pRRLsin-SV40 T antigen-IRES-mCherry

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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: pRRLsin-SV40 T antigen-IRES-mCherry
出品公司: Addgene
目录编号: 58993
存储实验室: Snorri Thorgeirsson

载体骨架基本信息
载体名称: pRRLsin.cPPT.PGK/GFP.WPRE
载体出品公司: --
空载体大小: 6110 bp
完整质粒大小: 11405 bp
载体修饰: 位于nt 924的NotI站点被删除
载体类型: 哺乳细胞表达,慢病毒
筛选标记: mCherry 红色荧光
载体抗性: 氨苄霉素
生长温度: 30 ℃
宿主菌: Stbl3
拷贝数: 高拷贝

插入基因信息
插入基因名称: CMV enhancer-chicken beta actin promoter
物种来源: --
插入大小: 1730 bp
基因突变: MCS 中的 EcoRV 站点转换为 BamHI 站点
GenBank编号 --
启动子: --
标签/融合蛋白: --
克隆方法: 限制性内切酶
5’克隆位点: SalI(克隆期间销毁)
3’克隆位点: BamHI(未破坏)
5’测序引物 --
3’测序引物 pCAG-F

文献引用方法
材料和方法部分: pRRLsin-SV40 T antigen-IRES-mCherry was a gift from Snorri Thorgeirsson (Addgene plasmid # 58993 ; http://n2t.net/addgene:58993 ; RRID:Addgene_58993)
参考文献部分: Modeling pathogenesis of primary liver cancer in lineage-specific mouse cell types. Holczbauer A, Factor VM, Andersen JB, Marquardt JU, Kleiner DE, Raggi C, Kitade M, Seo D, Akita H, Durkin ME, Thorgeirsson SS. Gastroenterology. 2013 Jul;145(1):221-31. doi: 10.1053/j.gastro.2013.03.013. Epub 2013 Mar 19. 10.1053/j.gastro.2013.03.013 PubMed 23523670

