LOCUS       sequence-266494-        9621 bp    DNA     circular SYN 14-APR-2025
DEFINITION  lysosome-targeted Sac1 catalytic domain C389S.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9621)
  AUTHORS   Lim CY, Davis OB, Shin HR, Zhang J, Berdan CA, Jiang X, Counihan JL,
            Ory DS, Nomura DK, Zoncu R
  TITLE     ER-lysosome contacts enable cholesterol sensing by mTORC1 and drive 
            aberrant growth signalling in Niemann-Pick type C.
  JOURNAL   Nat Cell Biol. 2019 Oct;21(10):1206-1218. doi: 
            10.1038/s41556-019-0391-5. Epub 2019 Sep 23.
  PUBMED    31548609
REFERENCE   2  (bases 1 to 9621)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/s41556-019-0391-5"; journalName: "Nat Cell Biol"; date: 
            "2019-10"; volume: "21"; issue: "10"; pages: "1206-1218"
COMMENT     Sequence Label: pLJM1-Lyso-FLAG-GFP-Sac1-Cat-CS
FEATURES             Location/Qualifiers
     source          1..9621
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        186..389
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     340..360
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             562..585
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             join(601..603,604..1314)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(601..603,604..1314)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(643..664)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(904..923)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1251..1272
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2759..2876
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2925..3435
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     3354..3372
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             3457..4056
                     /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(3457..3476)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3953..3973
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4184..4417
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4489..4610
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4526..4545)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4580..4599
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4650..4785
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4805..4824)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4806..4824)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4834..4851)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4834..4850)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4843..4865)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4992..5447
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5079..5098)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5289..5310
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5473..5577
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5578..5646,5647..6438)
                     /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(5796..5815)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6609..7197
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7098..7117
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7351..7368
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7485..7506
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7521..7526,7527..7544,7545..7551)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7559..7575
                     /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     7564..7586
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7583..7599
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7583..7599
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7618..7638
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7620..7638
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        7666..7892
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             7893..8073
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    8120..8245
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    8738..8971
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             9156..9200
                     /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"
     primer_bind     9498..9518
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
ORIGIN
        1 aataatttct tgggtagttt gcagttttaa aattatgttt taaaatggac tatcatatgc
       61 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
      121 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
      181 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
      241 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
      301 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
      361 tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc cgctagcgct
      421 accggtgcca tggggtgctg ctacagcagc gagaacgagg actcggacca ggaccgagag
      481 gagcggaagc tgctgctgga ccctagcagc ccccctacca aagctctcaa tggagccgag
      541 cccaacggcg gaggtggatc tgactacaag gatgacgatg acaagggcgg aggtggatct
      601 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
      661 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
      721 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
