pET28a_T7-ARG1

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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: pET28a_T7-ARG1 
出品公司: 爱迪基因
目录编号: 106473
存储实验室: Mikhail Shapiro

载体骨架基本信息
载体名称: pET28a
载体出品公司: Novagen
载体抗性: 卡那霉素
空载体大小: 5374 bp
完整质粒大小: 12129 bp
载体修饰: --
载体类型: 细菌表达
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: ARG1,Acoustic Reporter Gene 1,Hybrid Gas Vesicle gene cluster
物种来源: Anabaena flos-aquae and Bacillus megaterium
插入大小: 6755 bp
基因突变: --
GenBank编号 --
启动子 T7
标签/融合蛋白: --
克隆方法: Gibson克隆
5' 克隆位点: --
3' 克隆位点: --
5' 测序引物 T7_Forward
3' 测序引物 T7 terminator_Reverse

文献引用方法
材料和方法部分: pET28a_T7-ARG1 was a gift from Mikhail Shapiro (Addgene plasmid # 106473 ; http://n2t.net/addgene:106473 ; RRID:Addgene_106473)
参考文献部分: Acoustic reporter genes for noninvasive imaging of microorganisms in mammalian hosts. Bourdeau RW, Lee-Gosselin A, Lakshmanan A, Farhadi A, Kumar SR, Nety SP, Shapiro MG. Nature. 2018 Jan 3;553(7686):86-90. doi: 10.1038/nature25021. 10.1038/nature25021 PubMed 29300010

