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AKT-translocation-mStrawberry reporter

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AKT-translocation-mStrawberry reporter載體質粒基本信息

質粒名稱: AKT-translocation-mStrawberry reporter
出品公司: Addgene
目錄編號: 158685
存儲實驗室: Ravid Straussman

載體骨架基本信息
載體名稱: pLKO.1
載體出品公司: Clontech
空載體大小: 10887bp
完整質粒大小: 7728bp
載體類型: 哺乳細胞表達
篩選標記: Blasticidin
載體抗性: 氨芐霉素
生長溫度: 37 ℃
宿主菌: NEB Stable
拷貝數: 高拷貝

插入基因信息
插入基因名稱: NEB Stable
物種來源: H. sapiens (human)
插入大小: 3158 bp
啟動子: EF1a
克隆方法: Gibson Cloning

文獻引用方法
材料和方法部分: AKT-translocation-mStrawberry reporter was a gift from Ravid Straussman (Addgene plasmid # 158685 ; //n2t.net/addgene:158685 ; RRID:Addgene_158685)
參考文獻部分: Visual barcodes for clonal-multiplexing of live microscopy-based assays. Kaufman T, Nitzan E, Firestein N, Ginzberg MB, Iyengar S, Patel N, Ben-Hamo R, Porat Z, Hunter J, Hilfinger A, Rotter V, Kafri R, Straussman R. Nat Commun. 2022 May 18;13(1):2725. doi: 10.1038/s41467-022-30008-0. 10.1038/s41467-022-30008-0 PubMed 35585055

AKT-translocation-mStrawberry reporter質粒圖譜載體圖譜和AKT-translocation-mStrawberry reporter載體序列質粒序列多克隆位點信息

