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pLenti-PGK-flag-RAP1B-N17-puro

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pLenti-PGK-flag-RAP1B-N17-puro載體質粒基本信息

質粒名稱: pLenti-PGK-flag-RAP1B-N17-puro
出品公司: Addgene
目錄編號: 156169
存儲實驗室: Roland Friedel

載體骨架基本信息
載體名稱:
pLenti PGK Puro DEST (w529-2)
Backbone manufacturerCampeau lab (addgene #19068)
載體出品公司: Clontech
空載體大小: 9605bp
完整質粒大小: 8619bp
載體類型: 哺乳細胞表達, Lentiviral
篩選標記: Puromycin
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: RAP1B
物種來源: B. taurus (bovine)
插入大小: 609bp
基因突變: changed Serine 17 to Asparagine
GenBank編號 NM_175824
啟動子: human PGK
標簽/融合蛋白: 2xFLAG (N terminal on insert)
克隆方法: Gateway Cloning
3’克隆位點: NotI (未破壞)
5’測序引物 PGK_F (GTGTTCCGCATTCTGCAAG)
3’測序引物
WPRE_R (CATAGCGTAAAAGGAGCAACA)
(Common Sequencing Primers)

文獻引用方法
材料和方法部分: pLenti-PGK-flag-RAP1B-N17-puro was a gift from Roland Friedel (Addgene plasmid # 156169 ; //n2t.net/addgene:156169 ; RRID:Addgene_156169)
參考文獻部分: Plexin-B2 orchestrates collective stem cell dynamics via actomyosin contractility, cytoskeletal tension and adhesion. Junqueira Alves C, Dariolli R, Haydak J, Kang S, Hannah T, Wiener RJ, DeFronzo S, Tejero R, Gusella GL, Ramakrishnan A, Alves Dias R, Wojcinski A, Kesari S, Shen L, Sobie EA, Rodrigues Furtado de Mendonca JP, Azeloglu EU, Zou H, Friedel RH. Nat Commun. 2021 Oct 14;12(1):6019. doi: 10.1038/s41467-021-26296-7. 10.1038/s41467-021-26296-7 PubMed 34650052

pLenti-PGK-flag-RAP1B-N17-puro質粒圖譜載體圖譜和 pLenti-PGK-flag-RAP1B-N17-puro載體序列質粒序列多克隆位點信息

