LOCUS       sequence-258838-        9618 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  lysosome-targeted Sac1 catalytic domain C389S.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9618)
  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 9618)
  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: 
            "10.1038/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..9618
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        135..338
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     289..309
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             511..534
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             join(550..552,553..1263)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(550..552,553..1263)
                     /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(592..613)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(853..872)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1200..1221
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2705..2822
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2871..3381
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     3300..3318
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             3403..4002
                     /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(3403..3422)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3899..3919
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4130..4363
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4435..4556
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4472..4491)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4526..4545
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4596..4731
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4751..4770)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4752..4770)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4780..4797)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4780..4796)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4789..4811)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4938..5393
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5025..5044)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5235..5256
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5419..5523
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5524..5592,5593..6384)
                     /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(5742..5761)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6555..7143
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7044..7063
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7297..7314
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7431..7452
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7467..7472,7473..7490,7491..7497)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7505..7521
                     /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     7510..7532
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7529..7545
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7529..7545
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7564..7584
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7566..7584
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        7612..7838
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             7839..8019
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    8066..8191
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    8684..8917
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             9102..9146
                     /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     9444..9464
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
ORIGIN
        1 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
       61 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      121 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      181 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      241 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      301 gtaggcgtgt acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc
      361 gctagcgcta ccggtgccat ggggtgctgc tacagcagcg agaacgagga ctcggaccag
      421 gaccgagagg agcggaagct gctgctggac cctagcagcc cccctaccaa agctctcaat
      481 ggagccgagc ccaacggcgg aggtggatct gactacaagg atgacgatga caagggcgga
      541 ggtggatctg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
      601 ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc
      661 acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg
      721 cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac
      781 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
      841 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac
      901 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
      961 gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag
     1021 aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag
     1081 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     1141 aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac
     1201 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
     1261 aagggaggcg gtggatcggg cgggggtgga tcaaagtcga cgatgaaaaa gataaaagta
     1321 ggtgaatttt tcagtcatgt agtctggaaa gcaacagatt ttgatgtcct ttcttataag
     1381 aagacaatgt tgcacttaac tgatattcag ttacaagata ataaaacctt cctagcgatg
     1441 ctaaaccatg tcttgaatgt ggatggattt tacttttcaa caacatatga tttgacccat
     1501 actttgcagc ggctatccaa cactagtcct gaattccaag aaatgagtct cttggaaagg
     1561 gcagatcagc ggtttgtatg gaatggtcat cttctaagag aactttctgc acagccagag
     1621 gttcatcggt ttgcccttcc agtgttacat ggctttatta ccatgcattc atgttctatt
     1681 aatggaaaat actttgattg gattctcatc tcgaggagga gctgtttcag agctggtgtg
     1741 cgctattatg taagaggaat tgattcggaa ggccatgcag ctaactttgt agaaacagaa
     1801 caaattgtgc actacaatgg gagcaaagct tcgtttgtac agactcgagg atcaatacct
     1861 gttttctggt cccaaagacc aaacctcaag tacaaaccac tgccacagat cagcaaagta
     1921 gcaaatcaca tggacggttt ccaaaggcat tttgattccc aagtaattat ttatggaaaa
     1981 caagttataa tcaatctgat taaccagaag ggctcggaga agccacttga gcagacattt
     2041 gcaacaatgg tgtcttcctt gggaagtgga atgatgagat acattgcctt tgacttccat
     2101 aaggaatgta aaaatatgag atgggatcga ctaagtattt tattggatca ggtagcagaa
     2161 atgcaagatg aattaagtta ttttctagtg gactctgctg gccaggtggt ggcaaaccag
     2221 gaaggcgtgt tccgaagcaa tagcatggat tgtctagata gaaccaatgt gatccagagt
     2281 ttgttagctc gtcgttcact tcaggcccaa cttcagagac taggagtttt gcatgtggga
     2341 caaaagcttg aagaacaaga tgaatttgag aagatttaca aaaatgcctg ggctgacaac
     2401 gcaaatgctt gtgccaagca atatgcggga actggtgcct tgaagactga ctttaccaga
     2461 actggaaaga gaactcattt gggacttata atggatggct ggaactcaat gatacgatat
     2521 tataagaaca acttttccga tggatttaga caagattcca tagacttatt tcttggaaac
     2581 tattcagtgg atgaattaga atctcatagt cctttaagtg ttccaaggga ctggaaatga
     2641 gcggccgcta agtaagtaag gatcaattct cgacctcgag acaaatggca gtattcatcc
     2701 acaattttaa aagaaaaggg gggattgggg ggtacagtgc aggggaaaga atagtagaca
     2761 taatagcaac agacatacaa actaaagaat tacaaaaaca aattacaaaa attcaaaatt
     2821 ttcgggttta ttacagggac agcagagatc cactttggcc gcggctcgag ggggttgggg
     2881 ttgcgccttt tccaaggcag ccctgggttt gcgcagggac gcggctgctc tgggcgtggt
     2941 tccgggaaac gcagcggcgc cgaccctggg tctcgcacat tcttcacgtc cgttcgcagc
     3001 gtcacccgga tcttcgccgc tacccttgtg ggccccccgg cgacgcttcc tgctccgccc
     3061 ctaagtcggg aaggttcctt gcggttcgcg gcgtgccgga cgtgacaaac ggaagccgca
