pJBEI-6409

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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pJBEI6409载体基本信息

质粒名称: pJBEI-6409 
出品公司: 爱迪基因
目录编号: 47048
存储实验室: Taek Soon Lee

载体骨架基本信息
载体名称: pBbA5c
载体出品公司: --
载体抗性: 氯霉素
空载体大小: 3500 bp
完整质粒大小: 14932 bp
载体修饰: --
载体类型: 细菌表达,合成生物学;布格瑞克
筛选标记: --
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: Codon-optimized sequences for MEV pathway expression in E. coli to produce limonene
物种来源: S. cerevisiae (budding yeast); Mentha spicata, Abies grandis and Staphylococcus aureus
插入大小: 11500 bp
基因突变: --
GenBank编号 --
启动子 PlacUV5
标签/融合蛋白: --
克隆方法: 限制性内切酶
5' 克隆位点: EcoRI, BglII (未破坏)
3' 克隆位点: BamHI, XhoI (未破坏)
5' 测序引物 F5
3' 测序引物 R1

文献引用方法
材料和方法部分: pJBEI-6409 was a gift from Taek Soon Lee (Addgene plasmid # 47048 ; http://n2t.net/addgene:47048 ; RRID:Addgene_47048)
参考文献部分: Metabolic engineering of Escherichia coli for limonene and perillyl alcohol production. Alonso-Gutierrez J, Chan R, Batth TS, Adams PD, Keasling JD, Petzold CJ, Lee TS. Metab Eng. 2013 May 29;19C:33-41. doi: 10.1016/j.ymben.2013.05.004. 10.1016/j.ymben.2013.05.004 PubMed 23727191

pJBEI6409载体图谱质粒图谱和多克隆位点信息

pJBEI-6409质粒图谱

pJBEI6409载体简介

pJBEI6409载体序列

LOCUS       pJBEI-6409               14932 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  BglBrick plasmid 
            (=pBbA5c-MTSAe-T1f-MBI(f)-T1002i-Ptrc-trGPPS(co)-LS) coding for MEV 
            pathway enzymes to produce limonene from glucose in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    pJBEI-6409
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14932)
  AUTHORS   Alonso-Gutierrez J, Chan R, Batth TS, Adams PD, Keasling JD, Petzold
            CJ, Lee TS
  TITLE     Metabolic engineering of Escherichia coli for limonene and perillyl 
            alcohol production.
  JOURNAL   Metab Eng. 2013 May 29;19C:33-41. doi: 10.1016/j.ymben.2013.05.004.
  PUBMED    23727191
REFERENCE   2  (bases 1 to 14932)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..14932
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             complement(49..1131)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     1093..1112
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1132..1209)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1415..1434
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        1520..1550
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    1558..1574
                     /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)."
     CDS             complement(2309..2320)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     terminator      5324..5395
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5411..5438
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        5650..5679
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    5687..5703
                     /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)."
     CDS             8726..8737
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        10432..10461
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    10469..10485
                     /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)."
     terminator      13097..13168
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      13184..13211
