| 作者: | SV.Wesley （2001） |
|---|---|
| 载体名称: | pHELLSGATE8 |
| 质粒类型: | 基因沉默载体；Gateway载体；植物双元表达载体 |
| 高拷贝/低拷贝: | 高拷贝 |
| 克隆方法: | Gateway |
| 启动子: | CaMV 35S |
| 载体大小: | 17485 bp |
| 5' 测序引物及序列: | 35S promoter：5’CTATCCTTCGCAAGACCCTTC 3’ |
| 3' 测序引物及序列: | -- |
| 载体标签: | 无 |
| 载体抗性: | 壮观霉素和链霉素 |
| 筛选标记: | 卡那霉素 |
| 克隆菌株: | DB3.1 |
| 宿主细胞（系）: | 植物细胞、农杆菌 |
| 备注: | pHELLSGATE8载体是高效、高通量、有效的植物基因沉默载体。 |
| 产品目录号: | -- |
| 稳定性: | 稳表达 |
| 组成型/诱导型: | 组成型 |
| 病毒/非病毒: | 非病毒 |
买家导航
pHELLSGATE8载体质粒基本信息
pHELLSGATE8质粒图谱载体图谱和pHELLSGATE8载体序列质粒序列多克隆位点信息
pHELLSGATE8质粒载体简介
pHELLSGATE8载体简介
Post-transcriptional silencing of plant genes using anti-sense or co-suppression constructs usually results in only a modest proportion of silenced individuals. Recent work has demonstrated the potential for constructs encoding self-complementary 'hairpin' RNA (hpRNA) to efficiently silence genes. In this study we examine design rules for efficient gene silencing, in terms of both the proportion of independent transgenic plants showing silencing, and the degree of silencing. Using hpRNA constructs containing sense/anti-sense arms ranging from 98 to 853 nt gave efficient silencing in a wide range of plant species, and inclusion of an intron in these constructs had a consistently enhancing effect. Intron-containing constructs (ihpRNA) generally gave 90-100% of independent transgenic plants showing silencing. The degree of silencing with these constructs was much greater than that obtained using either co-suppression or anti-sense constructs. We have made a generic vector, pHANNIBAL, that allows a simple, single PCR product from a gene of interest to be easily converted into a highly effective ihpRNA silencing construct. We have also created a high-throughput vector, pHELLSGATE, that should facilitate the cloning of gene libraries or large numbers of defined genes, such as those in EST collections, using an in vitro recombinase system. This system may facilitate the large-scale determination and discovery of plant gene functions in the same way as RNAi is being used to examine gene function in Caenorhabditis elegans.
pHELLSGATE8质粒序列载体序列
LOCUS AF489904 17485 bp DNA circular SYN 14-NOV-2005
DEFINITION Cloning vector pHellsgate 8, complete sequence.
ACCESSION AF489904
VERSION AF489904.1 GI:33320493
KEYWORDS .
SOURCE Cloning vector pHellsgate 8
ORGANISM Cloning vector pHellsgate 8
other sequences; artificial sequences; vectors.
REFERENCE 1 (bases 1 to 17485)
AUTHORS Helliwell,C.A. III and Waterhouse,P.M.
TITLE High throughput vectors for gene silencing in plants
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 17485)
AUTHORS Helliwell,C.A. III and Waterhouse,P.M.
