pcDNA 3.1(Hygro) myc-hPolQ-Flag

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载体基本信息

质粒名称: pcDNA 3.1(Hygro) myc-hPolQ-Flag
出品公司: Addgene
目录编号: 73132
存储实验室: Agnel Sfeir

载体骨架基本信息
载体名称: pcDNA3.1
载体出品公司: --
空载体大小: 5607 bp
完整质粒大小: 13416 bp
载体修饰: --
载体类型: 哺乳细胞表达
筛选标记: 潮霉素
载体抗性: 氨苄霉素100 μg/mL
生长温度: 37 ℃
宿主菌: DH5a
拷贝数: 高拷贝

插入基因信息
插入基因名称: Polymerase theta
物种来源: H. sapiens (human)
插入大小: 7797 bp
基因突变: --
GenBank编号 --
启动子: cmv promoter
标签/融合蛋白:
Flag(插入时C端子)
myc(插接件上的N端子)
克隆方法: --
5’克隆位点: --
3’克隆位点: --
5’测序引物 TAATACGACTCACTATAGG
3’测序引物 ---

文献引用方法
材料和方法部分: pcDNA 3.1(Hygro) myc-hPolQ-Flag was a gift from Agnel Sfeir (Addgene plasmid # 73132 ; http://n2t.net/addgene:73132 ; RRID:Addgene_73132)
参考文献部分: Mammalian polymerase theta promotes alternative NHEJ and suppresses recombination. Mateos-Gomez PA, Gong F, Nair N, Miller KM, Lazzerini-Denchi E, Sfeir A. Nature. 2015 Feb 12;518(7538):254-7. doi: 10.1038/nature14157. Epub 2015 Feb 2. 10.1038/nature14157 PubMed 25642960

