LOCUS       sequence-267182-       10943 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mutant TPC2 with reduced Ca2+ selectivity, tagged on C terminus with
            mCherry..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10943)
  AUTHORS   Ruas M, Davis LC, Chen CC, Morgan AJ, Chuang KT, Walseth TF, Grimm 
            C, Garnham C, Powell T, Platt N, Platt FM, Biel M, Wahl-Schott C, 
            Parrington J, Galione A
  TITLE     Expression of Ca(2)(+)-permeable two-pore channels rescues NAADP 
            signalling in TPC-deficient cells.
  JOURNAL   EMBO J. 2015 Jul 2;34(13):1743-58. doi: 10.15252/embj.201490009. 
            Epub 2015 Apr 14.
  PUBMED    25872774
REFERENCE   2  (bases 1 to 10943)
  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: 
            "10.15252/embj.201490009"; journalName: "EMBO J"; date: "2015-07-2- 
            2"; volume: "34"; issue: "13"; pages: "1743-58"
COMMENT     Sequence Label: TPC2(E643A)-mCherry
FEATURES             Location/Qualifiers
     source          1..10943
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             130..763
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    810..935
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1432..1665
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1850..1894
                     /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"
     misc_feature    2157..2272
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1 (lacking the first T)"
     enhancer        2330..2633
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2634..2837
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2788..2808
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2834..2858
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      2967..2976
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5171..5878
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNLAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(5313..5331)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     5581..5600
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     promoter        5907..6406
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(5942..5965)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     6339..6358
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             6427..7026
                     /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(6427..6446)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6923..6943
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    7040..7628
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7093..7113)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7511..7522)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7835..8468
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(8597..8613)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8597..8613)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8610..8632)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8621..8637
                     /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)."
     promoter        complement(join(8645..8651,8652..8669,8670..8675))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8690..8711
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8828..8845)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8999..9587)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9079..9098)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9758..10549,10550..10618))
                     /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     10381..10400
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10619..10723)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    10771..10905
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10808..10827)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10862..10881
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 tcccacccca taccctatta ccactgccaa ttacctgtgg tttcatttac tctaaacctg
       61 tgattcctct gaattatttt cattttaaag aaattgtatt tgttaaatat gtactacaaa
      121 cttagtagtt ggaagggcta attcactccc aaagaagaca agatatcctt gatctgtgga
      181 tctaccacac acaaggctac ttccctgatt agcagaacta cacaccaggg ccaggggtca
      241 gatatccact gacctttgga tggtgctaca agctagtacc agttgagcca gataaggtag