TITLE Direct Submission
JOURNAL Submitted (13-FEB-2002) Plant Industry, CSIRO, GPO Box 1600,
Canberra, ACT 2601, Australia
FEATURES Location/Qualifiers
source 1..17485
/organism="Cloning vector pHellsgate 8"
/mol_type="genomic DNA"
/db_xref="taxon:198117"
/focus
/note="derivative of cloning vector pART27"
source 1..264
/organism="Escherichia coli K-12"
/mol_type="genomic DNA"
/strain="K-12"
/db_xref="taxon:83333"
source 265..448
/organism="Agrobacterium tumefaciens"
/mol_type="genomic DNA"
/db_xref="taxon:358"
source 449..1442
/organism="Escherichia coli"
/mol_type="genomic DNA"
/db_xref="taxon:562"
source 1443..7792
/organism="Agrobacterium tumefaciens"
/mol_type="genomic DNA"
/db_xref="taxon:358"
source 7793..9388
/organism="Escherichia coli"
/mol_type="genomic DNA"
/db_xref="taxon:562"
source 9389..11673
/organism="Agrobacterium tumefaciens"
/mol_type="genomic DNA"
/db_xref="taxon:358"
source 11674..13019
/organism="Cauliflower mosaic virus"
/mol_type="genomic DNA"
/db_xref="taxon:10641"
source 14474..16072
/organism="Flaveria trinervia"
/mol_type="genomic DNA"
/db_xref="taxon:4227"
source 16716..17485
/organism="Agrobacterium tumefaciens"
/mol_type="genomic DNA"
/db_xref="taxon:358"
regulatory 264..447
/regulatory_class="promoter"
/note="NOS promoter"
gene 448..1269
/gene="nptII"
CDS 448..1269
/gene="nptII"
/note="neomycin phosphotransferase II"
/codon_start=1
/product="kanamycin resistance protein"
/protein_id="AAQ05967.1"
/db_xref="GI:33320494"
/translation="MAITLSATSLPISARIRAGSPAAWVERLFGYDWAQQTIGCSDAA
VFRLSAQGRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLL
LGEVPGQDLLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAG
LVDQDDLDEEHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDC
GRLGVADRYQDIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
regulatory 1443..2148
/regulatory_class="terminator"
/note="NOS terminator"
repeat_region 2149..2706
/note="left border"
mobile_element 7793..9388
/mobile_element_type="transposon:Tn7"
gene 8600..9388
/gene="spec"
CDS 8600..9388
/gene="spec"
/codon_start=1
/product="spectinomycin resistance protein"
/protein_id="AAQ05968.1"
/db_xref="GI:33320495"
/translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKP
HSDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYP
AKRELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQ
DLFEALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPA
QYQPVILEARQAYLGQEDRLASRADQLEEFVHYVKGEITKVVGK"
repeat_region 10706..11324
/note="right border"
regulatory 11674..13019
/regulatory_class="promoter"
/note="35S promoter"
gene 14474..16019
/gene="pdk"
intron 14474..16019
/gene="pdk"
/note="pyruvate orthophosphate dikinase"
/number=2
regulatory 16716..17485
/regulatory_class="terminator"
/note="octopine esynthase (ocs) terminator"
ORIGIN
1 ggccgcacta gtgatatccc gcggccatgg cggccgggag catgcgacgt cgggcccaat
61 tcgccctata gtgagtcgta ttacaattca ctggccgtcg ttttacaacg tcgtgactgg
121 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
181 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
241 gaatggaaat tgtaaacgtt aatgggtttc tggagtttaa tgagctaagc acatacgtca
301 gaaaccatta ttgcgcgttc aaaagtcgcc taaggtcact atcagctagc aaatatttct
361 tgtcaaaaat gctccactga cgttccataa attcccctcg gtatccaatt agagtctcat
421 attcactctc aatccaaata atctgcaatg gcaattacct tatccgcaac ttctttacct