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(13239..13256)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(13373..13918)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(13453..13472)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      14032..14126
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(14147..14806)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     14721..14740
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(14807..14909)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
ORIGIN
        1 gacgtcggtg cctaatgagt gagctaactt acattaattg cgttgcgctc actgcccgct
       61 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
      121 ggcggtttgc gtattgggcg ccagggtggt ttttcttttc accagtgaga cgggcaacag
      181 ctgattgccc ttcaccgcct ggccctgaga gagttgcagc aagcggtcca cgctggtttg
      241 ccccagcagg cgaaaatcct gtttgatggt ggttaacggc gggatataac atgagctgtc
      301 ttcggtatcg tcgtatccca ctaccgagat gtccgcacca acgcgcagcc cggactcggt
      361 aatggcgcgc attgcgccca gcgccatctg atcgttggca accagcatcg cagtgggaac
      421 gatgccctca ttcagcattt gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc
      481 ttcccgttcc gctatcggct gaatttgatt gcgagtgaga tatttatgcc agccagccag
      541 acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt gctggtgacc
      601 caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga aaataatact
      661 gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag tgcaggcagc
      721 ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc cactgacgcg
      781 ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc gttctaccat
      841 cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg
      901 cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg actgtttgcc
      961 cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg ccgcttccac
     1021 tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg aaacggtctg
     1081 ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca cattcaccac
     1141 cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcgccattc
     1201 gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg aagcagccca
     1261 gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg
     1321 cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca
     1381 tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag
     1441 caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga
     1501 tcgtttaggc accccaggct ttacacttta tgcttccggc tcgtataatg tgtggaattg
     1561 tgagcggata acaatttcag aattcaaaag atcttaggag gaatataaaa tgaaaaattg
     1621 tgtcatcgtc agtgcggtac gtactgctat cggtagtttt aacggttcac tcgcttccac
     1681 cagcgccatc gacctggggg cgacagtaat taaagccgcc attgaacgtg caaaaatcga
     1741 ttcacaacac gttgatgaag tgattatggg taacgtgtta caagccgggc tggggcaaaa
     1801 tccggcgcgt caggcactgt taaaaagcgg gctggcagaa acggtgtgcg gattcacggt
     1861 caataaagta tgtggttcgg gtcttaaaag tgtggcgctt gccgcccagg ccattcaggc
     1921 aggtcaggcg cagagcattg tggcgggggg tatggaaaat atgagtttag ccccctactt
     1981 actcgatgca aaagcacgct ctggttatcg tcttggagac ggacaggttt atgacgtaat
     2041 cctgcgcgat ggcctgatgt gcgccaccca tggttatcat atggggatta ccgccgaaaa
     2101 cgtggctaaa gagtacggaa ttacccgtga aatgcaggat gaactggcgc tacattcaca
