| 质粒名称: | pLVX-EF1alpha-SARS-CoV-2-nsp5-2xStrep-IRES-Puro |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 141370 |
| 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: | 9834 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 |
| 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-nsp5-2xStrep-IRES-Puro was a gift from Nevan Krogan (Addgene plasmid # 141370 ; http://n2t.net/addgene:141370 ; RRID:Addgene_141370) |
| 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
pLVX-EF1alpha-SARS-CoV-2-nsp5-2xStrep-IRES-Puro载体质粒基本信息
pLVX-EF1alpha-SARS-CoV-2-nsp5-2xStrep-IRES-Puro质粒图谱载体图谱和pLVX-EF1alpha-SARS-CoV-2-nsp5-2xStrep-IRES-Puro载体序列质粒序列多克隆位点信息
pLVX-EF1alpha-SARS-CoV-2-nsp5-2xStrep-IRES-Puro质粒载体简介
pLVX-EF1alpha-SARS-CoV-2-nsp5-2xStrep-IRES-Puro质粒序列载体序列
LOCUS pLVX-EF1alpha-SARS-CoV-2-nsp5-2xStrep-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-nsp5-2xStrep-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"
promoter 1..1182
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
intron 231..1173
/label=EF-1-alpha intron A
/note="intron upstream of the start codon of human
EF-1-alpha"
primer_bind 1130..1150
/label=EF1a-F
/note="Human elongation factor-1a promoter, forward primer"
CDS 2152..2175
/codon_start=1
/product="peptide that binds Strep-Tactin(R), an engineered
form of streptavidin"
/label=Strep-Tag II
/translation="WSHPQFEK"
CDS 2212..2235
/codon_start=1
/product="peptide that binds Strep-Tactin(R), an engineered
form of streptavidin"
/label=Strep-Tag II
/translation="WSHPQFEK"
misc_feature 2247..2820
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
primer_bind complement(2414..2431)
/label=IRES reverse
/note="IRES internal ribosome entry site, reverse primer.
Also called pCDH-rev"
primer_bind 2641..2660
/label=IRES-F
/note="IRES internal ribosome entry site, forward primer"
CDS 2853..3452
/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(2853..2872)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 3349..3369
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
misc_feature 3466..4054
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(3519..3539)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(3937..3948)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
LTR 4261..4894
/label=3' LTR
/note="3' long terminal repeat (LTR) from HIV-1"
primer_bind complement(5022..5038)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5022..5038)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(5035..5057)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 5046..5062
/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(5070..5100)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5115..5136
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(5253..5270)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5424..6012)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5504..5523)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(6183..7043)
/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 6806..6825
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(7044..7148)
/gene="bla"
/label=AmpR promoter
polyA_signal 7196..7330
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(7233..7252)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 7287..7306
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
LTR 7498..8131
/label=3' LTR
/note="3' long terminal repeat (LTR) from HIV-1"
misc_feature 8178..8303
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 8800..9033
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 9218..9262
/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 9525..9640
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1 (lacking the first T)"
ORIGIN
1 gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg
61 gaggggtcgg caattgaacc ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg
121 atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag
181 tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag gtaagtgccg
241 tgtgtggttc ccgcgggcct ggcctcttta cgggttatgg cccttgcgtg ccttgaatta
301 cttccacgcc cctggctgca gtacgtgatt cttgatcccg agcttcgggt tggaagtggg
361 tgggagagtt cgaggccttg cgcttaagga gccccttcgc ctcgtgcttg agttgaggcc
421 tggcttgggc gctggggccg ccgcgtgcga atctggtggc accttcgcgc ctgtctcgct
481 gctttcgata agtctctagc catttaaaat ttttgatgac ctgctgcgac gctttttttc
541 tggcaagata gtcttgtaaa tgcgggccaa gatctgcaca ctggtatttc ggtttttggg
601 gccgcgggcg gcgacggggc ccgtgcgtcc cagcgcacat gttcggcgag gcggggcctg
661 cgagcgcggc caccgagaat cggacggggg tagtctcaag ctggccggcc tgctctggtg
721 cctggcctcg cgccgccgtg tatcgccccg ccctgggcgg caaggctggc ccggtcggca
781 ccagttgcgt gagcggaaag atggccgctt cccggccctg ctgcagggag ctcaaaatgg
841 aggacgcggc gctcgggaga gcgggcgggt gagtcaccca cacaaaggaa aagggccttt
901 ccgtcctcag ccgtcgcttc atgtgactcc acggagtacc gggcgccgtc caggcacctc
961 gattagttct cgagcttttg gagtacgtcg tctttaggtt ggggggaggg gttttatgcg
