LOCUS       sequence-412244-       12943 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducibly expresses mouse RipK3 fused to light activatable Cry2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12943)
  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 12943)
  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-mRipK3
FEATURES             Location/Qualifiers
     source          1..12943
                     /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             5174..5881
                     /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(5319..5337)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     5587..5606
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             5897..7390
                     /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="MKMDKKTIVWFRRDLRIEDNPALAAAAHEGSVFPVFIWCPEEEGQ
                     FYPGRASRWWMKQSLAHLSQSLKALGSDLTLIKTHNTISAILDCIRVTGATKVVFNHLY
                     DPVSLVRDHTVKEKLVERGISVQSYNGDLLYEPWEIYCEKGKPFTSFNSYWKKCLDMSI
                     ESVMLPPPWRLMPITAAAEAIWACSIEELGLENEAEKPSNALLTRAWSPGWSNADKLLN
                     EFIEKQLIDYAKNSKKVVGNSTSLLSPYLHFGEISVRHVFQCARMKQIIWARDKNSEGE
                     ESADLFLRGIGLREYSRYICFNFPFTHEQSLLSHLRFFPWDADVDKFKAWRQGRTGYPL
                     VDAGMRELWATGWMHNRIRVIVSSFAVKFLLLPWKWGMKYFWDTLLDADLECDILGWQY
                     ISGSIPDGHELDRLDNPALQGAKYDPEGEYIRQWLPELARLPTEWIHHPWDAPLTVLKA
                     SGVELGTNYAKPIVDIDTARELLAKAISRTRGAQIMIGAA"
     polyA_signal    9138..9272
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    9316..9431
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1 (lacking the first T)"
     CDS             complement(9694..9738)
                     /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    9923..10156
                     /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    10653..10778
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             10825..11458
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     polyA_signal    11626..11760
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        11808..11912
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(11913..11981,11982..12773)
                     /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(12131..12150)
                     /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 attatggtga gcaagggcga ggaggataac atggccatca tcaaggagtt
     5221 catgcgcttc aaggtgcaca tggagggctc cgtgaacggc cacgagttcg agatcgaggg
     5281 cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg
     5341 tggccccctg cccttcgcct gggacatcct gtcccctcag ttcatgtacg gctccaaggc
     5401 ctacgtgaag caccccgccg acatccccga ctacttgaag ctgtccttcc ccgagggctt
     5461 caagtgggag cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc
     5521 ctccctgcag gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc
     5581 cgacggcccc gtaatgcaga agaagaccat gggctgggag gcctcctccg agcggatgta
     5641 ccccgaggac ggcgccctga agggcgagat caagcagagg ctgaagctga aggacggcgg
     5701 ccactacgac gctgaggtca agaccaccta caaggccaag aagcccgtgc agctgcccgg
     5761 cgcctacaac gtcaacatca agttggacat cacctcccac aacgaggact acaccatcgt
     5821 ggaacagtac gaacgcgccg agggccgcca ctccaccggc ggcatggacg agctgtacaa
     5881 gggaggagga ggaatgatga agatggacaa aaagactata gtttggttta gaagagacct
     5941 aaggattgag gataatcctg cattagcagc agctgctcac gaaggatctg tttttcctgt
     6001 cttcatttgg tgtcctgaag aagaaggaca gttttatcct ggaagagctt caagatggtg
     6061 gatgaaacaa tcacttgctc acttatctca atccttgaag gctcttggat ctgacctcac
     6121 tttaatcaaa acccacaaca cgatttcagc gatcttggat tgtatccgcg ttaccggtgc
     6181 tacaaaagtc gtctttaacc acctctatga tcctgtttcg ttagttcggg accataccgt
     6241 aaaggagaag ctggtggaac gtgggatctc tgtgcaaagc tacaatggag atctattgta
     6301 tgaaccgtgg gagatatact gcgaaaaggg caaacctttt acgagtttca attcttactg
     6361 gaagaaatgc ttagatatgt cgattgaatc cgttatgctt cctcctcctt ggcggttgat
     6421 gccaataact gcagcggctg aagcgatttg ggcgtgttcg attgaagaac tagggctgga