AKT-translocation-mStrawberry

AKT-translocation-mStrawberry reporter質粒載體簡介

AKT-translocation-mStrawberry reporter質粒序列載體序列

LOCUS       AKT-translocation-mStrawberry reporter       10887 bp    DNA     circular SYN 14-APR-2025
DEFINITION  This plasmid is a AKT pathway reporter. It has 1EF1a promoter 
            driving the expression of truncated FoxO1 fused to 
            mStrawberry-pGK-BSD.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10887)
  AUTHORS   Kaufman T, Nitzan E, Firestein N, Ginzberg MB, Iyengar S, Patel N, 
            Ben-Hamo R, Porat Z, Hunter J, Hilfinger A, Rotter V, Kafri R, 
            Straussman R
  TITLE     Visual barcodes for clonal-multiplexing of live microscopy-based 
            assays.
  JOURNAL   Nat Commun. 2022 May 18;13(1):2725. doi: 10.1038/s41467-022-30008-0.
  PUBMED    35585055
REFERENCE   2  (bases 1 to 10887)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            //gt12365.cn
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2022-05-18"; volume: "13(1):2725. doi"; pages: " 
            10.1038/s41467-022-30008-0"
COMMENT     Sequence Label: AKT-translocation-mStrawberry reporter
FEATURES             Location/Qualifiers
     source          1..10887
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..212
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     intron          213..1151
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1108..1128
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             3153..3863
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent 
                     protein"
                     /label=mStrawberry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEENNMAIIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILTPNFTYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKMRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYIVGIKLDITSHNEDYTIVELYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(3298..3316)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3566..3585
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    3905..4022
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        4071..4581
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     4500..4518
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             4593..4988
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    5024..5612
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5077..5097)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5495..5506)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5684..5917
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5995..6116
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6032..6051)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6086..6105
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      6156..6291
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6311..6330)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6312..6330)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6340..6357)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6340..6356)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6349..6371)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6498..6953
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6585..6604)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6795..6816
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6979..7083
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7084..7152,7153..7944)
                     /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(7302..7321)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8115..8703
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8604..8623
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8857..8874
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8991..9012
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9027..9032,9033..9050,9051..9057)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9065..9081
                     /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     9070..9092
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9089..9105
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9089..9105
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     9124..9144
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        9126..9144
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        9172..9398
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             9399..9579
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    9626..9751
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    10244..10477
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             10662..10706
                     /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"
ORIGIN
        1 gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc ggcaattgaa
       61 ccggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg tactggctcc
      121 gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc
      181 tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc cgtgtgtggt tcccgcgggc
      241 ctggcctctt tacgggttat ggcccttgcg tgccttgaat tacttccacc tggctgcagt
      301 acgtgattct tgatcccgag cttcgggttg gaagtgggtg ggagagttcg aggccttgcg
      361 cttaaggagc cccttcgcct cgtgcttgag ttgaggcctg gcctgggcgc tggggccgcc
      421 gcgtgcgaat ctggtggcac cttcgcgcct gtctcgctgc tttcgataag tctctagcca
      481 tttaaaattt ttgatgacct gctgcgacgc tttttttctg gcaagatagt cttgtaaatg
      541 cgggccaaga tctgcacact ggtatttcgg tttttggggc cgcgggcggc gacggggccc
      601 gtgcgtccca gcgcacatgt tcggcgaggc ggggcctgcg agcgcggcca ccgagaatcg
      661 gacgggggta gtctcaagct ggccggcctg ctctggtgcc tggcctcgcg ccgccgtgta
      721 tcgccccgcc ctgggcggca aggctggccc ggtcggcacc agttgcgtga gcggaaagat
      781 ggccgcttcc cggccctgct gcagggagct caaaatggag gacgcggcgc tcgggagagc
      841 gggcgggtga gtcacccaca caaaggaaaa gggcctttcc gtcctcagcc gtcgcttcat
