| 质粒名称: | WT PBRM1- TRIPZ-neo |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 107406 |
| 存储实验室: | William Kaelin |
|
载体骨架基本信息 |
|
| 载体名称: | pTRIPZ-neo |
| 载体出品公司: | -- |
| 空载体大小: | -- |
| 完整质粒大小: | 14942 bp |
| 载体修饰: | -- |
| 载体类型: | 慢病毒载体 |
| 筛选标记: | -- |
| 载体抗性: | 卡那霉素，50μg/mL |
| 生长温度: | 30 ℃ |
| 宿主菌: | NEB Stable |
| 拷贝数: | -- |
|
插入基因信息 |
|
| 插入基因名称: | PBRM1型 |
| 物种来源: | H. sapiens (human) |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子: | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | AgeI(未知是否破坏) |
| 3’克隆位点: | ClaI (未知是否破坏) |
| 5’测序引物 | -- |
| 3’测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | WT PBRM1- TRIPZ-neo was a gift from William Kaelin (Addgene plasmid # 107406 ; http://n2t.net/addgene:107406 ; RRID:Addgene_107406) |
| 参考文献部分: | Inactivation of the PBRM1 tumor suppressor gene amplifies the HIF-response in VHL-/- clear cell renal carcinoma. Gao W, Li W, Xiao T, Liu XS, Kaelin WG Jr. Proc Natl Acad Sci U S A. 2017 Jan 31;114(5):1027-1032. doi: 10.1073/pnas.1619726114. Epub 2017 Jan 12. 10.1073/pnas.1619726114 PubMed 28082722 |
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 WT PBRM1- TRIPZ-neo 14924 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Mammalian expression, inducible PBRM1.
ACCESSION .
VERSION .
KEYWORDS WT PBRM1- TRIPZ-neo
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 14924)
AUTHORS Gao W, Li W, Xiao T, Liu XS, Kaelin WG Jr
TITLE Inactivation of the PBRM1 tumor suppressor gene amplifies the
HIF-response in VHL-/- clear cell renal carcinoma.
JOURNAL Proc Natl Acad Sci U S A. 2017 Jan 31;114(5):1027-1032. doi:
10.1073/pnas.1619726114. Epub 2017 Jan 12.
PUBMED 28082722
REFERENCE 2 (bases 1 to 14924)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..14924
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS complement(1013..1078)
/codon_start=1
/product="three tandem FLAG(R) epitope tags, followed by an
enterokinase cleavage site"
/label=3xFLAG
/translation="DYKDHDGDYKDHDIDYKDDDDK"
CDS complement(6005..6031)
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
CDS complement(6032..6055)
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
promoter complement(6099..6413)
/label=tight TRE promoter
/note="Tet-responsive promoter PTight, consisting of seven
tet operator sequences followed by the minimal CMV
promoter"
primer_bind complement(6102..6126)
/label=LNCX
/note="Human CMV promoter, forward primer"
protein_bind 6173..6191
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 6209..6227
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 6244..6262
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 6280..6298
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 6316..6334
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 6351..6369
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 6387..6405
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
misc_feature 6487..6604
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
CDS complement(6902..6946)
/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 7131..7364
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
misc_feature 7861..7986
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
LTR 8033..8213
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
promoter complement(8231..8429)
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind complement(8256..8276)
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
enhancer 8431..8734
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
rep_origin complement(8920..9508)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(9000..9019)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
polyA_signal 9837..9884
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
primer_bind 9909..9928
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
CDS complement(10116..10910)
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
promoter complement(10945..11302)
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 10959..11094
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 11259..11279
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter complement(11304..11408)
/gene="bla"
/label=AmpR promoter
rep_origin 11435..11793
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(11522..11541)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 11732..11753
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 11816..11835
/label=T7
/note="T7 promoter, forward primer"
promoter 11816..11834
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
rep_origin 11855..11990
/label=SV40 ori
/note="SV40 origin of replication"
polyA_signal 12030..12151
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(12041..12060)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 12095..12114
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
LTR 12223..12456
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
CDS complement(12528..13322)
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
misc_feature 13329..13902
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
primer_bind complement(13489..13508)
/label=IRES-F
/note="IRES internal ribosome entry site, forward primer"
primer_bind 13718..13735
/label=IRES reverse
/note="IRES internal ribosome entry site, reverse primer.