载体图谱质粒图谱和多克隆位点信息

pcDNA 3.1(Hygro) myc-hPolQ-Flag质粒图谱

载体简介

载体序列

LOCUS       pcDNA 3.1(Hygro) myc-hPolQ-Flag              13418 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  To express in mammalian cells  human PolQ.
ACCESSION   .
VERSION     .
KEYWORDS    pcDNA 3.1(Hygro) myc-hPolQ-Flag
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13418)
  AUTHORS   Mateos-Gomez PA, Gong F, Nair N, Miller KM, Lazzerini-Denchi E, 
            Sfeir A
  TITLE     Mammalian polymerase theta promotes alternative NHEJ and suppresses 
            recombination.
  JOURNAL   Nature. 2015 Feb 12;518(7538):254-7. doi: 10.1038/nature14157. Epub 
            2015 Feb 2.
  PUBMED    25642960
REFERENCE   2  (bases 1 to 13418)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..13418
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             946..975
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             8773..8796
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(8840..8857)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8846..9070
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      9116..9544
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9203..9222)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9413..9434
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9553..9573)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        9558..9887
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      9738..9873
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     9800..9819
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             9936..10961
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     polyA_signal    11091..11212
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11128..11147)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     11182..11201
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(11261..11277)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11261..11277)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11274..11296)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11285..11301
                     /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)."
     promoter        complement(11309..11339)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11354..11375
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11492..11509)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11663..12251)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11743..11762)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(12422..13282)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     13045..13064
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13283..13387)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atctgcggcc gcatggaaca aaagttgatt
      961 tctgaagaag atttgttaag aaagagaggg atcctgaatc ttctgcgtcg gagtgggaaa
     1021 cggcggcgtt cagaatcagg ctcagattcg ttctcgggaa gcggcggtga cagcagtgcc
     1081 agcccccagt tcctctccgg gtccgtgctg agcccgccgc ccggccttgg tcgctgcctg
     1141 aaggccgcag ctgcaggaga atgcaagcct acagttcctg actacgaaat agacaagcta
     1201 ctattggcaa actggggact tcctaaagca gttctggaaa aataccacag ttttggtgta
     1261 aaaaagatgt ttgaatggca ggcagagtgc cttttgcttg gacaagtcct ggaaggaaag
     1321 aatttagttt attcagctcc tacaagtgct gggaagactc ttgtggcaga attacttatt
     1381 ttgaagcggg ttttggaaat gcggaagaaa gctttgttta ttcttccctt tgtttctgtg
     1441 gctaaagaga agaaatacta cctccagagt ctgtttcagg aagtaggaat aaaagtagac
     1501 ggttatatgg gcagcacctc tccatcaagg catttctctt cattggatat tgcagtctgc
     1561 acaattgaga gagccaatgg tctgatcaat cgcctcatag aggaaaataa gatggatctg
     1621 ttaggaatgg tggttgtgga tgaattacat atgctgggag actctcaccg agggtatctg
     1681 ctggaacttt tgctgaccaa gatttgctat attactcgga aatcagcatc ttgtcaggca
     1741 gatctagcca gttctctgtc taatgctgtg caaatcgttg gcatgagtgc tacccttcct
     1801 aatttggagc ttgtggcttc ctggttgaat gctgaactct accataccga ctttcgccct
     1861 gtaccgcttt tggagtcagt aaaagttgga aattccatat atgactcttc aatgaaactt
     1921 gtgagggaat ttgagcccat gcttcaagtg aagggagatg aggaccatgt tgttagctta
     1981 tgttatgaga cgatttgtga taaccattca gtattacttt tttgtccatc aaagaaatgg
     2041 tgtgagaagc tggcagatat cattgctcga gagttttata atctacatca tcaagctgag
     2101 ggattggtga aaccctctga atgcccacca gtaattctgg aacaaaaaga actcctggaa
     2161 gtgatggatc agttaagacg gttgccttca ggactggact ctgtattaca gaaaactgta
     2221 ccatggggag tagcatttca tcatgcaggt cttacttttg aggagaggga tatcattgaa