     3121 cgtctcacta gtaccctcgc agacggacag cgccagggag caatggcagc gcgccgaccg
     3181 cgatgggctg tggccaatag cggctgctca gcagggcgcg ccgagagcag cggccgggaa
     3241 ggggcggtgc gggaggcggg gtgtggggcg gtagtgtggg ccctgttcct gcccgcgcgg
     3301 tgttccgcat tctgcaagcc tccggagcgc acgtcggcag tcggctccct cgttgaccga
     3361 atcaccgacc tctctcccca gggggatcca ccggagctta ccatgaccga gtacaagccc
     3421 acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg
     3481 ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc
     3541 accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc
     3601 gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg
     3661 gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag
     3721 caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc
     3781 accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc
     3841 ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc
     3901 aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa
     3961 ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gacgcccgcc ccacgacccg
     4021 cagcgcccga ccgaaaggag cgcacgaccc catgcatcgg tacctttaag accaatgact
     4081 tacaaggcag ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta
     4141 attcactccc aacgaagaca agatctgctt tttgcttgta ctgggtctct ctggttagac
     4201 cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa
     4261 agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag
     4321 agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat
     4381 cttattattc agtatttata acttgcaaag aaatgaatat cagagagtga gaggaacttg
     4441 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     4501 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     4561 gtctggctct agctatcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     4621 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     4681 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctagggacgt
     4741 acccaattcg ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg
     4801 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
     4861 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
     4921 cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     4981 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     5041 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     5101 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     5161 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     5221 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     5281 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     5341 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc
     5401 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     5461 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     5521 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     5581 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     5641 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     5701 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     5761 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     5821 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     5881 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     5941 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     6001 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     6061 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     6121 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     6181 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     6241 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     6301 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     6361 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     6421 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     6481 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     6541 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     6601 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     6661 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     6721 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     6781 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     6841 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     6901 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     6961 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     7021 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     7081 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     7141 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     7201 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     7261 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     7321 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     7381 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
     7441 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
     7501 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag
     7561 cgcgcaatta accctcacta aagggaacaa aagctggagc tgcaagctta atgtagtctt
     7621 atgcaatact cttgtagtct tgcaacatgg taacgatgag ttagcaacat gccttacaag
     7681 gagagaaaaa gcaccgtgca tgccgattgg tggaagtaag gtggtacgat cgtgccttat
     7741 taggaaggca acagacgggt ctgacatgga ttggacgaac cactgaattg ccgcattgca
     7801 gagatattgt atttaagtgc ctagctcgat acataaacgg gtctctctgg ttagaccaga
     7861 tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct
     7921 tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat
     7981 ccctcagacc cttttagtca gtgtggaaaa tctctagcag tggcgcccga acagggactt
     8041 gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg actcggcttg ctgaagcgcg
     8101 cacggcaaga ggcgaggggc ggcgactggt gagtacgcca aaaattttga ctagcggagg
     8161 ctagaaggag agagatgggt gcgagagcgt cagtattaag cgggggagaa ttagatcgcg
     8221 atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa tataaattaa aacatatagt
     8281 atgggcaagc agggagctag aacgattcgc agttaatcct ggcctgttag aaacatcaga
     8341 aggctgtaga caaatactgg gacagctaca accatccctt cagacaggat cagaagaact
     8401 tagatcatta tataatacag tagcaaccct ctattgtgtg catcaaagga tagagataaa
     8461 agacaccaag gaagctttag acaagataga ggaagagcaa aacaaaagta agaccaccgc
     8521 acagcaagcg gccgctgatc ttcagacctg gaggaggaga tatgagggac aattggagaa
     8581 gtgaattata taaatataaa gtagtaaaaa ttgaaccatt aggagtagca cccaccaagg
     8641 caaagagaag agtggtgcag agagaaaaaa gagcagtggg aataggagct ttgttccttg
     8701 ggttcttggg agcagcagga agcactatgg gcgcagcgtc aatgacgctg acggtacagg
     8761 ccagacaatt attgtctggt atagtgcagc agcagaacaa tttgctgagg gctattgagg
     8821 cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa gcagctccag gcaagaatcc
     8881 tggctgtgga aagataccta aaggatcaac agctcctggg gatttggggt tgctctggaa
     8941 aactcatttg caccactgct gtgccttgga atgctagttg gagtaataaa tctctggaac
     9001 agatttggaa tcacacgacc tggatggagt gggacagaga aattaacaat tacacaagct
     9061 taatacactc cttaattgaa gaatcgcaaa accagcaaga aaagaatgaa caagaattat
     9121 tggaattaga taaatgggca agtttgtgga attggtttaa cataacaaat tggctgtggt
     9181 atataaaatt attcataatg atagtaggag gcttggtagg tttaagaata gtttttgctg
     9241 tactttctat agtgaataga gttaggcagg gatattcacc attatcgttt cagacccacc
     9301 tcccaacccc gaggggaccc gacaggcccg aaggaataga agaagaaggt ggagagagag
     9361 acagagacag atccattcga ttagtgaacg gatctcgacg gtatcgatca cgagactagc
     9421 ctcgagcggc cgcccccttc accgagggcc tatttcccat gattccttca tatttgcata
     9481 tacgatacaa ggctgttaga gagataattg gaattaattt gactgtaaac acaaagatat
     9541 tagtacaaaa tacgtgacgt agaaagtaat aatttcttgg gtagtttgca gttttaaaat
     9601 tatgttttaa aatggact
//