     2161 gcgtaaagcg gcagccgcaa ttgagtccgg tgcttttaca gccgaaatcg tcccggtaaa
     2221 tgttgtcact cgaaagaaaa ccttcgtctt cagtcaagac gagttcccga aagcgaactc
     2281 aacggctgaa gcgttaggtg cattgcgccc ggccttcgat aaagcaggaa cagtcaccgc
     2341 tgggaacgcg tctggtatta acgacggtgc tgccgctctg gtgattatgg aagaatctgc
     2401 ggcgctggca gcaggcctta cccccctggc tcgcattaaa agttatgcca gcggtggcgt
     2461 gccccccgca ttgatgggta tggggccagt acctgccacg caaaaagcgt tacaactggc
     2521 ggggctgcaa ctggcggata ttgatctcat tgaggctaat gaagcatttg ctgcacagtt
     2581 ccttgccgtt gggaaaaacc tgggctttga ttctgagaaa gtgaatgtca acggcggggc
     2641 catcgcgctc gggcatccta tcggtgccag tggtgctcgt attctggtca cactattaca
     2701 tgccatgcag gcacgcgata aaacgctggg gctggcaaca ctgtgcattg gcggcggtca
     2761 gggaattgcg atggtgattg aacggttgaa ttgaggatct tgaattaagg aggacagcta
     2821 aatgacaata ggtatcgata aaataaactt ttacgttcca aagtactatg tagacatggc
     2881 taaattagca gaagcacgcc aagtagaccc aaacaaattt ttaattggaa ttggtcaaac
     2941 tgaaatggct gttagtcctg taaaccaaga catcgtttca atgggcgcta acgctgctaa
     3001 ggacattata acagacgaag acaaaaagaa aattggtatg gtaattgtgg caactgaatc
     3061 agcagttgat gctgctaaag cagccgctgt tcaaattcac aacttattag gtattcaacc
     3121 ttttgcacgc tgctttgaaa tgaaagaagc ttgttatgct gcaacaccag caattcaatt
     3181 agctaaagat tatttagcaa ctagaccgaa tgaaaaagta ttagttattg ctacagatac
     3241 agcacgttat ggattgaact caggcggcga gccaacacaa ggtgctggcg cagttgcgat
     3301 ggttattgca cataatccaa gcattttggc attaaatgaa gatgctgttg cttacactga
     3361 agacgtttat gatttctggc gtccaactgg acataaatat ccattagttg atggtgcatt
     3421 atctaaagat gcttatatcc gctcattcca acaaagctgg aatgaatacg caaaacgtca
     3481 aggtaagtcg ctagctgact tcgcatctct atgcttccat gttccattta caaaaatggg
     3541 taaaaaggca ttagagtcaa tcattgataa cgctgatgaa acaactcaag agcgtttacg
     3601 ttcaggatat gaagatgctg tagattataa ccgttatgtc ggtaatattt atactggatc
     3661 attatattta agcctaatat cattacttga aaatcgagat ttacaagctg gtgaaacaat
     3721 cggtttattc agttatggct caggttcagt tggtgaattt tatagtgcga cattagttga
     3781 aggctacaaa gatcatttag atcaagctgc acataaagca ttattaaata accgtactga
     3841 agtatctgtt gatgcatatg aaacattctt caaacgtttt gatgacgttg aatttgacga
     3901 agaacaagat gctgttcatg aagatcgtca tattttctac ttatcaaata ttgaaaataa
     3961 cgttcgcgaa tatcacagac cagagtaatt aggatctatt caggaaacag accatgtcca
     4021 tgcaaagttt agataagaat tttcgacatt tatctcgtaa agaaaagtta caacaattgg
     4081 ttgataagca atggttatca gaagaacaat tcgacatttt actgaatcat ccattaatcg
     4141 atgaagaagt agccaatagt ttaattgaaa atgtcatcgc gcaaggtgca ttacccgttg
     4201 gattattacc gaatatcatt gtggacgata aggcatatgt tgtacctatg atggtggaag
     4261 agccttcagt tgtcgctgca gctagttatg gtgcaaagct agtgaatcag actggcggat
     4321 ttaaaacggt atcttctgaa cgtattatga taggtcaaat cgtctttgat ggcgttgacg
     4381 atactgaaaa attatcagca gacattaaag ctttagaaaa gcaaattcat aaaattgcgg
     4441 atgaggcata tccttctatt aaagcgcgtg gtggtggtta ccaacgtata gcgattgata
     4501 catttcctga gcaacagtta ctatctttaa aagtatttgt tgatacgaaa gatgctatgg
     4561 gcgctaatat gcttaatacg attttagagg ccataactgc atttttaaaa aatgaatttc
     4621 cgcaaagcga cattttaatg agtattttat ccaatcatgc aacagcgtcc gttgttaaag
     4681 ttcaaggcga aattgatgtt aaagatttag caaggggcga gagaactgga gaagaggttg
