LOCUS       sequence-416905-       10415 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral construct expressing shRNA against LYCHOS and LYCHOS 
            WT-FLAG (resistant to shRNA).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10415)
  AUTHORS   Shin HR, Citron YR, Wang L, Tribouillard L, Goul CS, Stipp R, 
            Sugasawa Y, Jain A, Samson N, Lim CY, Davis OB, Castaneda-Carpio D, 
            Qian M, Nomura DK, Perera RM, Park E, Covey DF, Laplante M, Evers 
            AS, Zoncu R
  TITLE     Lysosomal GPCR-like protein LYCHOS signals cholesterol sufficiency 
            to mTORC1.
  JOURNAL   Science. 2022 Sep 16;377(6612):1290-1298. doi: 
            10.1126/science.abg6621. Epub 2022 Aug 25.
  PUBMED    36007018
REFERENCE   2  (bases 1 to 10415)
  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: 
            "10.1126/science.abg6621"; journalName: "Science"; date: 
            "2022-09-16- 16"; volume: "377"; issue: "6612"; pages: "1290-1298"
COMMENT     Sequence Label: pLKO shLYCHOS/LYCHOS WT-FLAG
FEATURES             Location/Qualifiers
     source          1..10415
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(245..289)
                     /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    474..707
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    1200..1325
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             1372..1552
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(1553..1779)
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     primer_bind     complement(1807..1827)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1807..1825)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1846..1862)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1846..1862)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1859..1881)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1870..1886
                     /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(1894..1900,1901..1918,1919..1924))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1939..1960
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2077..2094)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2248..2836)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2328..2347)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3007..3798,3799..3867))
                     /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     3630..3649
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3868..3972)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3998..4453)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4135..4156)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4347..4366
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4580..4602
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4594..4611
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4595..4611
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4621..4640
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4621..4639
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      4660..4795
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4835..4956
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4846..4865)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4900..4919
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     LTR             5028..5261
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     CDS             complement(5389..5988)
                     /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(5472..5492)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     5969..5988
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     misc_feature    6008..6581
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(6168..6187)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     6397..6414
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     CDS             complement(6678..6701)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             7420..7431
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     promoter        complement(9341..9851)
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     complement(9404..9422)
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     misc_feature    9900..10017
