| 质粒名称: | pLVX-EF1alpha-eGFP-2xStrep-IRES-Puro |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 141395 |
| 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: | eGFP |
| 物种来源: | jellyfish |
| 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-eGFP-2xStrep-IRES-Puro was a gift from Nevan Krogan (Addgene plasmid # 141395 ; http://n2t.net/addgene:141395 ; RRID:Addgene_141395) |
| 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-eGFP-2xStrep-IRES-Puro载体质粒基本信息
pLVX-EF1alpha-eGFP-2xStrep-IRES-Puro质粒图谱载体图谱和pLVX-EF1alpha-eGFP-2xStrep-IRES-Puro载体序列质粒序列多克隆位点信息
pLVX-EF1alpha-eGFP-2xStrep-IRES-Puro质粒载体简介
pLVX-EF1alpha-eGFP-2xStrep-IRES-Puro质粒序列载体序列
LOCUS pLVX-EF1alpha-eGFP-2xStrep-IRES-Puro 9642 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-eGFP-2xStrep-IRES-Puro
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9642)
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 9642)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..9642
/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 3544..4260
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
CDS 3544..4260
/codon_start=1
/product="the original enhanced GFP (Yang et al., 1996)"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(3589..3610)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(3850..3869)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 4197..4218
/label=EGFP-C
/note="EGFP, forward primer"
CDS 4288..4311
/codon_start=1
/product="peptide that binds Strep-Tactin(R), an engineered
form of streptavidin"
/label=Strep-Tag II
/translation="WSHPQFEK"
CDS 4348..4371
/codon_start=1
/product="peptide that binds Strep-Tactin(R), an engineered
form of streptavidin"
/label=Strep-Tag II
/translation="WSHPQFEK"
misc_feature 4392..4965
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
primer_bind complement(4559..4576)
/label=IRES reverse
/note="IRES internal ribosome entry site, reverse primer.
Also called pCDH-rev"
primer_bind 4786..4805
/label=IRES-F
/note="IRES internal ribosome entry site, forward primer"
CDS 4998..5597
/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(4998..5017)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 5494..5514
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
misc_feature 5611..6199
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(5664..5684)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(6082..6093)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
LTR 6406..7039
/label=3' LTR
/note="3' long terminal repeat (LTR) from HIV-1"
primer_bind complement(7167..7183)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(7167..7183)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(7180..7202)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 7191..7207
/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(7215..7245)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 7260..7281
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(7398..7415)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(7569..8157)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(7649..7668)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(8328..9188)
/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 8951..8970
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(9189..9293)
/gene="bla"
/label=AmpR promoter
polyA_signal 9341..9475
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(9378..9397)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 9432..9451
/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 accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
3601 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
3661 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
3721 accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg
3781 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
3841 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
3901 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
3961 cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga caagcagaag
4021 aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc
4081 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
4141 cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg
4201 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag
4261 gtgcggccgc tcgagggagg cggtggatgg agccaccccc agttcgagaa gggcggcggc
4321 agcggcggcg gcagcggcgg cggcagctgg agccaccccc agttcgagaa ggggcccgtt
4381 taaggatccc gcccctctcc ctcccccccc cctaacgtta ctggccgaag ccgcttggaa
4441 taaggccggt gtgcgtttgt ctatatgtta ttttccacca tattgccgtc ttttggcaat
4501 gtgagggccc ggaaacctgg ccctgtcttc ttgacgagca ttcctagggg tctttcccct
4561 ctcgccaaag gaatgcaagg tctgttgaat gtcgtgaagg aagcagttcc tctggaagct
4621 tcttgaagac aaacaacgtc tgtagcgacc ctttgcaggc agcggaaccc cccacctggc
4681 gacaggtgcc tctgcggcca aaagccacgt gtataagata cacctgcaaa ggcggcacaa
4741 ccccagtgcc acgttgtgag ttggatagtt gtggaaagag tcaaatggct ctcctcaagc
4801 gtattcaaca aggggctgaa ggatgcccag aaggtacccc attgtatggg atctgatctg
4861 gggcctcggt gcacatgctt tacatgtgtt tagtcgaggt taaaaaaacg tctaggcccc
4921 ccgaaccacg gggacgtggt tttcctttga aaaacacgat gataagcttg ccacaaccca
4981 caaggagacg accttccatg accgagtaca agcccacggt gcgcctcgcc acccgcgacg
5041 acgtcccccg ggccgtacgc accctcgccg ccgcgttcgc cgactacccc gccacgcgcc
5101 acaccgtcga cccggaccgc cacatcgagc gggtcaccga gctgcaagaa ctcttcctca
5161 cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg
5221 tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca
5281 tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggaaggc ctcctggcgc
5341 cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc
5401 agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg
5461 gggtgcccgc cttcctggag acctccgcgc cccgcaacct ccccttctac gagcggctcg
5521 gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc
5581 gcaagcccgg tgcctagacg cgtctggaac aatcaacctc tggattacaa aatttgtgaa
5641 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
5701 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
5761 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
5821 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
5881 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
5941 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
6001 gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat tctgcgcggg
6061 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
6121 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
6181 ctttgggccg cctccccgcc tggaattaat tctgcagtcg agacctagaa aaacatggag
6241 caatcacaag tagcaataca gcagctacca atgctgattg tgcctggcta gaagcacaag
6301 aggaggagga ggtgggtttt ccagtcacac ctcaggtacc tttaagacca atgacttaca
6361 aggcagctgt agatcttagc cactttttaa aagaaaagag gggactggaa gggctaattc
6421 actcccaacg aagacaagat atccttgatc tgtggatcta ccacacacaa ggctacttcc
6481 ctgattagca gaactacaca ccagggccag gggtcagata tccactgacc tttggatggt
6541 gctacaagct agtaccagtt gagccagata aggtagaaga ggccaataaa ggagagaaca
6601 ccagcttgtt acaccctgtg agcctgcatg ggatggatga cccggagaga gaagtgttag
6661 agtggaggtt tgacagccgc ctagcatttc atcacgtggc ccgagagctg catccggagt
6721 acttcaagaa ctgctgatat cgagcttgct acaagggact ttccgctggg gactttccag
6781 ggaggcgtgg cctgggcggg actggggagt ggcgagccct cagatcctgc atataagcag
6841 ctgctttttg cctgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct
6901 ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta
6961 gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca
7021 gtgtggaaaa tctctagcag tagtagttca tgtcatctta ttattcagta tttataactt
7081 gcaaagaaat gaatatcaga gagtgagagg ccttgacatt gctagcgttt accgtcgacc
7141 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
7201 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
7261 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
7321 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
7381 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
7441 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
7501 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
7561 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
7621 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
7681 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
7741 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
7801 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
7861 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
7921 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
7981 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
8041 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
8101 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
8161 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
8221 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
8281 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
8341 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
8401 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
8461 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
8521 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
8581 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
8641 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
8701 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
8761 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
8821 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
8881 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
8941 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
9001 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
9061 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
9121 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
9181 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
9241 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
9301 ccccgaaaag tgccacctga cgtcgacgga tcgggagatc aacttgttta ttgcagctta
9361 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
9421 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatcaactg
9481 gataactcaa gctaaccaaa atcatcccaa acttcccacc ccatacccta ttaccactgc
9541 caattacctg tggtttcatt tactctaaac ctgtgattcc tctgaattat tttcatttta
9601 aagaaattgt atttgttaaa tatgtactac aaacttagta gt
//
