LOCUS       sequence-367130-        8595 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tet/dox-repressible expression of C-terminus Flag tagged human 
            mitochondrial calcium uniporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8595)
  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 8595)
  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-MCU-Flag-TetOFF
FEATURES             Location/Qualifiers
     source          1..8595
                     /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             1592..1615
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        1692..2202
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     2121..2139
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             2212..2604
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MVPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTGV
                     NVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGIK
                     AIVKDSDGQPTAVGIRELLPSGYVWEG"
     CDS             2614..2667
                     /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(2668..3285,3286..3291,3292..3327,3328..3330,3331..3366,
                     3367..3369,3370..3405,3406..3414)
                     /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(3454..3474)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3872..3883)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4088..4321
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4393..4514
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4430..4449)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4484..4503
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4554..4689
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4689..4760
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4761..4829,4830..5621)
                     /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(4979..4998)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5792..6380
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6281..6300
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6534..6551
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6668..6689
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6704..6709,6710..6727,6728..6734)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6742..6758
                     /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     6747..6769
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6766..6782
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6766..6782
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6801..6821
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6803..6821
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6849..7075
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             7076..7256
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7303..7428
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7921..8154
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8339..8383
                     /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 gcatggcggc cgccgcaggt agatcgctcc tgctgctcct
      541 ctcctctcgg ggcggcggcg gcgggggcgc cggcggctgc ggggcgctga ctgccggctg
      601 cttccctggg ctgggcgtca gccgccaccg gcagcagcag caccaccgga cggtacacca
      661 gaggatcgct tcctggcaga atttgggagc tgtttattgc agcactgttg tgccctctga
      721 tgatgttaca gtggtttatc aaaatgggtt acctgtgata tctgtgaggc taccatcccg
      781 gcgtgaacgc tgtcagttca cactcaagcc tatctctgac tctgttggtg tatttttacg
      841 acaactgcaa gaagaggatc ggggaattga cagagttgct atctattcac cagatggtgt
      901 tcgcgttgct gcttcaacag gaatagacct cctcctcctt gatgacttta agctggtcat
      961 taatgactta acataccacg tacgaccacc aaaaagagac ctcttaagtc atgaaaatgc
     1021 agcaacgctg aatgatgtaa agacattggt ccagcaacta tacaccacac tgtgcattga
     1081 gcagcaccag ttaaacaagg aaagggagct tattgaaaga ctagaggatc tcaaagagca
     1141 gctggctccc ctggaaaagg tacgaattga gattagcaga aaagctgaga agaggaccac
     1201 tttggtgcta tggggtggcc ttgcctacat ggccacacag tttggcattt tggcccggct
     1261 tacctggtgg gaatattcct gggacatcat ggagccagta acatacttca tcacttatgg
     1321 aagtgccatg gcaatgtatg catattttgt aatgacacgc caggaatatg tttatccaga
     1381 agccagagac agacaatact tactattttt ccataaagga gccaaaaagt cacgttttga
     1441 cctagagaaa tacaatcaac tcaaggatgc aattgctcag gcagaaatgg accttaagag
     1501 actgagagac ccattacaag tacatctgcc tctccgacaa attggtgaaa aagattctag
     1561 aggtggatct ggtggatctg gtggatctat ggattacaag gatgacgatg acaagtaact
     1621 gcagcctatg gccactttgg tagggacagc ttcccatggg aagtgcccaa aaagcttaaa
     1681 tattgaggtc cggggttggg gttgcgcctt ttccaaggca gccctgggtt tgcgcaggga
     1741 cgcggctgct ctgggcgtgg ttccgggaaa cgcagcggcg ccgaccctgg gtctcgcaca
     1801 ttcttcacgt ccgttcgcag cgtcacccgg atcttcgccg ctacccttgt gggccccccg
     1861 gcgacgcttc ctgctccgcc cctaagtcgg gaaggttcct tgcggttcgc ggcgtgccgg
     1921 acgtgacaaa cggaagccgc acgtctcact agtaccctcg cagacggaca gcgccaggga
     1981 gcaatggcag cgcgccgacc gcgatgggct gtggccaata gcggctgctc agcagggcgc
     2041 gccgagagca gcggccggga aggggcggtg cgggaggcgg ggtgtggggc ggtagtgtgg
     2101 gccctgttcc tgcccgcgcg gtgttccgca ttctgcaagc ctccggagcg cacgtcggca
     2161 gtcggctccc tcgttgaccg aatcaccgac ctctctcccc agcaattcac catggttcct
     2221 ttgtctcaag aagaatccac cctcattgaa agagcaacgg ctacaatcaa cagcatcccc
     2281 atctctgaag actacagcgt cgccagcgca gctctctcta gcgacggccg catcttcact
     2341 ggtgtcaatg tatatcattt tactggggga ccttgtgcag aactcgtggt gctgggcact
     2401 gctgctgctg cggcagctgg caacctgact tgtatcgtcg cgatcggaaa tgagaacagg
     2461 ggcatcttga gcccctgcgg acggtgccga caggtgcttc tcgatctgca tcctgggatc
     2521 aaagccatag tgaaggacag tgatggacag ccgacggcag ttgggattcg tgaattgctg
     2581 ccctctggtt atgtgtggga gggccccggt gccgaaggta gaggttctct cctcacttgt
     2641 ggtgatgttg aagaaaaccc tggtccaatg tctagactgg acaagagcaa agtcataaac
     2701 tctgctctgg aattactcaa tgaagtcggt atcgaaggcc tgacgacaag gaaactcgct
     2761 caaaagctgg gagttgagca gcctaccctg tactggcacg tgaagaacaa gcgggccctg
     2821 ctcgatgccc tggcaatcga gatgctggac aggcatcata cccacttctg ccccctggaa
     2881 ggcgagtcat ggcaagactt tctgcggaac aacgccaagt cattccgctg tgctctcctc
     2941 tcacatcgcg acggggctaa agtgcatctc ggcacccgcc caacagagaa acagtacgaa
     3001 accctggaaa atcagctcgc gttcctgtgt cagcaaggct tctccctgga gaacgcactg
