LOCUS       sequence-412245-       12832 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducibly expresses mouse MLKL fused to light-activatable Cry2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12832)
  AUTHORS   Shkarina K, Hasel de Carvalho E, Santos JC, Ramos S, Leptin M, Broz 
            P
  TITLE     Optogenetic activators of apoptosis, necroptosis, and pyroptosis.
  JOURNAL   J Cell Biol. 2022 Jun 6;221(6). pii: 213145. doi: 
            10.1083/jcb.202109038. Epub 2022 Apr 14.
  PUBMED    35420640
REFERENCE   2  (bases 1 to 12832)
  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: "J Cell 
            Biol."; date: "2022-06-6"; volume: "221(6). pii: 213145. doi"; 
            pages: " 10.1083/jcb.202109038. Epub 2022 Apr 14"
COMMENT     Sequence Label: pLVX-opto-mMLKL
FEATURES             Location/Qualifiers
     source          1..12832
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    877..898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(913..918,919..936,937..943)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    951..967
                     /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     956..978
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     975..991
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     975..991
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     LTR             1119..1752
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    1959..2547
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             2065..2076
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     2474..2494
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2561..3160)
                     /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(2644..2664)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     3141..3160
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(3169..3498)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3183..3318
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             complement(3509..4255)
                     /codon_start=1
                     /product="modified rtTA protein that binds tightly to 
                     promoters containing the tet operator in the presence of 
                     doxycycline"
                     /label=Tet-On(R) 3G
                     /translation="MSRLDKSKVINSALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHSCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLKQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     promoter        complement(4274..4784)
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     complement(4337..4355)
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     promoter        4801..5168
                     /label=TRE3GS promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G, modified to eliminate
                     binding sites for endogenous mammalian transcription 
                     factors"
     protein_bind    4809..4827
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4845..4863
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4881..4899
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4917..4935
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4953..4971
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4989..5007
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5025..5043
