LOCUS       sequence-412265-       12379 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducibly expresses human Caspase1, fused to light-activatable Cry2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12379)
  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 12379)
  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-hCaspase-1
FEATURES             Location/Qualifiers
     source          1..12379
                     /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    8574..8708
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    8752..8867
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1 (lacking the first T)"
     CDS             complement(9130..9174)
                     /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    9359..9592
                     /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    10089..10214
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             10261..10894
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     polyA_signal    11062..11196
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        11244..11348
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(11349..11417,11418..12209)
                     /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(11567..11586)
                     /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 ggccgcccat 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 ggaggagatc aaacatctgg
     7441 aaattacctt aatatgcaag actctcaagg agtactttct tcctttccag ctcctcaggc
     7501 agtgcaggac aacccagcta tgcccacatc ctcaggctca gaagggaatg tcaagctttg
     7561 ctccctagaa gaagctcaaa ggatatggaa acaaaagtcg gcagagattt atccaataat
     7621 ggacaagtca agccgcacac gtcttgctct cattatctgc aatgaagaat ttgacagtat
     7681 tcctagaaga actggagctg aggttgacat cacaggcatg acaatgctgc tacaaaatct
     7741 ggggtacagc gtagatgtga aaaaaaatct cactgcttcg gacatgacta cagagctgga
     7801 ggcatttgca caccgcccag agcacaagac ctctgacagc acgttcctgg tgttcatgtc
     7861 tcatggtatt cgggaaggca tttgtgggaa gaaacactct gagcaagtcc cagatatact
     7921 acaactcaat gcaatcttta acatgttgaa taccaagaac tgcccaagtt tgaaggacaa
     7981 accgaaggtg atcatcatcc aggcctgccg tggtgacagc cctggtgtgg tgtggtttaa
     8041 agattcagta ggagtttctg gaaacctatc tttaccaact acagaagagt ttgaggatga
     8101 tgctgttaag aaagcccaca tagagaagga ttttatcgct ttctgctctt ccacaccaga
     8161 taatgtttct tggagacatc ccacaatggg ctctgttttt attggaagac tcattgaaca
     8221 tatgcaagaa tatgcctgtt cctgtgatgt ggaggaaatt ttccgcaagg ttcgattttc
     8281 atttgagcag ccagatggta gagcgcagat gcccaccact gaaagagtga ctttgacaag
     8341 atgtttctac ctcttcccag gacattaaaa ttcccgtata caccggtgcc ggcggatcca
     8401 gaccacctcc cctgcgagct aagctggaca gccaatgacg ggtaagagag tgacattttt
     8461 cactaaccta agacaggagg gccgtcagag ctactgccta atccaaagac gggtaaaagt
     8521 gataaaaatg tatcactcca acctaagaca ggcgcagctt ccgagggatt tgagatccag
     8581 acatgataag atacattgat gagtttggac aaaccaaaac tagaatgcag tgaaaaaaat
     8641 gccttatttg tgaaatttgt gatgctattg ccttatttgt aaccattata agctgcaata
     8701 aacaagttaa gtcgataaac tggatctctg ctgtccctgt aataaacccg aaaattttga
     8761 atttttgtaa tttgtttttg taattcttta gtttgtatgt ctgttgctat tatgtctact
     8821 attctttccc ctgcactgta ccccccaatc cccccttttc ttttaaaggc gataccgtcg
     8881 agatccgttc actaatcgaa tggatctgtc tctgtctctc tctccacctt cttcttctat
     8941 tccttcgggc ctgtcgggtc ccctcggggt tgggaggtgg gtctgaaacg ataatggtga
