LOCUS       sequence-370663-        8637 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Dox-repressible expression of the 0N4R Tau isoform .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8637)
  AUTHORS   Saha I, Yuste-Checa P, Da Silva Padilha M, Guo Q, Korner R, 
            Holthusen H, Trinkaus VA, Dudanova I, Fernandez-Busnadiego R, 
            Baumeister W, Sanders DW, Gautam S, Diamond MI, Hartl FU, Hipp MS
  TITLE     The AAA+ chaperone VCP disaggregates Tau fibrils and generates 
            aggregate seeds in a cellular system.
  JOURNAL   Nat Commun. 2023 Feb 2;14(1):560. doi: 10.1038/s41467-023-36058-2.
  PUBMED    36732333
REFERENCE   2  (bases 1 to 8637)
  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"; journalName: "Nat 
            Commun."; date: "2023-02-2"; volume: "14(1):560. doi"; pages: " 
            10.1038/s41467-023-36058-2"
COMMENT     Sequence Label: Tet-off Full length Tau (0N4R, P301L/V337M)
FEATURES             Location/Qualifiers
     source          1..8637
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..118
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        172..486
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    180..198
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    216..234
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    251..269
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    287..305
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    323..341
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    358..376
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    394..412
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     459..483
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      503..512
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        1734..2244
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     2163..2181
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             2254..2646
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MVPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTGV
                     NVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGIK
                     AIVKDSDGQPTAVGIRELLPSGYVWEG"
     CDS             2656..2709
                     /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(2710..3327,3328..3333,3334..3369,3370..3372,3373..3408,
                     3409..3411,3412..3447,3448..3456)
                     /codon_start=1
                     /product="improved tetracycline-controlled transactivator"
                     /label=tTA-Advanced
                     /note="In the Tet-Off(R) system, tTA-Advanced binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the absence of tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGGPADALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     primer_bind     complement(3496..3516)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3914..3925)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     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"
     promoter        4731..4802
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4803..4871,4872..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     complement(5021..5040)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5834..6422
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6323..6342
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6576..6593
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6710..6731
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6746..6751,6752..6769,6770..6776)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6784..6800
                     /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     6789..6811
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6808..6824
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6808..6824
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6843..6863
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6845..6863
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6891..7117
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             7118..7298
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7345..7470
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7963..8196
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8381..8425
                     /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"
ORIGIN
        1 ttttaaaaga aaagggggga ttggggggta cagtgcaggg gaaagaatag tagacataat
       61 agcaacagac atacaaacta aagaattaca aaaacaaatt acaaaaattc aaaattttcg
      121 ggtttattac agggacagca gagatccact ttggccgcga atcgatatgt cgagtttact
      181 ccctatcagt gatagagaac gtatgtcgag tttactccct atcagtgata gagaacgatg
      241 tcgagtttac tccctatcag tgatagagaa cgtatgtcga gtttactccc tatcagtgat
      301 agagaacgta tgtcgagttt actccctatc agtgatagag aacgtatgtc gagtttatcc
      361 ctatcagtga tagagaacgt atgtcgagtt tactccctat cagtgataga gaacgtatgt
      421 cgaggtaggc gtgtacggtg ggaggcctat ataagcagag ctcgtttagt gaaccgtcag
      481 atcgcctgga gaattggcta gcgccaccat ggctgagccc cgccaggagt tcgaagtgat
      541 ggaagatcac gctgggacgt acgggttggg ggacaggaaa gatcaggggg gctacaccat
      601 gcaccaagac caagagggtg acacggacgc tggcctgaaa gctgaagaag caggcattgg
      661 agacaccccc agcctggaag acgaagctgc tggtcacgtg acccaagctc gcatggtcag
      721 taaaagcaaa gacgggactg gaagcgatga caaaaaagcc aagggggctg atggtaaaac
      781 gaagatcgcc acaccgcggg gagcagcccc tccaggccag aagggccagg ccaacgccac
      841 caggattcca gcaaaaaccc cgcccgctcc aaagacacca cccagctctg gtgaacctcc
      901 aaaatcaggg gatcgcagcg gctacagcag ccccggctcc ccaggcactc ccggcagccg
      961 ctcccgcacc ccgtcccttc caaccccacc cacccgggag cccaagaagg tggcagtggt
     1021 ccgtactcca cccaagtcgc cgtcttccgc caagagccgc ctgcagacag cccccgtgcc
     1081 catgccagac ctgaagaatg tcaagtccaa gatcggctcc actgagaacc tgaagcacca
     1141 gccgggaggc gggaaggtgc agataattaa taagaagctg gatcttagca acgtccagtc
     1201 caagtgtggc tcaaaggata atatcaaaca cgtcctggga ggcggcagtg tgcaaatagt
     1261 ctacaaacca gttgacctga gcaaggtgac ctccaagtgt ggctcattag gcaacatcca
     1321 tcataaacca ggaggtggcc agatggaagt aaaatctgag aagcttgact tcaaggacag
     1381 agtccagtcg aagattgggt ccctggacaa tatcacccac gtccctggcg gaggaaataa
     1441 aaagattgaa acccacaagc tgaccttccg cgagaacgcc aaagccaaga cagaccacgg
     1501 ggcggagatc gtgtacaagt cgccagtggt gtctggggac acgtctccac ggcatctcag
     1561 caatgtctcc tccaccggca gcatcgacat ggtagactcg ccccagctcg ccacgctagc
     1621 tgacgaggtg tctgcctccc tggccaagca gggtttgtaa ctgcagccta tggccacttt
     1681 ggtagggaca gcttcccatg ggaagtgccc aaaaagctta aatattgagg tccggggttg
     1741 gggttgcgcc ttttccaagg cagccctggg tttgcgcagg gacgcggctg ctctgggcgt
     1801 ggttccggga aacgcagcgg cgccgaccct gggtctcgca cattcttcac gtccgttcgc
     1861 agcgtcaccc ggatcttcgc cgctaccctt gtgggccccc cggcgacgct tcctgctccg
     1921 cccctaagtc gggaaggttc cttgcggttc gcggcgtgcc ggacgtgaca aacggaagcc