     4741 ccaaacgaat ggaacgtgct tctgtattgg cccaagtaga tattcatcgt gcagcaacac
     4801 ataataaagg tgttatgaat ggcatacatg ctgttgtttt agcaacagga aatgatacgc
     4861 gtggtgcaga agcaagtgcg catgcatacg cgagtcgtga cggacagtat cgtggtattg
     4921 ctacatggcg ttacgatcaa gatcgtcaac gattgattgg tacaattgaa gtgcctatga
     4981 cattggcaat cgttggcggt ggtacaaaag tattaccaat tgctaaagct tcattagagc
     5041 tactaaatgt agagtcagca caagaattag gtcatgtagt tgctgccgtt ggtttagcgc
     5101 aaaactttgc agcatgtcgc gcgcttgtgt cagaaggtat tcaacaaggt catatgagtt
     5161 tacaatataa atcattagct atcgttgtag gggcaaaagg tgatgaaatt gctaaagtag
     5221 ctgaagcttt gaaaaaagaa ccccgtgcaa atacacaagc agcggaacat attttacaag
     5281 aaattagaca acaataagga tctttttaag gatctccagg catcaaataa aacgaaaggc
     5341 tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctctacta
     5401 gagtcacact ggctcacctt cgggtgggcc tttctgcgtt tatagcgaat tgatctggtt
     5461 tgacagctta tcatcgactg cacggtgcac caatgcttct ggcgtcaggc agccatcgga
     5521 agctgtggta tggctgtgca ggtcgtaaat cactgcataa ttcgtgtcgc tcaaggcgca
     5581 ctcccgttct ggataatgtt ttttgcgccg acatcataac ggttctggca aatattctga
     5641 aatgagctgt tgacaattaa tcatccggct cgtataatgt gtggaattgt gagcggataa
     5701 caatttcagg atctaggagg aaataaccat gtctctgcca ttcctgacgt ctgcgccagg
     5761 taaggtgatc atcttcggcg agcactctgc ggtgtacaat aagccggccg tcgccgcctc
     5821 tgtgtctgcg ttacgcacct acctgctgat cagcgaatct tctgcaccgg acacgatcga
     5881 gctggacttt ccggacatca gcttcaacca caagtggagc atcaacgact tcaacgcgat
     5941 cacggaggac caggtgaaca gccaaaagct ggccaaagcc cagcaagcaa ccgacggtct
     6001 gtctcaggag ctggtgtctc tgctggaccc gctgttagcg cagttaagcg agagcttcca
     6061 ttaccacgcc gcgttctgct tcctgtacat gttcgtttgc ctgtgcccgc acgcaaagaa
     6121 catcaagttc agcctgaaga gcacgctgcc gattggcgca ggcttaggct ctagcgcatc
     6181 tatcagcgtg agcctggcgc tggcgatggc ctatctgggt ggcctgattg gcagcaacga
     6241 cctggagaaa ctgagcgaaa acgacaagca catcgtgaac cagtgggcct ttatcggcga
     6301 gaagtgcatt catggcaccc cgagcggcat tgacaacgca gttgccacgt atggcaacgc
     6361 cctgctgttc gagaaagaca gccacaacgg cacgatcaac acgaacaact tcaagttcct
     6421 ggacgacttc ccggcgatcc cgatgattct gacctacacc cgtatcccac gcagcaccaa
     6481 ggatttagtc gcccgcgtgc gtgttttagt caccgaaaag ttcccggagg tgatgaagcc
     6541 gatcctggac gcgatgggcg agtgcgcgct gcagggtctg gagatcatga ccaagctgag
     6601 caagtgcaag ggcaccgacg atgaggcggt ggagaccaac aatgagctgt acgagcagct
     6661 gctggagctg atccgtatca atcacggcct gctggtctct atcggtgtgt ctcacccggg
     6721 cctggaactg atcaaaaacc tgagcgacga cctgcgcatt ggctctacga aattaacggg
     6781 tgcaggtggc ggtggctgct ctttaacgct gctgcgccgt gacattacgc aggagcaaat
     6841 cgacagcttc aagaagaagc tgcaggacga cttcagctac gagacgttcg agacggacct
     6901 gggcggcacg ggctgttgcc tgctgagcgc caaaaatctg aacaaggacc tgaagatcaa
     6961 aagcctggtg ttccagctgt tcgaaaacaa gacgaccacg aagcagcaga tcgacgacct
     7021 gttactgccg ggtaacacca atctgccgtg gacgtcttaa ggatctagga gggagatcat
     7081 atgagcgaat tacgtgcatt cagcgcgcca ggtaaggcac tgctggccgg tggctacctg
     7141 gtgttagaca ccaagtacga ggcgttcgtc gtcggcttat ctgcccgtat gcatgcagtt
     7201 gcccacccgt atggtagcct gcagggctct gacaagttcg aagtgcgtgt gaagagcaag