      301 aagaggccaa taaaggagag aacaccagct tgttacaccc tgtgagcctg catgggatgg
      361 atgacccgga gagagaagtg ttagagtgga ggtttgacag ccgcctagca tttcatcacg
      421 tggcccgaga gctgcatccg gagtacttca agaactgctg atatcgagct tgctacaagg
      481 gactttccgc tggggacttt ccagggaggc gtggcctggg cgggactggg gagtggcgag
      541 ccctcagatc ctgcatataa gcagctgctt tttgcctgta ctgggtctct ctggttagac
      601 cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa
      661 agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag
      721 agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc ccgaacaggg
      781 acttgaaagc gaaagggaaa ccagaggagc tctctcgacg caggactcgg cttgctgaag
      841 cgcgcacggc aagaggcgag gggcggcgac tggtgagtac gccaaaaatt ttgactagcg
      901 gaggctagaa ggagagagat gggtgcgaga gcgtcagtat taagcggggg agaattagat
      961 cgcgatggga aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa ttaaaacata
     1021 tagtatgggc aagcagggag ctagaacgat tcgcagttaa tcctggcctg ttagaaacat
     1081 cagaaggctg tagacaaata ctgggacagc tacaaccatc ccttcagaca ggatcagaag
     1141 aacttagatc attatataat acagtagcaa ccctctattg tgtgcatcaa aggatagaga
     1201 taaaagacac caaggaagct ttagacaaga tagaggaaga gcaaaacaaa agtaagacca
     1261 ccgcacagca agcggccggc cgctgatctt cagacctgga ggaggagata tgagggacaa
     1321 ttggagaagt gaattatata aatataaagt agtaaaaatt gaaccattag gagtagcacc
     1381 caccaaggca aagagaagag tggtgcagag agaaaaaaga gcagtgggaa taggagcttt
     1441 gttccttggg ttcttgggag cagcaggaag cactatgggc gcagcgtcaa tgacgctgac
     1501 ggtacaggcc agacaattat tgtctggtat agtgcagcag cagaacaatt tgctgagggc
     1561 tattgaggcg caacagcatc tgttgcaact cacagtctgg ggcatcaagc agctccaggc
     1621 aagaatcctg gctgtggaaa gatacctaaa ggatcaacag ctcctgggga tttggggttg
     1681 ctctggaaaa ctcatttgca ccactgctgt gccttggaat gctagttgga gtaataaatc
     1741 tctggaacag atttggaatc acacgacctg gatggagtgg gacagagaaa ttaacaatta
     1801 cacaagctta atacactcct taattgaaga atcgcaaaac cagcaagaaa agaatgaaca
     1861 agaattattg gaattagata aatgggcaag tttgtggaat tggtttaaca taacaaattg
     1921 gctgtggtat ataaaattat tcataatgat agtaggaggc ttggtaggtt taagaatagt
     1981 ttttgctgta ctttctatag tgaatagagt taggcaggga tattcaccat tatcgtttca
     2041 gacccacctc ccaaccccga ggggacccga caggcccgaa ggaatagaag aagaaggtgg
     2101 agagagagac agagacagat ccattcgatt agtgaacgga tctcgacggt atcgccttta
     2161 aaagaaaagg ggggattggg gggtacagtg caggggaaag aatagtagac ataatagcaa
     2221 cagacataca aactaaagaa ttacaaaaac aaattacaaa aattcaaaat tttcgggttt
     2281 attacaggga cagcagagat ccagtttatc gataagcttg ggagttccgc gttacataac
     2341 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
     2401 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
     2461 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
     2521 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat
     2581 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc
     2641 ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc
     2701 tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa
     2761 aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg
     2821 tctatataag cagagctcgt ttagtgaacc gtcagatcgc ctggagacgc catccacgct
     2881 gttttgacct ccatagaaga caccgactct actagaggat cgctagcgct accggactca
     2941 gatctcgagc tcaagcttcg aattcaccat ggcggcagaa gagcagcccc ttctgggccg
     3001 gggccgaggc agtggccagg tccactccgg tgctgcagcc gatcaggagc tctgcataga
     3061 ccaggctgtg gtcttcattg aagacgccat aaagtaccgc tccatctacc accgcatgga
     3121 tgctggttca ttgtggcttt accgctggta ttactcgaac gtctgccaac gtgtgctggg
     3181 cttcatcatt ttcctgatcc tgatcttggc ttttgtggag gtcccatctt ccttcacaaa
     3241 gactgcagat gtgcgctacc gttcccagcc ctggcagccg ccctgtggcc tgaccgagac
     3301 gatcgaggcg ttctgcttgc ttgccttcct ggttgacctc tctgtgaagg gctacttggt
     3361 tgggcaggcc cagttgcagc agaacctgtg gctgctggcc tactttatgg tgctggttgt
     3421 gtctgtggtg gactggatcg tttcgctgag ccttgcttgt gaggagcccc tgcggatgcg
     3481 ccggctgctc cgccccttct tcctgctgca gaattcctcc atgatgaaga agaccctgaa
     3541 gtgtatacgg tggtcgctac cagaaatggc cagtgtcggg ctgctgctgg ccatccacct
     3601 gtgtctcttc accatcattg ggatgctgct gttcaccatt ggtgagaagg atgaagcaca
     3661 ggaccaggag aggctggcct acttccggaa ccttccagaa gcactgacct cactcctagt
