pLV-egfp-U3-Osp.gRNA

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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: pLV-egfp-U3-Osp.gRNA
出品公司: Addgene
目录编号: 166034
存储实验室: Yujia Cai

载体骨架基本信息
载体名称: pCCL-PGK-egfp
载体出品公司: --
空载体大小: --
完整质粒大小: --
载体修饰: --
载体类型: 慢病毒,CRISPR
筛选标记: EGFP
载体抗性: 氨苄霉素
生长温度: 37 ℃
宿主菌: NEB Stable
拷贝数: 高拷贝

插入基因信息
插入基因名称: --
物种来源: --
插入大小: --
基因突变: --
GenBank编号 --
启动子: --
标签/融合蛋白: --
克隆方法: --
5’克隆位点: --
3’克隆位点: --
5’测序引物 --
3’测序引物 --

文献引用方法
材料和方法部分: pLV-egfp-U3-Osp.gRNA was a gift from Yujia Cai (Addgene plasmid # 166034 ; http://n2t.net/addgene:166034 ; RRID:Addgene_166034)
参考文献部分: Lentiviral delivery of co-packaged Cas9 mRNA and a Vegfa-targeting guide RNA prevents wet age-related macular degeneration in mice. Ling S, Yang S, Hu X, Yin D, Dai Y, Qian X, Wang D, Pan X, Hong J, Sun X, Yang H, Paludan SR, Cai Y. Nat Biomed Eng. 2021 Jan 4. pii: 10.1038/s41551-020-00656-y. doi: 10.1038/s41551-020-00656-y. 10.1038/s41551-020-00656-y PubMed 33398131