载体图谱质粒图谱和多克隆位点信息

pET28a_T7-ARG1质粒图谱

载体简介

载体序列

LOCUS       pET28a_T7-ARG1              12125 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Expresses acoustic reporter genes or gas vesicles (ARG1) in 
            Escherichia coli.
ACCESSION   .
VERSION     .
KEYWORDS    pET28a_T7-ARG1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12125)
  AUTHORS   Bourdeau RW, Lee-Gosselin A, Lakshmanan A, Farhadi A, Kumar SR, Nety
            SP, Shapiro MG
  TITLE     Acoustic reporter genes for noninvasive imaging of microorganisms in
            mammalian hosts.
  JOURNAL   Nature. 2018 Jan 3;553(7686):86-90. doi: 10.1038/nature25021.
  PUBMED    29300010
REFERENCE   2  (bases 1 to 12125)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..12125
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      12..467
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(99..118)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     309..330
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(559..1374)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     1282..1301
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1366..1385
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1496..2084
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1985..2004
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2238..2255
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2270..2409
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2335..2354)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2495..2517
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2511..2702)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3511..4593)
                     /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     4555..4574
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4594..4671)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     4877..4896
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     4980..4999
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4980..4998
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    4999..5023
                     /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             8305..8322
                     /codon_start=1
                     /product="N-myristoylation signal from Src kinase (Pellman 
                     et al., 1985; Kaplan et al., 1988)"
                     /label=myr
                     /translation="GSSKSK"
     primer_bind     complement(12039..12057)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      12053..12100
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
       61 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
      121 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
      181 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
      241 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
      301 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
      361 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
      421 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt
      481 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
      541 ccgctcatga attaattctt agaaaaactc atcgagcatc aaatgaaact gcaatttatt
      601 catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa
      661 ctcaccgagg cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg
      721 tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa
      781 atcaccatga gtgacgactg aatccggtga gaatggcaaa agtttatgca tttctttcca
      841 gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc
      901 gttattcatt cgtgattgcg cctgagcgag acgaaatacg cgatcgctgt taaaaggaca
      961 attacaaaca ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt
     1021 ttcacctgaa tcaggatatt cttctaatac ctggaatgct gttttcccgg ggatcgcagt
     1081 ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat
     1141 aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc
     1201 tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaatc gatagattgt
     1261 cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat
     1321 gttggaattt aatcgcggcc tagagcaaga cgtttcccgt tgaatatggc tcataacacc
     1381 ccttgtatta ctgtttatgt aagcagacag ttttattgtt catgaccaaa atcccttaac
     1441 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     1501 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     1561 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     1621 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