481 atttccgccc ggatccgggc aggttctccg gccgcttggg tggagaggct attcggctat
541 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
601 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
661 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
721 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
781 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
841 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
901 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
961 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
1021 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
1081 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
1141 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
1201 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
1261 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
1321 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
1381 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
1441 ccgatccaac acttacgttt gcaacgtcca agagcaaata gaccacgaac gccggaaggt
1501 tgccgcagcg tgtggattgc gtctcaattc tctcttgcag gaatgcaatg atgaatatga
1561 tactgactat gaaactttga gggaatactg cctagcaccg tcacctcata acgtgcatca
1621 tgcatgccct gacaacatgg aacatcgcta tttttctgaa gaattatgct cgttggagga
1681 tgtcgcggca attgcagcta ttgccaacat cgaactaccc ctcacgcatg cattcatcaa
1741 tattattcat gcggggaaag gcaagattaa tccaactggc aaatcatcca gcgtgattgg
1801 taacttcagt tccagcgact tgattcgttt tggtgctacc cacgttttca ataaggacga
1861 gatggtggag taaagaagga gtgcgtcgaa gcagatcgtt caaacatttg gcaataaagt
1921 ttcttaagat tgaatcctgt tgccggtctt gcgatgatta tcatataatt tctgttgaat
1981 tacgttaagc atgtaataat taacatgtaa tgcatgacgt tatttatgag atgggttttt
2041 atgattagag tcccgcaatt atacatttaa tacgcgatag aaaacaaaat atagcgcgca
2101 aactaggata aattatcgcg cgcggtgtca tctatgttac tagatcgaat taattccagg
2161 cggtgaaggg caatcagctg ttgcccgtct cactggtgaa aagaaaaacc accccagtac
2221 attaaaaacg tccgcaatgt gttattaagt tgtctaagcg tcaatttgtt tacaccacaa
2281 tatatcctgc caccagccag ccaacagctc cccgaccggc agctcggcac aaaatcacca
2341 ctcgatacag gcagcccatc agtccgggac ggcgtcagcg ggagagccgt tgtaaggcgg
2401 cagactttgc tcatgttacc gatgctattc ggaagaacgg caactaagct gccgggtttg
2461 aaacacggat gatctcgcgg agggtagcat gttgattgta acgatgacag agcgttgctg
2521 cctgtgatca aatatcatct ccctcgcaga gatccgaatt atcagccttc ttattcattt
2581 ctcgcttaac cgtgacaggc tgtcgatctt gagaactatg ccgacataat aggaaatcgc
2641 tggataaagc cgctgaggaa gctgagtggc gctatttctt tagaagtgaa cgttgacgat
2701 gtcgacggat cttttccgct gcataaccct gcttcggggt cattatagcg attttttcgg
2761 tatatccatc ctttttcgca cgatatacag gattttgcca aagggttcgt gtagactttc
2821 cttggtgtat ccaacggcgt cagccgggca ggataggtga agtaggccca cccgcgagcg
2881 ggtgttcctt cttcactgtc ccttattcgc acctggcggt gctcaacggg aatcctgctc
2941 tgcgaggctg gccggctacc gccggcgtaa cagatgaggg caagcggatg gctgatgaaa
3001 ccaagccaac caggggtgat gctgccaact tactgattta gtgtatgatg gtgtttttga
3061 ggtgctccag tggcttctgt ttctatcagc tgtccctcct gttcagctac tgacggggtg
3121 gtgcgtaacg gcaaaagcac cgccggacat cagcgctatc tctgctctca ctgccgtaaa
3181 acatggcaac tgcagttcac ttacaccgct tctcaacccg gtacgcacca gaaaatcatt
3241 gatatggcca tgaatggcgt tggatgccgg gcaacagccc gcattatggg cgttggcctc
3301 aacacgattt tacgtcactt aaaaaactca ggccgcagtc ggtaacctcg cgcatacagc
3361 cgggcagtga cgtcatcgtc tgcgcggaaa tggacgaaca gtggggctat gtcggggcta