Also called pCDH-rev"
protein_bind complement(13922..13955)
/label=loxP511
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GTATACAT). loxP511 sites are compatible with each
other, but incompatible with wild-type loxP sites."
CDS complement(13956..14663)
/codon_start=1
/product="reverse tetracycline transactivator 3"
/label=rtTA3
/note="activates expression from Tet-responsive promoters
in the presence of doxycycline"
/translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
DALDDFDLDMLPG"
protein_bind complement(14670..14703)
/label=loxP511
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GTATACAT). loxP511 sites are compatible with each
other, but incompatible with wild-type loxP sites."
promoter complement(14716..1003)
/label=UbC promoter
/note="human ubiquitin C promoter"
primer_bind complement(14860..14879)
/label=hUBCpro-F
/note="Human Ubiquitin C (UbC) promoter, forward primer"
ORIGIN
1 gcctcactta tccctcccct caccagaggt ccggcgcctg tcgattcagg agagcctacc
61 ctaggcccga accctgcgtc ctgcgacgga gaaaagccta ccgcacacct accggcaggt
121 ggccccaccc tgcattataa gccaacagaa cgggtgacgt cacgacacga cgagggcgcg
181 cgctcccaaa ggtacgggtg cactgcccaa cggcaccgcc ataactgccg cccccgcaac
241 agacgacaaa ccgagttctc cagtcagtga caaacttcac gtcagggtcc ccagatggtg
301 ccccagccca tctcacccga ataagagctt tcccgcatta gcgaaggcct caagaccttg
361 ggttcttgcc gcccaccatg ccccccacct tgtttcaacg acctcacagc ccgcctcaca
421 agcgtcttcc attcaagact cgggaacagc cgccattttg ctgcgctccc cccaaccccc
481 agttcagggc aaccttgctc gcggacccag actacagccc ttggcggtct ctccacacgc
541 ttccgtccca ccgagcggcc cggcggccac gaaagccccg gccagcccag cagcccgcta
601 ctcaccaagt gacgatcaca gcgatccaca aacaagaacc gcgacccaaa tcccggctgc
661 gacggaacta gctgtgccac acccggcgcg tccttatata atcatcggcg ttcaccgccc
721 cacggagatc cctccgcaga atcgccgaga agggactact tttcctcgcc tgttccgctc
781 tctggaaaga aaaccagtgc cctagagtca cccaagtccc gtcctaaaat gtccttctgc
841 tgatactggg gttctaaggc cgagtcttat gagcagcggg ccgctgtcct gagcgtccgg
901 gcggaaggat caggacgctc gctgcgccct tcgtctgacg tggcagcgct cgccgtgagg
961 aggggggcgc ccgcgggagg cgccaaaacc cggcgcggag gccacgcgtc tacttgtcat
1021 cgtcatcctt gtaatcgatg tcatgatctt tataatcacc gtcatggtct ttgtagtcac
1081 ctccctcgag ggccggcctt aattaaacat tttctaggtt gtatgcttgg cgaatgttga
1141 gggtgtcccg gagcatcaaa tctcgaaggc gccagagggc atctgccatg gtggtgtggg
1201 cccctttgct tttcagccag tgagagggta ggcggctctc ctgttctttc gacaaatgga
1261 cgtcgcgtct tcgagctgcc agtgtctgat cccacttgct aatgctgttg gactccgcac
1321 tgagtccttc aatgtatttc aggtaggcct ctgagtgaag aagccgctgg gtctttggtg
1381 ggggagctac aaacatgggt gttgttggct gctgtatgac agggggtcca gctggatgtg