     1681 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     1741 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     1801 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     1861 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     1921 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     1981 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     2041 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     2101 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     2161 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     2221 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt
     2281 attttctcct tacgcatctg tgcggtattt cacaccgcaa tggtgcactc tcagtacaat
     2341 ctgctctgat gccgcatagt taagccagta tacactccgc tatcgctacg tgactgggtc
     2401 atggctgcgc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     2461 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     2521 tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga
     2581 agcgattcac agatgtctgc ctgttcatcc gcgtccagct cgttgagttt ctccagaagc
     2641 gttaatgtct ggcttctgat aaagcgggcc atgttaaggg cggttttttc ctgtttggtc
     2701 actgatgcct ccgtgtaagg gggatttctg ttcatggggg taatgatacc gatgaaacga
     2761 gagaggatgc tcacgatacg ggttactgat gatgaacatg cccggttact ggaacgttgt
     2821 gagggtaaac aactggcggt atggatgcgg cgggaccaga gaaaaatcac tcagggtcaa
     2881 tgccagcgct tcgttaatac agatgtaggt gttccacagg gtagccagca gcatcctgcg
     2941 atgcagatcc ggaacataat ggtgcagggc gctgacttcc gcgtttccag actttacgaa
     3001 acacggaaac cgaagaccat tcatgttgtt gctcaggtcg cagacgtttt gcagcagcag
     3061 tcgcttcacg ttcgctcgcg tatcggtgat tcattctgct aaccagtaag gcaaccccgc
     3121 cagcctagcc gggtcctcaa cgacaggagc acgatcatgc gcacccgtgg ggccgccatg
     3181 ccggcgataa tggcctgctt ctcgccgaaa cgtttggtgg cgggaccagt gacgaaggct
     3241 tgagcgaggg cgtgcaagat tccgaatacc gcaagcgaca ggccgatcat cgtcgcgctc
     3301 cagcgaaagc ggtcctcgcc gaaaatgacc cagagcgctg ccggcacctg tcctacgagt
     3361 tgcatgataa agaagacagt cataagtgcg gcgacgatag tcatgccccg cgcccaccgg
     3421 aaggagctga ctgggttgaa ggctctcaag ggcatcggtc gagatcccgg tgcctaatga
     3481 gtgagctaac ttacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     3541 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     3601 cgccagggtg gtttttcttt tcaccagtga gacgggcaac agctgattgc ccttcaccgc
     3661 ctggccctga gagagttgca gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc
     3721 ctgtttgatg gtggttaacg gcgggatata acatgagctg tcttcggtat cgtcgtatcc
     3781 cactaccgag atatccgcac caacgcgcag cccggactcg gtaatggcgc gcattgcgcc
     3841 cagcgccatc tgatcgttgg caaccagcat cgcagtggga acgatgccct cattcagcat
     3901 ttgcatggtt tgttgaaaac cggacatggc actccagtcg ccttcccgtt ccgctatcgg
     3961 ctgaatttga ttgcgagtga gatatttatg ccagccagcc agacgcagac gcgccgagac
     4021 agaacttaat gggcccgcta acagcgcgat ttgctggtga cccaatgcga ccagatgctc
     4081 cacgcccagt cgcgtaccgt cttcatggga gaaaataata ctgttgatgg gtgtctggtc
     4141 agagacatca agaaataacg ccggaacatt agtgcaggca gcttccacag caatggcatc
     4201 ctggtcatcc agcggatagt taatgatcag cccactgacg cgttgcgcga gaagattgtg
     4261 caccgccgct ttacaggctt cgacgccgct tcgttctacc atcgacacca ccacgctggc
     4321 acccagttga tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc
     4381 cagactggag gtggcaacgc caatcagcaa cgactgtttg cccgccagtt gttgtgccac
     4441 gcggttggga atgtaattca gctccgccat cgccgcttcc actttttccc gcgttttcgc
     4501 agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc tgataagaga caccggcata
     4561 ctctgcgaca tcgtataacg ttactggttt cacattcacc accctgaatt gactctcttc
     4621 cgggcgctat catgccatac cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc
     4681 gacgctctcc cttatgcgac tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt
     4741 tgagcaccgc cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac agtcccccgg
     4801 ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag
     4861 cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc
     4921 cggtgatgcc ggccacgatg cgtccggcgt agaggatcga gatctcgatc ccgcgaaatt
     4981 aatacgactc actatagggg aattgtgagc ggataacaat tccaagcttc gccagggttt
     5041 tcccagtcga gctcatgact catatttata gttattctcc tcatttccca agtttccgac
     5101 aacaaataac aaggaattaa aaaaatggca gttgaaaaaa ccaattcttc ctccagcttg
     5161 gcagaagtta ttgatagaat cctcgacaaa ggtatcgtaa ttgacgcttg ggttcgtgtt
