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tet-pLKO.puro_shLuc

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tet-pLKO.puro_shLuc載體質粒基本信息

質粒名稱: tet-pLKO.puro_shLuc
出品公司: Addgene
目錄編號: 110422
存儲實驗室: Sandra Martha Gomes Dias

載體骨架基本信息
載體名稱: Tet-pLKO-puro
載體出品公司: Clontech
空載體大小: 10633bp
完整質粒大小: 8815bp
載體類型: 哺乳細胞表達, Lentiviral, RNAi
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: Luciferase
物種來源: Synthetic
啟動子: H1/TO (RNA PolIII)
克隆方法: 限制性內切酶
5’克隆位點: AgeI (destroyed during cloning)
3’克隆位點: EcoRI (not destroyed)
5’測序引物 GACAGAGACAGATCCATTCG

文獻引用方法
材料和方法部分: tet-pLKO.puro_shLuc was a gift from Sandra Martha Gomes Dias (Addgene plasmid # 110422 ; //n2t.net/addgene:110422 ; RRID:Addgene_110422)

tet-pLKO.puro_shLuc質粒圖譜載體圖譜和 tet-pLKO.puro_shLuc載體序列質粒序列多克隆位點信息

tet-pLKO.puro_shLuc載體圖譜

tet-pLKO.puro_shLuc質粒載體簡介

tet-pLKO.puro_shLuc質粒序列載體序列

LOCUS       tet-pLKO.puro_shLuc       8815 bp    DNA     circular SYN 14-APR-2025
DEFINITION  shLuc control, silence Luc gene, doxycycline inducible, puromycin 
            selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8815)
  TITLE     Dias lab diverse plasmid collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8815)
  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: 
            "Unpublished"
COMMENT     Sequence Label: tet-pLKO.puro_shLuc
FEATURES             Location/Qualifiers
     source          1..8815
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..227
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             228..408
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    455..580
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1073..1306
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1491..1535
                     /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"
     promoter        1824..1918
                     /label=H1-2O2 promoter
                     /note="doxycycline-inducible variant of the human H1 RNA 
                     promoter (Henriksen et al., 2007)"
     primer_bind     1835..1854
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     protein_bind    1872..1890
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1900..1918
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_feature    2018..2135
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2184..2694
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     2613..2631
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     intron          2703..3275
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     3283..3302
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     promoter        3330..3348
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3359..3979
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     complement(3432..3451)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     misc_feature    4014..4587
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4181..4198)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4408..4427
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4607..5206
                     /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(4607..4626)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5103..5123
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             5334..5567
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5639..5760
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5676..5695)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5730..5749
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5800..5935
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(5956..5974)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5984..6001)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5984..6000)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5993..6015)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6142..6597
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6229..6248)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6439..6460
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6623..6727
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6728..6796,6797..7588)
                     /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(6946..6965)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7759..8347
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8248..8267
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8501..8518
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8635..8656
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8671..8676,8677..8694,8695..8701)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8709..8725
                     /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     8714..8736
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8733..8749
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8733..8749
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8768..8788
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        8770..8788
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 tgtagtctta tgcaatactc ttgtagtctt gcaacatggt aacgatgagt tagcaacatg
       61 ccttacaagg agagaaaaag caccgtgcat gccgattggt ggaagtaagg tggtacgatc
      121 gtgccttatt aggaaggcaa cagacgggtc tgacatggat tggacgaacc actgaattgc
      181 cgcattgcag agatattgta tttaagtgcc tagctcgata cataaacggg tctctctggt
      241 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
      301 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
      361 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa
      421 cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga ctcggcttgc
      481 tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa aaattttgac
      541 tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc gggggagaat
      601 tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat ataaattaaa
      661 acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg gcctgttaga
      721 aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc agacaggatc
      781 agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc atcaaaggat
      841 agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa acaaaagtaa
      901 gaccaccgca cagcaagcgg ccgctgatct tcagacctgg aggaggagat atgagggaca
      961 attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac
     1021 ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt
     1081 tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga
     1141 cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg
     1201 ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg
     1261 caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt
     1321 gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat
     1381 ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt
     1441 acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac
     1501 aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt
     1561 ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag
     1621 tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc
     1681 agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg
     1741 gagagagaga cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgatcac
     1801 gagactagcc tcgagcggcc gcaatatttg catgtcgcta tgtgttctgg gaaatcacca
     1861 taaacgtgaa atccctatca gtgatagaga cttataagtt ccctatcagt gatagagaca
     1921 ccggcttacg ctgagtactt cgacctgacc catcgaagta ctcagcgtaa gtttttgaat
     1981 tctcgacctc gagacaaatg gcagtattca tccacaattt taaaagaaaa ggggggattg
     2041 gggggtacag tgcaggggaa agaatagtag acataatagc aacagacata caaactaaag
     2101 aattacaaaa acaaattaca aaaattcaaa attttcgggt ttattacagg gacagcagag
     2161 atccactttg gccgcggctc gagggggttg gggttgcgcc ttttccaagg cagccctggg
