pGFPGUSPlus

价格:1200元

联系方式:I47-825O-882O

相关技术服务:质粒构建    基因合成    质粒大提

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载体基本信息

质粒名称: pGFPGUSPlus 
出品公司: Addgene
目录编号: 64401
存储实验室: Claudia Vickers

载体骨架基本信息
载体名称: pCAMBIA1305.1
载体出品公司: CAMBIA
载体抗性: 卡那霉素
空载体大小: --
完整质粒大小: --
载体修饰: --
载体类型: 植物表达;植物转化的二元载体
筛选标记: 潮霉素
高拷贝/低拷贝: 高拷贝
生长条件: 37 ℃

插入基因信息
插入基因名称: GFP
物种来源: --
插入大小: --
基因突变: S65T
GenBank编号 ABQ22954.1
启动子 35S
标签/融合蛋白: --
克隆方法: 限制性内切酶
5' 克隆位点: HindIII(未破坏)
3' 克隆位点: EcoRI(未破坏)
5' 测序引物 M13-F20
3' 测序引物 M13-R

文献引用方法
材料和方法部分: pGFPGUSPlus was a gift from Claudia Vickers (Addgene plasmid # 64401 ; http://n2t.net/addgene:64401 ; RRID:Addgene_64401)
参考文献部分: pGFPGUSPlus, a new binary vector for gene expression studies and optimising transformation systems in plants. Vickers CE, Schenk PM, Li D, Mullineaux PM, Gresshoff PM. Biotechnol Lett. 2007 Nov;29(11):1793-6. Epub 2007 Aug 9. 10.1007/s10529-007-9467-6 PubMed 17687623