1441 ggccgggata tggaggtggt gcctgctgcc ctggaggccc caaaactccc acctgttgtc
1501 catatggact tccacccggt gctggagtcc ctaccatagg ggccactcct tggttcatca
1561 cacccggtgg tgggatgccc gggaggccag gcacacctgg cggaagatgg tggggtggag
1621 gccccccagg gtgaagtggc tgcatgctgc ccatgctaac aaggccatca actgggccct
1681 gcaaaggtgg aaggcctggc ggatagccac ccatcatgcc ttcatattct gctttcttgg
1741 ctgtctcaag atttctccat tctgtcccca ccaggcggct gagctccccg aaagagtagt
1801 ctgggtgttg ggccttaatc acagccctca tctcactgct gaacaggatg tagccactca
1861 tgttgatttt ccgtttggag ccttccttct ttgcactgcc tttggcagac tttggggtag
1921 actgtggggg tgtgtagggc atgaggtcca gctcactggc caggggggtc tgaagctgag
1981 gtagagaagg aggttcaatg acctcactat cttcttctcc catctcttca atatcatcat
2041 ctccaccttc taactcggca aatttagctt ctagcaactg gatcttcttt tccagcaaag
2101 gtgatggctc cttctgagga acaattggtt ttctgaagta gtaaatttca tcatctacca
2161 ctttagcaga gagtgaaaac ctcttcaatc ctttgaattt cttcatctgc ttgtcgctct
2221 cattgtagcg gctctcacaa agcagaatgt cattttctgg tatttcagtt ggcctgcagg
2281 agaggaagtc cttgaatgac aacacagcac actttccgag aatacatgtc atggggcagg
2341 tttcttccag attactcaga aatacttctt ttttgtagaa catttttgtg ggctcatgct
2401 ctgtttcttc agggtgaatg aagatggggc cataaaaata tgcagctcca tctcgaaccc
2461 atactttttc aattctgccc acacgaggac gcaccaggcc atgggacttg atgaagacac
2521 agtcgccaac cttcagccac atgtcattgt aatggagctg ctcaaagtag tggcaacctg
2581 gttcaccatt ggacatttcc acagggacat cttctttttc cttttccaag ttaaaattgt
2641 cttcagctcg actgtcctct gagttgtctg tattcttctc atcatcacct ttatctgcat
2701 ttgcaaatac agaggccacg cgaaccacag gcagaggcac atcccgaggg acaaacctga
2761 ctgagctgat gggcatggtc cacagtttaa ttttcttaaa agatttggtt ttggcagaat
2821 accgtgattc acagacaaaa acatcctcat ctcggaagtt ttctgggcat aacttaaagt
2881 attccttgac aaacatgacc acacacttgc ctagaatttt actaactgga actttgttgt
2941 aatagtcact cttaaaaact tctttttcta gaaattttcg tgtagccagg tggaatgttt
3001 catttggtcg gtaaaaccaa cagccataca accatttttc accagctgaa tcctcccaca
3061 gtctttcaat acagacgata tgtggttgta ggttggcctc tgcaggttcc acatagacgt
3121 aatctccaac atggtacatg ctgtttttaa agctacagtc ctggctgtat gtgcgatgta
3181 agcctgagag gcctgcagca ccagaggaat cttcactctt ttcagcttct cttttttctt
3241 cttctcgttt tagtttatct tcctctattt cttttggcaa tttttccttt ctctctttct
3301 ccacatcatt atgcaaatgt tttgtggtat agctgagtgc cggtgaaaga agaatctctc
3361 catttttgca gagttcatca cgaattttaa taaaaaactg ctgaagttct actgcatctt
3421 catatatttc tgaatctgtc cgattcatcc ttcttgctcg ttccaatact tcaaacatat
3481 gctcttgaaa taaatcaagc cgacggtagc gattattttc aacattcttc ctaattatgt
3541 caaatgtaag gggtggtttg ttaggaaagt tgggatccac agcaggaatt tctgctaaag
3601 aatcgctgta gcatcttccc tcatcatcct gatgactcat gactgacaca aaaagattgt
3661 ggataagctc ttgaatcagc aaagtcacat ttgggacatg agagtcctca tctccctcca