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             6596..8086
                     /codon_start=1
                     /product="N-terminal photolyase homology region of 
                     Arabidopsis thaliana cryptochrome 2"
                     /label=CRY2PHR
                     /note="binds to CIB1 upon exposure to blue light (Kennedy 
                     et al., 2010)"
                     /translation="KMDKKTIVWFRRDLRIEDNPALAAAAHEGSVFPVFIWCPEEEGQF
                     YPGRASRWWMKQSLAHLSQSLKALGSDLTLIKTHNTISAILDCIRVTGATKVVFNHLYD
                     PVSLVRDHTVKEKLVERGISVQSYNGDLLYEPWEIYCEKGKPFTSFNSYWKKCLDMSIE
                     SVMLPPPWRLMPITAAAEAIWACSIEELGLENEAEKPSNALLTRAWSPGWSNADKLLNE
                     FIEKQLIDYAKNSKKVVGNSTSLLSPYLHFGEISVRHVFQCARMKQIIWARDKNSEGEE
                     SADLFLRGIGLREYSRYICFNFPFTHEQSLLSHLRFFPWDADVDKFKAWRQGRTGYPLV
                     DAGMRELWATGWMHNRIRVIVSSFAVKFLLLPWKWGMKYFWDTLLDADLECDILGWQYI
                     SGSIPDGHELDRLDNPALQGAKYDPEGEYIRQWLPELARLPTEWIHHPWDAPLTVLKAS
                     GVELGTNYAKPIVDIDTARELLAKAISRTRGAQIMIGAA"
     regulatory      8105..8114
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             8111..8821
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(8256..8274)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     8524..8543
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    9027..9161
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    9205..9320
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1 (lacking the first T)"
     CDS             complement(9583..9627)
                     /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    9812..10045
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    10542..10667
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             10714..11347
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     polyA_signal    11515..11649
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        11697..11801
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(11802..11870,11871..12662)
                     /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(12020..12039)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      901 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      961 ataacaattt cacacaggaa acagctatga ccatgattac gccaagctct agctagaggt
     1021 cgacggtaaa cgctagcaat gtcaaggcct ctcactctct gatattcatt tctttgcaag
     1081 ttataaatac tgaataataa gatgacatga actactactg ctagagattt tccacactga
     1141 ctaaaagggt ctgagggatc tctagttacc agagtcacac aacagacggg cacacactac
     1201 ttgaagcact caaggcaagc tttattgagg cttaagcagt gggttcccta gttagccaga
     1261 gagctcccag gctcagatct ggtctaacca gagagaccca gtacaggcaa aaagcagctg
     1321 cttatatgca ggatctgagg gctcgccact ccccagtccc gcccaggcca cgcctccctg
     1381 gaaagtcccc agcggaaagt cccttgtagc aagctcgata tcagcagttc ttgaagtact
     1441 ccggatgcag ctctcgggcc acgtgatgaa atgctaggcg gctgtcaaac ctccactcta
     1501 acacttctct ctccgggtca tccatcccat gcaggctcac agggtgtaac aagctggtgt
     1561 tctctccttt attggcctct tctaccttat ctggctcaac tggtactagc ttgtagcacc
     1621 atccaaaggt cagtggatat ctgacccctg gccctggtgt gtagttctgc taatcaggga
     1681 agtagccttg tgtgtggtag atccacagat caaggatatc ttgtcttcgt tgggagtgaa
     1741 ttagcccttc cagtcccctc ttttctttta aaaagtggct aagatctaca gctgccttgt
     1801 aagtcattgg tcttaaaggt acctgaggtg tgactggaaa acccacctcc tcctcctctt