     2221 tttgcgcagg gacgcggctg ctctgggcgt ggttccggga aacgcagcgg cgccgaccct
     2281 gggtctcgca cattcttcac gtccgttcgc agcgtcaccc ggatcttcgc cgctaccctt
     2341 gtgggccccc cggcgacgct tcctgctccg cccctaagtc gggaaggttc cttgcggttc
     2401 gcggcgtgcc ggacgtgaca aacggaagcc gcacgtctca ctagtaccct cgcagacgga
     2461 cagcgccagg gagcaatggc agcgcgccga ccgcgatggg ctgtggccaa tagcggctgc
     2521 tcagcagggc gcgccgagag cagcggccgg gaaggggcgg tgcgggaggc ggggtgtggg
     2581 gcggtagtgt gggccctgtt cctgcccgcg cggtgttccg cattctgcaa gcctccggag
     2641 cgcacgtcgg cagtcggctc cctcgttgac cgaatcaccg acctctctcc ccagggggat
     2701 ctgtgagttt ggggaccctt gattgttctt tctttttcgc tattgtaaaa ttcatgttat
     2761 atggaggggg caaagttttc agggtgttgt ttagaatggg aagatgtccc ttgtatcacc
     2821 atggaccctc atgataattt tgtttctttc actttctact ctgttgacaa ccattgtctc
     2881 ctcttatttt cttttcattt tctgtaactt tttcgttaaa ctttagcttg catttgtaac
     2941 gaatttttaa attcactttt gtttatttgt cagattgtaa gtactttctc taatcacttt
     3001 tttttcaagg caatcagggt atattatatt gtacttcagc acagttttag agaacaattg
     3061 ttataattaa atgataaggt agaatatttc tgcatataaa ttctggctgg cgtggaaata
     3121 ttcttattgg tagaaacaac tacatcctgg tcatcatcct gcctttctct ttatggttac
     3181 aatgatatac actgtttgag atgaggataa aatactctga gtccaaaccg ggcccctctg
     3241 ctaaccatgt tcatgccttc ttctttttcc tacagctcct gggcaacgtg ctggttattg
     3301 tgctgtctca tcattttggc aaagaattgt aatacgactc actatagggc gaattgatat
     3361 gtctagatta gataaaagta aagtgattaa cagcgcatta gagctgctta atgaggtcgg
     3421 aatcgaaggt ttaacaaccc gtaaactcgc ccagaagcta ggtgtagagc agcctacatt
     3481 gtattggcat gtaaaaaata agcgggcttt gctcgacgcc ttagccattg agatgttaga
     3541 taggcaccat actcactttt gccctttaga aggggaaagc tggcaagatt ttttacgtaa
     3601 taacgctaaa agttttagat gtgctttact aagtcatcgc gatggagcaa aagtacattt
     3661 aggtacacgg cctacagaaa aacagtatga aactctcgaa aatcaattag cctttttatg
     3721 ccaacaaggt ttttcactag agaatgcatt atatgcactc agcgctgtgg ggcattttac
     3781 tttaggttgc gtattggaag atcaagagca tcaagtcgct aaagaagaaa gggaaacacc
     3841 tactactgat agtatgccgc cattattacg acaagctatc gaattatttg atcaccaagg
     3901 tgcagagcca gccttcttat tcggccttga attgatcata tgcggattag aaaaacaact
     3961 taaatgtgaa agtgggtccg cgtacagcgg ctcccgggag ttctagggat ctgcccctct
     4021 ccctcccccc cccctaacgt tactggccga agccgcttgg aataaggccg gtgtgcgttt
     4081 gtctatatgt tattttccac catattgccg tcttttggca atgtgagggc ccggaaacct
     4141 ggccctgtct tcttgacgag cattcctagg ggtctttccc ctctcgccaa aggaatgcaa
     4201 ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag acaaacaacg
     4261 tctgtagcga ccctttgcag gcagcggaac cccccacctg gcgacaggtg cctctgcggc
     4321 caaaagccac gtgtataaga tacacctgca aaggcggcac aaccccagtg ccacgttgtg
     4381 agttggatag ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg
     4441 aaggatgccc agaaggtacc ccattgtatg ggatctgatc tggggcctcg gtgcacatgc
     4501 tttacatgtg tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg
     4561 gttttccttt gaaaaacacg atgataagga tccaccggag cttaccatga ccgagtacaa
     4621 gcccacggtg cgcctcgcca cccgcgacga cgtccccagg gccgtacgca ccctcgccgc
     4681 cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg
     4741 ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg
     4801 ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg
     4861 ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc
     4921 gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct
     4981 ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct
     5041 ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc
     5101 ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc
     5161 cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctgacgcc cgccccacga
     5221 cccgcagcgc ccgaccgaaa ggagcgcacg accccatgca tcggtacctt taagaccaat
     5281 gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg
     5341 gctaattcac tcccaacgaa gacaagatct gctttttgct tgtactgggt ctctctggtt
     5401 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca
     5461 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa
     5521 ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta gtagttcatg
     5581 tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggaa
     5641 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     5701 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     5761 tcatgtctgg ctctagctat cccgccccta actccgccca tcccgcccct aactccgccc
     5821 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     5881 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggg
     5941 acgtacccaa ttcgccctat agtgagtcgt attacgcgcg ctcactggcc gtcgttttac
     6001 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc
     6061 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
     6121 gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     6181 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     6241 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     6301 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     6361 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     6421 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     6481 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     6541 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg
     6601 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     6661 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     6721 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     6781 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     6841 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     6901 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     6961 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     7021 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     7081 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     7141 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     7201 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     7261 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     7321 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     7381 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     7441 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     7501 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     7561 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     7621 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     7681 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     7741 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     7801 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     7861 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     7921 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     7981 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     8041 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     8101 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     8161 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     8221 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     8281 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     8341 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     8401 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     8461 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     8521 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
     8581 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
     8641 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
     8701 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc
     8761 caagcgcgca attaaccctc actaaaggga acaaaagctg gagctgcaag cttaa
//

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