3721 ggtctctgcg tgtttcaagc aggactttgt gtagaacaag agcatctttg tagatcaaag
3781 actccggctc attgtacgta caggcattat taaacatcat gacaaagtcc tcaaccatag
3841 agtcaatatc ttggtacttg ttggccatca tgtgacttcg aattttttcc atgtccatgg
3901 gctttttaat agtcagatag tagtcaggca actcagatct agagggaagc ctcagaaata
3961 tggcactgag gcggcgaccc ctcttatcag tatagttctt tacagcttca tagacctcat
4021 ttagtttctg ctgcattgga gtcatgtatt ttgatttttt aggagaaatg ccactcttcc
4081 tactcagctt gagtttggga gaagccatgt catcatcatc aggcagtggg cccagctctt
4141 tccttttctc cttgagtaac ttctccagga tatgtgcatc attataaacc tgggagccct
4201 cctcattata gtgcctggca ttccggaaca tcagcttcat gtcttctatc attccctctt
4261 caccagcata tttgtcattg cggatgttat gctcaattat tttcaagtcc attggctcca
4321 agatgatttt ataataatca ggatagtcct ttttggatgg tttaaccata aataggtcac
4381 aaagtcttct gcctgaacct ggctctcgag cttcaagaac aacattgaat aagattttca
4441 ttcgctgctt tcttatgttc tttttacttt ttcttttggc actaccagta tcagaggtgg
4501 ctgaagagat catgctgtct ccgtcctcga tatcgtctct cctggcaagc tctttcttct
4561 ttgcctgcat aacttgctgc aattttagaa ctcgcttgta gatggctgaa ttgggcacat
4621 tatagcgttt ggcattttca aacattaaat tcagatcaca ctccaaatga tctaaagttt
4681 catattcttg attcttcagt tttgttcgga tctgttgtag tgatatgggc attttaattt
4741 gctggtaata atcagggtat tttttctttg aaggcaaatg gtaaaaaggt tcagctatta
4801 gctgcccttg gttattccga caactcctaa ctgtgtcata aagctgataa aaaggatttg
4861 aaacatccat aaaggaagtg atgctttctg cttctgactc tccctcttca tagcgtgcag
4921 cagctaaagc agcatcttct tcactttctg agccatattg aagtgccatt gtaattgctg
4981 ataaacgacc tccttgtact gctcttttat tacggtaata cttgctagtg ggatttctct
5041 cttcaccaag gctgccttta ctgtgtgaag gacctgtcag tctggctgca gccaagttgg
5101 atggagtcct cattcgaaga cttgacttag ccatttcatg atgttcaatt tcagcctttt
5161 tcatataaaa tattttttta attgaatttg catccttgaa tacttgagag ccaggctcat
5221 tataagtttt ggcatttttt gcgaggagat ctatatcttt ggccattgca tgaatacttt
5281 tgtagcttcc attctgtatc ctctgggcaa tggtcttgag atctataggc tccttaatta
5341 ttgcataata atctggatat tgcactttag aaggcagttt ctgaaaaagt tcgctaatga
5401 gacgtcctga tggatttgta gctacaacta tggcttcaag aagctgctcc aggatctcct
5461 tcaagtaagc tggagaagat ccttcagtca ctgtgccctg attgtcttgc ccatcttcat
5521 catcatcttc gtcatctgct tctcctttct gaacaaactc atttcttgtt cgaaggtaca
5581 aatcccagag tttgcaagcg gctttatatt caggagaatc tggcttataa taggactttg
5641 cattgttaaa aagaagctgg aagtcagcag tcagcaaatt aacatcatca tactcttcca
5701 tttttagttt ctgttggatt ttcatcaagt caatgggctg agaaaccact tcataatagt
5761 ctggttgatt tcttcgcttt ggtgccctaa tgaagagctc acagagaagt ctgccctgtt
5821 catccttata gtctcggatg gtattataga gttcatggca cacggcaata ggatctacag
5881 ttggaagatt ggaaagtctc ctccttttcc tgcttgggcc tggtgttgac acagaatggt
