| 质粒名称: | pLVX-MPro-eGFP-2 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 184824 |
| 存储实验室: | Wenshe Liu |
|
载体骨架基本信息 |
|
| 载体名称: | PLVX-puro |
| 空载体大小: | 8102 bp |
| 完整质粒大小: | 10493 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: | 37 ℃ |
| 宿主菌: | NEB Stable |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | Mpro-eGFP conjugate / Mpro-eGFP |
| 物种来源: | SARS-CoV-2, jellyfish Aequorea Victoria |
| 插入大小: | 1706bp |
| 启动子: | EF-1alfa |
| 标签/融合蛋白: | 人细胞色素c氧化酶亚基VIII（N端） |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | XbaI(未破坏) |
| 3’克隆位点: | BamHI(未破坏) |
| 5’测序引物 | CTAGTTCTAGAatgAAAACAAGCGC |
| 3’测序引物 | GCGGGATCCGCGGC |
|
文献引用方法 |
|
| 材料和方法部分: | pLVX-MPro-eGFP-2 was a gift from Wenshe Liu (Addgene plasmid # 184824 ; http://n2t.net/addgene:184824 ; RRID:Addgene_184824) |
| 参考文献部分: | Evaluation of SARS-CoV-2 Main Protease Inhibitors Using a Novel Cell-Based Assay. Cao W, Cho CD, Geng ZZ, Shaabani N, Ma XR, Vatansever EC, Alugubelli YR, Ma Y, Chaki SP, Ellenburg WH, Yang KS, Qiao Y, Allen R, Neuman BW, Ji H, Xu S, Liu WR. ACS Cent Sci. 2022 Feb 23;8(2):192-204. doi: 10.1021/acscentsci.1c00910. Epub 2022 Feb 2. 10.1021/acscentsci.1c00910 PubMed 35229034 |
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-MPro-eGFP-2 10493 bp ds-DNA circular SYN 17-MAY-2022
DEFINITION Expresses Mpro-eGFP in mammalian cells.
ACCESSION .
VERSION .
KEYWORDS pLVX-MPro-eGFP-2
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10493)
AUTHORS Cao W, Cho CD, Geng ZZ, Shaabani N, Ma XR, Vatansever EC, Alugubelli
YR, Ma Y, Chaki SP, Ellenburg WH, Yang KS, Qiao Y, Allen R, Neuman
BW, Ji H, Xu S, Liu WR
TITLE Evaluation of SARS-CoV-2 Main Protease Inhibitors Using a Novel
Cell-Based Assay.
JOURNAL ACS Cent Sci. 2022 Feb 23;8(2):192-204. doi:
10.1021/acscentsci.1c00910. Epub 2022 Feb 2.
PUBMED 35229034
REFERENCE 2 (bases 1 to 10493)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 17, 2022 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..10493
/organism="synthetic DNA construct"
/mol_type="other DNA"
LTR 32..665
/label=3' LTR
/note="3' long terminal repeat (LTR) from HIV-1"
misc_feature 712..837
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1334..1567
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 1752..1796
/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 2059..2174
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1 (lacking the first T)"
promoter 2369..3550
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
intron 2599..3541
/label=EF-1-alpha intron A
/note="intron upstream of the start codon of human
EF-1-alpha"
primer_bind 3498..3518
/label=EF1a-F
/note="Human elongation factor-1a promoter, forward primer"
CDS 4538..5257
/codon_start=1
/product="the original enhanced GFP (Yang et al., 1996)"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
CDS 4538..5254
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(4583..4604)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(4844..4863)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 5191..5212
/label=EGFP-C
/note="EGFP, forward primer"
misc_feature 5274..5847
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
primer_bind complement(5441..5458)
/label=IRES reverse
/note="IRES internal ribosome entry site, reverse primer.
