| 质粒名称: | pLVX-EF1alpha-SARS-CoV-2-nsp13-2xStrep-IRES-Puro |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 141379 |
| 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-nsp13 |
| 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-nsp13-2xStrep-IRES-Puro was a gift from Nevan Krogan (Addgene plasmid # 141379 ; http://n2t.net/addgene:141379 ; RRID:Addgene_141379) |
| 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-nsp13-2xStrep-IRES-Puro 10719 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-nsp13-2xStrep-IRES-Puro
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10719)
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 10719)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..10719
/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 5374..5397
/codon_start=1
/product="peptide that binds Strep-Tactin(R), an engineered
form of streptavidin"
/label=Strep-Tag II
/translation="WSHPQFEK"
CDS 5434..5457
/codon_start=1
/product="peptide that binds Strep-Tactin(R), an engineered
form of streptavidin"
/label=Strep-Tag II
/translation="WSHPQFEK"
misc_feature 5469..6042
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
primer_bind complement(5636..5653)
/label=IRES reverse
/note="IRES internal ribosome entry site, reverse primer.
Also called pCDH-rev"
primer_bind 5863..5882
/label=IRES-F
/note="IRES internal ribosome entry site, forward primer"
CDS 6075..6674
/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(6075..6094)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 6571..6591
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
misc_feature 6688..7276
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(6741..6761)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(7159..7170)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
LTR 7483..8116
/label=3' LTR
/note="3' long terminal repeat (LTR) from HIV-1"
primer_bind complement(8244..8260)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(8244..8260)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(8257..8279)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 8268..8284
/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(8292..8322)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 8337..8358
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(8475..8492)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(8646..9234)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(8726..8745)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(9405..10265)
/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 10028..10047
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(10266..10370)
/gene="bla"
/label=AmpR promoter
polyA_signal 10418..10552
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(10455..10474)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 10509..10528
/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 tggctgttgg ggcgtgtgtg ttgtgcaaca gccaaacctc tctgcgatgc
3601 ggagcgtgca tacggaggcc atttctttgt tgtaaatgtt gttatgatca cgttattagt
3661 acgagccata agttggttct gagcgtcaac ccctatgtgt gtaacgctcc gggttgcgac
3721 gtaaccgatg tgacacaact ttacctgggg ggcatgtcat attactgcaa aagtcacaaa
3781 cccccaatct catttccgtt gtgtgcgaat ggccaggtct ttggccttta caaaaatact
3841 tgtgttggtt cagacaacgt aaccgatttt aacgcgattg ctacatgcga ttggacaaat
3901 gctggtgatt atattttggc taatacgtgc acggaaaggc tcaaattgtt tgcagcggag
3961 accctcaagg cgaccgagga aacctttaaa ctgagctacg gcattgcgac ggtcagagag
4021 gttctttccg accgagaatt gcacctctca tgggaggtcg gtaagccgcg cccgcctctg
4081 aataggaact atgtctttac aggctatcga gttacaaaaa acagtaaggt gcagataggg
4141 gaatatactt ttgagaaagg tgactatggg gacgccgtag tatacagggg gacaactacc
4201 tacaaactga atgtgggcga ttatttcgtg ttgactagcc atactgtcat gcccctctct
4261 gccccgaccc tcgttccaca ggaacattat gttcgcatta ctgggctgta ccccacgctc
4321 aacatatccg atgaatttag ctccaacgtg gcaaactacc agaaggtggg aatgcaaaag
4381 tatagcacgt tgcaaggccc ccctggcaca ggtaaatccc acttcgccat tggcttggcg
4441 ttgtactacc cgagtgcccg gatcgtctat actgcatgct ctcatgctgc ggtagacgca
4501 ttgtgtgaga aagccctgaa gtatctgccc attgacaagt gcagtcggat aatcccagct
4561 cgggcgcgag ttgaatgctt cgataagttt aaggtaaata gtacacttga gcagtatgta
4621 ttttgtaccg tcaacgctct ccccgaaacc actgcggaca tagtggtatt tgatgagata
4681 tctatggcta ccaattatga cctctcagta gtcaacgcac gcttgcgagc aaaacactat
4741 gtatacatcg gagacccagc gcaattgcct gctcctagga ctctcctcac caaagggact
4801 cttgagcctg aatatttcaa ctccgtctgc aggctgatga aaaccatcgg acccgatatg
4861 tttctcggga catgtcgaag atgtcctgcc gaaattgtag atactgttag cgccttggtg
4921 tacgataata agctgaaggc ccataaagac aagagcgccc agtgtttcaa gatgttctat
4981 aagggagtaa tcacacatga cgttagttct gccatcaacc gaccgcagat aggcgtggtc
5041 cgcgagttcc tgacccgaaa tccagcgtgg aggaaggccg tatttatatc accttacaat
5101 agccaaaatg ccgtggcgtc aaagattttg gggctcccga ctcagacggt ggatagctct
5161 caagggtcag aatacgatta cgttatattc acacagacga cagagacggc acacagttgc
5221 aacgtcaaca gatttaatgt cgctataacc cgcgccaagg ttgggattct ttgtataatg
5281 tctgacagag acctctacga caagctgcag ttcacgtcac ttgagattcc tcgcaggaac
5341 gttgcgacgc ttcaactcga aggcggcggg ggatggagcc atccacaatt cgagaaaggc
5401 ggtggttcag gaggaggtag cgggggtgga tcatggtcac atccgcagtt tgaaaagtaa
5461 ggatcccgcc cctctccctc ccccccccct aacgttactg gccgaagccg cttggaataa
5521 ggccggtgtg cgtttgtcta tatgttattt tccaccatat tgccgtcttt tggcaatgtg
5581 agggcccgga aacctggccc tgtcttcttg acgagcattc ctaggggtct ttcccctctc
5641 gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct ggaagcttct