      781 gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag
      841 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
      901 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
      961 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
     1021 ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc
     1081 atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac
     1141 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     1201 ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg
     1261 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagggaggc
     1321 ggtggatcgg gcgggggtgg atcaaagtcg acgatgacca aaaagataaa agtaggtgaa
     1381 tttttcagtc atgtagtctg gaaagcaaca gattttgatg tcctttctta taagaagaca
     1441 atgttgcact taactgatat tcagttacaa gataataaaa ccttcctagc gatgctaaac
     1501 catgtcttga atgtggatgg attttacttt tcaacaacat atgatttgac ccatactttg
     1561 cagcggctat ccaacactag tcctgaattc caagaaatga gtctcttgga aagggcagat
     1621 cagcggtttg tatggaatgg tcatcttcta agagaacttt ctgcacagcc ggaggttcat
     1681 cggtttgccc ttccagtgtt acatggcttt attaccatgc attcatgttc tattaatgga
     1741 aaatactttg attggattct catctcgagg aggagctgtt tcagagctgg tgtgcgctat
     1801 tatgtaagag gaattgattc ggaaggccat gcagctaact ttgtagaaac agaacaaatt
     1861 gtgcactaca atgggagcaa agcttcgttt gtacagactc gaggatcaat acctgttttc
     1921 tggtcccaaa gaccaaacct caagtacaaa ccactgccac agatcagcaa agtagcaaat
     1981 cacatggacg gtttccaaag gcattttgat tcccaagtaa ttatttatgg aaaacaagtt
     2041 ataatcaatc tgattaacca gaagggctcg gagaagccac ttgagcagac atttgcaaca
     2101 atggtgtctt ccttgggaag tggaatgatg agatacattg cctttgactt ccataaggaa
     2161 tgtaaaaata tgagatggga tcgactaagt attttattgg atcaggtagc agaaatgcaa
     2221 gatgaattaa gttattttct agtggactct gctggccagg tggtggcaaa ccaggaaggc
     2281 gtgttccgaa gcaatagcat ggattgtcta gatagaacca atgtgatcca gagtttgtta
     2341 gctcgtcgtt cacttcaggc ccaacttcag agactaggag ttttgcatgt gggacaaaag
     2401 cttgaagaac aagatgaatt tgagaagatt ttcaaaaatg cctgggctga caacgcaaat
     2461 gcttgtgcca agcaatatgc gggaactggt gccttgaaga ctgactttac cagaactgga
     2521 aagagaactc atttgggact tataatggat ggctggaact caatgatacg atattataag
     2581 aacaactttt ccgatggatt tagacaagat tccatagact tatttcttgg aaactattca
     2641 gtggatgaat tagaatctca tagtccttta agtgttccaa gggactggaa atgagcggcc
     2701 gctaagtaag taaggatcaa ttctcgacct cgagacaaat ggcagtattc atccacaatt
     2761 ttaaaagaaa aggggggatt ggggggtaca gtgcagggga aagaatagta gacataatag
     2821 caacagacat acaaactaaa gaattacaaa aacaaattac aaaaattcaa aattttcggg
     2881 tttattacag ggacagcaga gatccacttt ggccgcggct cgagggggtt ggggttgcgc
     2941 cttttccaag gcagccctgg gtttgcgcag ggacgcggct gctctgggcg tggttccggg
     3001 aaacgcagcg gcgccgaccc tgggtctcgc acattcttca cgtccgttcg cagcgtcacc
     3061 cggatcttcg ccgctaccct tgtgggcccc ccggcgacgc ttcctgctcc gcccctaagt
     3121 cgggaaggtt ccttgcggtt cgcggcgtgc cggacgtgac aaacggaagc cgcacgtctc
     3181 actagtaccc tcgcagacgg acagcgccag ggagcaatgg cagcgcgccg accgcgatgg
     3241 gctgtggcca atagcggctg ctcagcaggg cgcgccgaga gcagcggccg ggaaggggcg
     3301 gtgcgggagg cggggtgtgg ggcggtagtg tgggccctgt tcctgcccgc gcggtgttcc
     3361 gcattctgca agcctccgga gcgcacgtcg gcagtcggct ccctcgttga ccgaatcacc
     3421 gacctctctc cccaggggga tccaccggag cttaccatga ccgagtacaa gcccacggtg
     3481 cgcctcgcca cccgcgacga cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc
     3541 gactaccccg ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag
     3601 ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac
     3661 gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc
     3721 gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag
     3781 atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc
     3841 ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg
     3901 gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc
     3961 cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg
     4021 cgcacctggt gcatgacccg caagcccggt gcctgacgcc cgccccacga cccgcagcgc
     4081 ccgaccgaaa ggagcgcacg accccatgca tcggtacctt taagaccaat gacttacaag
     4141 gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctaattcac
     4201 tcccaacgaa gacaagatct gctttttgct tgtactgggt ctctctggtt agaccagatc
     4261 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
     4321 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
     4381 ctcagaccct tttagtcagt gtggaaaatc tctagcagta gtagttcatg tcatcttatt
     4441 attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggaa cttgtttatt
     4501 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     4561 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg
     4621 ctctagctat cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     4681 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     4741 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggg acgtacccaa
     4801 ttcgccctat agtgagtcgt attacgcgcg ctcactggcc gtcgttttac aacgtcgtga
     4861 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
     4921 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