载体图谱质粒图谱和多克隆位点信息

pGFPGUSPlus质粒图谱

载体简介

载体序列

LOCUS       pGFPGUSPlus               13702 bp ds-DNA     circular SYN 07-DEC-2018
DEFINITION  binary plant vector expressing GFP and GUS, both driven by a 35S 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    pGFPGUSPlus
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13702)
  AUTHORS   Vickers CE, Schenk PM, Li D, Mullineaux PM, Gresshoff PM
  TITLE     pGFPGUSPlus, a new binary vector for gene expression studies and 
            optimising transformation systems in plants.
  JOURNAL   Biotechnol Lett. 2007 Nov;29(11):1793-6. Epub 2007 Aug 9.
  PUBMED    17687623
REFERENCE   2  (bases 1 to 13702)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 7, 2018 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..13702
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(85..102)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(85..101)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(94..116)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        478..823
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     CDS             join(840..854,1045..2862)
                     /codon_start=1
                     /product="beta-glucuronidase"
                     /label=GUSPlus(TM)
                     /note="codon-optimized Staphylococcus gusA gene with a 
                     catalase intron to ensure expression in plants but not 
                     bacteria"
                     /translation="MVDLRNRRTSLYPINTETRGVFDLNGVWNFKLDYGKGLEEKWYES
                     KLTDTISMAVPSSYNDIGVTKEIRNHIGYVWYEREFTVPAYLKDQRIVLRFGSATHKAI
                     VYVNGELVVEHKGGFLPFEAEINNSLRDGMNRVTVAVDNILDDSTLPVGLYSERHEEGL
                     GKVIRNKPNFDFFNYAGLHRPVKIYTTPFTYVEDISVVTDFNGPTGTVTYTVDFQGKAE
                     TVKVSVVDEEGKVVASTEGLSGNVEIPNVILWEPLNTYLYQIKVELVNDGLTIDVYEEP
                     FGVRTVEVNDGKFLINNKPFYFKGFGKHEDTPINGRGFNEASNVMDFNILKWIGANSFR
                     TAHYPYSEELMRLADREGLVVIDETPAVGVHLNFMATTGLGEGSERVSTWEKIRTFEHH
                     QDVLRELVSRDKNHPSVVMWSIANEAATEEEGAYEYFKPLVELTKELDPQKRPVTIVLF
                     VMATPETDKVAELIDVIALNRYNGWYFDGGDLEAAKVHLRQEFHAWNKRCPGKPIMITE
                     YGADTVAGFHDIDPVMFTEEYQVEYYQANHVVFDEFENFVGEQAWNFADFATSQGVMRV
                     QGNKKGVFTRDRKPKLAAHVFRERWTNIPDFGYKN"
     intron          855..1044
                     /label=cat1 intron
                     /note="castor bean catalase intron, modified"
     CDS             2869..2886
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      2918..3170
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    3192..3216
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             4517..5146
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             5575..6648
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     rep_origin      6714..6908
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(7144..7166)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    7252..7392
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     7307..7326
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(7407..7424)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7578..8166)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7658..7677)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(8253..9047)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     misc_feature    9472..9496
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     polyA_signal    9574..9748
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(9789..10814)
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRSQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     K"
