LOCUS       sequence-367128-        8517 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tet/dox-repressible expression of N-terminus FRB tagged human 
            mitochondrial calcium uniporter with C-terminus Flag epitope.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8517)
  AUTHORS   Balderas E, Eberhardt DR, Lee S, Pleinis JM, Sommakia S, Balynas AM,
            Yin X, Parker MC, Maguire CT, Cho S, Szulik MW, Bakhtina A, Bia RD, 
            Friederich MW, Locke TM, Van Hove JLK, Drakos SG, Sancak Y, 
            Tristani-Firouzi M, Franklin S, Rodan AR, Chaudhuri D
  TITLE     Mitochondrial calcium uniporter stabilization preserves energetic 
            homeostasis during Complex I impairment.
  JOURNAL   Nat Commun. 2022 May 19;13(1):2769. doi: 10.1038/s41467-022-30236-4.
  PUBMED    35589699
REFERENCE   2  (bases 1 to 8517)
  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: "2022-05-19"; volume: "13(1):2769. doi"; pages: " 
            10.1038/s41467-022-30236-4"
COMMENT     Sequence Label: pCW57.1-FRB-MCU-Flag-TetOFF
FEATURES             Location/Qualifiers
     source          1..8517
                     /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"
     CDS             686..964
                     /codon_start=1
                     /product="T2098L mutant of FKBP-rapamycin binding domain of
                     human FRAP"
                     /label=FRB*
                     /note="binds synthetic ligands such as AP21967"
                     /translation="ILWHEMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTL
                     KETSFNQAYGRDLMEAQEWCRKYMKSGNVKDLLQAWDLYYHVFRRISK"
     CDS             1514..1537
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        1614..2124
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     2043..2061
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             2134..2526
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MVPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTGV
                     NVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGIK
                     AIVKDSDGQPTAVGIRELLPSGYVWEG"
     CDS             2536..2589
                     /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(2590..3207,3208..3213,3214..3249,3250..3252,3253..3288,
                     3289..3291,3292..3327,3328..3336)
                     /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(3376..3396)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3794..3805)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4010..4243
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4315..4436
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4352..4371)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4406..4425
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4476..4611
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4611..4682
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4683..4751,4752..5543)
                     /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(4901..4920)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5714..6302
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6203..6222
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6456..6473
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6590..6611
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6626..6631,6632..6649,6650..6656)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6664..6680
                     /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     6669..6691
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6688..6704
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6688..6704
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6723..6743
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6725..6743
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6771..6997
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6998..7178
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7225..7350
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7843..8076
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8261..8305
                     /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 gcatggcagc tgcagcaggt agatcgctac tgctactcct
      541 atcctctcgg ggaggtggag gtggaggtgc cggaggttgc gggacgctga ctgccggctg
      601 cttccctggg ctgggcgtca gccgccaccg gcagcagcag caccaccgga cggtacacca
      661 gaggatcgct tccggaggca ccggtatcct ctggcatgag atgtggcatg aaggcctgga
      721 agaggcatct cgtttgtact ttggggaaag gaacgtgaaa ggcatgtttg aggtgctgga
      781 gcccttgcat gctatgatgg aacggggccc ccagactctg aaggaaacat cctttaatca
      841 ggcctatggt cgagatttaa tggaggccca agagtggtgc aggaagtaca tgaaatcagg
      901 gaatgtcaag gacctcctcc aagcctggga cctctattat catgtgttcc gacgaatctc
      961 aaagggcgga tccggtggag gttacaccac actgcgcatt gagcagcacc agttaaacaa
     1021 ggaaagggag cttattgaaa gactagagga tctcaaagag cagctggctc ccctggaaaa
     1081 ggtacgaatt gagattagca gaaaagctga gaagaggacc acactagtgc tatggggtgg
     1141 ccttgcctac atggccacac agtttggcat tttggcccgg cttacctggt gggaatattc
     1201 ctgggacatc atggagccag taacatactt catcacttat ggaagtgcca tggcaatgta
     1261 tgcatatttt gtaatgacac gccaggaata tgtttatcca gaagccagag acagacaata
     1321 cttactattt ttccataaag gagccaaaaa gtcacgtttt gacctagaga aatacaatca
     1381 actcaaggat gcaattgctc aggcagaaat ggaccttaag agactgagag acccattaca
     1441 agtacatctg cctctccgac aaattggtga aaaagatccc gggggtggat ctggtggatc
     1501 tggtggatct atggattaca aggatgacga tgacaagtaa ctgcagccta tggccacttt
     1561 ggtagggaca gcttcccatg ggaagtgccc aaaaagctta aatattgagg tccggggttg
     1621 gggttgcgcc ttttccaagg cagccctggg tttgcgcagg gacgcggctg ctctgggcgt
     1681 ggttccggga aacgcagcgg cgccgaccct gggtctcgca cattcttcac gtccgttcgc
     1741 agcgtcaccc ggatcttcgc cgctaccctt gtgggccccc cggcgacgct tcctgctccg
     1801 cccctaagtc gggaaggttc cttgcggttc gcggcgtgcc ggacgtgaca aacggaagcc
     1861 gcacgtctca ctagtaccct cgcagacgga cagcgccagg gagcaatggc agcgcgccga
     1921 ccgcgatggg ctgtggccaa tagcggctgc tcagcagggc gcgccgagag cagcggccgg
     1981 gaaggggcgg tgcgggaggc ggggtgtggg gcggtagtgt gggccctgtt cctgcccgcg
     2041 cggtgttccg cattctgcaa gcctccggag cgcacgtcgg cagtcggctc cctcgttgac
     2101 cgaatcaccg acctctctcc ccagcaattc accatggttc ctttgtctca agaagaatcc
     2161 accctcattg aaagagcaac ggctacaatc aacagcatcc ccatctctga agactacagc
     2221 gtcgccagcg cagctctctc tagcgacggc cgcatcttca ctggtgtcaa tgtatatcat
     2281 tttactgggg gaccttgtgc agaactcgtg gtgctgggca ctgctgctgc tgcggcagct
     2341 ggcaacctga cttgtatcgt cgcgatcgga aatgagaaca ggggcatctt gagcccctgc
     2401 ggacggtgcc gacaggtgct tctcgatctg catcctggga tcaaagccat agtgaaggac
     2461 agtgatggac agccgacggc agttgggatt cgtgaattgc tgccctctgg ttatgtgtgg
