LOCUS       sequence-318522-        8769 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cas9 activity reporter with BFP and GFP..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8769)
  AUTHORS   Tzelepis K, Koike-Yusa H, De Braekeleer E, Li Y, Metzakopian E, 
            Dovey OM, Mupo A, Grinkevich V, Li M, Mazan M, Gozdecka M, Ohnishi 
            S, Cooper J, Patel M, McKerrell T, Chen B, Domingues AF, Gallipoli 
            P, Teichmann S, Ponstingl H, McDermott U, Saez-Rodriguez J, Huntly 
            BJ, Iorio F, Pina C, Vassiliou GS, Yusa K
  TITLE     A CRISPR Dropout Screen Identifies Genetic Vulnerabilities and 
            Therapeutic Targets in Acute Myeloid Leukemia.
  JOURNAL   Cell Rep. 2016 Oct 18;17(4):1193-1205. doi: 
            10.1016/j.celrep.2016.09.079.
  PUBMED    27760321
REFERENCE   2  (bases 1 to 8769)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.celrep.2016.09"; journalName: "Cell Rep"; date: 
            "2016-10-18- 18"; volume: "17"; issue: "4"; pages: "1193-1205"
COMMENT     Sequence Label: pKLV2-U6gRNA5(gGFP)-PGKBFP2AGFP-W
FEATURES             Location/Qualifiers
     source          1..8769
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..249
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1..21
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     172..191
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     promoter        378..877
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(413..436)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     810..829
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      901..910
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             907..1608
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MGSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIK
                     VVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMAL
                     KLVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARY
                     CDLPSKLGHKLN"
     CDS             1627..1680
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             join(1687..1689,1690..1692,1693..2406)
                     /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             join(1687..1689,1690..1692,1693..2403)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1732..1753)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1993..2012)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2340..2361
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2466..2486)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2884..2895)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3347..3364)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3353..3577
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3603..3623)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3603..3621)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3642..3658)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3642..3658)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3655..3677)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3666..3682
                     /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)."
     promoter        complement(join(3690..3696,3697..3714,3715..3720))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3735..3756
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3873..3890)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4044..4632)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4124..4143)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4803..5594,5595..5663))
                     /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     5426..5445
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5664..5768)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5794..6249)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5931..5952)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6143..6162
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6376..6398
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6390..6407
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6391..6407
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6417..6436
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6417..6435
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        6751..6949
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6904..6924
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             6967..7147
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7194..7319
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7818..8051
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8236..8280
                     /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    8578..8695
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag
       61 ataattagaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga
      121 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat
      181 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga
      241 cgaaacaccg aagttcgagg gcgacacccg tttaagagct atgctggaaa cagcatagca
      301 agtttaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt
      361 ttggatccaa ttctaccggg taggggaggc gcttttccca aggcagtctg gagcatgcgc
      421 tttagcagcc ccgctgggca cttggcgcta cacaagtggc ctctggcctc gcacacattc
      481 cacatccacc ggtaggcgcc aaccggctcc gttctttggt ggccccttcg cgccaccttc
      541 tactcctccc ctagtcagga agttcccccc cgccccgcag ctcgcgtcgt gcaggacgtg
      601 acaaatggaa gtagcacgtc tcactagtct cgtgcagatg gacagcaccg ctgagcaatg
      661 gaagcgggta ggcctttggg gcagcggcca atagcagctt tgctccttcg ctttctgggc
      721 tcagaggctg ggaaggggtg ggtccggggg cgggctcagg ggcgggctca ggggcggggc
      781 gggcgcccga aggtcctccg gaggcccggc attctgcacg cttcaaaagc gcacgtctgc
      841 cgcgctgttc tcctcttcct catctccggg cctttcgacc tgcatccatc tagatctcga
      901 gccaccatgg gcagcgagct gattaaggag aacatgcaca tgaagctgta catggagggc
      961 accgtggaca accatcactt caagtgcaca tccgagggcg aaggcaagcc ctacgagggc
     1021 acccagacca tgagaatcaa ggtggtcgag ggcggccctc tccccttcgc cttcgacatc
     1081 ctggctacta gcttcctcta cggcagcaag accttcatca accacaccca gggcatcccc
     1141 gacttcttca agcagtcctt ccctgagggc ttcacatggg agagagtcac cacatacgag
     1201 gacgggggcg tgctgaccgc tacccaggac accagcctcc aggacggctg cctcatctac
     1261 aacgtcaaga tcagaggggt gaacttcaca tccaacggcc ctgtgatgca gaagaaaaca
     1321 ctcggctggg aggccttcac cgagaccctg taccccgctg acggcggcct ggaaggcaga
     1381 aacgacatgg ccctgaagct cgtgggcggg agccatctga tcgcaaacat caagaccaca
     1441 tatagatcca agaaacccgc taagaacctc aagatgcctg gcgtctacta tgtggactac
     1501 agactggaaa gaatcaagga ggccaacaac gagacctacg tcgagcagca cgaggtggca
     1561 gtggccagat actgcgacct ccctagcaaa ctggggcaca agcttaatgg agggggcagc
     1621 ggcggggaag gaaggggctc cctgctcacc tgcggcgacg tggaagagaa ccccggacct
     1681 gaattcatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
     1741 ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc
     1801 acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg
     1861 cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac
     1921 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
     1981 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac
     2041 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
     2101 gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag
     2161 aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag
     2221 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     2281 aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac
     2341 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
     2401 aagtaagcgg ccgctcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     2461 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     2521 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
     2581 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
     2641 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
     2701 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     2761 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt
     2821 ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
     2881 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
     2941 cttccgcgtc tacgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
     3001 caggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa
     3061 gaaaaggggg gactggaagg gctaattcac tcccaaagaa gtcaagatct gctttttgcc
     3121 tgtactgggt ctctctggtt agaccagagt ctctctggtt agaccagatc tgagcctggg
     3181 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc
     3241 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct
     3301 tttagtcagt gtggaaaatc tctagcagtt taaacccgct gatcagcctc gactgtgcct
     3361 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     3421 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     3481 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagtc
     3541 aatagcaggc atgctgggga tgcggtgggc tctatgggcg gccgttaatt aagcttttgt
     3601 tccctttagt gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg
     3661 tgaaattgtt atccgctcac aattccacac aacatacgag ccgggagcat aaagtgtaaa
     3721 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     3781 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     3841 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     3901 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     3961 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     4021 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     4081 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     4141 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     4201 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     4261 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     4321 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     4381 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     4441 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     4501 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     4561 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     4621 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     4681 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     4741 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     4801 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     4861 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     4921 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     4981 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     5041 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     5101 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     5161 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     5221 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     5281 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     5341 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     5401 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     5461 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     5521 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     5581 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     5641 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     5701 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     5761 gttccgcgca catttccccg aaaagtgcca cctaaattgt aagcgttaat attttgttaa
     5821 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
     5881 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
     5941 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     6001 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
     6061 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     6121 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
     6181 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     6241 agggcgcgtc ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
     6301 cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg
     6361 taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgag cgcgcgtaat
     6421 acgactcact atagggcgaa ttgactagta cataacttac ggtaaatggc ccgcctggct
     6481 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
     6541 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
     6601 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
     6661 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     6721 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
     6781 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
     6841 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
     6901 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcagc gcgttttgcc
     6961 tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg
     7021 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt
     7081 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc
     7141 tctagcagtg gcgcccgaac agggacttga aagcgaaagg gaaaccagag gagctctctc
     7201 gacgcaggac tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg cgactggtga
     7261 gtacgccaaa aattttgact agcggaggct agaaggagag agatgggtgc gagagcgtca
     7321 gtattaagcg ggggagaatt agatcgcgat gggaaaaaat tcggttaagg ccagggggaa
     7381 agaaaaaata taaattaaaa catatagtat gggcaagcag ggagctagaa cgattcgcag
     7441 ttaatcctgg cctgttagaa acatcagaag gctgtagaca aatactggga cagctacaac
     7501 catcccttca gacaggatca gaagaactta gatcattata taatacagta gcaaccctct
     7561 attgtgtgca tcaaaggata gagataaaag acaccaagga agctttagac aagatagagg
     7621 aagagcaaaa caaaagtaag accaccgcac agcaagcggc cggccgcgct gatcttcaga
     7681 cctggaggag gagatatgag ggacaattgg agaagtgaat tatataaata taaagtagta
     7741 aaaattgaac cattaggagt agcacccacc aaggcaaaga gaagagtggt gcagagagaa
     7801 aaaagagcag tgggaatagg agctttgttc cttgggttct tgggagcagc aggaagcact
     7861 atgggcgcag cgtcaatgac gctgacggta caggccagac aattattgtc tggtatagtg
     7921 cagcagcaga acaatttgct gagggctatt gaggcgcaac agcatctgtt gcaactcaca
     7981 gtctggggca tcaagcagct ccaggcaaga atcctggctg tggaaagata cctaaaggat
     8041 caacagctcc tggggatttg gggttgctct ggaaaactca tttgcaccac tgctgtgcct
     8101 tggaatgcta gttggagtaa taaatctctg gaacagattt ggaatcacac gacctggatg
     8161 gagtgggaca gagaaattaa caattacaca agcttaatac actccttaat tgaagaatcg
     8221 caaaaccagc aagaaaagaa tgaacaagaa ttattggaat tagataaatg ggcaagtttg
     8281 tggaattggt ttaacataac aaattggctg tggtatataa aattattcat aatgatagta
     8341 ggaggcttgg taggtttaag aatagttttt gctgtacttt ctatagtgaa tagagttagg
     8401 cagggatatt caccattatc gtttcagacc cacctcccaa ccccgagggg acccgacagg
     8461 cccgaaggaa tagaagaaga aggtggagag agagacagag acagatccat tcgattagtg
     8521 aacggatcgg cactgcgtgc gccaattctg cagacaaatg gcagtattca tccacaattt
     8581 taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc
     8641 aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa attttcgggt
     8701 ttattacagg gacagcagag atccagtttg gttagtaccg ggccctacgc gtccaaggtc
     8761 gggcaggaa
//