载体图谱质粒图谱和多克隆位点信息

pRRLsin-SV40 T antigen-IRES-mCherry质粒图谱

载体简介

载体序列

LOCUS       pRRLsin-SV40 T antigen-IRES-mCherry               11418 bp ds-DNA     circular SYN 08-SEP-2021
DEFINITION  lentiviral expression of SV40 large T antigen and mCherry red 
            fluorescent reporter.
ACCESSION   .
VERSION     .
KEYWORDS    pRRLsin-SV40 T antigen-IRES-mCherry
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11418)
  AUTHORS   Holczbauer A, Factor VM, Andersen JB, Marquardt JU, Kleiner DE, 
            Raggi C, Kitade M, Seo D, Akita H, Durkin ME, Thorgeirsson SS
  TITLE     Modeling pathogenesis of primary liver cancer in lineage-specific 
            mouse cell types.
  JOURNAL   Gastroenterology. 2013 Jul;145(1):221-31. doi: 
            10.1053/j.gastro.2013.03.013. Epub 2013 Mar 19.
  PUBMED    23523670
REFERENCE   2  (bases 1 to 11418)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 8, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..11418
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        8..234
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             235..415
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    462..587
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1085..1318
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1503..1547
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    1814..1931
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        1953..2332
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          2611..3627
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     3551..3573
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     3635..3654
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             3726..5852
                     /codon_start=1
                     /product="large T antigen from SV40 (simian virus 40)"
                     /label=large T antigen
                     /translation="MDKVLNREESLQLMDLLGLERSAWGNIPLMRKAYLKKCKEFHPDK
                     GGDEEKMKKMNTLYKKMEDGVKYAHQPDFGGFWDATEIPTYGTDEWEQWWNAFNEENLF
                     CSEEMPSSDDEATADSQHSTPPKKKRKVEDPKDFPSELLSFLSHAVFSNRTLACFAIYT
                     TKEKAALLYKKIMEKYSVTFISRHNSYNHNILFFLTPHRHRVSAINNYAQKLCTFSFLI
                     CKGVNKEYLMYSALTRDPFSVIEESLPGGLKEHDFNPEEAEETKQVSWKLVTEYAMETK
                     CDDVLLLLGMYLEFQYSFEMCLKCIKKEQPSHYKYHEKHYANAAIFADSKNQKTICQQA
                     VDTVLAKKRVDSLQLTREQMLTNRFNDLLDRMDIMFGSTGSADIEEWMAGVAWLHCLLP
                     KMDSVVYDFLKCMVYNIPKKRYWLFKGPIDSGKTTLAAALLELCGGKALNVNLPLDRLN
                     FELGVAIDQFLVVFEDVKGTGGESRDLPSGQGINNLDNLRDYLDGSVKVNLEKKHLNKR
                     TQIFPPGIVTMNEYSVPKTLQARFVKQIDFRPKDYLKHCLERSEFLLEKRIIQSGIALL
                     LMLIWYRPVAEFAQSIQSRIVEWKERLDKEFSLSVYQKMKFNVAMGIGVLDWLRNSDDD
                     DEDSQENADKNEDGGEKNMEDSGHETGIDSQSQGSFQAPQSSQSVHDHNQPYHICRGFT
                     CFKKPPTPPPEPET"
     misc_feature    5921..6507
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(6088..6105)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     6315..6334
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             6508..7218
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(6653..6671)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     6921..6940
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(7238..7254)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7238..7254)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     misc_feature    7269..7857
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7322..7342)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7740..7751)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7944..8177
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    8249..8370
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8286..8305)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8340..8359
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      8410..8545
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     8472..8491
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     complement(8565..8584)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8566..8584)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8594..8611)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8594..8610)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8603..8625)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      8752..9207
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8839..8858)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9049..9070
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        9233..9337
                     /gene="bla"
                     /label=AmpR promoter
     CDS             9338..10198
                     /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     complement(9556..9575)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10369..10957
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10858..10877
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     11111..11128
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    11245..11266
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        11281..11311
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11319..11335
                     /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     11324..11346
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     11343..11359
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     11343..11359
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     11378..11398
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        11380..11398
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 agcttaatgt agtcttatgc aatactcttg tagtcttgca acatggtaac gatgagttag
       61 caacatgcct tacaaggaga gaaaaagcac cgtgcatgcc gattggtgga agtaaggtgg
      121 tacgatcgtg ccttattagg aaggcaacag acgggtctga catggattgg acgaaccact
      181 gaattgccgc attgcagaga tattgtattt aagtgcctag ctcgatacat aaacgggtct
      241 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      301 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      361 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
      421 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
      481 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
      541 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
      601 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