     5221 tccttagttg gtatcgaact actagcaatt gaagctcgga tcgttatcgc ttccgttgaa
     5281 acctacttga agtatgctga agcagttggt ttgacccaat cagcagcagt acctgcttaa
     5341 tttaattaag caagttacca agtaggagaa taaccatgac tcatatttat agttattctc
     5401 ctcatttccc aagtttccga caacaaataa caaggaatta aaaaaatggc agttgaaaaa
     5461 accaattctt cctccagctt ggcagaagtt attgatagaa tcctcgacaa aggtatcgta
     5521 attgacgctt gggttcgtgt ttccttagtt ggtatcgaac tactagcaat tgaagctcgg
     5581 atcgttatcg cttccgttga aacctacttg aagtatgctg aagcagttgg tttgacccaa
     5641 tcagcagcag tacctgctta atttaattaa gctccataat atcaggatta attaatcctg
     5701 atattggttc tgcttttttt aattatctag atatttggaa atcaacgctt ctcaaaggat
     5761 attacaaaaa agtacgaatc cttataatgt agtacctgga gaagcaattt ctgttaggtc
     5821 tgggagctcc catgggatcg ttatcccgcg gccaagcttg ctagcgctaa tttactattt
     5881 tttggagaac ttcatgattt ctttaatggc aaaaatccgg caagaacatc agtcaatagc
     5941 agagaaagtg gctgaactat ctcttgagac cagagaattc ttgtccgtca cgacagcgaa
     6001 aagacaagag caagctgaaa aacaagctca agaactgcaa gcattctaca aggatcttca
     6061 ggaaacaagt cagcagtttt tatcagaaac agcccaagcc agaattgctc aagctgaaaa
     6121 acaagctcaa gaactgttag cattccacaa agaacttcaa gaaacaagtc agcagttttt
     6181 atcagcaaca gcccaagcca gaattgctca agctgaaaaa caagcgcaag aactgttagc
     6241 attttatcaa gaagttcggg aaacaagtca gcagttttta tcagcaacag cccaagcaag
     6301 aattgctcaa gctgaaaaac aagctcaaga actgttagca ttccacaaag aacttcaaga
     6361 aacaagtcag cagtttttat cagcaacagc cgacgcaaga actgctcaag ctaaggaaca
     6421 gaaggaatct ctcctgaaat tccgtcagga tttgtttgtg agtatctttg gttaataaag
     6481 atctaactat tggaggctac taaaaatgga aattaaaaaa attatgcaag ccgtgaacga
     6541 ctttttcggt gaacacgtag ctcctcctca taaaattacc tcggtggaag ctactgaaga
     6601 tgaaggttgg agagttattg ttgaagtcat tgaagaacga gaatatatga aaaaatacgc
     6661 caaagatgaa atgctcggaa cgtacgagtg ctttgtaaat aaagaaaaag aagtcatttc
     6721 attcaaacga ctcgacgtca gatatagaag cgccattggc attgaagcat aaacgataag
     6781 atggcaggag gaacgtaaaa atgaccgtct taacagacaa aaggaaaaaa ggcagtggag
     6841 cttttataca agatgacgag acaaaagagg ttctttcaag agcgctgagc tatttaaaat
     6901 ccggctattc cattcatttt acaggtcctg ccggcggagg caaaacctct ttagcgcgag
     6961 cgcttgctaa aaagagaaag cgtcctgtaa tgctgatgca cgggaatcac gagctcaaca
     7021 acaaagattt aattggcgat tttacgggat acacgagcaa aaaagtaatc gaccagtacg
     7081 ttcgttctgt ctataaaaaa gatgaacagg tgagtgaaaa ctggcaggat ggccgattgc
     7141 ttgaagctgt aaaaaatggc tatacgctga tttacgacga atttactcgt tctaagcctg
     7201 cgacgaataa tatctttcta tcgatattag aagaaggcgt gctgccgctg tatggagtaa
     7261 aaatgaccga tccttttgtg cgcgtgcatc ccgatttccg cgtcatcttc acaagcaatc
     7321 cagctgagta tgccggcgta tatgatacgc aagatgcgct tctcgacagg ttaattacca
     7381 tgtttattga ttataaagac atcgacagag agacagcgat tttaacggag aaaacggacg
     7441 tagaagaaga tgaagcgcgc acaattgtaa cgctcgtagc aaacgtgcga aaccgctctg
     7501 gagacgaaaa cagcagcgga cttagcctgc gggcttcgct tatgatcgct acccttgcca
     7561 cgcagcaaga cattcctatc gatggaagtg acgaagattt tcaaacgtta tgtatcgata
     7621 ttttgcatca tccgcttacc aaatgtttgg atgaagaaaa tgcaaaaagc aaagccgaaa
     7681 aaatcatttt agaagaatgt aagaatatag acactgaaga aaagtaaagg agcttgaaaa
     7741 catgagtgaa acaaacgaaa caggtattta tatttttagc gccattcaaa cggataaaga
     7801 cgaagaattt ggcgccgtgg aagtagaagg aacaaaagct gaaacatttt tgattcgcta
     7861 caaagacgcg gctatggtag cagctgaagt accgatgaaa atttatcatc ctaatcgcca
     7921 aaatttatta atgcatcaaa acgcagtagc agcgattatg gacaagaacg atacggttat
     7981 tccaatcagc tttgggaatg tattcaaatc aaaagaagac gtaaaagttc ttttggaaaa
     8041 cctttatccg cagtttgaaa agctgtttcc agcgatcaaa ggaaaaattg aagtcggttt
     8101 aaaagtaatt gggaaaaaag aatggcttga gaaaaaagta aacgaaaatc ctgaacttga
     8161 gaaagtatca gcatccgtaa aaggaaaatc agaagcagcc ggttattatg agcgtattca
     8221 acttggagga atggctcaaa agatgtttac ttccctgcaa aaagaagtca agacagatgt
     8281 gttttctccg cttgaagaag cagcggaagc agcaaaagca aatgagccaa cgggcgaaac
     8341 gatgctttta aacgcgtctt tcttaattaa ccgagaagat gaagcgaagt ttgatgaaaa
     8401 agtaaatgaa gcgcatgaaa actggaaaga caaagccgat tttcattaca gcggtccttg
     8461 gcctgcttat aattttgtga acattcgcct aaaagtagaa gagaaataac gtgcttcaca
     8521 aattagtaac cgcacccatt aaccttgtag tgaaaatcgg cgaaaaagta caggaagaag
     8581 ctgataaaca gctatatgac cttccgacga ttcagcaaaa gctcattcag cttcaaatga
     8641 tgtttgagct tggtgaaatt ccagaagaag cgtttcaaga aaaagaagat gaattgttaa