pLenti-PGK-flag-RAP1B-N17-puro載體圖譜

pLenti-PGK-flag-RAP1B-N17-puro質粒載體簡介

pLenti-PGK-flag-RAP1B-N17-puro質粒序列載體序列

LOCUS        pLenti-PGK-flag-RAP1B-N17-puro       8619 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Lentiviral vector for expression of flag-RAP1B-N17, with puromycin 
            selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8619)
  AUTHORS   Junqueira Alves C, Dariolli R, Haydak J, Kang S, Hannah T, Wiener 
            RJ, DeFronzo S, Tejero R, Gusella GL, Ramakrishnan A, Alves Dias R, 
            Wojcinski A, Kesari S, Shen L, Sobie EA, Rodrigues Furtado de 
            Mendonca JP, Azeloglu EU, Zou H, Friedel RH
  TITLE     Plexin-B2 orchestrates collective stem cell dynamics via actomyosin 
            contractility, cytoskeletal tension and adhesion.
  JOURNAL   Nat Commun. 2021 Oct 14;12(1):6019. doi: 10.1038/s41467-021-26296-7.
  PUBMED    34650052
REFERENCE   2  (bases 1 to 8619)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            //gt12365.cn
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2021-10-14"; volume: "12(1):6019. doi"; pages: " 
            10.1038/s41467-021-26296-7"
COMMENT     Sequence Label: pLenti-PGK-flag-RAP1B-N17-puro
FEATURES             Location/Qualifiers
     source          1..8619
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        26..130
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(131..199,200..991)
                     /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(349..368)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1162..1750
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1651..1670
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1904..1921
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2038..2059
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2074..2079,2080..2097,2098..2104)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2112..2128
                     /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     2117..2139
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2136..2152
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2136..2152
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2171..2191
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2173..2191
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2219..2445
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             2446..2626
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    2673..2798
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    3291..3524
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             3709..3753
                     /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    4020..4137
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        4183..4693
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     4612..4630
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     protein_bind    4748..4772
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      4802..4811
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4811..4858
                     /codon_start=1
                     /product="two tandem FLAG(R) epitope tags"
                     /label=2xFLAG
                     /translation="DYKDDDDKDYKDDDDK"
     protein_bind    complement(5442..5466)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    5497..6085
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5550..5570)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5968..5979)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        6099..6598
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(6134..6157)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     6531..6550
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             6619..7218
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(6619..6638)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7115..7135
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             7356..7589
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    7661..7782
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7698..7717)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7752..7771
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      7822..7957
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(7977..7996)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7978..7996)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8006..8023)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8006..8022)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8015..8037)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      8164..8592
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8251..8270)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8461..8482
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
       61 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
      121 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
      181 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
      241 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
      301 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
      361 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
      421 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
      481 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
      541 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
      601 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
      661 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
      721 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
      781 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
      841 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
      901 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
      961 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     1021 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     1081 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     1141 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     1201 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     1261 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     1321 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     1381 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     1441 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     1501 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     1561 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     1621 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     1681 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     1741 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     1801 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     1861 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     1921 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     1981 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
     2041 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
     2101 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta
     2161 cgccaagcgc gcaattaacc ctcactaaag ggaacaaaag ctggagctgc aagcttaatg
     2221 tagtcttatg caatactctt gtagtcttgc aacatggtaa cgatgagtta gcaacatgcc
     2281 ttacaaggag agaaaaagca ccgtgcatgc cgattggtgg aagtaaggtg gtacgatcgt
     2341 gccttattag gaaggcaaca gacgggtctg acatggattg gacgaaccac tgaattgccg
     2401 cattgcagag atattgtatt taagtgccta gctcgataca taaacgggtc tctctggtta
     2461 gaccagatct gagcctggga gctctctggc taactaggga acccactgct taagcctcaa