Also called pCDH-rev"
primer_bind 5668..5687
/label=IRES-F
/note="IRES internal ribosome entry site, forward primer"
CDS 5880..6479
/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(5880..5899)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 6376..6396
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
misc_feature 6493..7081
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(6546..6566)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(6964..6975)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
LTR 7288..7921
/label=3' LTR
/note="3' long terminal repeat (LTR) from HIV-1"
primer_bind complement(8049..8065)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(8049..8065)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(8062..8084)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 8073..8089
/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(8097..8127)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 8142..8163
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(8280..8297)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(8451..9039)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(8531..8550)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(9210..10070)
/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 9833..9852
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(10071..10175)
/gene="bla"
/label=AmpR promoter
polyA_signal 10223..10357
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(10260..10279)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 10314..10333
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
ORIGIN
1 tttgttaaat atgtactaca aacttagtag ttggaagggc taattcactc ccaaagaaga
61 caagatatcc ttgatctgtg gatctaccac acacaaggct acttccctga ttagcagaac
121 tacacaccag ggccaggggt cagatatcca ctgacctttg gatggtgcta caagctagta
181 ccagttgagc cagataaggt agaagaggcc aataaaggag agaacaccag cttgttacac
241 cctgtgagcc tgcatgggat ggatgacccg gagagagaag tgttagagtg gaggtttgac
301 agccgcctag catttcatca cgtggcccga gagctgcatc cggagtactt caagaactgc
361 tgatatcgag cttgctacaa gggactttcc gctggggact ttccagggag gcgtggcctg
421 ggcgggactg gggagtggcg agccctcaga tcctgcatat aagcagctgc tttttgcctg
481 tactgggtct ctctggttag accagatctg agcctgggag ctctctggct aactagggaa
541 cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct
601 gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc
661 tagcagtggc gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga
721 cgcaggactc ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt
781 acgccaaaaa ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt
841 attaagcggg ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag
901 aaaaaatata aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt
961 aatcctggcc tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca
1021 tcccttcaga caggatcaga agaacttaga tcattatata atacagtagc aaccctctat
1081 tgtgtgcatc aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa
1141 gagcaaaaca aaagtaagac caccgcacag caagcggccg gccgctgatc ttcagacctg
1201 gaggaggaga tatgagggac aattggagaa gtgaattata taaatataaa gtagtaaaaa
1261 ttgaaccatt aggagtagca cccaccaagg caaagagaag agtggtgcag agagaaaaaa
1321 gagcagtggg aataggagct ttgttccttg ggttcttggg agcagcagga agcactatgg
1381 gcgcagcgtc aatgacgctg acggtacagg ccagacaatt attgtctggt atagtgcagc
1441 agcagaacaa tttgctgagg gctattgagg cgcaacagca tctgttgcaa ctcacagtct
1501 ggggcatcaa gcagctccag gcaagaatcc tggctgtgga aagataccta aaggatcaac
1561 agctcctggg gatttggggt tgctctggaa aactcatttg caccactgct gtgccttgga
1621 atgctagttg gagtaataaa tctctggaac agatttggaa tcacacgacc tggatggagt
1681 gggacagaga aattaacaat tacacaagct taatacactc cttaattgaa gaatcgcaaa
1741 accagcaaga aaagaatgaa caagaattat tggaattaga taaatgggca agtttgtgga
1801 attggtttaa cataacaaat tggctgtggt atataaaatt attcataatg atagtaggag
1861 gcttggtagg tttaagaata gtttttgctg tactttctat agtgaataga gttaggcagg
1921 gatattcacc attatcgttt cagacccacc tcccaacccc gaggggaccc gacaggcccg
1981 aaggaataga agaagaaggt ggagagagag acagagacag atccattcga ttagtgaacg
2041 gatctcgacg gtatcgcctt taaaagaaaa ggggggattg gggggtacag tgcaggggaa
2101 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca
2161 aaaattcaaa attttcgggt ttattacagg gacagcagag atccagttta tcgatgagta
2221 attcatacaa aaggactcgc ccctgccttg gggaatccca gggaccgtcg ttaaactccc
2281 actaacgtag aacccagaga tcgctgcgtt cccgccccct cacccgcccg ctctcgtcat
2341 cactgaggtg gagaagagca tgcgtgaggc tccggtgccc gtcagtgggc agagcgcaca