5941 gcccatcatc aaagtccccg ctgacactgc tggaagggga ggtagctctt cttctcttgg
6001 aaccagcgta atctggaaca tcgtatgggt acttgtcgtc atcgtctttg tagtccatga
6061 attcgtcgac aattcgatcc ggcagtctag attctccagg cgatctgacg gttcactaaa
6121 cgagctctgc ttatataggc ctcccaccgt acacgcctac ctcgacatac gttctctatc
6181 actgataggg agtaaactcg acatacgttc tctatcactg atagggataa actcgacata
6241 cgttctctat cactgatagg gagtaaactc gacatacgtt ctctatcact gatagggagt
6301 aaactcgaca tacgttctct atcactgata gggagtaaac tcgacatcgt tctctatcac
6361 tgatagggag taaactcgac atacgttctc tatcactgat agggagtaaa ctcgaatgaa
6421 gacgaaaggg cctcaggggc ccggtactaa ccaaactgga tctctgctgt ccctgtaata
6481 aacccgaaaa ttttgaattt ttgtaatttg tttttgtaat tctttagttt gtatgtctgt
6541 tgctattatg tctactattc tttcccctgc actgtacccc ccaatccccc cttttctttt
6601 aaaattgtgg atgaatactg ccatttgtct gcagaattgg cgcacgcagt gccgatccgt
6661 tcactaatcg aatggatctg tctctgtctc tctctccacc ttcttcttct attccttcgg
6721 gcctgtcggg tcccctcggg gttgggaggt gggtctgaaa cgataatggt gaatatccct
6781 gcctaactct attcactata gaaagtacag caaaaactat tcttaaacct accaagcctc
6841 ctactatcat tatgaataat tttatatacc acagccaatt tgttatgtta aaccaattcc
6901 acaaacttgc ccatttatct aattccaata attcttgttc attcttttct tgctggtttt
6961 gcgattcttc aattaaggag tgtattaagc ttgtgtaatt gttaatttct ctgtcccact
7021 ccatccaggt cgtgtgattc caaatctgtt ccagagattt attactccaa ctagcattcc
7081 aaggcacagc agtggtgcaa atgagttttc cagagcaacc ccaaatcccc aggagctgtt
7141 gatcctttag gtatctttcc acagccagga ttcttgcctg gagctgcttg atgccccaga
7201 ctgtgagttg caacagatgc tgttgcgcct caatagccct cagcaaattg ttctgctgct
7261 gcactatacc agacaataat tgtctggcct gtaccgtcag cgtcattgac gctgcgccca
7321 tagtgcttcc tgctgctccc aagaacccaa ggaacaaagc tcctattccc actgctcttt
7381 tttctctctg caccactctt ctctttgcct tggtgggtgc tactcctaat ggttcaattt
7441 ttactacttt atatttatat aattcacttc tccaattgtc cctcatatct cctcctccag
7501 gtctgaagat cagcggccgg ccgcttgctg tgcggtggtc ttacttttgt tttgctcttc
7561 ctctatcttg tctaaagctt ccttggtgtc ttttatctct atcctttgat gcacacaata
7621 gagggttgct actgtattat ataatgatct aagttcttct gatcctgtct gaagggatgg
7681 ttgtagctgt cccagtattt gtctacagcc ttctgatgtt tctaacaggc caggattaac
7741 tgcgaatcgt tctagctccc tgcttgccca tactatatgt tttaatttat attttttctt
7801 tccccctggc cttaaccgaa ttttttccca tcgcgatcta attctccccc gcttaatact
7861 gacgctctcg cacccatctc tctccttcta gcctccgcta gtcaaaattt ttggcgtact
7921 caccagtcgc cgcccctcgc ctcttgccgt gcgcgcttca gcaagccgag tcctgcgtcg
7981 agagagctcc tctggtttcc ctttcgcttt caagtccctg ttcgggcgcc actgctagag
8041 attttccaca ctgactaaaa gggtctgagg gatctctagt taccagagtc acacaacaga
8101 cgggcacaca ctacttgaag cactcaaggc aagctttatt gaggcttaag cagtgggttc