     2521 taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac
     2581 tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca
     2641 gggacttgaa agcgaaaggg aaaccagagg agctctctcg acgcaggact cggcttgctg
     2701 aagcgcgcac ggcaagaggc gaggggcggc gactggtgag tacgccaaaa attttgacta
     2761 gcggaggcta gaaggagaga gatgggtgcg agagcgtcag tattaagcgg gggagaatta
     2821 gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat aaattaaaac
     2881 atatagtatg ggcaagcagg gagctagaac gattcgcagt taatcctggc ctgttagaaa
     2941 catcagaagg ctgtagacaa atactgggac agctacaacc atcccttcag acaggatcag
     3001 aagaacttag atcattatat aatacagtag caaccctcta ttgtgtgcat caaaggatag
     3061 agataaaaga caccaaggaa gctttagaca agatagagga agagcaaaac aaaagtaaga
     3121 ccaccgcaca gcaagcggcc gctgatcttc agacctggag gaggagatat gagggacaat
     3181 tggagaagtg aattatataa atataaagta gtaaaaattg aaccattagg agtagcaccc
     3241 accaaggcaa agagaagagt ggtgcagaga gaaaaaagag cagtgggaat aggagctttg
     3301 ttccttgggt tcttgggagc agcaggaagc actatgggcg cagcgtcaat gacgctgacg
     3361 gtacaggcca gacaattatt gtctggtata gtgcagcagc agaacaattt gctgagggct
     3421 attgaggcgc aacagcatct gttgcaactc acagtctggg gcatcaagca gctccaggca
     3481 agaatcctgg ctgtggaaag atacctaaag gatcaacagc tcctggggat ttggggttgc
     3541 tctggaaaac tcatttgcac cactgctgtg ccttggaatg ctagttggag taataaatct
     3601 ctggaacaga tttggaatca cacgacctgg atggagtggg acagagaaat taacaattac
     3661 acaagcttaa tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa
     3721 gaattattgg aattagataa atgggcaagt ttgtggaatt ggtttaacat aacaaattgg
     3781 ctgtggtata taaaattatt cataatgata gtaggaggct tggtaggttt aagaatagtt
     3841 tttgctgtac tttctatagt gaatagagtt aggcagggat attcaccatt atcgtttcag
     3901 acccacctcc caaccccgag gggacccgac aggcccgaag gaatagaaga agaaggtgga
     3961 gagagagaca gagacagatc cattcgatta gtgaacggat ctcgacggta tcggttaact
     4021 tttaaaagaa aaggggggat tggggggtac agtgcagggg aaagaatagt agacataata
     4081 gcaacagaca tacaaactaa agaattacaa aaacaaatta caaaaattca aaattttatc
     4141 gatcacgaga ctagcctcga gaagcttgat atcgaattcc acggggttgg ggttgcgcct
     4201 tttccaaggc agccctgggt ttgcgcaggg acgcggctgc tctgggcgtg gttccgggaa
     4261 acgcagcggc gccgaccctg ggtctcgcac attcttcacg tccgttcgca gcgtcacccg
     4321 gatcttcgcc gctacccttg tgggcccccc ggcgacgctt cctgctccgc ccctaagtcg
     4381 ggaaggttcc ttgcggttcg cggcgtgccg gacgtgacaa acggaagccg cacgtctcac
     4441 tagtaccctc gcagacggac agcgccaggg agcaatggca gcgcgccgac cgcgatgggc
     4501 tgtggccaat agcggctgct cagcagggcg cgccgagagc agcggccggg aaggggcggt
     4561 gcgggaggcg gggtgtgggg cggtagtgtg ggccctgttc ctgcccgcgc ggtgttccgc
     4621 attctgcaag cctccggagc gcacgtcggc agtcggctcc ctcgttgacc gaatcaccga
     4681 cctctctccc cagggggatc caccggttcg tcgactagtc cagtgtggtg gaattctgca
     4741 gatatcaaca agtttgtaca aaaaagcagg ctccgcggcc gcccccttca ccctcggatc
     4801 cgccaccatg gactacaagg acgacgatga caaggactac aaggacgacg atgacaagaa
     4861 ttctcgtgag tataagctag tcgttcttgg ctctggaggc gttggaaaaa atgctctgac
     4921 tgtacagttt gttcaaggaa tttttgttga aaaatatgat cctacgatag aagattctta
     4981 tagaaagcaa gttgaagtag atgcacagca gtgtatgctt gaaatcttgg atactgcagg
     5041 aacggaacaa ttcacagcaa tgagggattt gtacatgaaa aatggacaag gctttgcgtt
     5101 agtttattcc atcacagcac agtccacatt taatgattta caagatctga gagaacagat
     5161 acttcgagtt aaagacacgg atgatgttcc aatgattctg gttggtaata agtgtgactt
     5221 ggaagatgaa agagttgtag gaaaggaaca aggtcaaaat ctagcaagac agtggaacaa
     5281 ctgtgcattc ttagaatcct ctgcaaaatc aaaaataaat gttaatgaga tcttttatga
     5341 cctagtgcga caaattaaca gaaaaactcc agtgcctggg aaggcccgca aaaagtcatc
     5401 atgtcagctg ctttaatcta gagggaaggg tgggcgcgcc gacccagctt tcttgtacaa
     5461 agtggttgat atccagcaca gtggcggccg ctcgacaatc aacctctgga ttacaaaatt
     5521 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     5581 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     5641 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc
     5701 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt
     5761 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc
     5821 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg
     5881 ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac ctggattctg
     5941 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc
     6001 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg
     6061 atctcccttt gggccgcctc cccgcctgga attctaccgg gtaggggagg cgcttttccc
     6121 aaggcagtct ggagcatgcg ctttagcagc cccgctgggc acttggcgct acacaagtgg
     6181 cctctggcct cgcacacatt ccacatccac cggtaggcgc caaccggctc cgttctttgg
     6241 tggccccttc gcgccacctt ctactcctcc cctagtcagg aagttccccc ccgccccgca
     6301 gctcgcgtcg tgcaggacgt gacaaatgga agtagcacgt ctcactagtc tcgtgcagat
     6361 ggacagcacc gctgagcaat ggaagcgggt aggcctttgg ggcagcggcc aatagcagct
     6421 ttgctccttc gctttctggg ctcagaggct gggaaggggt gggtccgggg gcgggctcag
     6481 gggcgggctc aggggcgggg cgggcgcccg aaggtcctcc ggaggcccgg cattctgcac
     6541 gcttcaaaag cgcacgtctg ccgcgctgtt ctcctcttcc tcatctccgg gcctttcgac
     6601 ctgcagccca agcttaccat gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac
     6661 gacgtcccca gggccgtacg caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc
     6721 cacaccgtcg atccggaccg ccacatcgag cgggtcaccg agctgcaaga actcttcctc
     6781 acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg
     6841 gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc
     6901 atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg cctcctggcg
     6961 ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac
     7021 cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc
     7081 ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta cgagcggctc
     7141 ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc
     7201 cgcaagcccg gtgcctgacg cccgccccac gacccgcagc gcccgaccga aaggagcgca
     7261 cgaccccatg catctcgagg gcccggtacc tttaagacca atgacttaca aggcagctgt
     7321 agatcttagc cactttttaa aagaaaaggg gggactggaa gggctagctc actcccaacg
     7381 aagacaagat ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg
     7441 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt
     7501 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc
     7561 cttttagtca gtgtggaaaa tctctagcag tagtagttca tgtcatctta ttattcagta
     7621 tttataactt gcaaagaaat gaatatcaga gagtgagagg aacttgttta ttgcagctta
     7681 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     7741 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggctctagct
     7801 atcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     7861 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     7921 ctattccaga agtagtgagg aggctttttt ggaggcctag ggacgtaccc aattcgccct
     7981 atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa
     8041 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta
     8101 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
     8161 gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     8221 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     8281 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     8341 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     8401 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     8461 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     8521 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     8581 ttaacgcgaa ttttaacaaa atattaacgc ttacaattt
//

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