     1861 gtgcttctag ccaggcacaa tcagcattgg tagctgctgt attgctactt gtgattgctc
     1921 catgtttttc taggtctcga ctgcagaatt aattccaggc ggggaggcgg cccaaaggga
     1981 gatccgactc gtctgagggc gaaggcgaag acgcggaaga ggccgcagag ccggcagcag
     2041 gccgcgggaa ggaaggtccg ctggattgag ggccgaaggg acgtagcaga aggacgtccc
     2101 gcgcagaatc caggtggcaa cacaggcgag cagccatgga aaggacgtca gcttccccga
     2161 caacaccacg gaattgtcag tgcccaacag ccgagcccct gtccagcagc gggcaaggca
     2221 ggcggcgatg agttccgccg tggcaatagg gagggggaaa gcgaaagtcc cggaaaggag
     2281 ctgacaggtg gtggcaatgc cccaaccagt gggggttgcg tcagcaaaca cagtgcacac
     2341 cacgccacgt tgcctgacaa cgggccacaa ctcctcataa agagacagca accaggattt
     2401 atacaaggag gagaaaatga aagccatacg ggaagcaata gcatgataca aaggcattaa
     2461 agcagcgtat ccacatagcg taaaaggagc aacatagtta agaataccag tcaatctttc
     2521 acaaattttg taatccagag gttgattgtt ccagacgcgt tcaggcaccg ggcttgcggg
     2581 tcatgcacca ggtgcgcggt ccttcgggca cctcgacgtc ggcggtgacg gtgaagccga
     2641 gccgctcgta gaaggggagg ttgcggggcg cggaggtctc caggaaggcg ggcaccccgg
     2701 cgcgctcggc cgcctccact ccggggagca cgacggcgct gcccagaccc ttgccctggt
     2761 ggtcgggcga gacgccgacg gtggccagga accacgcggg ctccttgggc cggtgcggcg
     2821 ccaggaggcc ttccatctgt tgctgcgcgg ccagccggga accgctcaac tcggccatgc
     2881 gcgggccgat ctcggcgaac accgcccccg cttcgacgct ctccggcgtg gtccagaccg
     2941 ccaccgcggc gccgtcgtcc gcgacccaca ccttgccgat gtcgagcccg acgcgcgtga
     3001 ggaagagttc ttgcagctcg gtgacccgct cgatgtggcg gtccgggtcg acggtgtggc
     3061 gcgtggcggg gtagtcggcg aacgcggcgg cgagggtgcg tacggcccgg gggacgtcgt
     3121 cgcgggtggc gaggcgcacc gtgggcttgt actcggtcat ggtcgcgttt tgcaaaagcc
     3181 taggcctcca aaaaagcctc ctcactactt ctggaatagc tcagaggccg aggcggcctc
     3241 ggcctctgca taaataaaaa aaattagtca gccatggggc ggagaatggg cggaactggg
     3301 cggagttagg ggcgggatgg gcggagttag gggcgggact atggttgctg actaattgag
     3361 atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac acctggttgc
     3421 tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg gggactttcc
     3481 acaccctaac tgacacacat tcgcgcgttt acccggggag catgtcaagg tcaaaatcgt
     3541 caagagcgtc agcaggcagc atatcaaggt caaagtcgtc aagggcatcg gctgggagca
     3601 tgtctaagtc aaaatcgtca agggcgtcgg tcggcccgcc gctttcgcac tttagctgtt
     3661 tctccaggcc acatatgatt agttccaggc cgaaaaggaa ggcaggttcg gctccctgcc
     3721 ggtcgaacag ctcaattgct tgtttcagaa gtgggggcat agaatcggtg gtaggtgtct
     3781 ctctttcctc ttttgctact tgatgctcct gttcctccaa tacgcagccc agtgtaaagt
     3841 ggcccacggc ggacagagcg tacagtgcgt tctccaggga gaagccttgc tgacacagga
     3901 acgcgagctg attttccagg gtttcgtact gtttctctgt tgggcgggtg ccgagatgca
     3961 ctttagcccc gtcgcgatgt gagaggagag cacagcggta tgacttggcg ttgttccgca
     4021 gaaagtcttg ccatgactcg ccttccaggg ggcaggagtg ggtatgatgc ctgtccagca
     4081 tctcgattgg cagggcatcg agcagggccc gcttgttctt cacgtgccag tacagggtag
     4141 gctgctcaac tcccagcttt tgagcgagtt tccttgtcgt caggccttcg ataccgactc
     4201 cattgagtaa ttccagagca gagtttatga ctttgctctt gtccagtcta gacatggtaa
     4261 ttcgatgatc cccctgggga gagaggtcgg tgattcggtc aacgagggag ccgactgccg
     4321 acgtgcgctc cggaggcttg cagaatgcgg aacaccgcgc gggcaggaac agggcccaca
     4381 ctaccgcccc acaccccgcc tcccgcaccg ccccttcccg gccgctgctc tcggcgcgcc
     4441 ctgctgagca gccgctattg gccacagccc atcgcggtcg gcgcgctgcc attgctccct
     4501 ggcgctgtcc gtctgcgagg gtactagtga gacgtgcggc ttccgtttgt cacgtccggc