     3721 gctgctgacg acttccaaca accctgatgt gatgatccct gcgtatactc agaaccgggc
     3781 ctttgctctc ttcttcatag tcttcacctt gataggaagc ttgtttctga tgaacctgct
     3841 gacagccatc atctataatc agttccgtgg ctacctaatg aaatctctac agacctcact
     3901 gttccggcgg cgactggggg cccgtgcagc ctacgaagtc ctcgcctcca gggccgggcc
     3961 agctggaacc acccctgagt tagttggggt gaatcctgag accttcctgc cagtgcttca
     4021 gaaaacgcag ctgaacaaaa cccacaaaca agccattatg cagaaggtgc agtcctatga
     4081 aggccgccca atgctggctg atgagttcca gaaactcttc gatgaggttg acaaaggtgt
     4141 ggccaaagag cgcccgctga agccccaata ccagtctcca tttctgcaga ctgcccagtt
     4201 catctttagc caccactact tcgactacct ggggaacctc gtcgctctgg gaaacctctt
     4261 gtctatttgt gtgttcctgg tgctggactc agacctgctg cctggggaac gtgatgactt
     4321 tgtcctgggg attcttgact acatcttcat cttgtactac ctgctggagc tgctgtttaa
     4381 ggtgttcgcg ctgggcctgc caggctacct gtcctaccat agcaatgtgt ttgatggcct
     4441 cctcaccatc atcctgctgg tttcggagat ttgcaccctg gctgtatacc gattgccgca
     4501 ctcaggatgg aagccagagc agtatggccc actgtcgctc tgggacatga cacgactgat
     4561 gaacacactg attgtgtttc gcttcctgcg catcatcccc aacataaagc caatggctga
     4621 ggtagccaac accatcctgg gcctaatccc taacttgagg gcatttggtg ggatcctggt
     4681 ggtggcatac tatgtgtttg ccatgatcgg gatcaacctg ttccgaggtg tcatcgtgcc
     4741 tcctggaaac agcagcctgg ttcccgacaa caactcagct gtatgtggaa gctttgagca
     4801 gctaggttac tggcccaaca actttgacga ctttgctgct gctttgatca cactgtggaa
     4861 cgtgatggtg gtgaacaatt ggcaggtgat cctggcagcc tataagcgct acgcaggccc
     4921 gtggtcaatg gtgtattttg tgctatggtg gctggtgtcc tctgttatct ggatcaacct
     4981 gtttctggct cttcttctgg agaactttct ccacagatgg gaccctcaag gtcataagca
     5041 gctccttgtt gggaccaagc agatgtctgt ggagctcatg ttcagggata tcctagaaga
     5101 gcccaaggag gaggaactga tggagaagct acacaaacac ccacacttgc atctgtgcag
     5161 ggcggccgcg gtgagcaagg gcgaggagga taacttagcc atcatcaagg agttcatgcg
     5221 cttcaaggtg cacatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg
     5281 cgagggccgc ccctacgagg gcacccagac cgccaagctg aaggtgacca agggtggccc
     5341 cctgcccttc gcctgggaca tcctgtcccc tcagttcatg tacggctcca aggcctacgt
     5401 gaagcacccc gccgacatcc ccgactactt gaagctgtcc ttccccgagg gcttcaagtg
     5461 ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc gtgacccagg actcctccct
     5521 gcaggacggc gagttcatct acaaggtgaa gctgcgcggc accaacttcc cctccgacgg
     5581 ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc tccgagcgga tgtaccccga
     5641 ggacggcgcc ctgaagggcg agatcaagca gaggctgaag ctgaaggacg gcggccacta
     5701 cgacgctgag gtcaagacca cctacaaggc caagaagccc gtgcagctgc ccggcgccta
     5761 caacgtcaac atcaagttgg acatcacctc ccacaacgag gactacacca tcgtggaaca
     5821 gtacgaacgc gccgagggcc gccactccac cggcggcatg gacgagctgt acaagtaggg
     5881 atcccgcgac tctagataat tctaccgggt aggggaggcg cttttcccaa ggcagtctgg
     5941 agcatgcgct ttagcagccc cgctgggcac ttggcgctac acaagtggcc tctggcctcg
     6001 cacacattcc acatccaccg gtaggcgcca accggctccg ttctttggtg gccccttcgc
     6061 gccaccttct actcctcccc tagtcaggaa gttccccccc gccccgcagc tcgcgtcgtg
     6121 caggacgtga caaatggaag tagcacgtct cactagtctc gtgcagatgg acagcaccgc
     6181 tgagcaatgg aagcgggtag gcctttgggg cagcggccaa tagcagcttt gctccttcgc
     6241 tttctgggct cagaggctgg gaaggggtgg gtccgggggc gggctcaggg gcgggctcag
     6301 gggcggggcg ggcgcccgaa ggtcctccgg aggcccggca ttctgcacgc ttcaaaagcg
     6361 cacgtctgcc gcgctgttct cctcttcctc atctccgggc ctttcgacct gcagcccaag
     6421 cttaccatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtccccagg
     6481 gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat
     6541 ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg
     6601 ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg
     6661 ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg
     6721 agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc
     6781 aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt
     6841 ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc
     6901 ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc
     6961 accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt
     7021 gcctgaccgc gtctggaaca atcaacctct ggattacaaa atttgtgaaa gattgactgg