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        complement(10114..10362)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(10172..10191)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(10342..10362)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
ORIGIN
        1 cgttcactaa tcgaatggat ctgtctctgt ctctctctcc accttcttct tctattcctt
       61 cgggcctgtc gggtcccctc ggggttggga ggtgggtctg aaacgataat ggtgaatatc
      121 cctgcctaac tctattcact atagaaagta cagcaaaaac tattcttaaa cctaccaagc
      181 ctcctactat cattatgaat aattttatat accacagcca atttgttatg ttaaaccaat
      241 tccacaaact tgcccattta tctaattcca ataattcttg ttcattcttt tcttgctggt
      301 tttgcgattc ttcaattaag gagtgtatta agcttgtgta attgttaatt tctctgtccc
      361 actccatcca ggtcgtgtga ttccaaatct gttccagaga tttattactc caactagcat
      421 tccaaggcac agcagtggtg caaatgagtt ttccagagca accccaaatc cccaggagct
      481 gttgatcctt taggtatctt tccacagcca ggattcttgc ctggagctgc ttgatgcccc
      541 agactgtgag ttgcaacaga tgctgttgcg cctcaatagc cctcagcaaa ttgttctgct
      601 gctgcactat accagacaat aattgtctgg cctgtaccgt cagcgtcatt gacgctgcgc
      661 ccatagtgct tcctgctgct cccaagaacc caaggaacaa agctcctatt cccactgctc
      721 ttttttctct ctgcaccact cttctctttg ccttggtggg tgctactcct aatggttcaa
      781 tttttactac tttatattta tataattcac ttctccaatt gtccctcata tctcctcctc
      841 caggtctgaa gatcagcggc cgcttgctgt gcggtggtct tacttttgtt ttgctcttcc
      901 tctatcttgt ctaaagcttc cttggtgtct tttatctcta tcctttgatg cacacaatag
      961 agggttgcta ctgtattata taatgatcta agttcttctg atcctgtctg aagggatggt
     1021 tgtagctgtc ccagtatttg tctacagcct tctgatgttt ctaacaggcc aggattaact
     1081 gcgaatcgtt ctagctccct gcttgcccat actatatgtt ttaatttata ttttttcttt
     1141 ccccctggcc ttaaccgaat tttttcccat cgcgatctaa ttctcccccg cttaatactg
     1201 acgctctcgc acccatctct ctccttctag cctccgctag tcaaaatttt tggcgtactc
     1261 accagtcgcc gcccctcgcc tcttgccgtg cgcgcttcag caagccgagt cctgcgtcga
     1321 gagagctcct ctggtttccc tttcgctttc aagtccctgt tcgggcgcca ctgctagaga
     1381 ttttccacac tgactaaaag ggtctgaggg atctctagtt accagagtca cacaacagac
     1441 gggcacacac tacttgaagc actcaaggca agctttattg aggcttaagc agtgggttcc
     1501 ctagttagcc agagagctcc caggctcaga tctggtctaa ccagagagac ccgtttatgt
     1561 atcgagctag gcacttaaat acaatatctc tgcaatgcgg caattcagtg gttcgtccaa
     1621 tccatgtcag acccgtctgt tgccttccta ataaggcacg atcgtaccac cttacttcca
     1681 ccaatcggca tgcacggtgc tttttctctc cttgtaaggc atgttgctaa ctcatcgtta
     1741 ccatgttgca agactacaag agtattgcat aagactacat taagcttgca gctccagctt
     1801 ttgttccctt tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc
     1861 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     1921 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     1981 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     2041 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     2101 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     2161 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     2221 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     2281 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     2341 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     2401 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     2461 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     2521 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     2581 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     2641 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     2701 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     2761 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     2821 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     2881 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     2941 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     3001 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     3061 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     3121 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     3181 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     3241 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     3301 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     3361 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     3421 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     3481 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     3541 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     3601 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     3661 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     3721 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     3781 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     3841 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     3901 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     3961 aggggttccg cgcacatttc cccgaaaagt gccacctaaa ttgtaagcgt taatattttg