     2521 gagggccccg gtgccgaagg tagaggttct ctcctcactt gtggtgatgt tgaagaaaac
     2581 cctggtccaa tgtctagact ggacaagagc aaagtcataa actctgctct ggaattactc
     2641 aatgaagtcg gtatcgaagg cctgacgaca aggaaactcg ctcaaaagct gggagttgag
     2701 cagcctaccc tgtactggca cgtgaagaac aagcgggccc tgctcgatgc cctggcaatc
     2761 gagatgctgg acaggcatca tacccacttc tgccccctgg aaggcgagtc atggcaagac
     2821 tttctgcgga acaacgccaa gtcattccgc tgtgctctcc tctcacatcg cgacggggct
     2881 aaagtgcatc tcggcacccg cccaacagag aaacagtacg aaaccctgga aaatcagctc
     2941 gcgttcctgt gtcagcaagg cttctccctg gagaacgcac tgtacgctct gtccgccgtg
     3001 ggccacttta cactgggctg cgtattggag gatcaggagc atcaagtagc aaaagaggaa
     3061 agagagacac ctaccaccga ttctatgccc ccacttctga gacaagcaat tgagctgttc
     3121 gaccatcagg gagccgaacc tgccttcctt ttcggcctgg aactaatcat atgtggcctg
     3181 gagaaacagc taaagtgcga aagcggcggg ccggccgacg cccttgacga ttttgactta
     3241 gacatgctcc cagccgatgc ccttgacgac tttgaccttg atatgctgcc tgctgacgct
     3301 cttgacgatt ttgaccttga catgctcccc gggtaactaa gtaaggatcg atccaagata
     3361 tcgtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     3421 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     3481 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     3541 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     3601 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     3661 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct
     3721 gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
     3781 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     3841 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     3901 cgcctccccg cctgtttcgc ctcggcgtcc ggactagagg tacctttaag accaatgact
     3961 tacaaggcag ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta
     4021 attcactccc aacgaagaca agatctgctt tttgcttgta ctgggtctct ctggttagac
     4081 cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa
     4141 agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag
     4201 agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat
     4261 cttattattc agtatttata acttgcaaag aaatgaatat cagagagtga gaggaacttg
     4321 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     4381 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     4441 gtctggctct agctatcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     4501 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     4561 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctttcaaata
     4621 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     4681 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     4741 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     4801 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     4861 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     4921 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     4981 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     5041 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     5101 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     5161 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     5221 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     5281 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     5341 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     5401 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     5461 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     5521 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     5581 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     5641 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     5701 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     5761 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     5821 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     5881 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     5941 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     6001 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     6061 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     6121 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     6181 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     6241 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     6301 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     6361 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     6421 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     6481 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
     6541 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
     6601 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
     6661 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc
     6721 gcgcaattaa ccctcactaa agggaacaaa agctggagct gcaagcttaa tgtagtctta
     6781 tgcaatactc ttgtagtctt gcaacatggt aacgatgagt tagcaacatg ccttacaagg
     6841 agagaaaaag caccgtgcat gccgattggt ggaagtaagg tggtacgatc gtgccttatt
     6901 aggaaggcaa cagacgggtc tgacatggat tggacgaacc actgaattgc cgcattgcag
     6961 agatattgta tttaagtgcc tagctcgata cataaacggg tctctctggt tagaccagat
     7021 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt
     7081 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc
     7141 cctcagaccc ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg
     7201 aaagcgaaag ggaaaccaga ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc
     7261 acggcaagag gcgaggggcg gcgactggtg agtacgccaa aaattttgac tagcggaggc
     7321 tagaaggaga gagatgggtg cgagagcgtc agtattaagc gggggagaat tagatcgcga
     7381 tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat ataaattaaa acatatagta
     7441 tgggcaagca gggagctaga acgattcgca gttaatcctg gcctgttaga aacatcagaa
     7501 ggctgtagac aaatactggg acagctacaa ccatcccttc agacaggatc agaagaactt
     7561 agatcattat ataatacagt agcaaccctc tattgtgtgc atcaaaggat agagataaaa
     7621 gacaccaagg aagctttaga caagatagag gaagagcaaa acaaaagtaa gaccaccgca
     7681 cagcaagcgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     7741 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     7801 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     7861 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     7921 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     7981 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     8041 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     8101 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     8161 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     8221 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     8281 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     8341 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     8401 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     8461 cccaaccccg aggggacaat tctcgacctc gagacaaatg gcagtattca tccacaa
//