3421 aatcgcgcca gcgctggctg ttttacgcgt atgacagtct ccggaagacg gttgttgcgc
3481 acgtattcgg tgaacgcact atggcgacgc tggggcgtct tatgagcctg ctgtcaccct
3541 ttgacgtggt gatatggatg acggatggct ggccgctgta tgaatcccgc ctgaagggaa
3601 agctgcacgt aatcagcaag cgatatacgc agcgaattga gcggcataac ctgaatctga
3661 ggcagcacct ggcacggctg ggacggaagt cgctgtcgtt ctcaaaatcg gtggagctgc
3721 atgacaaagt catcgggcat tatctgaaca taaaacacta tcaataagtt ggagtcatta
3781 cccaaccagg aagggcagcc cacctatcaa ggtgtactgc cttccagacg aacgaagagc
3841 gattgaggaa aaggcggcgg cggccggcat gagcctgtcg gcctacctgc tggccgtcgg
3901 ccagggctac aaaatcacgg gcgtcgtgga ctatgagcac gtccgcgagc tggcccgcat
3961 caatggcgac ctgggccgcc tgggcggcct gctgaaactc tggctcaccg acgacccgcg
4021 cacggcgcgg ttcggtgatg ccacgatcct cgccctgctg gcgaagatcg aagagaagca
4081 ggacgagctt ggcaaggtca tgatgggcgt ggtccgcccg agggcagagc catgactttt
4141 ttagccgcta aaacggccgg ggggtgcgcg tgattgccaa gcacgtcccc atgcgctcca
4201 tcaagaagag cgacttcgcg gagctggtat tcgtgcaggg caagattcgg aataccaagt
4261 acgagaagga cggccagacg gtctacggga ccgacttcat tgccgataag gtggattatc
4321 tggacaccaa ggcaccaggc gggtcaaatc aggaataagg gcacattgcc ccggcgtgag
4381 tcggggcaat cccgcaagga gggtgaatga atcggacgtt tgaccggaag gcatacaggc
4441 aagaactgat cgacgcgggg ttttccgccg aggatgccga aaccatcgca agccgcaccg
4501 tcatgcgtgc gccccgcgaa accttccagt ccgtcggctc gatggtccag caagctacgg
4561 ccaagatcga gcgcgacagc gtgcaactgg ctccccctgc cctgcccgcg ccatcggccg
4621 ccgtggagcg ttcgcgtcgt ctcgaacagg aggcggcagg tttggcgaag tcgatgacca
4681 tcgacacgcg aggaactatg acgaccaaga agcgaaaaac cgccggcgag gacctggcaa
4741 aacaggtcag cgaggccaag caggccgcgt tgctgaaaca cacgaagcag cagatcaagg
4801 aaatgcagct ttccttgttc gatattgcgc cgtggccgga cacgatgcga gcgatgccaa
4861 acgacacggc ccgctctgcc ctgttcacca cgcgcaacaa gaaaatcccg cgcgaggcgc
4921 tgcaaaacaa ggtcattttc cacgtcaaca aggacgtgaa gatcacctac accggcgtcg
4981 agctgcgggc cgacgatgac gaactggtgt ggcagcaggt gttggagtac gcgaagcgca
5041 cccctatcgg cgagccgatc accttcacgt tctacgagct ttgccaggac ctgggctggt
5101 cgatcaatgg ccggtattac acgaaggccg aggaatgcct gtcgcgccta caggcgacgg
5161 cgatgggctt cacgtccgac cgcgttgggc acctggaatc ggtgtcgctg ctgcaccgct
5221 tccgcgtcct ggaccgtggc aagaaaacgt cccgttgcca ggtcctgatc gacgaggaaa
5281 tcgtcgtgct gtttgctggc gaccactaca cgaaattcat atgggagaag taccgcaagc
5341 tgtcgccgac ggcccgacgg atgttcgact atttcagctc gcaccgggag ccgtacccgc
5401 tcaagctgga aaccttccgc ctcatgtgcg gatcggattc cacccgcgtg aagaagtggc
5461 gcgagcaggt cggcgaagcc tgcgaagagt tgcgaggcag cggcctggtg gaacacgcct
5521 gggtcaatga tgacctggtg cattgcaaac gctagggcct tgtggggtca gttccggctg
5581 ggggttcagc agccagcgct ttactggcat ttcaggaaca agcgggcact gctcgacgca
5641 cttgcttcgc tcagtatcgc tcgggacgca cggcgcgctc tacgaactgc cgataaacag
5701 aggattaaaa ttgacaattg tgattaaggc tcagattcga cggcttggag cggccgacgt
5761 gcaggatttc cgcgagatcc gattgtcggc cctgaagaaa gctccagaga tgttcgggtc
5821 cgtttacgag cacgaggaga aaaagcccat ggaggcgttc gctgaacggt tgcgagatgc
5881 cgtggcattc ggcgcctaca tcgacggcga gatcattggg ctgtcggtct tcaaacagga
5941 ggacggcccc aaggacgctc acaaggcgca tctgtccggc gttttcgtgg agcccgaaca
6001 gcgaggccga ggggtcgccg gtatgctgct gcgggcgttg ccggcgggtt tattgctcgt
6061 gatgatcgtc cgacagattc caacgggaat ctggtggatg cgcatcttca tcctcggcgc
6121 acttaatatt tcgctattct ggagcttgtt gtttatttcg gtctaccgcc tgccgggcgg
6181 ggtcgcggcg acggtaggcg ctgtgcagcc gctgatggtc gtgttcatct ctgccgctct