载体图谱质粒图谱和多克隆位点信息

pLV-egfp-U3-Osp.gRNA载体图谱

载体简介

载体序列

LOCUS      pLV-egfp-U3-Osp.gRNA               8119 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Empty backbone for expression of gRNA-expressing integration 
            deficient lentiviral particles..
ACCESSION   .
VERSION     .
KEYWORDS    pLV-egfp-U3-Osp.gRNA
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8119)
  AUTHORS   Ling S, Yang S, Hu X, Yin D, Dai Y, Qian X, Wang D, Pan X, Hong J, 
            Sun X, Yang H, Paludan SR, Cai Y
  TITLE     Lentiviral delivery of co-packaged Cas9 mRNA and a Vegfa-targeting 
            guide RNA prevents wet age-related macular degeneration in mice.
  JOURNAL   Nat Biomed Eng. 2021 Jan 4. pii: 10.1038/s41551-020-00656-y. doi: 
            10.1038/s41551-020-00656-y.
  PUBMED    33398131
REFERENCE   2  (bases 1 to 8119)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8119
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             219..399
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    446..571
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1063..1296
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1481..1525
                     /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    1792..1909
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1948..2447
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(1983..2006)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     2380..2399
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      2471..2480
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             2477..3196
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             2477..3193
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2522..2543)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2783..2802)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3130..3151
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3220..3808
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3273..3293)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3691..3702)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        3929..4169
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     primer_bind     3929..3949
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     4100..4119
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     LTR             4333..4513
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(4542..4560)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4543..4560)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(4806..4825)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4807..4825)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4835..4852)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4835..4851)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4844..4866)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4993..5448
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5080..5099)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5290..5311
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5474..5578
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5579..6439
                     /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(5797..5816)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6610..7198
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7099..7118
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7352..7369
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7486..7507
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        7522..7552
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7560..7576
                     /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     7565..7587
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7584..7600
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7584..7600
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7619..7639
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7621..7639
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     enhancer        7740..8119
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctcgttta gtgaaccggg gtctctctgg ttagaccaga
      241 tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct
      301 tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat
      361 ccctcagacc cttttagtca gtgtggaaaa tctctagcag tggcgcccga acagggactt
      421 gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg actcggcttg ctgaagcgcg
      481 cacggcaaga ggcgaggggc ggcgactggt gagtacgcca aaaattttga ctagcggagg
      541 ctagaaggag agagatgggt gcgagagcgt cagtattaag cggggagaat tagatcgcga
      601 tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat ataaattaaa acatatagta
      661 tgggcaagca gggagctaga acgattcgca gttaatcctg gcctgttaga aacatcagaa
      721 ggctgtagac aaatactggg acagctacaa ccatcccttc agacaggatc agaagaactt
      781 agatcattat ataatacagt agcaaccctc tattgtgtgc atcaaaggat agagataaaa
      841 gacaccaagg aagctttaga caagatagag gaagagcaaa acaaaagtaa gaccaccgca
      901 cagcaagcgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
      961 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1021 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1081 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1141 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1201 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1261 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1321 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1381 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1441 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1501 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1561 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1621 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1681 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1741 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcggttaa cttttaaaag
     1801 aaaagggggg attggggggt acagtgcagg ggaaagaata gtagacataa tagcaacaga
     1861 catacaaact aaagaattac aaaaacaaat tacaaaaatt caaaatttta tcgatcacga
     1921 gactagcctc gacgatggtc gagtaccggg taggggaggc gcttttccca aggcagtctg
     1981 gagcatgcgc tttagcagcc ccgctgggca cttggcgcta cacaagtggc ctctggcctc
     2041 gcacacattc cacatccacc ggtaggcgcc aaccggctcc gttctttggt ggccccttcg
     2101 cgccaccttc tactcctccc ctagtcagga agttcccccc cgccccgcag ctcgcgtcgt
     2161 gcaggacgtg acaaatggaa gtagcacgtc tcactagtct cgtgcagatg gacagcaccg
     2221 ctgagcaatg gaagcgggta ggcctttggg gcagcggcca atagcagctt tgctccttcg
     2281 ctttctgggc tcagaggctg ggaaggggtg ggtccggggg cgggctcagg ggcgggctca
     2341 ggggcggggc gggcgcccga aggtcctccg gaggcccggc attctgcacg cttcaaaagc
     2401 gcacgtctgc cgcgctgttc tcctcttcct catctccggg cctttcgacc tctagcggga
     2461 tccaccggtc gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
     2521 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga
     2581 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc
     2641 cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta
     2701 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
     2761 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
     2821 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
     2881 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc
     2941 cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg
     3001 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
     3061 gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa
     3121 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
     3181 cgagctgtac aagtaaagcg gcctcgaggg aattccgata atcaacctct ggattacaaa
     3241 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     3301 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     3361 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
     3421 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
     3481 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
     3541 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     3601 gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt
     3661 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
     3721 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
     3781 cggatctccc tttgggccgc ctccccgcat cgggaattcg agctcggtac ctttaagacc
     3841 aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga
     3901 agggctaatt cactcccaac gaagacaaga gggcctattt cccatgattc cttcatattt
     3961 gcatatacga tacaaggctg ttagagagat aattggaatt aatttgactg taaacacaaa
     4021 gatattagta caaaatacgt gacgtagaaa gtaataattt cttgggtagt ttgcagtttt
     4081 aaaattatgt tttaaaatgg actatcatat gcttaccgta acttgaaagt atttcgattt
     4141 cttggcttta tatatcttgt ggaaaggacg aaacaccaaa agcaggtgga catacaagtc
     4201 gacaatagca cctgcaaaag tttcagagct atgctggaaa cagcatagca agttgaaata
     4261 aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt tgatctgctt
     4321 tttgcttgta ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa
     4381 ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt
     4441 gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg
     4501 aaaatctcta gcagcatcta gctagaatta attccgtgta ttctatagtg tcacctaaat
     4561 cgtatgtgta tgatacataa ggttatgtat taattgtagc cgcgttctaa cgacaatatg
     4621 tacaagccta attgtgtagc atctggctta ctgaagcaga ccctatcatc tctctcgtaa
     4681 actgccgtca gagtcggttt ggttggacga accttctgag tttctggtaa cgccgtcccg
     4741 cacccggaaa tggtcagcga accaatcagc agggtcatcg ctagcctagg gacgtaccca
     4801 attcgcccta tagtgagtcg tattacgcgc gctcactggc cgtcgtttta caacgtcgtg
     4861 actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca
     4921 gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga
     4981 atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     5041 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     5101 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     5161 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     5221 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     5281 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     5341 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     5401 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag gtggcacttt
     5461 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     5521 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     5581 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     5641 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     5701 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     5761 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     5821 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     5881 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     5941 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     6001 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     6061 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     6121 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     6181 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     6241 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     6301 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     6361 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     6421 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     6481 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     6541 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     6601 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     6661 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     6721 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     6781 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     6841 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     6901 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     6961 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     7021 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     7081 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     7141 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     7201 cgccagcaac gcggtctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     7261 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     7321 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     7381 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
     7441 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
     7501 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
     7561 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccaagcgcgc
     7621 aattaaccct cactaaaggg aacaaaagct ggagctgcaa gcttggccat tgcatacgtt
     7681 gtatccatat cataatatgt acatttatat tggctcatgt ccaacattac cgccatgttg
     7741 acattgatta ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc
     7801 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
     7861 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
     7921 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
     7981 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
     8041 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
     8101 agtcatcgct attaccatg
//

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