LOCUS       sequence-269004-       10943 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses TPC2 with C-terminus mCherry tag in mammalian cells.
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-mCherry
FEATURES             Location/Qualifiers
     source          1..10943
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    172..287
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1 (lacking the first T)"
     enhancer        345..648
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        649..852
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     803..823
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     849..873
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      982..991
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3186..3893
                     /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(3328..3346)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3596..3615
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     promoter        3922..4421
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3957..3980)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     4354..4373
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             4442..5041
                     /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(4442..4461)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4938..4958
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    5055..5643
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5108..5128)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5526..5537)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5850..6483
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(6612..6628)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6612..6628)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6625..6647)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6636..6652
                     /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(6660..6666,6667..6684,6685..6690))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6705..6726
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6843..6860)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7014..7602)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7094..7113)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7773..8564,8565..8633))
                     /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     8396..8415
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8634..8738)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    8786..8920
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8823..8842)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8877..8896
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             9088..9721
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    9768..9893
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    10390..10623
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             10808..10852
                     /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 ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg tttcagaccc
       61 acctcccaac cccgagggga cccgacaggc ccgaaggaat agaagaagaa ggtggagaga
      121 gagacagaga cagatccatt cgattagtga acggatctcg acggtatcgc ctttaaaaga
      181 aaagggggga ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac
      241 atacaaacta aagaattaca aaaacaaatt acaaaaattc aaaattttcg ggtttattac
      301 agggacagca gagatccagt ttatcgataa gcttgggagt tccgcgttac ataacttacg
      361 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
      421 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
      481 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
      541 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
      601 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt
      661 tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
      721 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
      781 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
      841 ataagcagag ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgtttt
      901 gacctccata gaagacaccg actctactag aggatcgcta gcgctaccgg actcagatct
      961 cgagctcaag cttcgaattc accatggcgg cagaagagca gccccttctg ggccggggcc
     1021 gaggcagtgg ccaggtccac tccggtgctg cagccgatca ggagctctgc atagaccagg
     1081 ctgtggtctt cattgaagac gccataaagt accgctccat ctaccaccgc atggatgctg
     1141 gttcattgtg gctttaccgc tggtattact cgaacgtctg ccaacgtgtg ctgggcttca
     1201 tcattttcct gatcctgatc ttggcttttg tggaggtccc atcttccttc acaaagactg
     1261 cagatgtgcg ctaccgttcc cagccctggc agccgccctg tggcctgacc gagacgatcg
     1321 aggcgttctg cttgcttgcc ttcctggttg acctctctgt gaagggctac ttggttgggc
     1381 aggcccagtt gcagcagaac ctgtggctgc tggcctactt tatggtgctg gttgtgtctg
     1441 tggtggactg gatcgtttcg ctgagccttg cttgtgagga gcccctgcgg atgcgccggc
     1501 tgctccgccc cttcttcctg ctgcagaatt cctccatgat gaagaagacc ctgaagtgta
     1561 tacggtggtc gctaccagaa atggccagtg tcgggctgct gctggccatc cacctgtgtc
     1621 tcttcaccat cattgggatg ctgctgttca ccattggtga gaaggatgaa gcacaggacc
     1681 aggagaggct ggcctacttc cggaaccttc cagaagcact gacctcactc ctagtgctgc
     1741 tgacgacttc caacaaccct gatgtgatga tccctgcgta tactcagaac cgggcctttg
     1801 ctctcttctt catagtcttc accttgatag gaagcttgtt tctgatgaac ctgctgacag
     1861 ccatcatcta taatcagttc cgtggctacc taatgaaatc tctacagacc tcactgttcc