     7261 cagttcaagg acggcgagtg gctgtaccac attagcccaa agagcggctt catcccggtt
     7321 agcattggtg gcagcaagaa cccatttatc gagaaggtca ttgccaacgt cttcagctac
     7381 ttcaagccga atatggacga ttactgcaac cgcaacctgt tcgtcatcga cattttcagc
     7441 gacgacgcgt accacagcca agaggactct gttacggagc atcgtggtaa ccgccgcctg
     7501 agcttccaca gccatcgcat tgaggaggtg ccgaagacgg gtctgggttc tagcgccggt
     7561 ttagttaccg tcttaacgac ggcgttagcg agcttcttcg tgagcgacct ggagaacaac
     7621 gtggacaagt accgcgaagt gattcataac ctggcgcagg tggcacattg tcaggcccaa
     7681 ggtaagattg gctctggttt tgatgtggca gcggccgcct atggctctat ccgctatcgc
     7741 cgctttccgc cggccctgat cagcaatctg ccggacatcg gctctgcgac gtatggtagc
     7801 aaactggcgc atctggtgga cgaagaagac tggaacatca ccattaagtc taatcacctg
     7861 ccgagcggct taacgttatg gatgggcgat atcaagaacg gcagcgaaac ggttaagctg
     7921 gtgcagaaag tgaaaaactg gtacgacagc cacatgccgg aaagcctgaa gatttacacg
     7981 gagctggacc acgccaatag ccgtttcatg gatggtctga gcaagctgga ccgcctgcac
     8041 gaaacccacg acgactacag cgaccaaatc ttcgagagcc tggagcgcaa tgactgcacc
     8101 tgccagaagt acccggagat cacggaggtc cgcgatgccg tggcaacgat tcgccgtagc
     8161 ttccgcaaaa ttacgaagga gagcggcgcg gatatcgaac caccggtcca gacgtctctg
     8221 ctggacgact gtcaaacctt aaagggcgtg ttaacgtgcc tgattccggg cgcgggtggt
     8281 tacgacgcca ttgccgtcat cacgaaacag gacgtcgatc tgcgcgcaca aacggccaac
     8341 gacaaacgtt tcagcaaagt ccaatggctg gatgttacgc aggccgactg gggtgttcgc
     8401 aaggagaagg acccggaaac gtatctggat aagtgaggat ctaggaggat tatgagatga
     8461 ccgtttacac agcatccgtt accgcacccg tcaacatcgc aacccttaag tattggggga
     8521 aaagggacac gaagttgaat ctgcccacca attcgtccat atcagtgact ttatcgcaag
     8581 atgacctcag aacgttgacc tctgcggcta ctgcacctga gtttgaacgc gacactttgt
     8641 ggttaaatgg agaaccacac agcatcgaca atgaaagaac tcaaaattgt ctgcgcgacc
     8701 tacgccaatt aagaaaggaa atggaatcga aggacgcctc attgcccaca ttatctcaat
     8761 ggaaactcca cattgtctcc gaaaataact ttcctacagc agctggttta gcttcctccg
     8821 ctgctggctt tgctgcattg gtctctgcaa ttgctaagtt ataccaatta ccacagtcaa
     8881 cttcagaaat atctagaata gcaagaaagg ggtctggttc agcttgtaga tcgttgtttg
     8941 gcggatacgt ggcctgggaa atgggaaaag ctgaagatgg tcatgattcc atggcagtac
     9001 aaatcgcaga cagctctgac tggcctcaga tgaaagcttg tgtcctagtt gtcagcgata
     9061 ttaaaaagga tgtgagttcc actcagggta tgcaattgac cgtggcaacc tccgaactat
     9121 ttaaagaaag aattgaacat gtcgtaccaa agagatttga agtcatgcgt aaagccattg
     9181 ttgaaaaaga tttcgccacc tttgcaaagg aaacaatgat ggattccaac tctttccatg
     9241 ccacatgttt ggactctttc cctccaatat tctacatgaa tgacacttcc aagcgtatca
     9301 tcagttggtg ccacaccatt aatcagtttt acggagaaac aatcgttgca tacacgtttg
     9361 atgcaggtcc aaatgctgtg ttgtactact tagctgaaaa tgagtcgaaa ctctttgcat
     9421 ttatctataa attgtttggc tctgttcctg gatgggacaa gaaatttact actgagcagc
     9481 ttgaggcttt caaccatcaa tttgaatcat ctaactttac tgcacgtgaa ttggatcttg
     9541 agttgcaaaa ggatgttgcc agagtgattt taactcaagt cggttcaggc ccacaagaaa
     9601 caaacgaatc tttgattgac gcaaagactg gtctaccaaa ggaataagga tctaggaggt
     9661 aatgataatg caaacggaac acgtcatttt attgaatgca cagggagttc ccacgggtac
     9721 gctggaaaag tatgccgcac acacggcaga cacccgctta catctcgcgt tctccagttg
     9781 gctgtttaat gccaaaggac aattattagt tacccgccgc gcactgagca aaaaagcatg