     promoter        complement(10881..11557)
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     protein_bind    11748..11769
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        11784..11814
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11822..11838
                     /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)."
     primer_bind     11827..11849
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     11846..11862
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     11846..11862
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     terminator      11879..12131
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(12177..12896)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(12222..12243)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     12571..12590
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     12830..12851
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     promoter        complement(12927..13272)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
ORIGIN
        1 aagcctctct aaccatctgt gggttagcat tctttctgaa attgaaaagg ctaatctggg
       61 gacctgcagg catgcaagct tggcactggc cgtcgtttta caacgtcgtg actgggaaaa
      121 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa
      181 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
      241 ctagagcagc ttgagcttgg atcagattgt cgtttcccgc cttcagttta gcttcatgga
      301 gtcaaagatt caaatagagg acctaacaga actcgccgta aagactggcg aacagttcat
      361 acagagtctc ttacgactca atgacaagaa gaaaatcttc gtcaacatgg tggagcacga
      421 cacacttgtc tactccaaaa atatcaaaga tacagtctca gaagaccaaa gggcaattga
      481 gacttttcaa caaagggtaa tatccggaaa cctcctcgga ttccattgcc cagctatctg
      541 tcactttatt gtgaagatag tggaaaagga aggtggctcc tacaaatgcc atcattgcga
      601 taaaggaaag gccatcgttg aagatgcctc tgccgacagt ggtcccaaag atggaccccc
      661 acccacgagg agcatcgtgg aaaaagaaga cgttccaacc acgtcttcaa agcaagtgga
      721 ttgatgtgat atctccactg acgtaaggga tgacgcacaa tcccactatc cttcgcaaga
      781 cccttcctct atataaggaa gttcatttca tttggagaga acacggggga ctcttgacca
      841 tggtagatct gagggtaaat ttctagtttt tctccttcat tttcttggtt aggacccttt
      901 tctcttttta tttttttgag ctttgatctt tctttaaact gatctatttt ttaattgatt
      961 ggttatggtg taaatattac atagctttaa ctgataatct gattacttta tttcgtgtgt
     1021 ctatgatgat gatgatagtt acagaaccga cgaactagtc tgtacccgat caacaccgag
     1081 acccgtggcg tcttcgacct caatggcgtc tggaacttca agctggacta cgggaaagga
     1141 ctggaagaga agtggtacga aagcaagctg accgacacta ttagtatggc cgtcccaagc
     1201 agttacaatg acattggcgt gaccaaggaa atccgcaacc atatcggata tgtctggtac
     1261 gaacgtgagt tcacggtgcc ggcctatctg aaggatcagc gtatcgtgct ccgcttcggc
     1321 tctgcaactc acaaagcaat tgtctatgtc aatggtgagc tggtcgtgga gcacaagggc
     1381 ggattcctgc cattcgaagc ggaaatcaac aactcgctgc gtgatggcat gaatcgcgtc
     1441 accgtcgccg tggacaacat cctcgacgat agcaccctcc cggtggggct gtacagcgag
     1501 cgccacgaag agggcctcgg aaaagtcatt cgtaacaagc cgaacttcga cttcttcaac
     1561 tatgcaggcc tgcaccgtcc ggtgaaaatc tacacgaccc cgtttacgta cgtcgaggac
     1621 atctcggttg tgaccgactt caatggccca accgggactg tgacctatac ggtggacttt
     1681 caaggcaaag ccgagaccgt gaaagtgtcg gtcgtggatg aggaaggcaa agtggtcgca
     1741 agcaccgagg gcctgagcgg taacgtggag attccgaatg tcatcctctg ggaaccactg
     1801 aacacgtatc tctaccagat caaagtggaa ctggtgaacg acggactgac catcgatgtc
     1861 tatgaagagc cgttcggcgt gcggaccgtg gaagtcaacg acggcaagtt cctcatcaac
     1921 aacaaaccgt tctacttcaa gggctttggc aaacatgagg acactcctat caacggccgt
     1981 ggctttaacg aagcgagcaa tgtgatggat ttcaatatcc tcaaatggat cggcgccaac