     4981 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     5041 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     5101 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     5161 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     5221 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     5281 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     5341 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     5401 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt
     5461 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     5521 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     5581 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     5641 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     5701 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     5761 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     5821 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     5881 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     5941 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     6001 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     6061 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     6121 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     6181 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     6241 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     6301 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     6361 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     6421 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     6481 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     6541 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     6601 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     6661 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     6721 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     6781 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     6841 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     6901 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     6961 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     7021 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     7081 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     7141 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     7201 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     7261 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     7321 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     7381 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
     7441 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
     7501 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
     7561 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagcgcgca
     7621 attaaccctc actaaaggga acaaaagctg gagctgcaag cttaatgtag tcttatgcaa
     7681 tactcttgta gtcttgcaac atggtaacga tgagttagca acatgcctta caaggagaga
     7741 aaaagcaccg tgcatgccga ttggtggaag taaggtggta cgatcgtgcc ttattaggaa
     7801 ggcaacagac gggtctgaca tggattggac gaaccactga attgccgcat tgcagagata
     7861 ttgtatttaa gtgcctagct cgatacataa acgggtctct ctggttagac cagatctgag
     7921 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt
     7981 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca
     8041 gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc ccgaacaggg acttgaaagc
     8101 gaaagggaaa ccagaggagc tctctcgacg caggactcgg cttgctgaag cgcgcacggc
     8161 aagaggcgag gggcggcgac tggtgagtac gccaaaaatt ttgactagcg gaggctagaa
     8221 ggagagagat gggtgcgaga gcgtcagtat taagcggggg agaattagat cgcgatggga
     8281 aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa ttaaaacata tagtatgggc
     8341 aagcagggag ctagaacgat tcgcagttaa tcctggcctg ttagaaacat cagaaggctg
     8401 tagacaaata ctgggacagc tacaaccatc ccttcagaca ggatcagaag aacttagatc
     8461 attatataat acagtagcaa ccctctattg tgtgcatcaa aggatagaga taaaagacac
     8521 caaggaagct ttagacaaga tagaggaaga gcaaaacaaa agtaagacca ccgcacagca
     8581 agcggccgct gatcttcaga cctggaggag gagatatgag ggacaattgg agaagtgaat
     8641 tatataaata taaagtagta aaaattgaac cattaggagt agcacccacc aaggcaaaga
     8701 gaagagtggt gcagagagaa aaaagagcag tgggaatagg agctttgttc cttgggttct
     8761 tgggagcagc aggaagcact atgggcgcag cgtcaatgac gctgacggta caggccagac
     8821 aattattgtc tggtatagtg cagcagcaga acaatttgct gagggctatt gaggcgcaac
     8881 agcatctgtt gcaactcaca gtctggggca tcaagcagct ccaggcaaga atcctggctg
     8941 tggaaagata cctaaaggat caacagctcc tggggatttg gggttgctct ggaaaactca
     9001 tttgcaccac tgctgtgcct tggaatgcta gttggagtaa taaatctctg gaacagattt
     9061 ggaatcacac gacctggatg gagtgggaca gagaaattaa caattacaca agcttaatac
     9121 actccttaat tgaagaatcg caaaaccagc aagaaaagaa tgaacaagaa ttattggaat
     9181 tagataaatg ggcaagtttg tggaattggt ttaacataac aaattggctg tggtatataa
     9241 aattattcat aatgatagta ggaggcttgg taggtttaag aatagttttt gctgtacttt
     9301 ctatagtgaa tagagttagg cagggatatt caccattatc gtttcagacc cacctcccaa
     9361 ccccgagggg acccgacagg cccgaaggaa tagaagaaga aggtggagag agagacagag
     9421 acagatccat tcgattagtg aacggatctc gacggtatcg atcacgagac tagcctcgag
     9481 cggccgcccc cttcaccgag ggcctatttc ccatgattcc ttcatatttg catatacgat
     9541 acaaggctgt tagagagata attggaatta atttgactgt aaacacaaag atattagtac
     9601 aaaatacgtg acgtagaaag t
//
