| 质粒名称: | pLVX-EF1alpha-SARS-CoV-2-orf10-2xStrep-IRES-Puro |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 141389 |
| 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-orf10 |
| 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-orf10-2xStrep-IRES-Puro was a gift from Nevan Krogan (Addgene plasmid # 141394 ; http://n2t.net/addgene:141394 ; RRID:Addgene_141394) |
| 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-orf10-2xStrep-IRES-Puro 9027 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-orf10-2xStrep-IRES-Puro
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9027)
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 9027)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..9027
/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"
regulatory 3544..3553
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 3682..3705
/codon_start=1
/product="peptide that binds Strep-Tactin(R), an engineered
form of streptavidin"
/label=Strep-Tag II
/translation="WSHPQFEK"
CDS 3742..3765
/codon_start=1
/product="peptide that binds Strep-Tactin(R), an engineered
form of streptavidin"
/label=Strep-Tag II
/translation="WSHPQFEK"
misc_feature 3777..4350
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
primer_bind complement(3944..3961)
/label=IRES reverse
/note="IRES internal ribosome entry site, reverse primer.
Also called pCDH-rev"
primer_bind 4171..4190
/label=IRES-F
/note="IRES internal ribosome entry site, forward primer"
CDS 4383..4982
/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(4383..4402)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 4879..4899
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
misc_feature 4996..5584
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(5049..5069)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(5467..5478)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
LTR 5791..6424
/label=3' LTR
/note="3' long terminal repeat (LTR) from HIV-1"
primer_bind complement(6552..6568)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(6552..6568)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(6565..6587)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 6576..6592
/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(6600..6630)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 6645..6666
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(6783..6800)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(6954..7542)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(7034..7053)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(7713..8573)
/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 8336..8355
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(8574..8678)
/gene="bla"
/label=AmpR promoter
polyA_signal 8726..8860
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(8763..8782)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 8817..8836
/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 tgggctacat caatgtcttc gcctttcctt tcaccatcta ttctctcttg
3601 ttgtgtcgca tgaactctag gaattacata gctcaggtcg atgttgtaaa cttcaatctc
3661 acactcgaag gcggcggggg atggagccat ccacaattcg agaaaggcgg tggttcagga
3721 ggaggtagcg ggggtggatc atggtcacat ccgcagtttg aaaagtaagg atcccgcccc
3781 tctccctccc ccccccctaa cgttactggc cgaagccgct tggaataagg ccggtgtgcg
3841 tttgtctata tgttattttc caccatattg ccgtcttttg gcaatgtgag ggcccggaaa
3901 cctggccctg tcttcttgac gagcattcct aggggtcttt cccctctcgc caaaggaatg
3961 caaggtctgt tgaatgtcgt gaaggaagca gttcctctgg aagcttcttg aagacaaaca
4021 acgtctgtag cgaccctttg caggcagcgg aaccccccac ctggcgacag gtgcctctgc
4081 ggccaaaagc cacgtgtata agatacacct gcaaaggcgg cacaacccca gtgccacgtt
4141 gtgagttgga tagttgtgga aagagtcaaa tggctctcct caagcgtatt caacaagggg
4201 ctgaaggatg cccagaaggt accccattgt atgggatctg atctggggcc tcggtgcaca
4261 tgctttacat gtgtttagtc gaggttaaaa aaacgtctag gccccccgaa ccacggggac
4321 gtggttttcc tttgaaaaac acgatgataa gcttgccaca acccacaagg agacgacctt
4381 ccatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc ccccgggccg
4441 tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgacccgg
4501 accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg
4561 acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg
4621 agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg
4681 gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg
4741 agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg
4801 gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc
4861 tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg
4921 ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct
4981 agacgcgtct ggaacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt
5041 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
5101 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
5161 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
5221 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
5281 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
5341 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc
5401 tttccatggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac
5461 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
5521 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
5581 ccgcctggaa ttaattctgc agtcgagacc tagaaaaaca tggagcaatc acaagtagca
5641 atacagcagc taccaatgct gattgtgcct ggctagaagc acaagaggag gaggaggtgg
5701 gttttccagt cacacctcag gtacctttaa gaccaatgac ttacaaggca gctgtagatc
5761 ttagccactt tttaaaagaa aagaggggac tggaagggct aattcactcc caacgaagac
5821 aagatatcct tgatctgtgg atctaccaca cacaaggcta cttccctgat tagcagaact
5881 acacaccagg gccaggggtc agatatccac tgacctttgg atggtgctac aagctagtac
5941 cagttgagcc agataaggta gaagaggcca ataaaggaga gaacaccagc ttgttacacc
6001 ctgtgagcct gcatgggatg gatgacccgg agagagaagt gttagagtgg aggtttgaca
6061 gccgcctagc atttcatcac gtggcccgag agctgcatcc ggagtacttc aagaactgct
6121 gatatcgagc ttgctacaag ggactttccg ctggggactt tccagggagg cgtggcctgg
6181 gcgggactgg ggagtggcga gccctcagat cctgcatata agcagctgct ttttgcctgt
6241 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
6301 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
6361 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
6421 agcagtagta gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata
6481 tcagagagtg agaggccttg acattgctag cgtttaccgt cgacctctag ctagagcttg
6541 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
6601 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
6661 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
6721 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
6781 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
6841 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
6901 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
6961 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
7021 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
7081 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
7141 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
7201 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
7261 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
7321 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
7381 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
7441 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
7501 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
7561 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
7621 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
7681 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
7741 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
7801 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
7861 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
7921 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
7981 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
8041 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
8101 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
8161 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
8221 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
8281 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
8341 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
8401 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
8461 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
8521 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
8581 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
8641 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
8701 cctgacgtcg acggatcggg agatcaactt gtttattgca gcttataatg gttacaaata
8761 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
8821 tttgtccaaa ctcatcaatg tatcttatca tgtctggatc aactggataa ctcaagctaa
8881 ccaaaatcat cccaaacttc ccaccccata ccctattacc actgccaatt acctgtggtt
8941 tcatttactc taaacctgtg attcctctga attattttca ttttaaagaa attgtatttg
9001 ttaaatatgt actacaaact tagtagt
//
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