6241 gctaggtagc ccgatacgat tgatggcggt cctgggggct atttgcggaa ctgcgggcgt
6301 ggcgctgttg gtgttgacac caaacgcagc gctagatcct gtcggcgtcg cagcgggcct
6361 ggcgggggcg gtttccatgg cgttcggaac cgtgctgacc cgcaagtggc aacctcccgt
6421 gcctctgctc acctttaccg cctggcaact ggcggccgga ggacttctgc tcgttccagt
6481 agctttagtg tttgatccgc caatcccgat gcctacagga accaatgttc tcggcctggc
6541 gtggctcggc ctgatcggag cgggtttaac ctacttcctt tggttccggg ggatctcgcg
6601 actcgaacct acagttgttt ccttactggg ctttctcagc cgggatggcg ctaagaagct
6661 attgccgccg atcttcatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
6721 cgcatcaggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
6781 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
6841 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
6901 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
6961 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
7021 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
7081 ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg
7141 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
7201 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
7261 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
7321 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
7381 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
7441 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatat
7501 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
7561 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
7621 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
7681 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
7741 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
7801 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
7861 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
7921 aaacaagtgg cagcaacgga ttcgcaaacc tgtcacgcct tttgtgccaa aagccgcgcc
7981 aggtttgcga tccgctgtgc caggcgttag gcgtcatatg aagatttcgg tgatccctga
8041 gcaggtggcg gaaacattgg atgctgagaa ccatttcatt gttcgtgaag tgttcgatgt
8101 gcacctatcc gaccaaggct ttgaactatc taccagaagt gtgagcccct accggaagga
8161 ttacatctcg gatgatgact ctgatgaaga ctctgcttgc tatggcgcat tcatcgacca
8221 agagcttgtc gggaagattg aactcaactc aacatggaac gatctagcct ctatcgaaca
8281 cattgttgtg tcgcacacgc accgaggcaa aggagtcgcg cacagtctca tcgaatttgc
8341 gaaaaagtgg gcactaagca gacagctcct tggcatacga ttagagacac aaacgaacaa
8401 tgtacctgcc tgcaatttgt acgcaaaatg tggctttact ctcggcggca ttgacctgtt
8461 cacgtataaa actagacctc aagtctcgaa cgaaacagcg atgtactggt actggttctc
8521 gggagcacag gatgacgcct aacaattcat tcaagccgac accgcttcgc ggcgcggctt
8581 aattcaggag ttaaacatca tgagggaagc ggtgatcgcc gaagtatcga ctcaactatc
8641 agaggtagtt ggcgtcatcg agcgccatct cgaaccgacg ttgctggccg tacatttgta
8701 cggctccgca gtggatggcg gcctgaagcc acacagtgat attgatttgc tggttacggt
8761 gaccgtaagg cttgatgaaa caacgcggcg agctttgatc aacgaccttt tggaaacttc
8821 ggcttcccct ggagagagcg agattctccg cgctgtagaa gtcaccattg ttgtgcacga
8881 cgacatcatt ccgtggcgtt atccagctaa gcgcgaactg caatttggag aatggcagcg
8941 caatgacatt cttgcaggta tcttcgagcc agccacgatc gacattgatc tggctatctt
9001 gctgacaaaa gcaagagaac atagcgttgc cttggtaggt ccagcggcgg aggaactctt
9061 tgatccggtt cctgaacagg atctatttga ggcgctaaat gaaaccttaa cgctatggaa
9121 ctcgccgccc gactgggctg gcgatgagcg aaatgtagtg cttacgttgt cccgcatttg
9181 gtacagcgca gtaaccggca aaatcgcgcc gaaggatgtc gctgccgact gggcaatgga