     6481 gaatgaggcc gagaaaccga gcaatgcgtt gttaactaga gcttggtctc caggatggag
     6541 caatgctgat aagttactaa atgagttcat cgagaagcag ttgatagatt atgcaaagaa
     6601 cagcaagaaa gttgttggga attctacttc actactttct ccgtatctcc atttcgggga
     6661 aataagcgtc agacacgttt tccagtgtgc ccggatgaaa caaattatat gggcaagaga
     6721 taagaacagt gaaggagaag aaagtgcaga tctttttctt aggggaatcg gtttaagaga
     6781 gtattctcgg tatatatgtt tcaacttccc gtttactcac gagcaatcgt tgttgagtca
     6841 tcttcggttt ttcccttggg atgctgatgt tgataagttc aaggcctgga gacaaggcag
     6901 gaccggttat ccgttggtgg atgccggaat gagagagctt tgggctaccg gatggatgca
     6961 taacagaata agagtgattg tttcaagctt tgctgtgaag tttcttctcc ttccatggaa
     7021 atggggaatg aagtatttct gggatacact tttggatgct gatttggaat gtgacatcct
     7081 tggctggcag tatatctctg ggagtatccc cgatggccac gagcttgatc gcttggacaa
     7141 tcccgcgtta caaggcgcca aatatgaccc agaaggtgag tacataaggc aatggcttcc
     7201 cgagcttgcg agattgccaa ctgaatggat ccatcatcca tgggacgctc ctttaaccgt
     7261 actcaaagct tctggtgtgg aactcggaac aaactatgcg aaacccattg tagacatcga
     7321 cacagctcgt gagctactag ctaaagctat ttcaagaacc cgtggagcac agatcatgat
     7381 cggagcagca gcccgggatc caccggtcgc caccggagga ggaggaagcg gaggaggagg
     7441 aagcggagga ggaggaagcg gaggaggagg aatgtcttct gtcaagttat ggcctactgg
     7501 tgcgtcagcg gttcctctgg tgagccgtga agaactgaag aagctggagt ttgtgggcaa
     7561 aggagggttc ggagtcgtgt tccgggcaca ccacagaaca tggaaccatg atgtagcagt
     7621 caagatcgtg aactcgaaga agatatcctg ggaggtgaag gctatggtta atcttcgtaa
     7681 tgagaacgtt ctgctcctgc tgggggtcac tgaggacctc cagtgggact tcgtgtccgg
     7741 gcaggctctg gtgacaagat tcatggagaa tggctccctc gcagggctgc tgcaacccga
     7801 gtgccctcgg ccctggccac tcctctgtcg cctgctgcag gaagtggtgc tggggatgtg
     7861 ctacctacac agcttggacc ctccgctcct gcaccgggac ctcaagccct ctaacattct
     7921 gctggatcca gagctccacg ccaagctagc agattttggc ctgtccacgt ttcagggagg
     7981 gtcccagtca gggtcaggat caggatcagg atccagggac tctgggggca ccctagcgta
     8041 cttggaccca gagctgttat ttaaagtcaa cctgaaggct tctaaagcga gtgatgtcta
     8101 cagctttggg atcctcgtgt gggcagtgct ggctggcaga gaagctgagt tggtagacaa
     8161 gacttcacta atccgggaaa cagtgtgtga caggcagagt cgtcctccac tgacagagct
     8221 gcctccaggt agccctgaga ctcccggctt ggaaaaactg aaggagttaa tgattcattg
     8281 ctggggttcc cagtccgaaa acaggccatc cttccaggac tgcgaaccaa aaaccaatga
     8341 agtttacaat ctggtaaagg acaaggtaga tgctgctgtc tccgaggtaa agcattatct
     8401 gtctcagcac agaagcagtg gcagaaactt gtctgccaga gagccaagcc aaagaggcac
     8461 agaaatggat tgcccgaggg aaaccatggt ttctaaaatg ctggaccgcc tgcatttgga
     8521 ggaaccctcc ggaccagttc ctggaaaatg tcctgagagg caagcacagg acacatcagt
     8581 tgggcctgcc acaccagcaa ggacatcttc tgaccccgtg gctggcactc ctcagattcc
     8641 acatacttta cccttcagag gcacaacacc tgggccagtc tttactgaga ctcccggtcc
     8701 tcacccccaa aggaatcagg gagatggaag acacggcact ccttggtatc cctggacccc
     8761 accgaatcca atgacagggc caccggctct cgtcttcaac aactgttctg aagtggcggc
     8821 tgcggcctac aactccttgg tagcaccacc aagaactact gcctcaagtt cggccaagta
     8881 tgaccaagca cagttcggca ggggtagggg ctggcagccc ttccacaagt agaattcccg
     8941 tatacaccgg tgccggcgga tccagaccac ctcccctgcg agctaagctg gacagccaat
     9001 gacgggtaag agagtgacat ttttcactaa cctaagacag gagggccgtc agagctactg
     9061 cctaatccaa agacgggtaa aagtgataaa aatgtatcac tccaacctaa gacaggcgca
     9121 gcttccgagg gatttgagat ccagacatga taagatacat tgatgagttt ggacaaacca
     9181 aaactagaat gcagtgaaaa aaatgcctta tttgtgaaat ttgtgatgct attgccttat
     9241 ttgtaaccat tataagctgc aataaacaag ttaagtcgat aaactggatc tctgctgtcc
     9301 ctgtaataaa cccgaaaatt ttgaattttt gtaatttgtt tttgtaattc tttagtttgt
     9361 atgtctgttg ctattatgtc tactattctt tcccctgcac tgtacccccc aatcccccct
     9421 tttcttttaa aggcgatacc gtcgagatcc gttcactaat cgaatggatc tgtctctgtc
     9481 tctctctcca ccttcttctt ctattccttc gggcctgtcg ggtcccctcg gggttgggag
     9541 gtgggtctga aacgataatg gtgaatatcc ctgcctaact ctattcacta tagaaagtac