     2041 agcttccgga ccgcacacta tccgtactct gaagagttga tgcgtcttgc ggatcgcgag
     2101 ggtctggtcg tgatcgacga gactccggca gttggcgtgc acctcaactt catggccacc
     2161 acgggactcg gcgaaggcag cgagcgcgtc agtacctggg agaagattcg gacgtttgag
     2221 caccatcaag acgttctccg tgaactggtg tctcgtgaca agaaccatcc aagcgtcgtg
     2281 atgtggagca tcgccaacga ggcggcgact gaggaagagg gcgcgtacga gtacttcaag
     2341 ccgttggtgg agctgaccaa ggaactcgac ccacagaagc gtccggtcac gatcgtgctg
     2401 tttgtgatgg ctaccccgga gacggacaaa gtcgccgaac tgattgacgt catcgcgctc
     2461 aatcgctata acggatggta cttcgatggc ggtgatctcg aagcggccaa agtccatctc
     2521 cgccaggaat ttcacgcgtg gaacaagcgt tgcccaggaa agccgatcat gatcactgag
     2581 tacggcgcag acaccgttgc gggctttcac gacattgatc cagtgatgtt caccgaggaa
     2641 tatcaagtcg agtactacca ggcgaaccac gtcgtgttcg atgagtttga gaacttcgtg
     2701 ggtgagcaag cgtggaactt cgcggacttc gcgacctctc agggcgtgat gcgcgtccaa
     2761 ggaaacaaga agggcgtgtt cactcgtgac cgcaagccga agctcgccgc gcacgtcttt
     2821 cgcgagcgct ggaccaacat tccagatttc ggctacaaga acgctagcca tcaccatcac
     2881 catcacgtgt gaattggtga ccagctcgaa tttccccgat cgttcaaaca tttggcaata
     2941 aagtttctta agattgaatc ctgttgccgg tcttgcgatg attatcatat aatttctgtt
     3001 gaattacgtt aagcatgtaa taattaacat gtaatgcatg acgttattta tgagatgggt
     3061 ttttatgatt agagtcccgc aattatacat ttaatacgcg atagaaaaca aaatatagcg
     3121 cgcaaactag gataaattat cgcgcgcggt gtcatctatg ttactagatc gggaattaaa
     3181 ctatcagtgt ttgacaggat atattggcgg gtaaacctaa gagaaaagag cgtttattag
     3241 aataatcgga tatttaaaag ggcgtgaaaa ggtttatccg ttcgtccatt tgtatgtgca
     3301 tgccaaccac agggttcccc tcgggatcaa agtactttga tccaacccct ccgctgctat
     3361 agtgcagtcg gcttctgacg ttcagtgcag ccgtcttctg aaaacgacat gtcgcacaag
     3421 tcctaagtta cgcgacaggc tgccgccctg cccttttcct ggcgttttct tgtcgcgtgt
     3481 tttagtcgca taaagtagaa tacttgcgac tagaaccgga gacattacgc catgaacaag
     3541 agcgccgccg ctggcctgct gggctatgcc cgcgtcagca ccgacgacca ggacttgacc
     3601 aaccaacggg ccgaactgca cgcggccggc tgcaccaagc tgttttccga gaagatcacc
     3661 ggcaccaggc gcgaccgccc ggagctggcc aggatgcttg accacctacg ccctggcgac
     3721 gttgtgacag tgaccaggct agaccgcctg gcccgcagca cccgcgacct actggacatt
     3781 gccgagcgca tccaggaggc cggcgcgggc ctgcgtagcc tggcagagcc gtgggccgac
     3841 accaccacgc cggccggccg catggtgttg accgtgttcg ccggcattgc cgagttcgag
     3901 cgttccctaa tcatcgaccg cacccggagc gggcgcgagg ccgccaaggc ccgaggcgtg
     3961 aagtttggcc cccgccctac cctcaccccg gcacagatcg cgcacgcccg cgagctgatc
     4021 gaccaggaag gccgcaccgt gaaagaggcg gctgcactgc ttggcgtgca tcgctcgacc
     4081 ctgtaccgcg cacttgagcg cagcgaggaa gtgacgccca ccgaggccag gcggcgcggt
     4141 gccttccgtg aggacgcatt gaccgaggcc gacgccctgg cggccgccga gaatgaacgc
     4201 caagaggaac aagcatgaaa ccgcaccagg acggccagga cgaaccgttt ttcattaccg
     4261 aagagatcga ggcggagatg atcgcggccg ggtacgtgtt cgagccgccc gcgcacgtct
     4321 caaccgtgcg gctgcatgaa atcctggccg gtttgtctga tgccaagctg gcggcctggc
     4381 cggccagctt ggccgctgaa gaaaccgagc gccgccgtct aaaaaggtga tgtgtatttg
     4441 agtaaaacag cttgcgtcat gcggtcgctg cgtatatgat gcgatgagta aataaacaaa
     4501 tacgcaaggg gaacgcatga aggttatcgc tgtacttaac cagaaaggcg ggtcaggcaa
     4561 gacgaccatc gcaacccatc tagcccgcgc cctgcaactc gccggggccg atgttctgtt
     4621 agtcgattcc gatccccagg gcagtgcccg cgattgggcg gccgtgcggg aagatcaacc
     4681 gctaaccgtt gtcggcatcg accgcccgac gattgaccgc gacgtgaagg ccatcggccg
     4741 gcgcgacttc gtagtgatcg acggagcgcc ccaggcggcg gacttggctg tgtccgcgat
     4801 caaggcagcc gacttcgtgc tgattccggt gcagccaagc ccttacgaca tatgggccac
     4861 cgccgacctg gtggagctgg ttaagcagcg cattgaggtc acggatggaa ggctacaagc