9241 gcgcctgccg gcccagtatc agcccgtcat acttgaagct aggcaggctt atcttggaca
9301 agaagatcgc ttggcctcgc gcgcagatca gttggaagaa tttgttcact acgtgaaagg
9361 cgagatcacc aaggtagtcg gcaaataatg tctaacaatt cgttcaagcc gacgccgctt
9421 cgcggcgcgg cttaactcaa gcgttagaga gctggggaag actatgcgcg atctgttgaa
9481 ggtggttcta agcctcgtac ttgcgatggc atcggggcag gcacttgctg acctgccaat
9541 tgttttagtg gatgaagctc gtcttcccta tgactactcc ccatccaact acgacatttc
9601 tccaagcaac tacgacaact ccataagcaa ttacgacaat agtccatcaa attacgacaa
9661 ctctgagagc aactacgata atagttcatc caattacgac aatagtcgca acggaaatcg
9721 taggcttata tatagcgcaa atgggtctcg cactttcgcc ggctactacg tcattgccaa
9781 caatgggaca acgaacttct tttccacatc tggcaaaagg atgttctaca ccccaaaagg
9841 ggggcgcggc gtctatggcg gcaaagatgg gagcttctgc ggggcattgg tcgtcataaa
9901 tggccaattt tcgcttgccc tgacagataa cggcctgaag atcatgtatc taagcaacta
9961 gcctgctctc taataaaatg ttaggagctt ggctgccatt tttggggtga ggccgttcgc
10021 ggccgagggg cgcagcccct ggggggatgg gaggcccgcg ttagcgggcc gggagggttc
10081 gagaaggggg ggcacccccc ttcggcgtgc gcggtcacgc gccagggcgc agccctggtt
10141 aaaaacaagg tttataaata ttggtttaaa agcaggttaa aagacaggtt agcggtggcc
10201 gaaaaacggg cggaaaccct tgcaaatgct ggattttctg cctgtggaca gcccctcaaa
10261 tgtcaatagg tgcgcccctc atctgtcagc actctgcccc tcaagtgtca aggatcgcgc
10321 ccctcatctg tcagtagtcg cgcccctcaa gtgtcaatac cgcagggcac ttatccccag
10381 gcttgtccac atcatctgtg ggaaactcgc gtaaaatcag gcgttttcgc cgatttgcga
10441 ggctggccag ctccacgtcg ccggccgaaa tcgagcctgc ccctcatctg tcaacgccgc
10501 gccgggtgag tcggcccctc aagtgtcaac gtccgcccct catctgtcag tgagggccaa
10561 gttttccgcg aggtatccac aacgccggcg gccggccgcg gtgtctcgca cacggcttcg
10621 acggcgtttc tggcgcgttt gcagggccat agacggccgc cagcccagcg gcgagggcaa
10681 ccagcccggt gagcgtcgga aagggtcgac atcttgctgc gttcggatat tttcgtggag
10741 ttcccgccac agacccggat tgaaggcgag atccagcaac tcgcgccaga tcatcctgtg
10801 acggaacttt ggcgcgtgat gactggccag gacgtcggcc gaaagagcga caagcagatc
10861 acgattttcg acagcgtcgg atttgcgatc gaggattttt cggcgctgcg ctacgtccgc
10921 gaccgcgttg agggatcaag ccacagcagc ccactcgacc ttctagccga cccagacgag
10981 ccaagggatc tttttggaat gctgctccgt cgtcaggctt tccgacgttt gggtggttga
11041 acagaagtca ttatcgtacg gaatgccagc actcccgagg ggaaccctgt ggttggcatg
11101 cacatacaaa tggacgaacg gataaacctt ttcacgccct tttaaatatc cgttattcta
11161 ataaacgctc ttttctctta ggtttacccg ccaatatatc ctgtcaaaca ctgatagttt
11221 aaactgaagg cgggaaacga caatctgatc atgagcggag aattaaggga gtcacgttat
11281 gacccccgcc gatgacgcgg gacaagccgt tttacgtttg gaactgacag aaccgcaacg
11341 attgaaggag ccactcagcc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
11401 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
11461 ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc
11521 gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg
11581 attacgccaa gctatttagg tgacactata gaatactcaa gctatgcatc caacgcgttg
11641 ggagctctcc catatcgacc tgcaggcggc cgctcgacga attaattcca atcccacaaa
11701 aatctgagct taacagcaca gttgctcctc tcagagcaga atcgggtatt caacaccctc
11761 atatcaacta ctacgttgtg tataacggtc cacatgccgg tatatacgat gactggggtt
11821 gtacaaaggc ggcaacaaac ggcgttcccg gagttgcaca caagaaattt gccactatta
11881 cagaggcaag agcagcagct gacgcgtaca caacaagtca gcaaacagac aggttgaact
11941 tcatccccaa aggagaagct caactcaagc ccaagagctt tgctaaggcc ctaacaagcc