     4561 acgccgcgaa ccgcaaggaa ccttcccgac ttaggggcgg agcaggaagc gtcgccgggg
     4621 ggcccacaag ggtagcggcg aagatccggg tgacgctgcg aacggacgtg aagaatgtgc
     4681 gagacccagg gtcggcgccg ctgcgtttcc cggaaccacg cccagagcag ccgcgtccct
     4741 gcgcaaaccc agggctgcct tggaaaaggc gcaaccccaa ccccgtggaa ttatcacctc
     4801 gagtttactc cctatcagtg atagagaacg tatgaagagt ttactcccta tcagtgatag
     4861 agaacgtatg cagactttac tccctatcag tgatagagaa cgtataagga gtttactccc
     4921 tatcagtgat agagaacgta tgaccagttt actccctatc agtgatagag aacgtatcta
     4981 cagtttactc cctatcagtg atagagaacg tatatccagt ttactcccta tcagtgatag
     5041 agaacgtata agctttgctt atgtaaacca gggcgcctat aaaagagtgc tgattttttg
     5101 agtaaacttc aattccacaa cacttttgtc ttataccaac tttccgtacc acttcctacc
     5161 ctcgtaaaga attatggata aattgggaca gatcatcaag ttaggccagc tcatctatga
     5221 acagtgtgaa aagatgaaat actgccggaa acaatgccag cgtctaggaa accgtgtgca
     5281 cggcctgcta cagcctctcc agagactcca ggcccaagga aagaagaacc tgcccgatga
     5341 cattactgct gccctgggcc gttttgatga agtcctgaag gaggctaacc agcagataga
     5401 aaagttcagc aagaagtccc atatttggaa gtttgtgagt gtgggcaatg ataagatcct
     5461 cttccatgaa gtgaatgaga agctgagaga cgtctgggag gagctgttgc tgctgcttca
     5521 ggtttatcat tggaataccg tttcagatgt cagccagcca gcatcctggc agcaggaaga
     5581 tcgacaggat gcagaggaag acggaaatga aaatatgaaa gttatcctga tgcagttgca
     5641 aattagcgtg gaagaaatca acaaaaccct gaagcaatgc tcactaaaac ccacacagga
     5701 gatcccacaa gatctccaaa tcaaggagat tccaaaggaa catcttggac ctccgtggac
     5761 caaactgaag acaagtaaaa tgagcaccat ttatagagga gagtatcaca gatctccagt
     5821 taccatcaaa gtattcaaca acccccaggc cgaaagtgtt ggaatagtga ggttcacttt
     5881 caatgacgag atcaaaacca tgaagaaatt cgattctccc aacatcttgc gtatatttgg
     5941 gatttgcatt gatcaaacag tgaagccccc tgagttctcc attgtcatgg agtactgtga
     6001 acttggaacc ctgagggaac tgctggatag agaaaaagac ctcacaatga gtgtgcgcag
     6061 cctcctagtc ctgagggcag ccagaggctt atacaggcta caccattcgg aaacactcca
     6121 cagaaacatc agcagctcca gtttcctcgt agccggaggc taccaagtaa agcttgcagg
     6181 atttgagtta agcaaaacac agaattccat cagccggaca gcaaagagca ctaaagcaga
     6241 gagatccagt tcaacgatat atgtctcccc tgagagactg aaaaatccat tttgccttta
     6301 tgacataaaa gctgaaatat atagctttgg aattgtactc tgggaaattg ccactggaaa
     6361 gatcccattt gaaggctgtg attctaagaa gatccgggag ctggtggctg aggacaagaa
     6421 gcaggaacca gtgggtcagg attgccctga gttgttgcgg gaaatcatta atgagtgtcg
     6481 tgcccatgag ccctcccaac ggccctctgt ggacggaatc ttggagagac tgtctgcggt
     6541 tgaagaatcc acggacaaga aggtgggagg tggaggctcc ggcggaggcg gttcaaagat
     6601 ggacaaaaag actatagttt ggtttagaag agacctaagg attgaggata atcctgcatt
     6661 agcagcagct gctcacgaag gatctgtttt tcctgtcttc atttggtgtc ctgaagaaga
     6721 aggacagttt tatcctggaa gagcttcaag atggtggatg aaacaatcac ttgctcactt
     6781 atctcaatcc ttgaaggctc ttggatctga cctcacttta atcaaaaccc acaacacgat
     6841 ttcagcgatc ttggattgta tccgcgttac cggtgctaca aaagtcgtct ttaaccacct
     6901 ctatgatcct gtttcgttag ttcgggacca taccgtaaag gagaagctgg tggaacgtgg
     6961 gatctctgtg caaagctaca atggagatct attgtatgaa ccgtgggaga tatactgcga
     7021 aaagggcaaa ccttttacga gtttcaattc ttactggaag aaatgcttag atatgtcgat
     7081 tgaatccgtt atgcttcctc ctccttggcg gttgatgcca ataactgcag cggctgaagc
     7141 gatttgggcg tgttcgattg aagaactagg gctggagaat gaggccgaga aaccgagcaa
     7201 tgcgttgtta actagagctt ggtctccagg atggagcaat gctgataagt tactaaatga
