgaca ttgctagcgt
7321 ttaccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
7381 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
7441 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
7501 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
7561 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
7621 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
7681 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
7741 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
7801 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
7861 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
7921 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
7981 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
8041 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
8101 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
8161 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
8221 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
8281 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
8341 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
8401 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
8461 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
8521 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
8581 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
8641 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
8701 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
8761 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
8821 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
8881 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
8941 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
9001 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
9061 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
9121 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
9181 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
9241 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
9301 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
9361 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
9421 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
9481 ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tcaacttgtt
9541 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
9601 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
9661 ctggatcaac tggataactc aagctaacca aaatcatccc aaacttccca ccccataccc
9721 tattaccact gccaattacc tgtggtttca tttactctaa acctgtgatt cctctgaatt
9781 attttcattt taaagaaatt gtatttgtta aatatgtact acaaacttag tagt
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