1021 atggagtttc cccacactga gtgggtggag actgaagtta ggccagcttg gcacttgatg
1081 taattctcct tggaatttgc cctttttgag tttggatctt ggttcattct caagcctcag
1141 acagtggttc aaagtttttt tcttccattt caggtgtcgt gaggatctat ttccggtgaa
1201 ttcgccgcca ccatgtctgg ttttaggaaa atggcgttcc ccagcggtaa agttgaagga
1261 tgtatggtcc aagtaacctg tggtaccact acccttaatg ggctttggtt ggacgacgta
1321 gtctactgcc cccgacacgt aatctgcact agtgaggata tgcttaatcc caattacgaa
1381 gaccttttga ttcggaaatc caatcacaac ttcctggtcc aagcgggcaa cgtccaactc
1441 agggttattg gacatagtat gcagaattgc gtactgaagc tcaaagtcga tactgcaaac
1501 cccaagacgc ccaagtataa attcgtccga atccaaccag gccaaacatt ttccgtattg
1561 gcttgctata atggaagccc cagcggtgtc taccaatgtg caatgagacc aaactttacg
1621 ataaagggtt catttctgaa cggctcttgc ggttccgttg gttttaacat cgactatgac
1681 tgtgtatcct tttgctacat gcaccatatg gaactcccta ccggtgtcca cgccggtaca
1741 gatctggaag gaaatttcta cggtccgttc gttgaccggc aaaccgcgca agcggctgga
1801 accgacacaa cgattacagt gaatgtgctc gcgtggctgt acgcagcagt cataaacgga
1861 gacaggtggt ttctgaaccg atttacgacg actctcaatg acttcaacct tgttgcgatg
1921 aagtacaatt acgagccact cacccaggac catgttgata tcctgggtcc cctcagtgcc
1981 cagacaggga tcgcagttct cgatatgtgc gcgtcactga aggagcttct ccaaaatgga
2041 atgaatgggc ggaccatact tggttccgca ctcctcgaag atgaatttac tccatttgac
2101 gtggtcagac aatgcagtgg ggtcactttc cagctcgaag gcggcggggg atggagccat
2161 ccacaattcg agaaaggcgg tggttcagga ggaggtagcg ggggtggatc atggtcacat
2221 ccgcagtttg aaaagtaagg atcccgcccc tctccctccc ccccccctaa cgttactggc
2281 cgaagccgct tggaataagg ccggtgtgcg tttgtctata tgttattttc caccatattg
2341 ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg tcttcttgac gagcattcct
2401 aggggtcttt cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt gaaggaagca
2461 gttcctctgg aagcttcttg aagacaaaca acgtctgtag cgaccctttg caggcagcgg
2521 aaccccccac ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata agatacacct
2581 gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa
2641 tggctctcct caagcgtatt caacaagggg ctgaaggatg cccagaaggt accccattgt
2701 atgggatctg atctggggcc tcggtgcaca tgctttacat gtgtttagtc gaggttaaaa
2761 aaacgtctag gccccccgaa ccacggggac gtggttttcc tttgaaaaac acgatgataa
2821 gcttgccaca acccacaagg agacgacctt ccatgaccga gtacaagccc acggtgcgcc
2881 tcgccacccg cgacgacgtc ccccgggccg tacgcaccct cgccgccgcg ttcgccgact
2941 accccgccac gcgccacacc gtcgacccgg accgccacat cgagcgggtc accgagctgc
3001 aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg
3061 gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg
3121 agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg
3181 aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg
3241 tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg
3301 cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct
3361 tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca
3421 cctggtgcat gacccgcaag cccggtgcct agacgcgtct ggaacaatca acctctggat
3481 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
3541 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
3601 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg
3661 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc
3721 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa
3781 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
3841 tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg tgttgccacc
3901 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt
3961 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag
4021 acgagtcgga tctccctttg ggccgcctcc ccgcctggaa ttaattctgc agtcgagacc
4081 tagaaaaaca tggagcaatc acaagtagca atacagcagc taccaatgct gattgtgcct
4141 ggctagaagc acaagaggag gaggaggtgg gttttccagt cacacctcag gtacctttaa
4201 gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa aagaggggac
4261 tggaagggct aattcactcc caacgaagac aagatatcct tgatctgtgg atctaccaca
4321 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
4381 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
4441 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
4501 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
4561 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
4621 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
4681 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
4741 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
4801 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
4861 agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca tcttattatt
4921 cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggccttg acattgctag
4981 cgtttaccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg
5041 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
5101 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
5161 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
5221 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
5281 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
5341 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