     1921 ggcggcgact gggggcccgt gcagcctacg aagtcctcgc ctccagggcc gggccagctg
     1981 gaaccacccc tgagttagtt ggggtgaatc ctgagacctt cctgccagtg cttcagaaaa
     2041 cgcagctgaa caaaacccac aaacaagcca ttatgcagaa ggtgcagtcc tatgaaggcc
     2101 gcccaatgct ggctgatgag ttccagaaac tcttcgatga ggttgacaaa ggtgtggcca
     2161 aagagcgccc gctgaagccc caataccagt ctccatttct gcagactgcc cagttcatct
     2221 ttagccacca ctacttcgac tacctgggga acctcgtcgc tctgggaaac ctcttgtcta
     2281 tttgtgtgtt cctggtgctg gactcagacc tgctgcctgg ggaacgtgat gactttgtcc
     2341 tggggattct tgactacatc ttcatcttgt actacctgct ggagctgctg tttaaggtgt
     2401 tcgcgctggg cctgccaggc tacctgtcct accatagcaa tgtgtttgat ggcctcctca
     2461 ccatcatcct gctggtttcg gagatttgca ccctggctgt ataccgattg ccgcactcag
     2521 gatggaagcc agagcagtat ggcccactgt cgctctggga catgacacga ctgatgaaca
     2581 cactgattgt gtttcgcttc ctgcgcatca tccccaacat aaagccaatg gctgaggtag
     2641 ccaacaccat cctgggccta atccctaact tgagggcatt tggtgggatc ctggtggtgg
     2701 catactatgt gtttgccatg atcgggatca acctgttccg aggtgtcatc gtgcctcctg
     2761 gaaacagcag cctggttccc gacaacaact cagctgtatg tggaagcttt gagcagctag
     2821 gttactggcc caacaacttt gacgactttg ctgctgcttt gatcacactg tggaacgtga
     2881 tggtggtgaa caattggcag gtgatcctgg aagcctataa gcgctacgca ggcccgtggt
     2941 caatggtgta ttttgtgcta tggtggctgg tgtcctctgt tatctggatc aacctgtttc
     3001 tggctcttct tctggagaac tttctccaca gatgggaccc tcaaggtcat aagcagctcc
     3061 ttgttgggac caagcagatg tctgtggagc tcatgttcag ggatatccta gaagagccca
     3121 aggaggagga actgatggag aagctacaca aacacccaca cttgcatctg tgcagggcgg
     3181 ccgcggtgag caagggcgag gaggataact tagccatcat caaggagttc atgcgcttca
     3241 aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc gagggcgagg
     3301 gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt ggccccctgc
     3361 ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc
     3421 accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc aagtgggagc
     3481 gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg
     3541 acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc gacggccccg
     3601 taatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac cccgaggacg
     3661 gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc cactacgacg
     3721 ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc gcctacaacg
     3781 tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg
     3841 aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag tagggatccc
     3901 gcgactctag ataattctac cgggtagggg aggcgctttt cccaaggcag tctggagcat
     3961 gcgctttagc agccccgctg ggcacttggc gctacacaag tggcctctgg cctcgcacac
     4021 attccacatc caccggtagg cgccaaccgg ctccgttctt tggtggcccc ttcgcgccac
     4081 cttctactcc tcccctagtc aggaagttcc cccccgcccc gcagctcgcg tcgtgcagga
     4141 cgtgacaaat ggaagtagca cgtctcacta gtctcgtgca gatggacagc accgctgagc
     4201 aatggaagcg ggtaggcctt tggggcagcg gccaatagca gctttgctcc ttcgctttct
     4261 gggctcagag gctgggaagg ggtgggtccg ggggcgggct caggggcggg ctcaggggcg
     4321 gggcgggcgc ccgaaggtcc tccggaggcc cggcattctg cacgcttcaa aagcgcacgt
     4381 ctgccgcgct gttctcctct tcctcatctc cgggcctttc gacctgcagc ccaagcttac
     4441 catgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt
     4501 acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga
     4561 ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga
     4621 catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga
     4681 gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg
     4741 ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga
     4801 gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg
     4861 cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct
     4921 ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc
     4981 cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg
     5041 accgcgtctg gaacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc
     5101 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg
     5161 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc
     5221 tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg
     5281 acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg
     5341 ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga
     5401 caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct
     5461 ttccatggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg
     5521 tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc
     5581 ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc
     5641 cgcctggaat taattctgca gtcgagacct agaaaaacat ggagcaatca caagtagcaa
     5701 tacagcagct accaatgctg attgtgcctg gctagaagca caagaggagg aggaggtggg
     5761 ttttccagtc acacctcagg tacctttaag accaatgact tacaaggcag ctgtagatct
     5821 tagccacttt ttaaaagaaa agaggggact ggaagggcta attcactccc aacgaagaca
     5881 agatatcctt gatctgtgga tctaccacac acaaggctac ttccctgatt agcagaacta
     5941 cacaccaggg ccaggggtca gatatccact gacctttgga tggtgctaca agctagtacc
     6001 agttgagcca gataaggtag aagaggccaa taaaggagag aacaccagct tgttacaccc
     6061 tgtgagcctg catgggatgg atgacccgga gagagaagtg ttagagtgga ggtttgacag
     6121 ccgcctagca tttcatcacg tggcccgaga gctgcatccg gagtacttca agaactgctg
     6181 atatcgagct tgctacaagg gactttccgc tggggacttt ccagggaggc gtggcctggg
     6241 cgggactggg gagtggcgag ccctcagatc ctgcatataa gcagctgctt tttgcctgta