8161 cctagttagc cagagagctc ccaggctcag atctggtcta accagagaga cccagtacag
8221 gcaaaacgcg ctgcttatat agacctccca ccgtacacgc ctaccgccca tttgcgtcaa
8281 tggggcggag ttgttacgac attttggaaa gtcccgttga ttttggtgcc aaaacaaact
8341 cccattgacg tcaatggggt ggagacttgg aaatccccgt gagtcaaacc gctatccacg
8401 cccattgatg tactgccaaa accgcatcac catggtaata gcgatgacta atacgtagat
8461 gtactgccaa gtaggaaagt cccataaggt catgtactgg gcataatgcc aggcgggcca
8521 tttaccgtca ttgacgtcaa tagggggcgt acttggcata tgatacactt gatgtactgc
8581 caagtgggca gtttaccgta aatactccac ccattgacgt caatggaaag tccctattgg
8641 cgttactatg ggaacatacg tcattattga cgtcaatggg cgggggtcgt tgggcggtca
8701 gccaggcggg ccatttaccg taagttatgt aacgcggaac tccatatatg ggctatgaac
8761 taatgacccc gtaattgatt actattaata actaatgcat ggcggtaata cggttatcca
8821 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagca
8881 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
8941 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
9001 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
9061 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
9121 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
9181 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
9241 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
9301 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
9361 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
9421 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
9481 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
9541 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
9601 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
9661 gtaacctgag gctatggcag ggcctgccgc cccgacgttg gctgcgagcc ctgggccttc
9721 acccgaactt ggggggtggg gtggggaaaa ggaagaaacg cgggcgtatt ggccccaatg
9781 gggtctcggt ggggtatcga cagagtgcca gccctgggac cgaaccccgc gtttatgaac
9841 aaacgaccca acaccgtgcg ttttattctg tctttttatt gccgtcatag cgcgggttcc
9901 ttccggtatt gtctccttcc gtgtttcagt tagcctcccc ctagggtggg cgaagaactc
9961 cagcatgaga tccccgcgct ggaggatcat ccagccggcg tcccggaaaa cgattccgaa
10021 gcccaacctt tcatagaagg cggcggtgga atcgaaatct cgtgatggca ggttgggcgt
10081 cgcttggtcg gtcatttcga accccagagt cccgctcaga agaactcgtc aagaaggcga
10141 tagaaggcga tgcgctgcga atcgggagcg gcgataccgt aaagcacgag gaagcggtca
10201 gcccattcgc cgccaagctc ttcagcaata tcacgggtag ccaacgctat gtcctgatag
10261 cggtccgcca cacccagccg gccacagtcg atgaatccag aaaagcggcc attttccacc
10321 atgatattcg gcaagcaggc atcgccatgg gtcacgacga gatcctcgcc gtcgggcatg
10381 ctcgccttga gcctggcgaa cagttcggct ggcgcgagcc cctgatgctc ttcgtccaga
10441 tcatcctgat cgacaagacc ggcttccatc cgagtacgtg ctcgctcgat gcgatgtttc