     2281 ggagcctttc gtcaaggtct cattcgagtc ttggcggcaa cttctactct ttcttctggg
     2341 gtgaatttac ctgcacgtcg tgtgattatt cgaaccccta tttttggtgg tcgacctcta
     2401 gatattctta cttataagca gatggttggt cgtgctggca ggaaaggagt ggacacagta
     2461 ggcgagagta tcttaatttg taagaactct gagaaatcaa aaggcatagc tctccttcag
     2521 ggttctctaa agcctgttcg cagctgtctg caaagacgag aaggagaaga agtaactggc
     2581 agcatgatac gagctattct ggagataata gttggtggag tggcaagtac atcacaagat
     2641 atgcatactt atgctgcctg cacatttttg gctgcaagta tgaaagaagg gaagcaagga
     2701 attcagagaa atcaagagtc tgttcagctt ggagcgattg aggcctgtgt gatgtggcta
     2761 ctagaaaatg aattcatcca gagtacagaa gccagtgatg gaacagaagg aaaggtgtat
     2821 catccaacac atcttggttc ggccactctt tcttcttcac tttctccagc tgatacttta
     2881 gatatttttg ctgacctgca aagagcaatg aagggctttg ttttagagaa tgatcttcat
     2941 attctctatc tggttacacc tatgtttgag gattggacta ctattgattg gtatcgattt
     3001 ttctgtttat gggagaagtt gccaacttca atgaaaaggg tggcagagct agtgggagtt
     3061 gaagaggggt tcttggcccg ttgtgtgaaa ggaaaagtag tagccagaac tgagagacag
     3121 catcgacaaa tggccatcca taaaaggttt ttcaccagtc ttgtgctatt agatttaatc
     3181 agtgaagttc ccttaaggga aataaatcag aaatatggat gcaatcgtgg gcagattcaa
     3241 tctttgcaac agtcagctgc tgtttatgca gggatgatta cagtattttc caaccgtctg
     3301 ggctggcaca acatggaact actactttcc caatttcaga agcgtcttac gtttggcatc
     3361 cagagggagc tgtgtgacct ggttcgggta tccttactaa atgctcagag agccagggtt
     3421 ctctatgctt ctggctttca tactgtggca gaccttgcta gagcaaatat tgtggaggtg
     3481 gaggtgattc tgaaaaatgc tgtgcctttc aaaagtgccc ggaaggcagt ggatgaggaa
     3541 gaggaagcag ttgaagaacg tcgcaatatg cgaactatct gggtgactgg cagaaaaggt
     3601 ttaactgaaa gggaagcagc agcccttata gtggaagaag ccagaatgat tctgcagcag
     3661 gacttagttg aaatgggagt gcaatggaat ccatgtgccc tgttacattc tagtacatgc
     3721 tcattgactc atagtgagtc cgaagtaaag gaacacacat ttatatccca aactaagagt
     3781 tcttataaaa aattaacatc aaagaacaaa agtaacacaa tatttagtga ttcttatatt
     3841 aagcattcac caaatatagt gcaagactta aataaaagta gagagcatac aagttccttt
     3901 aattgtaatt tccagaatgg gaatcaagaa catcagagat gttccatttt cagagcaaga
     3961 aaacgggcct ctttagatat aaataaagag aagccaggag cctctcagaa tgaggggaaa
     4021 acaagtgata agaaagttgt tcagactttt tcacagaaaa caaaaaaggc acctttgaat
     4081 ttcaattcag aaaagatgag cagaagtttt cgatcttgga aacgtagaaa gcatctaaag
     4141 cgatctaggg acagcagccc cctgaaagac tctggagcgt gtagaatcca tttacaagga
     4201 cagactctgt ctaatcctag tctttgtgaa gacccgttta ccttagatga gaagaaaacg
     4261 gaatttagaa attcagggcc atttgctaaa aatgtatctt tgagtggtaa ggaaaaagat
     4321 aataaaacat cattcccatt acaaataaag caaaattgtt catggaacat aacactaact
     4381 aatgataatt ttgtggagca tattgtcaca ggatctcaga gtaaaaatgt gacttgtcag
     4441 gccactagtg tggttagtga aaagggcaga ggagtagctg ttgaggcaga aaaaataaat
     4501 gaagtgctga tacaaaatgg ttcaaaaaac cagaatgttt atatgaaaca ccatgacatc
     4561 catccaatta accagtacct gcgaaagcaa tctcatgaac agacaagcac tattaccaaa
     4621 cagaaaaata taatagagag acaaatgccc tgtgaagcag tcagtagtta cataaataga
     4681 gactcaaatg ttactatcaa ttgtgaaagg ataaagctta atacagagga aaataaacca
     4741 agtcattttc aggcattagg agatgatata agcagaactg tgatacccag tgaagtactt
     4801 ccatcagctg gagcatttag caaatcagaa ggccagcatg agaattttct aaatatttct
     4861 agactacaag aaaaaacagg tacttataca acaaacaaaa ctaaaaataa tcatgtttct
     4921 gacttaggtt tagtcctctg tgattttgaa gatagtttct atctggatac tcagtcagag
     4981 aaaataatac aacagatggc aactgaaaat gccaaactag gagcaaagga caccaacctg
     5041 gcagcaggga taatgcagaa gagcttagtc caacagaact caatgaactc ttttcagaag
     5101 gagtgtcaca ttccttttcc tgctgaacag caccctctag gagcgactaa gatagatcat
     5161 ttggacctta agactgtagg tactatgaaa caaagcagtg attcacatgg ggttgatatc
     5221 ctgactccag aaagcccgat tttccattct ccaatactat tggaggaaaa tggtcttttt
     5281 ttaaaaaaga atgaagtttc tgttactgat tcacaattaa atagttttct tcaaggttat
     5341 caaacacaag aaactgtgaa accagttata cttctgattc ctcaaaagag aactcccact
     5401 ggtgtagaag gagaatgtct tccagttcct gaaacaagtt tgaatatgag tgatagttta
     5461 ctatttgata gcttcagtga tgactatcta gtaaaagaac aattacctga tatgcaaatg
     5521 aaagaacccc ttccttcaga agtaacatca aaccatttta gtgattctct gtgtctacaa
     5581 gaagacctaa ttaaaaaatc aaatgtaaat gagaatcaag atacccacca gcagttgact
     5641 tgttccaatg atgaatctat tatattttca gaaatggatt ctgttcagat ggttgaagct
     5701 ttggacaatg tggatatatt tcctgtccaa gagaagaatc atactgtagt atctcctaga
     5761 gcattagaac taagtgatcc agtacttgat gagcaccacc aaggtgatca agatggagga