5701 tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc ggaacccccc acctggcgac
5761 aggtgcctct gcggccaaaa gccacgtgta taagatacac ctgcaaaggc ggcacaaccc
5821 cagtgccacg ttgtgagttg gatagttgtg gaaagagtca aatggctctc ctcaagcgta
5881 ttcaacaagg ggctgaagga tgcccagaag gtaccccatt gtatgggatc tgatctgggg
5941 cctcggtgca catgctttac atgtgtttag tcgaggttaa aaaaacgtct aggccccccg
6001 aaccacgggg acgtggtttt cctttgaaaa acacgatgat aagcttgcca caacccacaa
6061 ggagacgacc ttccatgacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg
6121 tcccccgggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca
6181 ccgtcgaccc ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc
6241 gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct
6301 ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg
6361 ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc
6421 accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg
6481 gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg
6541 tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct
6601 tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca
6661 agcccggtgc ctagacgcgt ctggaacaat caacctctgg attacaaaat ttgtgaaaga
6721 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
6781 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
6841 tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc
6901 actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt
6961 tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt
7021 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg
7081 aagctgacgt cctttccatg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg
7141 tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg
7201 ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt
7261 tgggccgcct ccccgcctgg aattaattct gcagtcgaga cctagaaaaa catggagcaa
7321 tcacaagtag caatacagca gctaccaatg ctgattgtgc ctggctagaa gcacaagagg
7381 aggaggaggt gggttttcca gtcacacctc aggtaccttt aagaccaatg acttacaagg
7441 cagctgtaga tcttagccac tttttaaaag aaaagagggg actggaaggg ctaattcact
7501 cccaacgaag acaagatatc cttgatctgt ggatctacca cacacaaggc tacttccctg
7561 attagcagaa ctacacacca gggccagggg tcagatatcc actgaccttt ggatggtgct
7621 acaagctagt accagttgag ccagataagg tagaagaggc caataaagga gagaacacca
7681 gcttgttaca ccctgtgagc ctgcatggga tggatgaccc ggagagagaa gtgttagagt
7741 ggaggtttga cagccgccta gcatttcatc acgtggcccg agagctgcat ccggagtact
7801 tcaagaactg ctgatatcga gcttgctaca agggactttc cgctggggac tttccaggga
7861 ggcgtggcct gggcgggact ggggagtggc gagccctcag atcctgcata taagcagctg
7921 ctttttgcct gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc
7981 taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg
8041 tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg
8101 tggaaaatct ctagcagtag tagttcatgt catcttatta ttcagtattt ataacttgca
8161 aagaaatgaa tatcagagag tgagaggcct tgacattgct agcgtttacc gtcgacctct
8221 agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
8281 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
8341 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
8401 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
8461 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
8521 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
8581 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
8641 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
8701 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
8761 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
8821 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
8881 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
8941 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
9001 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
9061 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
9121 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
9181 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
9241 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
9301 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
9361 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
9421 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
9481 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
9541 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
9601 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
9661 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
9721 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
9781 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
9841 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
9901 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
9961 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
10021 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
10081 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
10141 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
10201 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
10261 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
10321 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
10381 cgaaaagtgc cacctgacgt cgacggatcg ggagatcaac ttgtttattg cagcttataa
10441 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
10501 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga tcaactggat
10561 aactcaagct aaccaaaatc atcccaaact tcccacccca taccctatta ccactgccaa
10621 ttacctgtgg tttcatttac tctaaacctg tgattcctct gaattatttt cattttaaag
10681 aaattgtatt tgttaaatat gtactacaaa cttagtagt
//
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