2401 tcgcccacag tccccgagaa gttgggggga ggggtcggca attgaaccgg tgcctagaga
2461 aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag
2521 ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgcaacggg
2581 tttgccgcca gaacacaggt aagtgccgtg tgtggttccc gcgggcctgg cctctttacg
2641 ggttatggcc cttgcgtgcc ttgaattact tccacgcccc tggctgcagt acgtgattct
2701 tgatcccgag cttcgggttg gaagtgggtg ggagagttcg aggccttgcg cttaaggagc
2761 cccttcgcct cgtgcttgag ttgaggcctg gcttgggcgc tggggccgcc gcgtgcgaat
2821 ctggtggcac cttcgcgcct gtctcgctgc tttcgataag tctctagcca tttaaaattt
2881 ttgatgacct gctgcgacgc tttttttctg gcaagatagt cttgtaaatg cgggccaaga
2941 tctgcacact ggtatttcgg tttttggggc cgcgggcggc gacggggccc gtgcgtccca
3001 gcgcacatgt tcggcgaggc ggggcctgcg agcgcggcca ccgagaatcg gacgggggta
3061 gtctcaagct ggccggcctg ctctggtgcc tggcctcgcg ccgccgtgta tcgccccgcc
3121 ctgggcggca aggctggccc ggtcggcacc agttgcgtga gcggaaagat ggccgcttcc
3181 cggccctgct gcagggagct caaaatggag gacgcggcgc tcgggagagc gggcgggtga
3241 gtcacccaca caaaggaaaa gggcctttcc gtcctcagcc gtcgcttcat gtgactccac
3301 ggagtaccgg gcgccgtcca ggcacctcga ttagttctcg agcttttgga gtacgtcgtc
3361 tttaggttgg ggggaggggt tttatgcgat ggagtttccc cacactgagt gggtggagac
3421 tgaagttagg ccagcttggc acttgatgta attctccttg gaatttgccc tttttgagtt
3481 tggatcttgg ttcattctca agcctcagac agtggttcaa agtttttttc ttccatttca
3541 ggtgtcgtga ggatctattt ccggtgaatt cctcgagact agttctagaa tgaaaacaag
3601 cgcggtgctc cagtcagggt ttcgcaagat ggcctttcca agcggtaagg ttgaaggttg
3661 catggtacaa gtaacgtgcg gtacgactac gttgaacggc ctgtggttgg acgatgtggt
3721 atattgcccc cgccacgtca tctgcacctc cgaagacatg ttaaacccca attacgagga
3781 tttgcttatc cgtaaaagca accataactt ccttgtgcaa gcaggcaatg tgcaattacg
3841 tgttattggc cactccatgc agaattgcgt acttaaatta aaggtggata cggccaaccc
3901 taaaacgccc aaatataaat ttgtccgcat tcaaccgggt caaaccttct ctgtattggc
3961 ctgttacaac ggttccccca gtggtgttta tcagtgtgca atgcgcccga acttcacaat
4021 caagggatcg ttcctgaatg ggagttgtgg ttcggttgga ttcaatatcg actacgactg
4081 cgtctctttt tgctatatgc accatatgga attaccgacc ggcgtacatg ccgggaccga
4141 tcttgaagga aatttctacg gcccgtttgt cgatcgtcag actgctcaag cagcaggtac
4201 ggatacgaca attaccgtga acgtccttgc atggctgtat gccgcggtca ttaacggaga
4261 tcgttggttt cttaaccgtt ttactaccac attaaacgat ttcaacttgg tggccatgaa
4321 gtacaattat gagcctctta cgcaagatca tgtagatatt ctgggacctt tatcggccca
4381 aacaggaatc gctgtgctgg acatgtgcgc atcattgaaa gagttgctgc agaatggtat
4441 gaacggacgc acaattctgg ggtcggcgtt gctggaggat gaattcaccc cgtttgatgt
4501 tgtgcgtcag tgttccggcg taacttttgg caagcttatg gtgagcaagg gcgaggagct
4561 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
4621 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat
4681 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg
4741 cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc
4801 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa
4861 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
4921 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag
4981 ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat
5041 ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc
5101 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct
5161 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
5221 cgggatcact ctcggcatgg acgagctgta caagtaagcg gccgcggatc ccgcccctct
5281 ccctcccccc cccctaacgt tactggccga agccgcttgg aataaggccg gtgtgcgttt
5341 gtctatatgt tattttccac catattgccg tcttttggca atgtgagggc ccggaaacct
5401 ggccctgtct tcttgacgag cattcctagg ggtctttccc ctctcgccaa aggaatgcaa
5461 ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag acaaacaacg
5521 tctgtagcga ccctttgcag gcagcggaac cccccacctg gcgacaggtg cctctgcggc
5581 caaaagccac gtgtataaga tacacctgca aaggcggcac aaccccagtg ccacgttgtg
5641 agttggatag ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg
5701 aaggatgccc agaaggtacc ccattgtatg ggatctgatc tggggcctcg gtgcacatgc
5761 tttacatgtg tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg
5821 gttttccttt gaaaaacacg atgataagct tgccacaacc cacaaggaga cgaccttcca
5881 tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc cgggccgtac
5941 gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gacccggacc
6001 gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca
6061 tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga
6121 gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt
6181 cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc
6241 ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca
6301 gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg
6361 agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg
6421 acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctaga
6481 cgcgtctgga acaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
6541 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
6601 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
6661 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
6721 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
6781 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
6841 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct gacgtccttt
6901 ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
6961 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
7021 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
7081 cctggaatta attctgcagt cgagacctag aaaaacatgg agcaatcaca agtagcaata
7141 cagcagctac caatgctgat tgtgcctggc tagaagcaca agaggaggag gaggtgggtt
7201 ttccagtcac acctcaggta cctttaagac caatgactta caaggcagct gtagatctta
7261 gccacttttt aaaagaaaag aggggactgg aagggctaat tcactcccaa cgaagacaag
7321 atatccttga tctgtggatc taccacacac aaggctactt ccctgattag cagaactaca
7381 caccagggcc aggggtcaga tatccactga cctttggatg gtgctacaag ctagtaccag
7441 ttgagccaga taaggtagaa gaggccaata aaggagagaa caccagcttg ttacaccctg
7501 tgagcctgca tgggatggat gacccggaga gagaagtgtt agagtggagg tttgacagcc
7561 gcctagcatt tcatcacgtg gcccgagagc tgcatccgga gtacttcaag aactgctgat
7621 atcgagcttg ctacaaggga ctttccgctg gggactttcc agggaggcgt ggcctgggcg
7681 ggactgggga gtggcgagcc ctcagatcct gcatataagc agctgctttt tgcctgtact
7741 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
7801 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
7861 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
7921 agtagtagtt catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca
7981 gagagtgaga ggccttgaca ttgctagcgt ttaccgtcga cctctagcta gagcttggcg
8041 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
8101 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
8161 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
8221 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
8281 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
8341 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
8401 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
8461 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
8521 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
8581 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
8641 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
8701 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
8761 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
8821 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
8881 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
8941 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
9001 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
9061 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
9121 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
9181 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
9241 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
9301 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
9361 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
9421 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
9481 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
9541 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
9601 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
9661 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
9721 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
9781 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
9841 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
9901 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
9961 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
10021 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
10081 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
10141 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
10201 gacgtcgacg gatcgggaga tcaacttgtt tattgcagct tataatggtt acaaataaag
10261 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
10321 gtccaaactc atcaatgtat cttatcatgt ctggatcaac tggataactc aagctaacca
10381 aaatcatccc aaacttccca ccccataccc tattaccact gccaattacc tgtggtttca
10441 tttactctaa acctgtgatt cctctgaatt attttcattt taaagaaatt gta
//
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