10501 gcttggtggt cgaatgggca ggtagccgga tcaagcgtat gcagccgccg cattgcatca
10561 gccatgatgg atactttctc ggcaggagca aggtgagatg acaggagatc ctgccccggc
10621 acttcgccca atagcagcca gtcccttccc gcttcagtga caacgtcgag cacagctgcg
10681 caaggaacgc ccgtcgtggc cagccacgat agccgcgctg cctcgtcttg cagttcattc
10741 agggcaccgg acaggtcggt cttgacaaaa agaaccgggc gcccctgcgc tgacagccgg
10801 aacacggcgg catcagagca gccgattgtc tgttgtgccc agtcatagcc gaatagcctc
10861 tccacccaag cggccggaga acctgcgtgc aatccatctt gttcaatcat gcgaaacgat
10921 cctcatcctg tctcttgatc gatctttgca aaagcctagg cctccaaaaa agcctcctca
10981 ctacttctgg aatagctcag aggccgaggc ggcctcggcc tctgcataaa taaaaaaaat
11041 tagtcagcca tggggcggag aatgggcgga actgggcgga gttaggggcg ggatgggcgg
11101 agttaggggc gggactatgg ttgctgacta attgagatgc atgctttgca tacttctgcc
11161 tgctggggag cctggggact ttccacacct ggttgctgac taattgagat gcatgctttg
11221 catacttctg cctgctgggg agcctgggga ctttccacac cctaactgac acacattcca
11281 cagctggttc tttccgcctc aggactcttc ctttttcaat attattgaag catttatcag
11341 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
11401 gttccgcgca catttccccg aaaagtgcca cctgacgcgc cctgtagcgg cgcattaagc
11461 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
11521 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
11581 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
11641 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
11701 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
11761 ctcaacccta tctcggtcta ttcttttgat ttatgctggc cttttgctca catgctaata
11821 cgactcacta tagggcgaat tgggtacgtc cctaggcctc caaaaaagcc tcctcactac
11881 ttctggaata gctcagaggc cgaggcggcc tcggcctctg cataaataaa aaaaattagt
11941 cagccatggg gcggagaatg ggcggaactg ggcggagtta ggggcgggat gggcggagtt
12001 aggggcggga tagctagagc cagacatgat aagatacatt gatgagtttg gacaaaccac
12061 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
12121 tgtaaccatt ataagctgca ataaacaagt tcctctcact ctctgatatt catttctttg
12181 caagttataa atactgaata ataagatgac atgaactact actgctagag attttccaca
12241 ctgactaaaa gggtctgagg gatctctagt taccagagtc acacaacaga cgggcacaca
12301 ctacttgaag cactcaaggc aagctttatt gaggcttaag cagtgggttc cctagttagc
12361 cagagagctc ccaggctcag atctggtcta accagagaga cccagtacaa gcaaaaagca
12421 gatcttgtct tcgttgggag tgaattagcc cttccagtcc ccccttttct tttaaaaagt
12481 ggctaagatc tacagctgcc ttgtaagtca ttggtcttaa aggtacctca gaagaactcg
12541 tcaagaaggc gatagaaggc gatgcgctgc gaatcgggag cggcgatacc gtaaagcacg
12601 aggaagcggt cagcccattc gccgccaagc tcttcagcaa tatcacgggt agccaacgct
12661 atgtcctgat agcggtccgc cacacccagc cggccacagt cgatgaatcc agaaaagcgg
12721 ccattttcca ccatgatatt cggcaagcag gcatcgccat gggtcacgac gagatcctcg