12001 caccaaagca aaaagcccac tggctcacgc taggaaccaa aaggcccagc agtgatccag
12061 ccccaaaaga gatctccttt gccccggaga ttacaatgga cgatttcctc tatctttacg
12121 atctaggaag gaagttcgaa ggtgaaggtg acgacactat gttcaccact gataatgaga
12181 aggttagcct cttcaatttc agaaagaatg ctgacccaca gatggttaga gaggcctacg
12241 cagcaggtct catcaagacg atctacccga gtaacaatct ccaggagatc aaataccttc
12301 ccaagaaggt taaagatgca gtcaaaagat tcaggactaa ttgcatcaag aacacagaga
12361 aagacatatt tctcaagatc agaagtacta ttccagtatg gacgattcaa ggcttgcttc
12421 ataaaccaag gcaagtaata gagattggag tctctaaaaa ggtagttcct actgaatcta
12481 aggccatgca tggagtctaa gattcaaatc gaggatctaa cagaactcgc cgtgaagact
12541 ggcgaacagt tcatacagag tcttttacga ctcaatgaca agaagaaaat cttcgtcaac
12601 atggtggagc acgacactct ggtctactcc aaaaatgtca aagatacagt ctcagaagac
12661 caaagggcta ttgagacttt tcaacaaagg ataatttcgg gaaacctcct cggattccat
12721 tgcccagcta tctgtcactt catcgaaagg acagtagaaa aggaaggtgg ctcctacaaa
12781 tgccatcatt gcgataaagg aaaggctatc attcaagatc tctctgccga cagtggtccc
12841 aaagatggac ccccacccac gaggagcatc gtggaaaaag aagacgttcc aaccacgtct
12901 tcaaagcaag tggattgatg tgacatctcc actgacgtaa gggatgacgc acaatcccac
12961 tatccttcgc aagacccttc ctctatataa ggaagttcat ttcatttgga gaggacacgc
13021 tcgaggacaa gtttgtacaa aaaagctgaa cgagaaacgt aaaatgatat aaatatcaat
13081 atattaaatt agattttgca taaaaaacag actacataat actgtaaaac acaacatatc
13141 cagtcactat gaatcaacta cttagatggt attagtgacc tgtagtcgac cgacagcctt
13201 ccaaatgttc ttcgggtgat gctgccaact tagtcgaccg acagccttcc aaatgttctt
13261 ctcaaacgga atcgtcgtat ccagcctact cgctattgtc ctcaatgccg tattaaatca
13321 taaaaagaaa taagaaaaag aggtgcgagc ctcttttttg tgtgacaaaa taaaaacatc
13381 tacctattca tatacgctag tgtcatagtc ctgaaaatca tctgcatcaa gaacaatttc
13441 acaactctta tacttttctc ttacaagtcg ttcggcttca tctggatttt cagcctctat
13501 acttactaaa cgtgataaag tttctgtaat ttctactgta tcgacctgca gactggctgt
13561 gtataaggga gcctgacatt tatattcccc agaacatcag gttaatggcg tttttgatgt
13621 cattttcgcg gtggctgaga tcagccactt cttccccgat aacggagacc ggcacactgg
13681 ccatatcggt ggtcatcatg cgccagcttt catccccgat atgcaccacc gggtaaagtt
13741 cacgggagac tttatctgac agcagacgtg cactggccag ggggatcacc atccgtcgcc
13801 cgggcgtgtc aataatatca ctctgtacat ccacaaacag acgataacgg ctctctcttt
13861 tataggtgta aaccttaaac tgcatttcac cagtccctgt tctcgtcagc aaaagagccg
13921 ttcatttcaa taaaccgggc gacctcagcc atcccttcct gattttccgc tttccagcgt
13981 tcggcacgca gacgacgggc ttcattctgc atggttgtgc ttaccagacc ggagatattg
14041 acatcatata tgccttgagc aactgatagc tgtcgctgtc aactgtcact gtaatacgct
14101 gcttcatagc acacctcttt ttgacatact tcgggtagtg ccgatcaacg tctcattttc
14161 gccaaaagtt ggcccagggc ttcccggtat caacagggac accaggattt atttattctg
14221 cgaagtgatc ttccgtcaca ggtatttatt cggcgcaaag tgcgtcgggt gatgctgcca
14281 acttagtcga ctacaggtca ctaataccat ctaagtagtt gattcatagt gactggatat
14341 gttgtgtttt acagtattat gtagtctgtt ttttatgcaa aatctaattt aatatattga
14401 tatttatatc attttacgtt tctcgttcag ctttcttgta caaagtggtc tcgaggaatt
14461 cggtacccca gcttggtaag gaaataatta ttttcttttt tccttttagt ataaaatagt
14521 taagtgatgt taattagtat gattataata atatagttgt tataattgtg aaaaaataat
14581 ttataaatat attgtttaca taaacaacat agtaatgtaa aaaaatatga caagtgatgt
14641 gtaagacgaa gaagataaaa gttgagagta agtatattat ttttaatgaa tttgatcgaa
14701 catgtaagat gatatactag cattaatatt tgttttaatc ataatagtaa ttctagctgg