     4921 ggcctttgtc gtgtcgcggg cgatcaaagg cacgcgcatc ggcggtgagg ttgccgaggc
     4981 gctggccggg tacgagctgc ccattcttga gtcccgtatc acgcagcgcg tgagctaccc
     5041 aggcactgcc gccgccggca caaccgttct tgaatcagaa cccgagggcg acgctgcccg
     5101 cgaggtccag gcgctggccg ctgaaattaa atcaaaactc atttgagtta atgaggtaaa
     5161 gagaaaatga gcaaaagcac aaacacgcta agtgccggcc gtccgagcgc acgcagcagc
     5221 aaggctgcaa cgttggccag cctggcagac acgccagcca tgaagcgggt caactttcag
     5281 ttgccggcgg aggatcacac caagctgaag atgtacgcgg tacgccaagg caagaccatt
     5341 accgagctgc tatctgaata catcgcgcag ctaccagagt aaatgagcaa atgaataaat
     5401 gagtagatga attttagcgg ctaaaggagg cggcatggaa aatcaagaac aaccaggcac
     5461 cgacgccgtg gaatgcccca tgtgtggagg aacgggcggt tggccaggcg taagcggctg
     5521 ggttgcctgc cggccctgca atggcactgg aacccccaag cccgaggaat cggcgtgagc
     5581 ggtcgcaaac catccggccc ggtacaaatc ggcgcggcgc tgggtgatga cctggtggag
     5641 aagttgaagg ccgcgcaggc cgcccagcgg caacgcatcg aggcagaagc acgccccggt
     5701 gaatcgtggc aagcggccgc tgatcgaatc cgcaaagaat cccggcaacc gccggcagcc
     5761 ggtgcgccgt cgattaggaa gccgcccaag ggcgacgagc aaccagattt tttcgttccg
     5821 atgctctatg acgtgggcac ccgcgatagt cgcagcatca tggacgtggc cgttttccgt
     5881 ctgtcgaagc gtgaccgacg agctggcgag gtgatccgct acgagcttcc agacgggcac
     5941 gtagaggttt ccgcagggcc ggccggcatg gccagtgtgt gggattacga cctggtactg
     6001 atggcggttt cccatctaac cgaatccatg aaccgatacc gggaagggaa gggagacaag
     6061 cccggccgcg tgttccgtcc acacgttgcg gacgtactca agttctgccg gcgagccgat
     6121 ggcggaaagc agaaagacga cctggtagaa acctgcattc ggttaaacac cacgcacgtt
     6181 gccatgcagc gtacgaagaa ggccaagaac ggccgcctgg tgacggtatc cgagggtgaa
     6241 gccttgatta gccgctacaa gatcgtaaag agcgaaaccg ggcggccgga gtacatcgag
     6301 atcgagctag ctgattggat gtaccgcgag atcacagaag gcaagaaccc ggacgtgctg
     6361 acggttcacc ccgattactt tttgatcgat cccggcatcg gccgttttct ctaccgcctg
     6421 gcacgccgcg ccgcaggcaa ggcagaagcc agatggttgt tcaagacgat ctacgaacgc
     6481 agtggcagcg ccggagagtt caagaagttc tgtttcaccg tgcgcaagct gatcgggtca
     6541 aatgacctgc cggagtacga tttgaaggag gaggcggggc aggctggccc gatcctagtc
     6601 atgcgctacc gcaacctgat cgagggcgaa gcatccgccg gttcctaatg tacggagcag
     6661 atgctagggc aaattgccct agcaggggaa aaaggtcgaa aaggtctctt tcctgtggat
     6721 agcacgtaca ttgggaaccc aaagccgtac attgggaacc ggaacccgta cattgggaac
     6781 ccaaagccgt acattgggaa ccggtcacac atgtaagtga ctgatataaa agagaaaaaa
     6841 ggcgattttt ccgcctaaaa ctctttaaaa cttattaaaa ctcttaaaac ccgcctggcc
     6901 tgtgcataac tgtctggcca gcgcacagcc gaagagctgc aaaaagcgcc tacccttcgg
     6961 tcgctgcgct ccctacgccc cgccgcttcg cgtcggccta tcgcggccgc tggccgctca
     7021 aaaatggctg gcctacggcc aggcaatcta ccagggcgcg gacaagccgc gccgtcgcca
     7081 ctcgaccgcc ggcgcccaca tcaaggcacc ctgcctcgcg cgtttcggtg atgacggtga
     7141 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
     7201 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat
     7261 gacccagtca cgtagcgata gcggagtgta tactggctta actatgcggc atcagagcag
     7321 attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa
     7381 taccgcatca ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     7441 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     7501 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     7561 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     7621 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     7681 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     7741 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     7801 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     7861 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     7921 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     7981 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