     5821 gatcaagatg aaagggctga aaaatcaaaa ttaactggga ccaggcaaaa tcattcattc
     5881 atttggtcag gggcatcatt tgatctaagt ccaggactgc aaaggatttt agataaagta
     5941 tccagtcctc tagaaaatga aaagctaaaa tcaatgacta taaacttttc cagtttgaat
     6001 agaaaaaata cagagttaaa tgaagaacaa gaagttattt caaacttgga gacaaaacaa
     6061 gtgcagggaa tttcattttc ttctaataat gaagtaaaaa gcaagattga gatgctagaa
     6121 aacaatgcca atcatgatga aacctcatcc ctcttacctc gtaaagaaag taatatagtt
     6181 gatgataatg gtctcattcc tcctacaccc attccaacat ctgcttctaa gctgacattt
     6241 ccagggattc ttgaaacacc tgtaaaccct tggaaaacta ataatgtttt acaacctggt
     6301 gaaagttatt tatttggctc accttcagat attaaaaacc acgatttaag tccagggagt
     6361 agaaatgggt tcaaagacaa cagccctatt agtgacacaa gcttttcact tcagttatca
     6421 caggatggat tacagttaac tccagcctca agcagttcag aaagtttgtc cataattgat
     6481 gtagcaagtg accaaaatct tttccaaaca ttcattaagg agtggcggtg caaaaagcga
     6541 ttttccatct cactggcttg tgaaaagatt agaagtttga catcttctaa aactgctact
     6601 attggcagta ggtttaagca agctagctca cctcaggaaa ttcctattag agatgatgga
     6661 tttcccatta aaggttgtga tgacaccttg gtggttggac tggcagtatg ctggggtgga
     6721 agggatgcct attatttttc actgcagaag gaacaaaagc attctgaaat tagtgccagt
     6781 ttggttccac cttctttaga tccaagcctg actttgaaag acaggatgtg gtaccttcaa
     6841 tcttgcttgc gaaaggaatc tgataaagaa tgttctgttg tcatctatga cttcatccag
     6901 agctataaaa ttcttcttct ttcttgtggc atctccttgg agcaaagtta tgaagatcct
     6961 aaggtggcat gctggttact agatccagat tctcaggagc cgactcttca tagcatagtt
     7021 accagttttc ttcctcatga gcttccactc ctagaaggga tggagaccag ccaagggatt
     7081 caaagcctgg ggctaaatgc tggcagtgag cattctgggc gatacagagc atctgtggag
     7141 tccattctca tcttcaactc tatgaatcag ctcaactctt tgttgcagaa ggaaaacctt
     7201 caagatgttt tccgtaaggt ggaaatgccc tctcagtact gcttggcctt gctagaacta
     7261 aatggaattg gctttagtac tgcagaatgt gaaagtcaga aacatataat gcaagccaag
     7321 ctggatgcaa ttgagaccca ggcctatcaa ctagctggcc acagtttttc tttcaccagt
     7381 tcagatgaca tcgctgaggt tttatttttg gaattgaagt tgcccccaaa tagagagatg
     7441 aaaaaccaag gcagcaagaa aactctgggt tctaccagaa gagggattga caatggacgc
     7501 aagctaaggc tgggaagaca gttcagcact agtaaggacg ttttaaataa attaaaggca
     7561 ttacatcctt taccaggctt gatattagaa tggagaagaa tcactaatgc tattaccaaa
     7621 gtggtctttc cccttcagcg ggaaaagtgt cttaatcctt ttcttggaat ggaaagaatc
     7681 tatcctgtat cacagtcgca cactgctaca ggacgaataa cctttacaga accaaatatt
     7741 cagaatgtgc caagagattt tgaaatcaaa atgccaacac tagtaggaga aagcccacct
     7801 tctcaagctg taggcaaagg cctacttccc atgggcagag gaaaatataa gaagggtttc
     7861 agcgtgaatc ctagatgcca ggcacagatg gaggagagag ctgcagacag aggaatgcca
     7921 ttttcaatta gcatgcgaca tgcctttgtg cctttcccag gtggttcaat actggctgct
     7981 gactactctc agcttgaact gaggatcttg gctcatttat cccatgatcg tcgtctcatt
     8041 caagtgttaa acactggagc tgatgttttc aggagcattg cagcagagtg gaagatgatt
     8101 gagccagagt ctgttgggga tgatctgagg cagcaggcaa aacagatttg ctatgggatc
     8161 atttatggaa tgggagctaa atctttggga gagcagatgg gcattaaaga aaatgatgct
     8221 gcatgctata ttgactcctt caaatccaga tacacaggga ttaatcaatt catgacagag
     8281 acagtgaaga attgtaaaag agacggattt gttcagacca ttttgggaag gcgtagatat
     8341 ttgccaggaa tcaaagacaa caacccttat cgtaaagctc acgctgagcg tcaagctatc
     8401 aacacaatag tccaaggatc agcagctgat attgtcaaaa tagccacagt taacattcag
     8461 aagcaattag agaccttcca ctcaaccttc aaatcccatg gtcatcgaga gggtatgctc
     8521 caaagtgacc gaacaggatt gtcacgaaag agaaaactgc aagggatgtt ctgcccaatc
     8581 agaggaggct tcttcatcct tcaactccat gatgaactcc tatatgaagt ggcagaagaa
     8641 gatgttgttc aggtagctca gattgtcaag aatgaaatgg aaagtgctgt aaaactgtct
     8701 gtgaaattga aagtgaaagt gaaaataggc gccagctggg gagagctaaa ggactttgat
     8761 gtgcccggga tggactacaa agacgatgac gacaagtaat agctcgagtc tagagggccc
     8821 gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     8881 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     8941 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     9001 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     9061 ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg
     9121 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     9181 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     9241 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     9301 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     9361 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     9421 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     9481 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     9541 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     9601 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     9661 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     9721 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     9781 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