14761 tttgatgaat taaatatcaa tgataaaata ctatagtaaa aataagaata aataaattaa
14821 aataatattt ttttatgatt aatagtttat tatataatta aatatctata ccattactaa
14881 atattttagt ttaaaagtta ataaatattt tgttagaaat tccaatctgc ttgtaattta
14941 tcaataaaca aaatattaaa taacaagcta aagtaacaaa taatatcaaa ctaatagaaa
15001 cagtaatcta atgtaacaaa acataatcta atgctaatat aacaaagcgc aagatctatc
15061 attttatata gtattatttt caatcaacat tcttattaat ttctaaataa tacttgtagt
15121 tttattaact tctaaatgga ttgactatta attaaatgaa ttagtcgaac atgaataaac
15181 aaggtaacat gatagatcat gtcattgtgt tatcattgat cttacatttg gattgattac
15241 agttgggaaa ttgggttcga aatcgataag cttggatcct ctagaccact ttgtacaaga
15301 aagctgaacg agaaacgtaa aatgatataa atatcaatat attaaattag attttgcata
15361 aaaaacagac tacataatac tgtaaaacac aacatatcca gtcactatga atcaactact
15421 tagatggtat tagtgacctg tagtcgacta agttggcagc atcacccgac gcactttgcg
15481 ccgaataaat acctgtgacg gaagatcact tcgcagaata aataaatcct ggtgtccctg
15541 ttgataccgg gaagccctgg gccaactttt ggcgaaaatg agacgttgat cggcactacc
15601 catttcacaa ctcttatact tttctcttac aagtcgttcg gcttcatctg gattttcagc
15661 ctctatactt actaaacgtg ataaagtttc tgtaatttct actgtatcga cctgcagact
15721 ggctgtgtat aagggagcct gacatttata ttccccagaa catcaggtta atggcgtttt
15781 tgatgtcatt ttcgcggtgg ctgagatcag ccacttcttc cccgataacg gagaccggca
15841 cactggccat atcggtggtc atcatgcgcc agctttcatc cccgatatgc accaccgggt
15901 aaagttcacg ggagacttta tctgacagca gacgtgcact ggccaggggg atcaccatcc
15961 gtcgcccggg cgtgtcaata atatcactct gtacatccac aaacagacga taacggctct
16021 ctcttttata ggtgtaaacc ttaaactgca tttcaccagt ccctgttctc gtcagcaaaa
16081 gagccgttca tttcaataaa ccgggcgacc tcagccatcc cttcctgatt ttccgctttc
16141 cagcgttcgg cacgcagacg acgggcttca ttctgcatgg ttgtgcttac cagaccggag
16201 atattgacat catatatgcc ttgagcaact gatagctgtc gctgtcaact gtcactgtaa
16261 tacgctgctt catagcacac ctctttttga catacttctg ttcttgatgc agatgatttt
16321 caggactatg acactagcgt atatgaatag gtagatgttt ttattttgtc acacaaaaaa
16381 gaggctcgca cctctttttc ttatttcttt ttatgattta atacggcatt gaggacaata
16441 gcgagtaggc tggatacgac gattccgttt gagaagaaca tttggaaggc tgtcggtcga
16501 ctaagttggc agcatcaccc gaagaacatt tggaaggctg tcggtcgact acaggtcact
16561 aataccatct aagtagttga ttcatagtga ctggatatgt tgtgttttac agtattatgt
16621 agtctgtttt ttatgcaaaa tctaatttaa tatattgata tttatatcat tttacgtttc
16681 tcgttcagct tttttgtaca aacttgtcta gagtcctgct ttaatgagat atgcgagacg
16741 cctatgatcg catgatattt gctttcaatt ctgttgtgca cgttgtaaaa aacctgagca
16801 tgtgtagctc agatccttac cgccggtttc ggttcattct aatgaatata tcacccgtta
16861 ctatcgtatt tttatgaata atattctccg ttcaatttac tgattgtacc ctactactta
16921 tatgtacaat attaaaatga aaacaatata ttgtgctgaa taggtttata gcgacatcta
16981 tgatagagcg ccacaataac aaacaattgc gttttattat tacaaatcca attttaaaaa
17041 aagcggcaga accggtcaaa cctaaaagac tgattacata aatcttattc aaatttcaaa
17101 aggccccagg ggctagtatc tacgacacac cgagcggcga actaataacg ttcactgaag
17161 ggaactccgg ttccccgccg gcgcgcatgg gtgagattcc ttgaagttga gtattggccg
17221 tccgctctac cgaaagttac gggcaccatt caacccggtc cagcacggcg gccgggtaac
17281 cgacttgctg ccccgagaat tatgcagcat ttttttggtg tatgtgggcc ccaaatgaag
17341 tgcaggtcaa accttgacag tgacgacaaa tcgttgggcg ggtccagggc gaattttgcg
17401 acaacatgtc gaggctcagc aggacctgca ggcatgcaag ctagcttact agtgatgcat
17461 attctatagt gtcacctaaa tctgc
//
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