     7081 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     7141 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
     7201 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
     7261 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
     7321 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     7381 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac
     7441 gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg
     7501 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
     7561 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
     7621 ctccccgcct ggaattaatt ctgcagtcga gacctagaaa aacatggagc aatcacaagt
     7681 agcaatacag cagctaccaa tgctgattgt gcctggctag aagcacaaga ggaggaggag
     7741 gtgggttttc cagtcacacc tcaggtacct ttaagaccaa tgacttacaa ggcagctgta
     7801 gatcttagcc actttttaaa agaaaagagg ggactggaag ggctaattca ctcccaacga
     7861 agacaagata tccttgatct gtggatctac cacacacaag gctacttccc tgattagcag
     7921 aactacacac cagggccagg ggtcagatat ccactgacct ttggatggtg ctacaagcta
     7981 gtaccagttg agccagataa ggtagaagag gccaataaag gagagaacac cagcttgtta
     8041 caccctgtga gcctgcatgg gatggatgac ccggagagag aagtgttaga gtggaggttt
     8101 gacagccgcc tagcatttca tcacgtggcc cgagagctgc atccggagta cttcaagaac
     8161 tgctgatatc gagcttgcta caagggactt tccgctgggg actttccagg gaggcgtggc
     8221 ctgggcggga ctggggagtg gcgagccctc agatcctgca tataagcagc tgctttttgc
     8281 ctgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg
     8341 gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg
     8401 tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat
     8461 ctctagcagt agtagttcat gtcatcttat tattcagtat ttataacttg caaagaaatg
     8521 aatatcagag agtgagaggc cttgacattg ctagcgtttt accgtcgacc tctagctaga
     8581 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     8641 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     8701 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     8761 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     8821 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     8881 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     8941 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     9001 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     9061 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     9121 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     9181 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     9241 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     9301 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     9361 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     9421 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     9481 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     9541 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     9601 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     9661 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     9721 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     9781 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     9841 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     9901 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     9961 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
    10021 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
    10081 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
    10141 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
    10201 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
    10261 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
    10321 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
    10381 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
    10441 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
    10501 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
    10561 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
    10621 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
    10681 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
    10741 tgccacctga cgtcgacgga tcgggagatc aacttgttta ttgcagctta taatggttac
    10801 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
    10861 tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatcaactg gataactcaa
    10921 gctaaccaaa atcatcccaa act
//