     7261 gttcatcgag aagcagttga tagattatgc aaagaacagc aagaaagttg ttgggaattc
     7321 tacttcacta ctttctccgt atctccattt cggggaaata agcgtcagac acgttttcca
     7381 gtgtgcccgg atgaaacaaa ttatatgggc aagagataag aacagtgaag gagaagaaag
     7441 tgcagatctt tttcttaggg gaatcggttt aagagagtat tctcggtata tatgtttcaa
     7501 cttcccgttt actcacgagc aatcgttgtt gagtcatctt cggtttttcc cttgggatgc
     7561 tgatgttgat aagttcaagg cctggagaca aggcaggacc ggttatccgt tggtggatgc
     7621 cggaatgaga gagctttggg ctaccggatg gatgcataac agaataagag tgattgtttc
     7681 aagctttgct gtgaagtttc ttctccttcc atggaaatgg ggaatgaagt atttctggga
     7741 tacacttttg gatgctgatt tggaatgtga catccttggc tggcagtata tctctgggag
     7801 tatccccgat ggccacgagc ttgatcgctt ggacaatccc gcgttacaag gcgccaaata
     7861 tgacccagaa ggtgagtaca taaggcaatg gcttcccgag cttgcgagat tgccaactga
     7921 atggatccat catccatggg acgctccttt aaccgtactc aaagcttctg gtgtggaact
     7981 cggaacaaac tatgcgaaac ccattgtaga catcgacaca gctcgtgagc tactagctaa
     8041 agctatttca agaacccgtg gagcacagat catgatcgga gcagcagccc gggatccacc
     8101 ggtcgccacc atggtgagca agggcgagga ggataacatg gccatcatca aggagttcat
     8161 gcgcttcaag gtgcacatgg agggctccgt gaacggccac gagttcgaga tcgagggcga
     8221 gggcgagggc cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggtgg
     8281 ccccctgccc ttcgcctggg acatcctgtc ccctcagttc atgtacggct ccaaggccta
     8341 cgtgaagcac cccgccgaca tccccgacta cttgaagctg tccttccccg agggcttcaa
     8401 gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc aggactcctc
     8461 cctgcaggac ggcgagttca tctacaaggt gaagctgcgc ggcaccaact tcccctccga
     8521 cggccccgta atgcagaaga agaccatggg ctgggaggcc tcctccgagc ggatgtaccc
     8581 cgaggacggc gccctgaagg gcgagatcaa gcagaggctg aagctgaagg acggcggcca
     8641 ctacgacgct gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc
     8701 ctacaacgtc aacatcaagt tggacatcac ctcccacaac gaggactaca ccatcgtgga
     8761 acagtacgaa cgcgccgagg gccgccactc caccggcggc atggacgagc tgtacaagta
     8821 aaattcccgt atacaccggt gccggcggat ccagaccacc tcccctgcga gctaagctgg
     8881 acagccaatg acgggtaaga gagtgacatt tttcactaac ctaagacagg agggccgtca
     8941 gagctactgc ctaatccaaa gacgggtaaa agtgataaaa atgtatcact ccaacctaag
     9001 acaggcgcag cttccgaggg atttgagatc cagacatgat aagatacatt gatgagtttg
     9061 gacaaaccaa aactagaatg cagtgaaaaa aatgccttat ttgtgaaatt tgtgatgcta
     9121 ttgccttatt tgtaaccatt ataagctgca ataaacaagt taagtcgata aactggatct
     9181 ctgctgtccc tgtaataaac ccgaaaattt tgaatttttg taatttgttt ttgtaattct
     9241 ttagtttgta tgtctgttgc tattatgtct actattcttt cccctgcact gtacccccca
     9301 atcccccctt ttcttttaaa ggcgataccg tcgagatccg ttcactaatc gaatggatct
     9361 gtctctgtct ctctctccac cttcttcttc tattccttcg ggcctgtcgg gtcccctcgg
     9421 ggttgggagg tgggtctgaa acgataatgg tgaatatccc tgcctaactc tattcactat
     9481 agaaagtaca gcaaaaacta ttcttaaacc taccaagcct cctactatca ttatgaataa
     9541 ttttatatac cacagccaat ttgttatgtt aaaccaattc cacaaacttg cccatttatc
     9601 taattccaat aattcttgtt cattcttttc ttgctggttt tgcgattctt caattaagga
     9661 gtgtattaag cttgtgtaat tgttaatttc tctgtcccac tccatccagg tcgtgtgatt
     9721 ccaaatctgt tccagagatt tattactcca actagcattc caaggcacag cagtggtgca
     9781 aatgagtttt ccagagcaac cccaaatccc caggagctgt tgatccttta ggtatctttc
     9841 cacagccagg attcttgcct ggagctgctt gatgccccag actgtgagtt gcaacagatg
     9901 ctgttgcgcc tcaatagccc tcagcaaatt gttctgctgc tgcactatac cagacaataa