     1981 gcacgtctca ctagtaccct cgcagacgga cagcgccagg gagcaatggc agcgcgccga
     2041 ccgcgatggg ctgtggccaa tagcggctgc tcagcagggc gcgccgagag cagcggccgg
     2101 gaaggggcgg tgcgggaggc ggggtgtggg gcggtagtgt gggccctgtt cctgcccgcg
     2161 cggtgttccg cattctgcaa gcctccggag cgcacgtcgg cagtcggctc cctcgttgac
     2221 cgaatcaccg acctctctcc ccagcaattc accatggttc ctttgtctca agaagaatcc
     2281 accctcattg aaagagcaac ggctacaatc aacagcatcc ccatctctga agactacagc
     2341 gtcgccagcg cagctctctc tagcgacggc cgcatcttca ctggtgtcaa tgtatatcat
     2401 tttactgggg gaccttgtgc agaactcgtg gtgctgggca ctgctgctgc tgcggcagct
     2461 ggcaacctga cttgtatcgt cgcgatcgga aatgagaaca ggggcatctt gagcccctgc
     2521 ggacggtgcc gacaggtgct tctcgatctg catcctggga tcaaagccat agtgaaggac
     2581 agtgatggac agccgacggc agttgggatt cgtgaattgc tgccctctgg ttatgtgtgg
     2641 gagggccccg gtgccgaagg tagaggttct ctcctcactt gtggtgatgt tgaagaaaac
     2701 cctggtccaa tgtctagact ggacaagagc aaagtcataa actctgctct ggaattactc
     2761 aatgaagtcg gtatcgaagg cctgacgaca aggaaactcg ctcaaaagct gggagttgag
     2821 cagcctaccc tgtactggca cgtgaagaac aagcgggccc tgctcgatgc cctggcaatc
     2881 gagatgctgg acaggcatca tacccacttc tgccccctgg aaggcgagtc atggcaagac
     2941 tttctgcgga acaacgccaa gtcattccgc tgtgctctcc tctcacatcg cgacggggct
     3001 aaagtgcatc tcggcacccg cccaacagag aaacagtacg aaaccctgga aaatcagctc
     3061 gcgttcctgt gtcagcaagg cttctccctg gagaacgcac tgtacgctct gtccgccgtg
     3121 ggccacttta cactgggctg cgtattggag gatcaggagc atcaagtagc aaaagaggaa
     3181 agagagacac ctaccaccga ttctatgccc ccacttctga gacaagcaat tgagctgttc
     3241 gaccatcagg gagccgaacc tgccttcctt ttcggcctgg aactaatcat atgtggcctg
     3301 gagaaacagc taaagtgcga aagcggcggg ccggccgacg cccttgacga ttttgactta
     3361 gacatgctcc cagccgatgc ccttgacgac tttgaccttg atatgctgcc tgctgacgct
     3421 cttgacgatt ttgaccttga catgctcccc gggtaactaa gtaaggatcg atccaagata
     3481 tcgtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     3541 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     3601 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     3661 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     3721 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     3781 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct
     3841 gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
     3901 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     3961 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     4021 cgcctccccg cctgtttcgc ctcggcgtcc ggactagagg 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 ctttcaaata
     4741 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     4801 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     4861 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     4921 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     4981 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     5041 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     5101 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     5161 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     5221 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     5281 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     5341 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     5401 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     5461 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     5521 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     5581 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     5641 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     5701 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     5761 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     5821 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     5881 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     5941 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     6001 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     6061 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     6121 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     6181 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     6241 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     6301 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     6361 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     6421 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     6481 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     6541 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     6601 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
     6661 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
     6721 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
     6781 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc
     6841 gcgcaattaa ccctcactaa agggaacaaa agctggagct gcaagcttaa tgtagtctta
     6901 tgcaatactc ttgtagtctt gcaacatggt aacgatgagt tagcaacatg ccttacaagg
     6961 agagaaaaag caccgtgcat gccgattggt ggaagtaagg tggtacgatc gtgccttatt
     7021 aggaaggcaa cagacgggtc tgacatggat tggacgaacc actgaattgc cgcattgcag
     7081 agatattgta tttaagtgcc tagctcgata cataaacggg tctctctggt tagaccagat
     7141 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt
     7201 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc
     7261 cctcagaccc ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg
     7321 aaagcgaaag ggaaaccaga ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc
     7381 acggcaagag gcgaggggcg gcgactggtg agtacgccaa aaattttgac tagcggaggc
     7441 tagaaggaga gagatgggtg cgagagcgtc agtattaagc gggggagaat tagatcgcga
     7501 tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat ataaattaaa acatatagta
     7561 tgggcaagca gggagctaga acgattcgca gttaatcctg gcctgttaga aacatcagaa
     7621 ggctgtagac aaatactggg acagctacaa ccatcccttc agacaggatc agaagaactt
     7681 agatcattat ataatacagt agcaaccctc tattgtgtgc atcaaaggat agagataaaa
     7741 gacaccaagg aagctttaga caagatagag gaagagcaaa acaaaagtaa gaccaccgca
     7801 cagcaagcgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     7861 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     7921 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     7981 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     8041 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     8101 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     8161 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     8221 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     8281 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     8341 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     8401 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     8461 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     8521 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     8581 cccaaccccg aggggacaat tctcgacctc gagacaaatg gcagtattca tccacaa
//