      901 gtgactccac ggagtaccgg gcgccgtcca ggcacctcga ttagttctcg agcttttgga
      961 gtacgtcgtc tttaggttgg ggggaggggt tttatgcgat ggagtttccc cacactgagt
     1021 gggtggagac tgaagttagg ccagcttggc acttgatgta attctccttg gaatttgccc
     1081 tttttgagtt tggatcttgg ttcattctca agcctcagac agtggttcaa agtttttttc
     1141 ttccatttca ggtgtcgtga ggaatttcga catttaaatt taattaaacc atggcagagg
     1201 ctccccaggt ggtggagacc gacccggact tcgagccgct gccccggcag cgctcctgta
     1261 cctggccgct gcccaggccg gagtttaacc agtccaactc gaccacctcc agtccggcgc
     1321 cgtcgggcgg cgcggccgcc aaccccgacg ccgcggcgag cctggcctcg gcgtccgctg
     1381 tcagcaccga ctttatgagc aacctgagcc tgctggagga gagtgaggac ttcgcgcggg
     1441 cgccaggctg cgtggccgtg gcggcggcgg ctgcggccag caggggcctg tgcggggact
     1501 tccagggccc cgaggcgggc tgcgtgcacc cagcgccgcc acagccccca ccgaccgggc
     1561 cgctgtcgca gcccccaccc gtgcctccct ccgctgccgc cgccgcgggg ccactcgcgg
     1621 gacagccgcg caagaccagc tcgtcgcgcc gcaacgcgtg gggcaacctg tcgtacgccg
     1681 acctcatcac caaggccatc gagagctcag ccgagaagag gctcaccctg tcgcagatct
     1741 acgagtggat ggtgaagagc gtgccctact tcaaggataa gggcgacagc aacagctcgg
     1801 cgggctggaa gaatgcaatt cgccgcaatc tagcccttca cagcaagttt attcgagtgc
     1861 agaatgaagg aactggaaag agttcttggt ggatgctcaa tccagaggga ggcaagagcg
     1921 gaaaatcacc ccggagaaga gctgcgtcca tggacaacaa cagtaaattt gctaagagcc
     1981 gagggcgggc tgctaagaaa aaagcatctc tccggtctgg gcaagagggt cctggagaca
     2041 gccctgggtc tcagttttct aagtggcctg cgagtcctgg gtcccacagc aacgatgact
     2101 ttgataactg gagtacattt cgtcctcgaa ccagctcaaa tgctagtacc atcagtggga
     2161 gactttctcc catcatgaca gagcaggatg acctgggaga tggggacgtg cattccctgg
     2221 tgtatccacc ctctgctgcc aagatggcgt ctacgctgcc cagtctgtct gaaatcagca
     2281 atccagaaaa catggagaac cttctggata atctcaacct tctctcgtcc ccaacatctt
     2341 taactgtgtc cactcagtcc tcgcctggca gcatgatgca gcagacacca tgctattcgt
     2401 ttgcaccgcc aaacaccagt ctaaattcac ccagtccaaa ctactcaaag tacacatacg
     2461 gccaatccag catgagccct ttgccccaga tgcctatgca gacacttcag gacagcaaat
     2521 caagttacgg aggattgaac cagtataact gtgccccagg actcttgaaa gagttgttga
     2581 cttctgactc tcctccccac aatgacatta tgtcaccggt tgatcccgga gtggcccaac
     2641 ccaacagtcg ggtcctgggc caaaatgtaa tgatgggccc taattcggtc atgccagcgt
     2701 atggcagcca ggcatctcat aacaaaatga tgtaccccag ctcccacacc caccctggac
     2761 atgcacagca aacggcttcg gtcaacggcc gtaccctgcc ccatgtggtg aacaccatgc
     2821 ctcacacatc tgccatgaac cgcttgaccc ccgtgaagac acctttacaa gtgcctctgt
     2881 cccaccccat gcagatgagt gccctgggca actactcctc ggtgagcagc tgcaatggct
     2941 atggtaggat gggtgtcctc caccaggaga agctcccaag tgacttggat ggcatgttta
     3001 ttgagcgctt ggactgtgac atggagtcca tcattcggaa tgaccccatg gatggagata
     3061 ccttggattt taactttgat aatgtgttgc ccaaccaaag cttcccacac agtgtcaaga
     3121 ctacaacaca cagctgggtg tcaggaggac ccatggtgag caagggcgag gagaataaca
     3181 tggccatcat caaggagttc atgcgcttca aggtgcgcat ggagggctcc gtgaacggcc
     3241 acgagttcga gatcgagggc gagggcgagg gccgccccta cgagggcacc cagaccgcca
     3301 agctgaaggt gaccaagggt ggccccctgc ccttcgcctg ggacatccta acccccaact
     3361 tcacctacgg ctccaaggcc tacgtgaagc accccgccga catccccgac tacttgaagc
     3421 tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg
     3481 tgaccgtgac ccaggactcc tccctgcagg acggcgagtt catctacaag gtgaagctgc
     3541 gcggcaccaa cttcccctcc gacggccccg taatgcagaa gaagaccatg ggctgggagg
     3601 cctcctccga gcggatgtac cccgaggacg gcgccctgaa gggcgagatc aagatgaggc
     3661 tgaagctgaa ggacggcggc cactacgacg ctgaggtcaa gaccacctac aaggccaaga
     3721 agcccgtgca gctgcccggc gcctacatcg tcggcatcaa gttggacatc acctcccaca
     3781 acgaggacta caccatcgtg gaactgtacg aacgcgccga gggccgccac tccaccggcg
     3841 gcatggacga gctgtacaag taagaattct agatcttgag acaaatggca gtattcatcc
     3901 acaattttaa aagaaaaggg gggattgggg ggtacagtgc aggggaaaga atagtagaca
     3961 taatagcaac agacatacaa actaaagaat tacaaaaaca aattacaaaa attcaaaatt
     4021 ttcgggttta ttacagggac agcagagatc cactttggcg ccggctcgag ggggcccggg
     4081 ttgcgccttt tccaaggcag ccctgggttt gcgcagggac gcggctgctc tgggcgtggt
     4141 tccgggaaac gcagcggcgc cgaccctggg tctcgcacat tcttcacgtc cgttcgcagc
     4201 gtcacccgga tcttcgccgc tacccttgtg ggccccccgg cgacgcttcc tgctccgccc
     4261 ctaagtcggg aaggttcctt gcggttcgcg gcgtgccgga cgtgacaaac ggaagccgca
     4321 cgtctcacta gtaccctcgc agacggacag cgccagggag caatggcagc gcgccgaccg
     4381 cgatgggctg tggccaatag cggctgctca gcagggcgcg ccgagagcag cggccgggaa
     4441 ggggcggtgc gggaggcggg gtgtggggcg gtagtgtggg ccctgttcct gcccgcgcgg
     4501 tgttccgcat tctgcaagcc tccggagcgc acgtcggcag tcggctccct cgttgaccga
     4561 atcaccgacc tctctcccca gggggtacca ccatggccaa gcctttgtct caagaagaat
     4621 ccaccctcat tgaaagagca acggctacaa tcaacagcat ccccatctct gaagactaca
     4681 gcgtcgccag cgcagctctc tctagcgacg gccgcatctt cactggtgtc aatgtatatc
     4741 attttactgg gggaccttgt gcagaactcg tggtgctggg cactgctgct gctgcggcag
     4801 ctggcaacct gacttgtatc gtcgcgatcg gaaatgagaa caggggcatc ttgagcccct
     4861 gcggacggtg ccgacaggtg cttctcgatc tgcatcctgg gatcaaagcc atagtgaagg
     4921 acagtgatgg acagccgacg gcagttggga ttcgtgaatt gctgccctct ggttatgtgt
     4981 gggagggcct gcagctgcag tagtaaagac gcgttaagtc gacaatcaac ctctggatta
     5041 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     5101 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     5161 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca
     5221 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac
     5281 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact
     5341 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     5401 cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg
     5461 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc
     5521 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac
     5581 gagtcggatc tccctttggg ccgcctcccc gcgtcgactt taagaccaat gacttacaag
     5641 gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctaattcac
     5701 tcccaacgaa gacaagatct gctttttgct tgtactgggt ctctctggtt agaccagatc
     5761 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
     5821 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
     5881 ctcagaccct tttagtcagt gtggaaaatc tctagcagta cgtatagtag ttcatgtcat
     5941 cttattattc agtatttata acttgcaaag aaatgaatat cagagagtga gaggaacttg
     6001 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     6061 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     6121 gtctggctct agctatcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     6181 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     6241 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctagggacgt
     6301 acccaattcg ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg
     6361 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
     6421 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
     6481 cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     6541 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     6601 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     6661 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     6721 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     6781 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     6841 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     6901 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc
     6961 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     7021 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     7081 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     7141 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     7201 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     7261 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     7321 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     7381 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     7441 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     7501 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     7561 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     7621 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     7681 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     7741 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     7801 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     7861 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     7921 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     7981 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     8041 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     8101 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     8161 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     8221 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     8281 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     8341 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     8401 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     8461 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     8521 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     8581 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     8641 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     8701 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     8761 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     8821 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     8881 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     8941 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
     9001 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
     9061 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag
     9121 cgcgcaatta accctcacta aagggaacaa aagctggagc tgcaagctta atgtagtctt
     9181 atgcaatact cttgtagtct tgcaacatgg taacgatgag ttagcaacat gccttacaag
     9241 gagagaaaaa gcaccgtgca tgccgattgg tggaagtaag gtggtacgat cgtgccttat
     9301 taggaaggca acagacgggt ctgacatgga ttggacgaac cactgaattg ccgcattgca
     9361 gagatattgt atttaagtgc ctagctcgat acataaacgg gtctctctgg ttagaccaga
     9421 tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct
     9481 tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat
     9541 ccctcagacc cttttagtca gtgtggaaaa tctctagcag tggcgcccga acagggactt
     9601 gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg actcggcttg ctgaagcgcg
     9661 cacggcaaga ggcgaggggc ggcgactggt gagtacgcca aaaattttga ctagcggagg
     9721 ctagaaggag agagatgggt gcgagagcgt cagtattaag cgggggagaa ttagatcgcg
     9781 atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa tataaattaa aacatatagt
     9841 atgggcaagc agggagctag aacgattcgc agttaatcct ggcctgttag aaacatcaga
     9901 aggctgtaga caaatactgg gacagctaca accatccctt cagacaggat cagaagaact
     9961 tagatcatta tataatacag tagcaaccct ctattgtgtg catcaaagga tagagataaa
    10021 agacaccaag gaagctttag acaagataga ggaagagcaa aacaaaagta agaccaccgc
    10081 acagcaagcg gccgctgatc ttcagacctg gaggaggaga tatgagggac aattggagaa
    10141 gtgaattata taaatataaa gtagtaaaaa ttgaaccatt aggagtagca cccaccaagg
    10201 caaagagaag agtggtgcag agagaaaaaa gagcagtggg aataggagct ttgttccttg
    10261 ggttcttggg agcagcagga agcactatgg gcgcagcgtc aatgacgctg acggtacagg
    10321 ccagacaatt attgtctggt atagtgcagc agcagaacaa tttgctgagg gctattgagg
    10381 cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa gcagctccag gcaagaatcc
    10441 tggctgtgga aagataccta aaggatcaac agctcctggg gatttggggt tgctctggaa
    10501 aactcatttg caccactgct gtgccttgga atgctagttg gagtaataaa tctctggaac
    10561 agatttggaa tcacacgacc tggatggagt gggacagaga aattaacaat tacacaagct
    10621 taatacactc cttaattgaa gaatcgcaaa accagcaaga aaagaatgaa caagaattat
    10681 tggaattaga taaatgggca agtttgtgga attggtttaa cataacaaat tggctgtggt
    10741 atataaaatt attcataatg atagtaggag gcttggtagg tttaagaata gtttttgctg
    10801 tactttctat agtgaataga gttaggcagg gatattcacc attatcgttt cagacccacc
    10861 tcccaacccc gaggggtgcc cgtcagt
//

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