     9601 agcaaaaact attcttaaac ctaccaagcc tcctactatc attatgaata attttatata
     9661 ccacagccaa tttgttatgt taaaccaatt ccacaaactt gcccatttat ctaattccaa
     9721 taattcttgt tcattctttt cttgctggtt ttgcgattct tcaattaagg agtgtattaa
     9781 gcttgtgtaa ttgttaattt ctctgtccca ctccatccag gtcgtgtgat tccaaatctg
     9841 ttccagagat ttattactcc aactagcatt ccaaggcaca gcagtggtgc aaatgagttt
     9901 tccagagcaa ccccaaatcc ccaggagctg ttgatccttt aggtatcttt ccacagccag
     9961 gattcttgcc tggagctgct tgatgcccca gactgtgagt tgcaacagat gctgttgcgc
    10021 ctcaatagcc ctcagcaaat tgttctgctg ctgcactata ccagacaata attgtctggc
    10081 ctgtaccgtc agcgtcattg acgctgcgcc catagtgctt cctgctgctc ccaagaaccc
    10141 aaggaacaaa gctcctattc ccactgctct tttttctctc tgcaccactc ttctctttgc
    10201 cttggtgggt gctactccta atggttcaat ttttactact ttatatttat ataattcact
    10261 tctccaattg tccctcatat ctcctcctcc aggtctgaag atcagcggcc ggccgcttgc
    10321 tgtgcggtgg tcttactttt gttttgctct tcctctatct tgtctaaagc ttccttggtg
    10381 tcttttatct ctatcctttg atgcacacaa tagagggttg ctactgtatt atataatgat
    10441 ctaagttctt ctgatcctgt ctgaagggat ggttgtagct gtcccagtat ttgtctacag
    10501 ccttctgatg tttctaacag gccaggatta actgcgaatc gttctagctc cctgcttgcc
    10561 catactatat gttttaattt atattttttc tttccccctg gccttaaccg aattttttcc
    10621 catcgcgatc taattctccc ccgcttaata ctgacgctct cgcacccatc tctctccttc
    10681 tagcctccgc tagtcaaaat ttttggcgta ctcaccagtc gccgcccctc gcctcttgcc
    10741 gtgcgcgctt cagcaagccg agtcctgcgt cgagagagct cctctggttt ccctttcgct
    10801 ttcaagtccc tgttcgggcg ccactgctag agattttcca cactgactaa aagggtctga
    10861 gggatctcta gttaccagag tcacacaaca gacgggcaca cactacttga agcactcaag
    10921 gcaagcttta ttgaggctta agcagtgggt tccctagtta gccagagagc tcccaggctc
    10981 agatctggtc taaccagaga gacccagtac aggcaaaaag cagctgctta tatgcaggat
    11041 ctgagggctc gccactcccc agtcccgccc aggccacgcc tccctggaaa gtccccagcg
    11101 gaaagtccct tgtagcaagc tcgatatcag cagttcttga agtactccgg atgcagctct
    11161 cgggccacgt gatgaaatgc taggcggctg tcaaacctcc actctaacac ttctctctcc
    11221 gggtcatcca tcccatgcag gctcacaggg tgtaacaagc tggtgttctc tcctttattg
    11281 gcctcttcta ccttatctgg ctcaactggt actagcttgt agcaccatcc aaaggtcagt
    11341 ggatatctga cccctggccc tggtgtgtag ttctgctaat cagggaagta gccttgtgtg
    11401 tggtagatcc acagatcaag gatatcttgt cttctttggg agtgaattag cccttccaac
    11461 tactaagttt gtagtacata tttaacaaat acaatttctt taaaatgaaa ataattcaga
    11521 ggaatcacag gtttagagta aatgaaacca caggtaattg gcagtggtaa tagggtatgg
    11581 ggtgggaagt ttgggatgat tttggttagc ttgagttatc cagttgatcc agacatgata
    11641 agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt
    11701 tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt
    11761 gatctcccga tccgtcgacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
    11821 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
    11881 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
    11941 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
    12001 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
    12061 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
    12121 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
    12181 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
    12241 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
    12301 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
    12361 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
    12421 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
    12481 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
    12541 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
    12601 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
    12661 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
    12721 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
    12781 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
    12841 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
    12901 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttc
//