     9961 ttgtctggcc tgtaccgtca gcgtcattga cgctgcgccc atagtgcttc ctgctgctcc
    10021 caagaaccca aggaacaaag ctcctattcc cactgctctt ttttctctct gcaccactct
    10081 tctctttgcc ttggtgggtg ctactcctaa tggttcaatt tttactactt tatatttata
    10141 taattcactt ctccaattgt ccctcatatc tcctcctcca ggtctgaaga tcagcggccg
    10201 gccgcttgct gtgcggtggt cttacttttg ttttgctctt cctctatctt gtctaaagct
    10261 tccttggtgt cttttatctc tatcctttga tgcacacaat agagggttgc tactgtatta
    10321 tataatgatc taagttcttc tgatcctgtc tgaagggatg gttgtagctg tcccagtatt
    10381 tgtctacagc cttctgatgt ttctaacagg ccaggattaa ctgcgaatcg ttctagctcc
    10441 ctgcttgccc atactatatg ttttaattta tattttttct ttccccctgg ccttaaccga
    10501 attttttccc atcgcgatct aattctcccc cgcttaatac tgacgctctc gcacccatct
    10561 ctctccttct agcctccgct agtcaaaatt tttggcgtac tcaccagtcg ccgcccctcg
    10621 cctcttgccg tgcgcgcttc agcaagccga gtcctgcgtc gagagagctc ctctggtttc
    10681 cctttcgctt tcaagtccct gttcgggcgc cactgctaga gattttccac actgactaaa
    10741 agggtctgag ggatctctag ttaccagagt cacacaacag acgggcacac actacttgaa
    10801 gcactcaagg caagctttat tgaggcttaa gcagtgggtt ccctagttag ccagagagct
    10861 cccaggctca gatctggtct aaccagagag acccagtaca ggcaaaaagc agctgcttat
    10921 atgcaggatc tgagggctcg ccactcccca gtcccgccca ggccacgcct ccctggaaag
    10981 tccccagcgg aaagtccctt gtagcaagct cgatatcagc agttcttgaa gtactccgga
    11041 tgcagctctc gggccacgtg atgaaatgct aggcggctgt caaacctcca ctctaacact
    11101 tctctctccg ggtcatccat cccatgcagg ctcacagggt gtaacaagct ggtgttctct
    11161 cctttattgg cctcttctac cttatctggc tcaactggta ctagcttgta gcaccatcca
    11221 aaggtcagtg gatatctgac ccctggccct ggtgtgtagt tctgctaatc agggaagtag
    11281 ccttgtgtgt ggtagatcca cagatcaagg atatcttgtc ttctttggga gtgaattagc
    11341 ccttccaact actaagtttg tagtacatat ttaacaaata caatttcttt aaaatgaaaa
    11401 taattcagag gaatcacagg tttagagtaa atgaaaccac aggtaattgg cagtggtaat
    11461 agggtatggg gtgggaagtt tgggatgatt ttggttagct tgagttatcc agttgatcca
    11521 gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa
    11581 tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat
    11641 aaacaagttg atctcccgat ccgtcgacgt caggtggcac ttttcgggga aatgtgcgcg
    11701 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
    11761 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
    11821 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
    11881 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
    11941 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
    12001 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
    12061 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
    12121 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
    12181 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
    12241 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
    12301 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
    12361 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
    12421 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
    12481 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
    12541 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
    12601 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
    12661 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
    12721 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
    12781 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tc
//