     4021 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     4081 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     4141 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     4201 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     4261 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     4321 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     4381 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     4441 ctacagggcg cgtcccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
     4501 cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt
     4561 tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgagcgcgcg
     4621 taatacgact cactataggg cgaattgggt acgtccctag gcctccaaaa aagcctcctc
     4681 actacttctg gaatagctca gaggccgagg cggcctcggc ctctgcataa ataaaaaaaa
     4741 ttagtcagcc atggggcgga gaatgggcgg aactgggcgg agttaggggc gggatgggcg
     4801 gagttagggg cgggatagct agagccagac atgataagat acattgatga gtttggacaa
     4861 accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct
     4921 ttatttgtaa ccattataag ctgcaataaa caagttcctc tcactctctg atattcattt
     4981 ctttgcaagt tataaatact gaataataag atgacatgaa ctactactgc tagagatttt
     5041 ccacactgac taaaagggtc tgagggatct ctagttacca gagtcacaca acagacgggc
     5101 acacactact tgaagcactc aaggcaagct ttattgaggc ttaagcagtg ggttccctag
     5161 ttagccagag agctcccagg ctcagatctg gtctaaccag agagacccag tacaagcaaa
     5221 aagcagatct tgtcttcgtt gggagtgaat tagcccttcc agtcccccct tttcttttaa
     5281 aaagtggcta agatctacag ctgccttgta agtcattggt cttaaaggta ccgatgcatg
     5341 gggtcgtgcg ctcctttcgg tcgggcgctg cgggtcgtgg ggcgggcgtc aggcaccggg
     5401 cttgcgggtc atgcaccagg tgcgcggtcc ttcgggcacc tcgacgtcgg cggtgacggt
     5461 gaagccgagc cgctcgtaga aggggaggtt gcggggcgcg gaggtctcca ggaaggcggg
     5521 caccccggcg cgctcggccg cctccactcc ggggagcacg acggcgctgc ccagaccctt
     5581 gccctggtgg tcgggcgaga cgccgacggt ggccaggaac cacgcgggct ccttgggccg
     5641 gtgcggcgcc aggaggcctt ccatctgttg ctgcgcggcc agccgggaac cgctcaactc
     5701 ggccatgcgc gggccgatct cggcgaacac cgcccccgct tcgacgctct ccggcgtggt
     5761 ccagaccgcc accgcggcgc cgtcgtccgc gacccacacc ttgccgatgt cgagcccgac
     5821 gcgcgtgagg aagagttctt gcagctcggt gacccgctcg atgtggcggt ccggatcgac
     5881 ggtgtggcgc gtggcggggt agtcggcgaa cgcggcggcg agggtgcgta cggccctggg
     5941 gacgtcgtcg cgggtggcga ggcgcaccgt gggcttgtac tcggtcatgg taagctccgg
     6001 tggatcttta tcatcgtgtt tttcaaagga aaaccacgtc cccgtggttc ggggggccta
     6061 gacgtttttt taacctcgac taaacacatg taaagcatgt gcaccgaggc cccagatcag
     6121 atcccataca atggggtacc ttctgggcat ccttcagccc cttgttgaat acgcttgagg
     6181 agagccattt gactctttcc acaactatcc aactcacaac gtggcactgg ggttgtgccg
     6241 cctttgcagg tgtatcttat acacgtggct tttggccgca gaggcacctg tcgccaggtg
     6301 gggggttccg ctgcctgcaa agggtcgcta cagacgttgt ttgtcttcaa gaagcttcca
     6361 gaggaactgc ttccttcacg acattcaaca gaccttgcat tcctttggcg agaggggaaa
     6421 gacccctagg aatgctcgtc aagaagacag ggccaggttt ccgggccctc acattgccaa
     6481 aagacggcaa tatggtggaa aataacatat agacaaacgc acaccggcct tattccaagc
     6541 ggcttcggcc agtaacgtta gggggggggg agggagaggg gcgcccgggt ctagatcaag
     6601 aggccatttt gtgcactgcc accgtgatgc catcgtgttt ctggatcata atgttcccat
     6661 tatctgattc tagattactt gtcatcgtca tccttgtagt caccaccacc accaccggtc
     6721 ttaggggaat gtgagggtgg ggatgaatgc tcaatttctt tatatctttc ctgttggaga
     6781 gtgtttgcat taatagcagg aggactctgt tcaggactct tttgaagaaa tctgtaaaac
     6841 aagtactcat cccggaattc atactcattg gtaatatgtt ggatgactcc cccttgtacc
     6901 agcctgtctc cgtatatcac agcttcacca cggtcggagg caaggccgac ttcaattagc
     6961 cagctcacca ggtcacagcc acagaaagtt ccagcagaag tctttgcacc acaccttctt