     8701 tgaggtacga aattgcgaaa cgcagagaaa ttgaacaatg ggaagagcta acacaaaaaa
     8761 gaaatgagga atcctagatg ggagaattac tgtatttata cggtttaatt ccaacaaaag
     8821 aagcagcagc catagagccg tttccatctt ataaggggtt tgacggagaa cattcactgt
     8881 acccaattgc gtttgatcag gtgacggctg tagtttctaa gctggatgct gacacctatt
     8941 cagaaaaagt gattcaagaa aaaatggagc aggatatgag ctggctgcag gaaaaagcat
     9001 ttcatcatca cgaaacggta gccgctttgt acgaagaatt tacgatcatt ccattaaaat
     9061 tttgcaccat ttataaaggt gaagaaagtc tgcaggcagc tattgagatt aacaaagaaa
     9121 agatagagaa ttcactgacg ctgcttcaag gaaatgaaga gtggaatgtg aaaatttact
     9181 gtgatgatac agagcttaaa aaaggaatca gcgaaacgaa tgaaagcgtg aaagcgaaaa
     9241 aacaagaaat tagtcactta tcaccaggaa gacagttttt tgaaaagaaa aaaatagatc
     9301 agctgattga aaaagaatta gagcttcaca aaaacaaagt gtgtgaagag atacatgaca
     9361 agctaaaaga attatcgctt tatgactctg ttaaaaagaa ttggagcaaa gacgtaactg
     9421 gcgcagctga acagatggcg tggaacagcg tgtttcttct cccgtctctg cagattacta
     9481 agttcgtaaa cgaaatagaa gagcttcagc aaaggcttga aaataaaggc tggaagtttg
     9541 aagtgacggg accatggccg ccctatcatt tctcgagctt tgcgtaaagt gaggaattaa
     9601 cattatgtct cttaaacaat ccatggagaa taaagatatt gctcttattg atattttaga
     9661 tgtcatttta gataaaggag tcgccattaa aggagactta atcatttcca tagctggcgt
     9721 cgatttagtg tatttggatt tgcgggtgct tatttcttcg gttgaaacgc ttgtgcaagc
     9781 aaaagaagga aatcacaaac caatcacttc tgaacaattt gataaacaaa aggaggaatt
     9841 aatggatgca accggtcagc caagcaaatg gacgaatcca cttggatcct gatcaagctg
     9901 aacaaggctt agcgcagctt gtgatgacag ttattgagct attgaggcaa atcgttgaac
     9961 gtcatgccat gaggcgggtg gagggtggaa cgttgacgga cgaacaaatt gaaaacttag
    10021 gaattgcact aatgaactta gaagaaaaaa tggacgagtt gaaagaggtg ttcggtctgg
    10081 atgcagaaga tttaaatatt gatcttggac cgctaggcag cctgctttaa gcggtcagta
    10141 ggaggaacag tatggcagtc gaacataata tgcagtcaag tacgattgta gatgtgctcg
    10201 aaaagatttt ggataaagga gtcgttatag cgggggacat caccgtagga attgcagatg
    10261 tcgagctatt aacgataaag atccgcttga ttgtggcttc ggttgataag gcaaaagaaa
    10321 tcggcatgga ctggtgggaa aatgatccgt atctcagttc aaaaggagcc aataacaaag
    10381 cgctcgaaga agaaaataaa atgctgcatg agcggttaaa aacgcttgaa gaaaaaatag
    10441 aaacgaaacg ttaaaaactg tacgctactt aaaaaatgga gggatttaca atggcaactg
    10501 aaacaaaatt agataacaca caggcagaaa acaaggaaaa taaaaatgcg gaaaacggtt
    10561 caaaagaaaa gaacggttca aaagcaagca aaacaacaag cagcgggcca atcaaacgag
    10621 cggtagcagg aggcatcatc ggtgcaacga ttggatatgt atcgactcct gaaaatcgaa
    10681 aaagtctcct tgaccgcatt gatacagacg aattaaaaag caaagcatct gatttaggaa
    10741 caaaggtaaa agaaaaatca aaaagcagcg tggccagcct gaaaacatct gcgggaagct
    10801 tgtttaaaaa agataaagat aaatcaaaag atgatgaaga aaacgtaaat tcttctagta
    10861 gcgaaacaga agacgataac gttcaagagt acgacgagtt aaaagaagaa aatcaaactc
    10921 ttcaagatcg cttatcacag cttgaagaaa aaatgaacat gcttgttgag cttagcctca
    10981 ataaaaatca agacgaagaa gcggaagata cagattccga cgaagaagag aacgatgaga
    11041 acgatgaaaa cgatgaaaac gagcaggacg atgaaaacga agaagaaaca tctaagccac
    11101 gtaaaaagga taaaaaagaa gctgaggaag aagaaagtga aagtgacgaa gacagcgagg
    11161 aagaagagga agattctcgc tcaaacaaaa aaaataaaaa agtaaaaaca gaagaagaag
    11221 acgaagatga aagcgaagaa gaaaaaaagg aagcgaaacc aaaaaagtca acagctaaaa
    11281 aatcaaaaaa tacaaaagca aagaaaaaca cggacgaaga agatgatgaa gcaacatctc
    11341 tttctagtga agacgataca acagcctaag acgtaaagga ggaaagaaag acatgagtac
    11401 aggcccttct ttttcaacta aagacaatac gcttgaatac tttgtgaaag cttctaataa
    11461 acacggcttt tcacttgata tttcattaaa tgtaaacggc gctgtgattt ccggtaccat
    11521 gatttcagca aaagaatact ttgattactt aagcgaaacg tttgaagaag gcagtgaagt
    11581 ggctcaggcg ctaagcgaac aattctcttt agcaagcgaa gcgagcgaat caaacggaga
    11641 agcagaagcc cattttattc atttgaaaaa tacaaagatt tactgtggag acagtaaatc
    11701 tactccttct aaaggcaaaa tcttttggag agggaaaata gcagaagtag acgggttttt
    11761 cttaggaaag atttctgatg caaaatcaac gagtaaaaag agttcataaa aaacggcggg
    11821 gtgattgccc cgccgttttt tagtgatgtg atgagatgtg cagcttcttt tttcgataca
    11881 tgagcggagt catcttcatt tcttttcgaa actgattaat aaaatagctt gtactattaa
    11941 accctacttg ataggcgact tccgtcacat ttgcttctgc ttgctgcagg atccggctgc
    12001 taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactagcata
    12061 accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag gaactatatc
    12121 cggat
//

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