     9841 gcctggcgtg tggactaact cggtttgtgg gcacccacaa ctgggagaaa gcaacgaaga
     9901 cgcagtgatc cgccgttgcc gttatgagct tggcgtggaa attacgcctc ctgaatctat
     9961 ctatcctgac tttcgctacc gcgccaccga tccgagtggc attgtggaaa atgaagtgtg
    10021 tccggtattt gccgcacgca ccactagtgc gttacagatc aatgatgatg aagtgatgga
    10081 ttatcaatgg tgtgatttag cagatgtatt acacggtatt gatgccacgc cgtgggcgtt
    10141 cagtccgtgg atggtgatgc aggcgacaaa tcgcgaagcc agaaaacgat tatctgcatt
    10201 tacccagctt aaataaggat ctcgcaaaaa accccgcttc ggcggggttt tttcgccgac
    10261 tgcacggtgc accaatgctt ctggcgtcag gcagccatcg gaagctgtgg tatggctgtg
    10321 caggtcgtaa atcactgcat aattcgtgtc gctcaaggcg cactcccgtt ctggataatg
    10381 ttttttgcgc cgacatcata acggttctgg caaatattct gaaatgagct gttgacaatt
    10441 aatcatccgg ctcgtataat gtgtggaatt gtgagcggat aacaatttca ggatctttta
    10501 agaaggagat atacatggaa tttgacttca acaaatacat ggactccaaa gcgatgacgg
    10561 taaatgaagc actgaacaaa gcgatccctc tgcgttatcc gcagaaaatc tacgaaagca
    10621 tgcgttacag cctgctggca ggcggcaagc gtgttcgtcc ggttctgtgt attgccgcat
    10681 gtgaactggt aggtggtacc gaagaactgg cgatcccgac cgcgtgcgca attgaaatga
    10741 tccacacgat gtccctgatg cacgatgatc tgccgtgtat cgacaacgac gatctgcgtc
    10801 gcggtaaacc gactaaccac aaaattttcg gtgaggatac cgcagtgact gctggtaacg
    10861 cactgcactc ttacgccttc gagcatatcg cggtttctac ttctaaaacc gttggtgctg
    10921 accgcatcct gcgtatggtg tccgagctgg gtcgtgctac tggctctgaa ggtgttatgg
    10981 gtggtcagat ggtagacatc gcatccgaag gcgatccgtc tatcgacctg cagaccctgg
    11041 aatggattca catccacaaa accgcaatgc tgctggaatg ctccgttgtt tgcggtgcaa
    11101 tcattggcgg tgccagcgaa atcgtaatcg aacgtgcccg tcgctacgcc cgctgtgttg
    11161 gtctgctgtt ccaggtagtt gatgacattc tggacgtaac taaaagcagc gacgaactgg
    11221 gtaagactgc gggcaaggac ctgatctctg ataaagccac ctacccaaag ctgatgggtc
    11281 tggaaaaggc caaggagttc tccgatgaac tgctgaaccg tgcgaagggt gaactgtcct
    11341 gcttcgaccc agttaaagcc gctccgctgc tgggcctggc agactacgtg gcatttcgtc
    11401 agaattaagg atcttttaag aaggagatat acatgcgtcg cagtggtaat tacaacccgt
    11461 cacgttggga tgtgaatttt atccaatccc tgctgagcga ttacaaagag gacaaacacg
    11521 tcattcgggc aagcgaactg gttacactgg ttaaaatgga actggaaaaa gaaacagacc
    11581 aaattcgcca attagagctg attgacgact tacagcgcat ggggctgagc gatcactttc
    11641 agaatgagtt caaagagatt ctgtcatcta tctatctgga ccaccattac tataaaaacc
    11701 cttttccgaa agaggaacgt gatctgtata gcacaagcct ggcctttcgg ttactgcgtg
    11761 agcatggctt ccaagtagca caagaggtat tcgatagctt caagaatgaa gagggcgagt
    11821 tcaaagaatc actgtccgat gatactcgtg gcttattaca actgtatgaa gcgagctttc
    11881 tgctgaccga gggggagacg actctggaaa gcgcacggga atttgcgacc aaattcttag
    11941 aagagaaagt aaatgaaggc ggcgtagatg gcgatctgtt aacccgcatc gcgtattctc
    12001 tggacattcc gttacactgg cgtattaaac ggccaaacgc gccggtgtgg attgagtggt
    12061 atcgcaaacg tccagacatg aacccggtgg tgctggaatt agcaatttta gacttaaaca
    12121 tcgtgcaggc tcaatttcag gaagaactga aagaatcttt tcgctggtgg cgcaacaccg
    12181 gcttcgttga aaaattaccg tttgcgcgcg atcgtctggt tgaatgttat ttctggaata
    12241 ccggcattat cgaaccgcgc cagcacgcga gcgcccgtat tatgatgggt aaggtcaatg
    12301 cgctgattac tgtcattgac gatatttacg atgtatacgg cactctggaa gaactggagc
    12361 agtttacaga tttaatccgc cggtgggaca tcaatagcat cgaccagctg ccagattaca