     7021 tctttgacaa tgtttcggat acagaggtca cggtgataat ggataaattg ttgacaggtc
     7081 atttttattt cctctgaaac aggagaatcc ctgttttctg ctgtgtcttt attgttccat
     7141 aggaattcaa gtcttctttt gaaaggcagg atgattaaat gtttatctaa tccaaagatt
     7201 ccaaaggaaa taaatccctg accaaagtta aacacggcac agaaaaactg taactcaaca
     7261 taaagtcttc caggctcttg gttgaatagc caccataaac aactggaaag attagcgaac
     7321 aggccaatga tgagaagtaa acacagcaac acatgtcggg tcagttgctg gtctccactc
     7381 tgtagatact gttcttcttc ctgggctaat atgcagctct gggagttaca gcgactcaca
     7441 caatgattgt ttttctcaaa cgaaggaatc acaggctcac tggtgcttgt gtttgctatt
     7501 ggctctattg atgggcagtg taattcacca tttcccatgg agcaggagca gcaacttgtt
     7561 tctggaatag agcttgtaaa aagttctggt tcatttactg gtgctggact ctcctcaaaa
     7621 gcagtatttc caggctccac cactttgtgg ctctgagact gatcgaaacc ttcatagctt
     7681 cctgcttggg cagtctggtt catgcacatg agggatatgc cagctatcag gatgctgcag
     7741 aacagggtga ctgctgtggt gatcatctgt tcttttccat aaaagaaggc tgagtcaatg
     7801 ctatctccat tgtgttttcc agttatcaaa agaacaccaa caaggagagc aggaattccc
     7861 cagccagata ttatgattat tccaacagga atttgtaccc tctctcgctt tttcaaaaga
     7921 aacaaagaaa ttgctagaag gcctgtccac aggtaggtgc tatagaggga gctgtacaat
     7981 agaacaaaca ccaaaatttg tccaacaaaa tttttttctt taacaaaatt ccatatcatc
     8041 attccagcac agacaataga ctgagcaatg agtaaattag ttgtaagcat atgaggaagc
     8101 tgtttatatt tcttactcaa aagaagaata gccagagacc agatcaagga gatcaggctg
     8161 acaatactta tatcaaaact aacattctgg atggcatatg ccaatggctt agggtccata
     8221 gtgggaaagg tcagtaacca ggcagaaacg tacatgatgg gagcagacac aaatgtgctt
     8281 atcaccatcc ctgaggttat aatttctact tccatgttga attgtgttgc aaagatagcc
     8341 actcctggtg ctacaggaaa tacaccatac agaaatgcat aatttgataa acttgtatgg
     8401 ttcaccacac tgtcgccctt gtccaagagt tccaccattt ctctgcacag aagtggcagc
     8461 accagaagtt tagctgtgat gagaagaatt agtactacaa atgccgactt cttcagtctc
     8521 tttatttttc ccaccatcgt gagaccaaga taaaataggg ctgatccaga aaaagaattt
     8581 ccaagtccat caagaaaatt ttcgacatat acaggtacct ttcgatcaag aataaaattg
     8641 aaggcgatgc caatgaagac cataaatact attgggttct gtaatacacg caggagtccg
     8701 agtcccacaa tttttatttt attttgagaa gcattttgag tgtctttcca cttttggatt
     8761 tcacagaaaa taaaccctat agggtttaac atcataagag atattggtgc caccaaataa
     8821 atgtactgga gatattctgg gtatgtagtt tgatataaag cttcaactat agggtatccc
     8881 aatgcaaagt cattactttg tgtagcaaaa atagggaata gtccagcttt gctaaatcga
     8941 ctatcaggac tggcaaccaa taaggttaat acacatacaa tgaaaaatac agaagctttg
     9001 gcaattaaga tactatatag gaaggaccag tccacattgg aaaaattaag tacaaccatg
     9061 tttttgaata ataaagctgg aagtgcaaat ctggagacaa aatttcctag tcctttggcc
     9121 tgggttgatg ttatgacatt ggcccttcct gctatgtagc cacaaaggac aatgccaaag
     9181 cattccagta aggctggaaa aagccttgta attgacattg aaggtgggtc attagtggaa
     9241 ttaaatccat gcgttactgc tgtaggcaaa gtcttggtca tgtttacggc gatagtcaga
     9301 ttctctgcag gtaaattaga attcatggtg gcggatcccc ctggggagag aggtcggtga
     9361 ttcggtcaac gagggagccg actgccgacg tgcgctccgg aggcttgcag aatgcggaac
     9421 accgcgcggg caggaacagg gcccacacta ccgccccaca ccccgcctcc cgcaccgccc
     9481 cttcccggcc gctgctctcg gcgcgccctg ctgagcagcc gctattggcc acagcccatc
     9541 gcggtcggcg cgctgccatt gctccctggc gctgtccgtc tgcgagggta ctagtgagac
     9601 gtgcggcttc cgtttgtcac gtccggcacg ccgcgaaccg caaggaacct tcccgactta
     9661 ggggcggagc aggaagcgtc gccggggggc ccacaagggt agcggcgaag atccgggtga
     9721 cgctgcgaac ggacgtgaag aatgtgcgag acccagggtc ggcgccgctg cgtttcccgg
     9781 aaccacgccc agagcagccg cgtccctgcg caaacccagg gctgccttgg aaaaggcgca
     9841 accccaaccc cctcgagccg cggccaaagt ggatctctgc tgtccctgta ataaacccga
     9901 aaattttgaa tttttgtaat ttgtttttgt aattctttag tttgtatgtc tgttgctatt
     9961 atgtctacta ttctttcccc tgcactgtac cccccaatcc ccccttttct tttaaaattg
    10021 tggatgaata ctgccatttg tctcgaggtc gagaattcaa aaacttaacc attgcagtca
    10081 atatctcgag atattgactg caatggttaa gccggtgttt cgtcctttcc acaagatata
    10141 taaagccaag aaatcgaaat actttcaagt tacggtaagc atatgatagt ccattttaaa
    10201 acataatttt aaaactgcaa actacccaag aaattattac tttctacgtc acgtattttg
    10261 tactaatatc tttgtgttta cagtcaaatt aattccaatt atctctctaa cagccttgta
    10321 tcgtatatgc aaatatgaag gaatcatggg aaataggccc tcggtgaagg gggcggccgc
    10381 tcgaggctag tctcgtgatc gataccgtcg agatc
//