     8041 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     8101 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     8161 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     8221 cgttaaggga ttttggtcat gcattctagg tactaaaaca attcatccag taaaatataa
     8281 tattttattt tctcccaatc aggcttgatc cccagtaagt caaaaaatag ctcgacatac
     8341 tgttcttccc cgatatcctc cctgatcgac cggacgcaga aggcaatgtc ataccacttg
     8401 tccgccctgc cgcttctccc aagatcaata aagccactta ctttgccatc tttcacaaag
     8461 atgttgctgt ctcccaggtc gccgtgggaa aagacaagtt cctcttcggg cttttccgtc
     8521 tttaaaaaat catacagctc gcgcggatct ttaaatggag tgtcttcttc ccagttttcg
     8581 caatccacat cggccagatc gttattcagt aagtaatcca attcggctaa gcggctgtct
     8641 aagctattcg tatagggaca atccgatatg tcgatggagt gaaagagcct gatgcactcc
     8701 gcatacagct cgataatctt ttcagggctt tgttcatctt catactcttc cgagcaaagg
     8761 acgccatcgg cctcactcat gagcagattg ctccagccat catgccgttc aaagtgcagg
     8821 acctttggaa caggcagctt tccttccagc catagcatca tgtccttttc ccgttccaca
     8881 tcataggtgg tccctttata ccggctgtcc gtcattttta aatataggtt ttcattttct
     8941 cccaccagct tatatacctt agcaggagac attccttccg tatcttttac gcagcggtat
     9001 ttttcgatca gttttttcaa ttccggtgat attctcattt tagccattta ttatttcctt
     9061 cctcttttct acagtattta aagatacccc aagaagctaa ttataacaag acgaactcca
     9121 attcactgtt ccttgcattc taaaacctta aataccagaa aacagctttt tcaaagttgt
     9181 tttcaaagtt ggcgtataac atagtatcga cggagccgat tttgaaaccg cggtgatcac
     9241 aggcagcaac gctctgtcat cgttacaatc aacatgctac cctccgcgag atcatccgtg
     9301 tttcaaaccc ggcagcttag ttgccgttct tccgaatagc atcggtaaca tgagcaaagt
     9361 ctgccgcctt acaacggctc tcccgctgac gccgtcccgg actgatgggc tgcctgtatc
     9421 gagtggtgat tttgtgccga gctgccggtc ggggagctgt tggctggctg gtggcaggat
     9481 atattgtggt gtaaacaaat tgacgcttag acaacttaat aacacattgc ggacgttttt
     9541 aatgtactga attaacgccg aattaattcg ggggatctgg attttagtac tggattttgg
     9601 ttttaggaat tagaaatttt attgatagaa gtattttaca aatacaaata catactaagg
     9661 gtttcttata tgctcaacac atgagcgaaa ccctatagga accctaattc ccttatctgg
     9721 gaactactca cacattatta tggagaaact cgagcttgtc gatcgacaga tcccggtcgg
     9781 catctactct atttctttgc cctcggacga gtgctggggc gtcggtttcc actatcggcg
     9841 agtacttcta cacagccatc ggtccagacg gccgcgcttc tgcgggcgat ttgtgtacgc
     9901 ccgacagtcc cggctccgga tcggacgatt gcgtcgcatc gaccctgcgc ccaagctgca
     9961 tcatcgaaat tgccgtcaac caagctctga tagagttggt caagaccaat gcggagcata
    10021 tacgcccgga gtcgtggcga tcctgcaagc tccggatgcc tccgctcgaa gtagcgcgtc
    10081 tgctgctcca tacaagccaa ccacggcctc cagaagaaga tgttggcgac ctcgtattgg
    10141 gaatccccga acatcgcctc gctccagtca atgaccgctg ttatgcggcc attgtccgtc
    10201 aggacattgt tggagccgaa atccgcgtgc acgaggtgcc ggacttcggg gcagtcctcg
    10261 gcccaaagca tcagctcatc gagagcctgc gcgacggacg cactgacggt gtcgtccatc
    10321 acagtttgcc agtgatacac atggggatca gcaatcgcgc atatgaaatc acgccatgta
    10381 gtgtattgac cgattccttg cggtccgaat gggccgaacc cgctcgtctg gctaagatcg
    10441 gccgcagcga tcgcatccat agcctccgcg accggttgta gaacagcggg cagttcggtt
    10501 tcaggcaggt cttgcaacgt gacaccctgt gaacggcggg agatgcaata ggtcaggctc
    10561 tcgctaaact ccccaatgtc aagcacttcc ggaatcggga gcgcggccga tgcaaagtgc
    10621 cgataaacat aacgatcttt gtagaaacca tcggcgcagc tatttacccg caggacatat
    10681 ccacgccctc ctacatcgaa gctgaaagca cgagattctt cgccctccga gagctgcatc
    10741 aggtcggaga cgctgtcgaa cttttcgatc agaaacttct cgacagacgt cgcggtgagt
    10801 tcaggctttt tcatatctca ttgccccccg gatctgcgaa agctcgagag agatagattt