     6301 ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc
     6361 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt
     6421 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta
     6481 gcagtagtag ttcatgtcat cttattattc agtatttata acttgcaaag aaatgaatat
     6541 cagagagtga gaggccttga cattgctagc gttttaccgt cgacctctag ctagagcttg
     6601 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     6661 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     6721 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     6781 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     6841 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     6901 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     6961 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     7021 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     7081 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     7141 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     7201 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     7261 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     7321 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     7381 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     7441 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     7501 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     7561 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     7621 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     7681 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     7741 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     7801 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     7861 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     7921 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     7981 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     8041 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     8101 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     8161 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     8221 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     8281 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     8341 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     8401 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     8461 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     8521 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     8581 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     8641 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     8701 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     8761 cctgacgtcg acggatcggg agatcaactt gtttattgca gcttataatg gttacaaata
     8821 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     8881 tttgtccaaa ctcatcaatg tatcttatca tgtctggatc aactggataa ctcaagctaa
     8941 ccaaaatcat cccaaacttc ccaccccata ccctattacc actgccaatt acctgtggtt
     9001 tcatttactc taaacctgtg attcctctga attattttca ttttaaagaa attgtatttg
     9061 ttaaatatgt actacaaact tagtagttgg aagggctaat tcactcccaa agaagacaag
     9121 atatccttga tctgtggatc taccacacac aaggctactt ccctgattag cagaactaca
     9181 caccagggcc aggggtcaga tatccactga cctttggatg gtgctacaag ctagtaccag
     9241 ttgagccaga taaggtagaa gaggccaata aaggagagaa caccagcttg ttacaccctg
     9301 tgagcctgca tgggatggat gacccggaga gagaagtgtt agagtggagg tttgacagcc
     9361 gcctagcatt tcatcacgtg gcccgagagc tgcatccgga gtacttcaag aactgctgat
     9421 atcgagcttg ctacaaggga ctttccgctg gggactttcc agggaggcgt ggcctgggcg
     9481 ggactgggga gtggcgagcc ctcagatcct gcatataagc agctgctttt tgcctgtact
     9541 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
     9601 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
     9661 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
     9721 agtggcgccc gaacagggac ttgaaagcga aagggaaacc agaggagctc tctcgacgca
     9781 ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc
     9841 caaaaatttt gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta
     9901 agcgggggag aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa
     9961 aatataaatt aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc
    10021 ctggcctgtt agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc
    10081 ttcagacagg atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg
    10141 tgcatcaaag gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc
    10201 aaaacaaaag taagaccacc gcacagcaag cggccggccg ctgatcttca gacctggagg
    10261 aggagatatg agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga
    10321 accattagga gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc
    10381 agtgggaata ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc
    10441 agcgtcaatg acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca
    10501 gaacaatttg ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg
    10561 catcaagcag ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct
    10621 cctggggatt tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc
    10681 tagttggagt aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga
    10741 cagagaaatt aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca
    10801 gcaagaaaag aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg
    10861 gtttaacata acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt
    10921 ggtaggttta agaatagttt ttg
//