     3061 tacgctctgt ccgccgtggg ccactttaca ctgggctgcg tattggagga tcaggagcat
     3121 caagtagcaa aagaggaaag agagacacct accaccgatt ctatgccccc acttctgaga
     3181 caagcaattg agctgttcga ccatcaggga gccgaacctg ccttcctttt cggcctggaa
     3241 ctaatcatat gtggcctgga gaaacagcta aagtgcgaaa gcggcgggcc ggccgacgcc
     3301 cttgacgatt ttgacttaga catgctccca gccgatgccc ttgacgactt tgaccttgat
     3361 atgctgcctg ctgacgctct tgacgatttt gaccttgaca tgctccccgg gtaactaagt
     3421 aaggatcgat ccaagatatc gtattcttaa ctatgttgct ccttttacgc tatgtggata
     3481 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     3541 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     3601 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     3661 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     3721 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     3781 ggtgttgtcg gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat
     3841 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     3901 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     3961 tcggatctcc ctttgggccg cctccccgcc tgtttcgcct cggcgtccgg actagaggta
     4021 cctttaagac caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag
     4081 gggggactgg aagggctaat tcactcccaa cgaagacaag atctgctttt tgcttgtact
     4141 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
     4201 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
     4261 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
     4321 agtagtagtt catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca
     4381 gagagtgaga ggaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     4441 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     4501 atcaatgtat cttatcatgt ctggctctag ctatcccgcc cctaactccg cccatcccgc
     4561 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     4621 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     4681 ttggaggcct ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     4741 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     4801 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     4861 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     4921 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     4981 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     5041 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     5101 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     5161 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     5221 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     5281 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     5341 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     5401 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     5461 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     5521 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     5581 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     5641 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     5701 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     5761 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     5821 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     5881 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     5941 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     6001 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     6061 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     6121 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     6181 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     6241 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     6301 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     6361 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     6421 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     6481 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     6541 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     6601 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     6661 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     6721 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     6781 accatgatta cgccaagcgc gcaattaacc ctcactaaag ggaacaaaag ctggagctgc
     6841 aagcttaatg tagtcttatg caatactctt gtagtcttgc aacatggtaa cgatgagtta
     6901 gcaacatgcc ttacaaggag agaaaaagca ccgtgcatgc cgattggtgg aagtaaggtg
     6961 gtacgatcgt gccttattag gaaggcaaca gacgggtctg acatggattg gacgaaccac
     7021 tgaattgccg cattgcagag atattgtatt taagtgccta gctcgataca taaacgggtc
     7081 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct
     7141 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga
     7201 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg
     7261 cgcccgaaca gggacttgaa agcgaaaggg aaaccagagg agctctctcg acgcaggact
     7321 cggcttgctg aagcgcgcac ggcaagaggc gaggggcggc gactggtgag tacgccaaaa
     7381 attttgacta gcggaggcta gaaggagaga gatgggtgcg agagcgtcag tattaagcgg
     7441 gggagaatta gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat
     7501 aaattaaaac atatagtatg ggcaagcagg gagctagaac gattcgcagt taatcctggc
     7561 ctgttagaaa catcagaagg ctgtagacaa atactgggac agctacaacc atcccttcag
     7621 acaggatcag aagaacttag atcattatat aatacagtag caaccctcta ttgtgtgcat
     7681 caaaggatag agataaaaga caccaaggaa gctttagaca agatagagga agagcaaaac
     7741 aaaagtaaga ccaccgcaca gcaagcggcc gctgatcttc agacctggag gaggagatat
     7801 gagggacaat tggagaagtg aattatataa atataaagta gtaaaaattg aaccattagg
     7861 agtagcaccc accaaggcaa agagaagagt ggtgcagaga gaaaaaagag cagtgggaat
     7921 aggagctttg ttccttgggt tcttgggagc agcaggaagc actatgggcg cagcgtcaat
     7981 gacgctgacg gtacaggcca gacaattatt gtctggtata gtgcagcagc agaacaattt
     8041 gctgagggct attgaggcgc aacagcatct gttgcaactc acagtctggg gcatcaagca
     8101 gctccaggca agaatcctgg ctgtggaaag atacctaaag gatcaacagc tcctggggat
     8161 ttggggttgc tctggaaaac tcatttgcac cactgctgtg ccttggaatg ctagttggag
     8221 taataaatct ctggaacaga tttggaatca cacgacctgg atggagtggg acagagaaat
     8281 taacaattac acaagcttaa tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa
     8341 gaatgaacaa gaattattgg aattagataa atgggcaagt ttgtggaatt ggtttaacat
     8401 aacaaattgg ctgtggtata taaaattatt cataatgata gtaggaggct tggtaggttt
     8461 aagaatagtt tttgctgtac tttctatagt gaatagagtt aggcagggat attcaccatt
     8521 atcgtttcag acccacctcc caaccccgag gggacaattc tcgacctcga gacaaatggc
     8581 agtattcatc cacaa
//