5401 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
5461 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
5521 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
5581 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
5641 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
5701 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
5761 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
5821 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
5881 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
5941 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
6001 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
6061 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
6121 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
6181 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
6241 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
6301 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
6361 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
6421 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
6481 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
6541 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
6601 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
6661 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
6721 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
6781 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
6841 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
6901 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
6961 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
7021 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
7081 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
7141 gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg agatcaactt
7201 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
7261 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
7321 tgtctggatc aactggataa ctcaagctaa ccaaaatcat cccaaacttc ccaccccata
7381 ccctattacc actgccaatt acctgtggtt tcatttactc taaacctgtg attcctctga
7441 attattttca ttttaaagaa attgtatttg ttaaatatgt actacaaact tagtagttgg
7501 aagggctaat tcactcccaa agaagacaag atatccttga tctgtggatc taccacacac
7561 aaggctactt ccctgattag cagaactaca caccagggcc aggggtcaga tatccactga
7621 cctttggatg gtgctacaag ctagtaccag ttgagccaga taaggtagaa gaggccaata
7681 aaggagagaa caccagcttg ttacaccctg tgagcctgca tgggatggat gacccggaga
7741 gagaagtgtt agagtggagg tttgacagcc gcctagcatt tcatcacgtg gcccgagagc
7801 tgcatccgga gtacttcaag aactgctgat atcgagcttg ctacaaggga ctttccgctg
7861 gggactttcc agggaggcgt ggcctgggcg ggactgggga gtggcgagcc ctcagatcct
7921 gcatataagc agctgctttt tgcctgtact gggtctctct ggttagacca gatctgagcc
7981 tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga
8041 gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga
8101 cccttttagt cagtgtggaa aatctctagc agtggcgccc gaacagggac ttgaaagcga
8161 aagggaaacc agaggagctc tctcgacgca ggactcggct tgctgaagcg cgcacggcaa
8221 gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt gactagcgga ggctagaagg
8281 agagagatgg gtgcgagagc gtcagtatta agcgggggag aattagatcg cgatgggaaa
8341 aaattcggtt aaggccaggg ggaaagaaaa aatataaatt aaaacatata gtatgggcaa
8401 gcagggagct agaacgattc gcagttaatc ctggcctgtt agaaacatca gaaggctgta
8461 gacaaatact gggacagcta caaccatccc ttcagacagg atcagaagaa cttagatcat
8521 tatataatac agtagcaacc ctctattgtg tgcatcaaag gatagagata aaagacacca
8581 aggaagcttt agacaagata gaggaagagc aaaacaaaag taagaccacc gcacagcaag
8641 cggccggccg ctgatcttca gacctggagg aggagatatg agggacaatt ggagaagtga
8701 attatataaa tataaagtag taaaaattga accattagga gtagcaccca ccaaggcaaa
8761 gagaagagtg gtgcagagag aaaaaagagc agtgggaata ggagctttgt tccttgggtt
8821 cttgggagca gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg tacaggccag
8881 acaattattg tctggtatag tgcagcagca gaacaatttg ctgagggcta ttgaggcgca
8941 acagcatctg ttgcaactca cagtctgggg catcaagcag ctccaggcaa gaatcctggc
9001 tgtggaaaga tacctaaagg atcaacagct cctggggatt tggggttgct ctggaaaact
9061 catttgcacc actgctgtgc cttggaatgc tagttggagt aataaatctc tggaacagat
9121 ttggaatcac acgacctgga tggagtggga cagagaaatt aacaattaca caagcttaat
9181 acactcctta attgaagaat cgcaaaacca gcaagaaaag aatgaacaag aattattgga
9241 attagataaa tgggcaagtt tgtggaattg gtttaacata acaaattggc tgtggtatat
9301 aaaattattc ataatgatag taggaggctt ggtaggttta agaatagttt ttgctgtact
9361 ttctatagtg aatagagtta ggcagggata ttcaccatta tcgtttcaga cccacctccc
9421 aaccccgagg ggacccgaca ggcccgaagg aatagaagaa gaaggtggag agagagacag
9481 agacagatcc attcgattag tgaacggatc tcgacggtat cgcctttaaa agaaaagggg
9541 ggattggggg gtacagtgca ggggaaagaa tagtagacat aatagcaaca gacatacaaa
9601 ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat tacagggaca
9661 gcagagatcc agtttatcga tgagtaattc atacaaaagg actcgcccct gccttgggga
9721 atcccaggga ccgtcgttaa actcccacta acgtagaacc cagagatcgc tgcgttcccg
9781 ccccctcacc cgcccgctct cgtcatcact gaggtggaga agagcatgcg tgag
//