12781 ccgtcgggca tgcgcgcctt gagcctggcg aacagttcgg ctggcgcgag cccctgatgc
12841 tcttcgtcca gatcatcctg atcgacaaga ccggcttcca tccgagtacg tgctcgctcg
12901 atgcgatgtt tcgcttggtg gtcgaatggg caggtagccg gatcaagcgt atgcagccgc
12961 cgcattgcat cagccatgat ggatactttc tcggcaggag caaggtgaga tgacaggaga
13021 tcctgccccg gcacttcgcc caatagcagc cagtcccttc ccgcttcagt gacaacgtcg
13081 agcacagctg cgcaaggaac gcccgtcgtg gccagccacg atagccgcgc tgcctcgtcc
13141 tgcagttcat tcagggcacc ggacaggtcg gtcttgacaa aaagaaccgg gcgcccctgc
13201 gctgacagcc ggaacacggc ggcatcagag cagccgattg tctgttgtgc ccagtcatag
13261 ccgaatagcc tctccaccca agcggccgga gaacctgcgt gcaatccatc ttgttcaatc
13321 atggatcctt atcatcgtgt ttttcaaagg aaaaccacgt ccccgtggtt cggggggcct
13381 agacgttttt ttaacctcga ctaaacacat gtaaagcatg tgcaccgagg ccccagatca
13441 gatcccatac aatggggtac cttctgggca tccttcagcc ccttgttgaa tacgcttgag
13501 gagagccatt tgactctttc cacaactatc caactcacaa cgtggcactg gggttgtgcc
13561 gcctttgcag gtgtatctta tacacgtggc ttttggccgc agaggcacct gtcgccaggt
13621 ggggggttcc gctgcctgca aagggtcgct acagacgttg tttgtcttca agaagcttcc
13681 agaggaactg cttccttcac gacattcaac agaccttgca ttcctttggc gagaggggaa
13741 agacccctag gaatgctcgt caagaagaca gggccaggtt tccgggccct cacattgcca
13801 aaagacggca atatggtgga aaataacata tagacaaacg cacaccggcc ttattccaag
13861 cggcttcggc cagtaacgtt aggggggggg gagggagagg ggcggatgcg gccgcggatc
13921 cataacttcg tataatgtat actatacgaa gttatttacc cggggagcat gtcaaggtca
13981 aaatcgtcaa gggcatcggc tgggagcatg tctaagtcaa aatcgtcaag ggcgtcggtc
14041 ggcccgccgc tttcgcactt tagctgtttc tccaggccac atatgattag ttccaggccg
14101 aaaaggaagg caggttcggc tccctgccgg tcgaacagct caattgcttg tctcagaagt
14161 gggggcatag aatcggtggt aggtgtctct ctttcctctt ttgctacttg atgctcctgt
14221 tcctccaata cgcagcccag tgtaaagtgg cccacggcgg acagagcgta cagtgcgttc
14281 tccagggaga agccttgctg acacaggaac gcgagctgat tttccagggt ttcgtactgt
14341 ttctctgttg ggcgggtgcc gagatgcact ttagccccgt cgcgatgtga gaggagagca
14401 cagcggtatg acttggcgtt gttccgcaga aagtcttgcc atgactcgcc ttccaggggg
14461 cagaagtggg tatgatgcct gtccagcatc tcgattggca gggcatcgag cagggcccgc
14521 ttgttcttca cgtgccagta cagggtaggc tgctcaactc ccagcttttg agcgagtttc
14581 cttgtcgtca ggccttcgat accgacacca ttgagtaatt ccagagctcc gtttatgact
14641 ttgctcttgt ccagcctaga catggtggca taacttcgta taatgtatac tatacgaagt
14701 tatggatccg ctagcgtcta acaaaaaagc caaaaacggc cagaatttag cggacaattt
14761 actagtctaa cactgaaaat tacatattga cccaaatgat tacatttcaa aaggtgccta
14821 aaaaacttca caaaacacac tcgccaaccc cgagcgcata gttcaaaacc ggagcttcag
14881 ctacttaaga agataggtac ataaaaccga ccaaagaaac tgac
//
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