      661 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
      721 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
      781 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
      841 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
      901 aaagtaagac caccgcacag caagcggcct ggccgctgat cttcagacct ggaggaggag
      961 atatgaggga caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat
     1021 taggagtagc acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg
     1081 gaataggagc tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt
     1141 caatgacgct gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca
     1201 atttgctgag ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca
     1261 agcagctcca ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg
     1321 ggatttgggg ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt
     1381 ggagtaataa atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag
     1441 aaattaacaa ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag
     1501 aaaagaatga acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta
     1561 acataacaaa ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag
     1621 gtttaagaat agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac
     1681 cattatcgtt tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag
     1741 aagaagaagg tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac
     1801 ggtatcggtt aacttttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa
     1861 tagtagacat aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa
     1921 ttcaaaattt tatcgatcac gagactagcc tcgacattga ttattgacta gttattaata
     1981 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
     2041 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
     2101 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
     2161 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
     2221 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
     2281 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtcgaggt
     2341 gagccccacg ttctgcttca ctctccccat ctcccccccc tccccacccc caattttgta
     2401 tttatttatt ttttaattat tttgtgcagc gatgggggcg gggggggggg gggnsgcgcg
     2461 ccrggsgggg sggggsgggg sgrggggsgg ggsggggsgr ggsggrgagg tgcggcggca
     2521 gccaatcaga gcggcgcgct ccgaaagttt ccttttatgg cgaggcggcg gcggcggcgg
     2581 ccctataaaa agcgaagcgc gcggcgggcg ggagtcgctg cgcgctgcct tcgccccgtg
     2641 ccccgctccg ccgccgcctc gcgccgcccg ccccggctct gactgaccgc gttactccca
     2701 caggtgagcg ggcgggacgg cccttctcct ccgggctgta attagcgctt ggtttaatga
     2761 cggcttgttt cttttctgtg gctgcgtgaa agccttgagg ggctccggga gggccctttg
     2821 tgcgggggga gcggctcggg gggtgcgtgc gtgtgtgtgt gcgtggggag cgccgcgtgc
     2881 ggctccgcgc tgcccggcgg ctgtgagcgc tgcgggcgcg gcgcggggct ttgtgcgctc
     2941 cgcagtgtgc gcgaggggag cgcggccggg ggcggtgccc cgcggtgcgg ggggggctgc
     3001 gaggggaaca aaggctgcgt gcggggtgtg tgcgtggggg ggtgagcagg gggtgtgggc
     3061 gcgtcggtcg ggctgcaacc ccccctgcac ccccctcccc gagttgctga gcacggcccg
     3121 gcttcgggtg cggggctccg tacggggcgt ggcgcggggc tcgccgtgcc gggcgggggg
     3181 tggcggcagg tgggggtgcc gggcggggcg gggccgcctc gggccggggr gggctcgggg
     3241 grggggcgcg gcggcccccg gagcgccggc ggctgtcgag gcgcggcgag ccgcagccat
     3301 tgccttttat ggtaatcgtg cgagagggcg cagggacttc ctttgtccca aatctgtgcg
     3361 gagccgaaat ctgggaggcg ccgccgcacc ccctctagcg ggcgcggggc gaagcggtgc
     3421 ggcgccggca ggaaggaaat gggcggggag ggccttcgtg cgtcgccgcg ccgccgtccc
     3481 cttctccctc tccagcctcg gggctgtccg cggggggacg gctgccttcg ggggggacgg
     3541 ggcagggcgg ggttcggctt ctggcgtgtg accggcggct ctagagcctc tgctaaccat
     3601 gttcatgcct tcttcttttt cctacagctc ctgggcaacg tgctggttat tgtgctgtct
     3661 catcattttg gcaaagaatt cctcgaggat gaggatcccg cgaccggtcg accagctttg
     3721 caaagatgga taaagtttta aacagagagg aatctttgca gctaatggac cttctaggtc
     3781 ttgaaaggag tgcctggggg aatattcctc tgatgagaaa ggcatattta aaaaaatgca
     3841 aggagtttca tcctgataaa ggaggagatg aagaaaaaat gaagaaaatg aatactctgt
     3901 acaagaaaat ggaagatgga gtaaaatatg ctcatcaacc tgactttgga ggcttctggg
     3961 atgcaactga gattccaacc tatggaactg atgaatggga gcagtggtgg aatgccttta
     4021 atgaggaaaa cctgttttgc tcagaagaaa tgccatctag tgatgatgag gctactgctg
     4081 actctcaaca ttctactcct ccaaaaaaga agagaaaggt agaagacccc aaggactttc
     4141 cttcagaatt gctaagtttt ttgagtcatg ctgtgtttag taatagaact cttgcttgct
     4201 ttgctattta caccacaaag gaaaaagctg cactgctata caagaaaatt atggaaaaat
     4261 attctgtaac ctttataagt aggcataaca gttataatca taacatactg ttttttctta
     4321 ctccacacag gcatagagtg tctgctatta ataactatgc tcaaaaattg tgtaccttta
     4381 gctttttaat ttgtaaaggg gttaataagg aatatttgat gtatagtgcc ttgactagag
     4441 atccattttc tgttattgag gaaagtttgc caggtgggtt aaaggagcat gattttaatc
     4501 cagaagaagc agaggaaact aaacaagtgt cctggaagct tgtaacagag tatgcaatgg
     4561 aaacaaaatg tgatgatgtg ttgttattgc ttgggatgta cttggaattt cagtacagtt
     4621 ttgaaatgtg tttaaaatgt attaaaaaag aacagcccag ccactataag taccatgaaa
     4681 agcattatgc aaatgctgct atatttgctg acagcaaaaa ccaaaaaacc atatgccaac
     4741 aggctgttga tactgtttta gctaaaaagc gggttgatag cctacaatta actagagaac
     4801 aaatgttaac aaacagattt aatgatcttt tggataggat ggatataatg tttggttcta
     4861 caggctctgc tgacatagaa gaatggatgg ctggagttgc ttggctacac tgtttgttgc
     4921 ccaaaatgga ttcagtggtg tatgactttt taaaatgcat ggtgtacaac attcctaaaa
     4981 aaagatactg gctgtttaaa ggaccaattg atagtggtaa aactacatta gcagctgctt
     5041 tgcttgaatt atgtgggggg aaagctttaa atgttaattt gcccttggac aggctgaact
     5101 ttgagctagg agtagctatt gaccagtttt tagtagtttt tgaggatgta aagggcactg
     5161 gaggggagtc cagagatttg ccttcaggtc agggaattaa taacctggac aatttaaggg
     5221 attatttgga tggcagtgtt aaggtaaact tagaaaagaa acacctaaat aaaagaactc
     5281 aaatatttcc ccctggaata gtcaccatga atgagtacag tgtgcctaaa acactgcagg
     5341 ccagatttgt aaaacaaata gattttaggc ccaaagatta tttaaagcat tgcctggaac
     5401 gcagtgagtt tttgttagaa aagagaataa ttcaaagtgg cattgctttg cttcttatgt
     5461 taatttggta cagacctgtg gctgagtttg ctcaaagtat tcagagcaga attgtggagt
     5521 ggaaagagag attggacaaa gagtttagtt tgtcagtgta tcaaaaaatg aagtttaatg
     5581 tggctatggg aattggagtt ttagattggc taagaaacag tgatgatgat gatgaagaca
     5641 gccaggaaaa tgctgataaa aatgaagatg gtggggagaa gaacatggaa gactcagggc
     5701 atgaaacagg cattgattca cagtcccaag gctcatttca ggcccctcag tcctcacagt
     5761 ctgttcatga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac
     5821 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacggg
     5881 gatcctcatc gcggccgcta cgtaaattcc gccccccccc cccctctccc tccccccccc
     5941 ctaacgttac tggccgaagc cgcttggaat aaggccggtg tgcgtttgtc tatatgttat
     6001 tttccaccat attgccgtct tttggcaatg tgagggcccg gaaacctggc cctgtcttct
     6061 tgacgagcat tcctaggggt ctttcccctc tcgccaaagg aatgcaaggt ctgttgaatg
     6121 tcgtgaagga agcagttcct ctggaagctt cttgaagaca aacaacgtct gtagcgaccc
     6181 tttgcaggca gcggaacccc ccacctggcg acaggtgcct ctgcggccaa aagccacgtg
     6241 tataagatac acctgcaaag gcggcacaac cccagtgcca cgttgtgagt tggatagttg
     6301 tggaaagagt caaatggctc tcctcaagcg tattcaacaa ggggctgaag gatgcccaga
     6361 aggtacccca ttgtatggga tctgatctgg ggcctcggtg cacatgcttt acatgtgttt
     6421 agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg ggacgtggtt ttcctttgaa
     6481 aaacacgatg ataatatggc cacaaccatg gtgagcaagg gcgaggagga taacatggcc
     6541 atcatcaagg agttcatgcg cttcaaggtg cacatggagg gctccgtgaa cggccacgag
     6601 ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg
     6661 aaggtgacca agggtggccc cctgcccttc gcctgggaca tcctgtcccc tcagttcatg
     6721 tacggctcca aggcctacgt gaagcacccc gccgacatcc ccgactactt gaagctgtcc
     6781 ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc
     6841 gtgacccagg actcctccct gcaggacggc gagttcatct acaaggtgaa gctgcgcggc
     6901 accaacttcc cctccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc
     6961 tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg agatcaagca gaggctgaag
     7021 ctgaaggacg gcggccacta cgacgctgag gtcaagacca cctacaaggc caagaagccc
     7081 gtgcagctgc ccggcgccta caacgtcaac atcaagttgg acatcacctc ccacaacgag
     7141 gactacacca tcgtggaaca gtacgaacgc gccgagggcc gccactccac cggcggcatg
     7201 gacgagctgt acaagtaaga attcctgcag cccgggggat ccactagttc tagagcggcc
     7261 gcgtcgacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact
     7321 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg
     7381 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg
     7441 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa
     7501 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc
     7561 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
     7621 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaagctgacg tcctttccat
     7681 ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt
     7741 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc
     7801 cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg
     7861 gaattcgagc tcggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca
     7921 ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa gacaagatct
     7981 gctttttgct tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg
     8041 ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt
     8101 gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt
     8161 gtggaaaatc tctagcagta gtagttcatg tcatcttatt attcagtatt tataacttgc
     8221 aaagaaatga atatcagaga gtgagaggaa cttgtttatt gcagcttata atggttacaa
     8281 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     8341 tggtttgtcc aaactcatca atgtatctta tcatgtctgg ctctagctat cccgccccta
     8401 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     8461 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     8521 tagtgaggag gcttttttgg aggcctaggg acgtacccaa ttcgccctat agtgagtcgt
     8581 attacgcgcg ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta
     8641 cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg
     8701 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg gacgcgccct
     8761 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     8821 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     8881 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     8941 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     9001 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     9061 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     9121 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     9181 ttaacaaaat attaacgctt acaatttagg tggcactttt cggggaaatg tgcgcggaac
     9241 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     9301 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     9361 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     9421 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     9481 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     9541 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     9601 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     9661 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     9721 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     9781 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     9841 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     9901 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     9961 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
    10021 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
    10081 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
    10141 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
    10201 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
    10261 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
    10321 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
    10381 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
    10441 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
    10501 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
    10561 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
    10621 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
    10681 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
    10741 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
    10801 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
    10861 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
    10921 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
    10981 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
    11041 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
    11101 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
    11161 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
    11221 agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc
    11281 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
    11341 cacaggaaac agctatgacc atgattacgc caagcgcgca attaaccctc actaaaggga
    11401 acaaaagctg gagctgca
//

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