    10861 gtagagagag actggtgatt tcagcgtgtc ctctccaaat gaaatgaact tccttatata
    10921 gaggaagggt cttgcgaagg atagtgggat tgtgcgtcat cccttacgtc agtggagata
    10981 tcacatcaat ccacttgctt tgaagacgtg gttggaacgt cttctttttc cacgatgctc
    11041 ctcgtgggtg ggggtccatc tttgggacca ctgtcggcag aggcatcttg aacgatagcc
    11101 tttcctttat cgcaatgatg gcatttgtag gtgccacctt ccttttctac tgtccttttg
    11161 atgaagtgac agatagctgg gcaatggaat ccgaggaggt ttcccgatat taccctttgt
    11221 tgaaaagtct caatagccct ttggtcttct gagactgtat ctttgatatt cttggagtag
    11281 acgagagtgt cgtgctccac catgttcaca tcaatccact tgctttgaag acgtggttgg
    11341 aacgtcttct ttttccacga tgctcctcgt gggtgggggt ccatctttgg gaccactgtc
    11401 ggcagaggca tcttgaacga tagcctttcc tttatcgcaa tgatggcatt tgtaggtgcc
    11461 accttccttt tctactgtcc ttttgatgaa gtgacagata gctgggcaat ggaatccgag
    11521 gaggtttccc gatattaccc tttgttgaaa agtctcaata gccctttggt cttctgagac
    11581 tgtatctttg atattcttgg agtagacgag agtgtcgtgc tccaccatgt tggcaagctg
    11641 ctctagccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
    11701 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
    11761 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
    11821 attgtgagcg gataacaatt tcacacagga aacagctatg acatgattac gaattcccga
    11881 tctagtaaca tagatgacac cgcgcgcgat aatttatcct agtttgcgcg ctatattttg
    11941 ttttctatcg cgtattaaat gtataattgc gggactctaa tcataaaaac ccatctcata
    12001 aataacgtca tgcattacat gttaattatt acatgcttaa cgtaattcaa cagaaattat
    12061 atgataatca tcgcaagacc ggcaacagga ttcaatctta agaaacttta ttgccaaatg
    12121 tttgaacgat cggggaaatt cgagctcgga tcaattcctg cagccgggcg gccgctttac
    12181 ttgtacagct cgtccatgcc gtgagtgatc ccggcggcgg tcacgaactc cagcaggacc
    12241 atgtgatcgc gcttctcgtt ggggtctttg ctcagggcgg actgggtgct caggtagtgg
    12301 ttgtcgggca gcagcacggg gccgtcgccg atgggggtgt tctgctggta gtggtcggcg
    12361 agctgcacgc tgccgtcctc gatgttgtgg cggatcttga agttcacctt gatgccgttc
    12421 ttctgcttgt cggccatgat atagacgttg tggctgttgt agttgtactc cagcttgtgc
    12481 cccaggatgt tgccgtcctc cttgaagtcg atgcccttca gctcgatgcg gttcaccagg
    12541 gtgtcgccct cgaacttcac ctcggcgcgg gtcttgtagt tgccgtcgtc cttgaagaag
    12601 atggtgcgct cctggacgta gccttcgggc atggcggact tgaagaagtc gtgctgcttc
    12661 atgtggtcgg ggtagcggct gaagcactgc acgccgtagg tgaaggtggt cacgagggtg
    12721 ggccagggca cgggcagctt gccggtggtg cagatgaact tcagggtcag cttgccgtag
    12781 gtggcatcgc cctcgccctc gccggacacg ctgaacttgt ggccgtttac gtcgccgtcc
    12841 agctcgacca ggatgggcac caccccggtg aacagctcct cgcccttgct caccatggga
    12901 tcgaattgat cctctagagt cccccgtgtt ctctccaaat gaaatgaact tccttatata
    12961 gaggaagggt cttgcgaagg atagtgggat tgtgcgtcat cccttacgtc agtggagata
    13021 tcacatcaat ccacttgctt tgaagacgtg gttggaacgt cttctttttc cacgatgctc
    13081 ctcgtgggtg ggggtccatc tttgggacca ctgtcggcag aggcatcttc aacgatggcc
    13141 tttcctttat cgcaatgatg gcatttgtag gagccacctt ccttttccac tatcttcaca
    13201 ataaagtgac agatagctgg gcaatggaat ccgaggaggt ttccggatat taccctttgt
    13261 tgaaaagtct caattgccct ttggtcttct gagactgtat ctttgatatt tttggagtag
    13321 acaagtgtgt cgtgctccac catgttgacg aagattttct tcttgtcatt gagtcgtaag
    13381 agactctgta tgaactgttc gccagtcttt acggcgagtt ctgttaggtc ctctatttga
    13441 atctttgact ccatggcctt tgattcagtg ggaactacct ttttagagac tccaatctct
    13501 attacttgcc ttggtttgtg aagcaagcct tgaatcgtcc atactggaat agtacttctg
    13561 atcttgagaa atatatcttt ctctgtgttc ttgatgcagt tagtcctgaa tcttttgact
    13621 gcatctttaa ccttcttggg aaggtatttg atttcctgga gattattgct cgggtagatc
    13681 gtcttgatga gacctgctgc gt
//

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