    12421 tgcagctgtg ttttctggcg ctgaataact ttgtggacga cacctcttat gacgtcatga
    12481 aagaaaaagg cgtgaatgtt atcccttatc tgcgtcagag ttgggtggat ctggccgata
    12541 aatatatggt tgaggcacgc tggttctacg gtgggcacaa accgagttta gaagaatatc
    12601 tggagaactc atggcagagt atcagtgggc catgcatgct gacgcatatc ttctttcgcg
    12661 tcacggattc atttaccaag gagacggtcg attccttata taaataccat gacctggttc
    12721 gttggtcatc tttcgtcctg cggctggcgg acgatctggg tacgtctgtg gaagaggtgt
    12781 ctcgtgggga tgttccgaaa tccttacagt gctacatgtc cgattataac gcaagtgaag
    12841 ctgaagctcg caaacatgtg aagtggctga tcgccgaagt gtggaagaag atgaacgctg
    12901 aacgtgtttc caaggatagt ccgtttggta aggattttat tggttgcgcc gttgatctgg
    12961 gtcgtatggc ccagctgatg tatcataacg gtgatggtca tgggacccag catcctatta
    13021 ttcatcagca gatgacgcgt accctgtttg cacctttcgc ctaaggatcc aaactcgagt
    13081 aaggatctcc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt
    13141 tatctgttgt ttgtcggtga acgctctcta ctagagtcac actggctcac cttcgggtgg
    13201 gcctttctgc gtttatacct agggatatat tccgcttcct cgctcactga ctcgctacgc
    13261 tcggtcgttc gactgcggcg agcggaaatg gcttacgaac ggggcggaga tttcctggaa
    13321 gatgccagga agatacttaa cagggaagtg agagggccgc ggcaaagccg tttttccata
    13381 ggctccgccc ccctgacaag catcacgaaa tctgacgctc aaatcagtgg tggcgaaacc
    13441 cgacaggact ataaagatac caggcgtttc cccctggcgg ctccctcgtg cgctctcctg
    13501 ttcctgcctt tcggtttacc ggtgtcattc cgctgttatg gccgcgtttg tctcattcca
    13561 cgcctgacac tcagttccgg gtaggcagtt cgctccaagc tggactgtat gcacgaaccc
    13621 cccgttcagt ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggaa
    13681 agacatgcaa aagcaccact ggcagcagcc actggtaatt gatttagagg agttagtctt
    13741 gaagtcatgc gccggttaag gctaaactga aaggacaagt tttggtgact gcgctcctcc
    13801 aagccagtta cctcggttca aagagttggt agctcagaga accttcgaaa aaccgccctg
    13861 caaggcggtt ttttcgtttt cagagcaaga gattacgcgc agaccaaaac gatctcaaga
    13921 agatcatctt attaatcaga taaaatattt ctagatttca gtgcaattta tctcttcaaa
    13981 tgtagcacct gaagtcagcc ccatacgata taagttgtta ctagtgcttg gattctcacc
    14041 aataaaaaac gcccggcggc aaccgagcgt tctgaacaaa tccagatgga gttctgaggt
    14101 cattactgga tctatcaaca ggagtccaag cgagctcgat atcaaattac gccccgccct
    14161 gccactcatc gcagtactgt tgtaattcat taagcattct gccgacatgg aagccatcac
    14221 aaacggcatg atgaacctga atcgccagcg gcatcagcac cttgtcgcct tgcgtataat
    14281 atttgcccat ggtgaaaacg ggggcgaaga agttgtccat attggccacg tttaaatcaa
    14341 aactggtgaa actcacccag ggattggctg agacgaaaaa catattctca ataaaccctt
    14401 tagggaaata ggccaggttt tcaccgtaac acgccacatc ttgcgaatat atgtgtagaa
    14461 actgccggaa atcgtcgtgg tattcactcc agagcgatga aaacgtttca gtttgctcat
    14521 ggaaaacggt gtaacaaggg tgaacactat cccatatcac cagctcaccg tctttcattg
    14581 ccatacgaaa ttccggatga gcattcatca ggcgggcaag aatgtgaata aaggccggat
    14641 aaaacttgtg cttatttttc tttacggtct ttaaaaaggc cgtaatatcc agctgaacgg
    14701 tctggttata ggtacattga gcaactgact gaaatgcctc aaaatgttct ttacgatgcc
    14761 attgggatat atcaacggtg gtatatccag tgattttttt ctccatttta gcttccttag
    14821 ctcctgaaaa tctcgataac tcaaaaaata cgcccggtag tgatcttatt tcattatggt
    14881 gaaagttgga acctcttacg tgccgatcaa cgtctcattt tcgccagata tc
//

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