     9841 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
     9901 cttgtatatc cattttcgga tctgatcagc acgtgatgaa aaagcctgaa ctcaccgcga
     9961 cgtctgtcga gaagtttctg atcgaaaagt tcgacagcgt ctccgacctg atgcagctct
    10021 cggagggcga agaatctcgt gctttcagct tcgatgtagg agggcgtgga tatgtcctgc
    10081 gggtaaatag ctgcgccgat ggtttctaca aagatcgtta tgtttatcgg cactttgcat
    10141 cggccgcgct cccgattccg gaagtgcttg acattgggga attcagcgag agcctgacct
    10201 attgcatctc ccgccgtgca cagggtgtca cgttgcaaga cctgcctgaa accgaactgc
    10261 ccgctgttct gcagccggtc gcggaggcca tggatgcgat cgctgcggcc gatcttagcc
    10321 agacgagcgg gttcggccca ttcggaccgc aaggaatcgg tcaatacact acatggcgtg
    10381 atttcatatg cgcgattgct gatccccatg tgtatcactg gcaaactgtg atggacgaca
    10441 ccgtcagtgc gtccgtcgcg caggctctcg atgagctgat gctttgggcc gaggactgcc
    10501 ccgaagtccg gcacctcgtg cacgcggatt tcggctccaa caatgtcctg acggacaatg
    10561 gccgcataac agcggtcatt gactggagcg aggcgatgtt cggggattcc caatacgagg
    10621 tcgccaacat cttcttctgg aggccgtggt tggcttgtat ggagcagcag acgcgctact
    10681 tcgagcggag gcatccggag cttgcaggat cgccgcggct ccgggcgtat atgctccgca
    10741 ttggtcttga ccaactctat cagagcttgg ttgacggcaa tttcgatgat gcagcttggg
    10801 cgcagggtcg atgcgacgca atcgtccgat ccggagccgg gactgtcggg cgtacacaaa
    10861 tcgcccgcag aagcgcggcc gtctggaccg atggctgtgt agaagtactc gccgatagtg
    10921 gaaaccgacg ccccagcact cgtccgaggg caaaggaata gcacgtgcta cgagatttcg
    10981 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
    11041 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
    11101 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
    11161 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
    11221 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
    11281 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
    11341 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
    11401 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
    11461 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
    11521 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
    11581 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
    11641 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
    11701 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
    11761 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
    11821 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
    11881 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
    11941 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
    12001 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
    12061 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
    12121 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
    12181 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
    12241 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
    12301 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
    12361 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
    12421 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
    12481 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
    12541 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
    12601 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
    12661 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
    12721 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
    12781 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
    12841 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
    12901 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
    12961 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
    13021 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
    13081 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
    13141 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
    13201 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
    13261 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
    13321 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
    13381 ttccgcgcac atttccccga aaagtgccac ctgacgtc
//

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