     9001 atatccctgc ctaactctat tcactataga aagtacagca aaaactattc ttaaacctac
     9061 caagcctcct actatcatta tgaataattt tatataccac agccaatttg ttatgttaaa
     9121 ccaattccac aaacttgccc atttatctaa ttccaataat tcttgttcat tcttttcttg
     9181 ctggttttgc gattcttcaa ttaaggagtg tattaagctt gtgtaattgt taatttctct
     9241 gtcccactcc atccaggtcg tgtgattcca aatctgttcc agagatttat tactccaact
     9301 agcattccaa ggcacagcag tggtgcaaat gagttttcca gagcaacccc aaatccccag
     9361 gagctgttga tcctttaggt atctttccac agccaggatt cttgcctgga gctgcttgat
     9421 gccccagact gtgagttgca acagatgctg ttgcgcctca atagccctca gcaaattgtt
     9481 ctgctgctgc actataccag acaataattg tctggcctgt accgtcagcg tcattgacgc
     9541 tgcgcccata gtgcttcctg ctgctcccaa gaacccaagg aacaaagctc ctattcccac
     9601 tgctcttttt tctctctgca ccactcttct ctttgccttg gtgggtgcta ctcctaatgg
     9661 ttcaattttt actactttat atttatataa ttcacttctc caattgtccc tcatatctcc
     9721 tcctccaggt ctgaagatca gcggccggcc gcttgctgtg cggtggtctt acttttgttt
     9781 tgctcttcct ctatcttgtc taaagcttcc ttggtgtctt ttatctctat cctttgatgc
     9841 acacaataga gggttgctac tgtattatat aatgatctaa gttcttctga tcctgtctga
     9901 agggatggtt gtagctgtcc cagtatttgt ctacagcctt ctgatgtttc taacaggcca
     9961 ggattaactg cgaatcgttc tagctccctg cttgcccata ctatatgttt taatttatat
    10021 tttttctttc cccctggcct taaccgaatt ttttcccatc gcgatctaat tctcccccgc
    10081 ttaatactga cgctctcgca cccatctctc tccttctagc ctccgctagt caaaattttt
    10141 ggcgtactca ccagtcgccg cccctcgcct cttgccgtgc gcgcttcagc aagccgagtc
    10201 ctgcgtcgag agagctcctc tggtttccct ttcgctttca agtccctgtt cgggcgccac
    10261 tgctagagat tttccacact gactaaaagg gtctgaggga tctctagtta ccagagtcac
    10321 acaacagacg ggcacacact acttgaagca ctcaaggcaa gctttattga ggcttaagca
    10381 gtgggttccc tagttagcca gagagctccc aggctcagat ctggtctaac cagagagacc
    10441 cagtacaggc aaaaagcagc tgcttatatg caggatctga gggctcgcca ctccccagtc
    10501 ccgcccaggc cacgcctccc tggaaagtcc ccagcggaaa gtcccttgta gcaagctcga
    10561 tatcagcagt tcttgaagta ctccggatgc agctctcggg ccacgtgatg aaatgctagg
    10621 cggctgtcaa acctccactc taacacttct ctctccgggt catccatccc atgcaggctc
    10681 acagggtgta acaagctggt gttctctcct ttattggcct cttctacctt atctggctca
    10741 actggtacta gcttgtagca ccatccaaag gtcagtggat atctgacccc tggccctggt
    10801 gtgtagttct gctaatcagg gaagtagcct tgtgtgtggt agatccacag atcaaggata
    10861 tcttgtcttc tttgggagtg aattagccct tccaactact aagtttgtag tacatattta
    10921 acaaatacaa tttctttaaa atgaaaataa ttcagaggaa tcacaggttt agagtaaatg
    10981 aaaccacagg taattggcag tggtaatagg gtatggggtg ggaagtttgg gatgattttg
    11041 gttagcttga gttatccagt tgatccagac atgataagat acattgatga gtttggacaa
    11101 accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct
    11161 ttatttgtaa ccattataag ctgcaataaa caagttgatc tcccgatccg tcgacgtcag
    11221 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
    11281 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
    11341 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
    11401 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
    11461 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
    11521 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
    11581 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
    11641 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
    11701 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
    11761 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
    11821 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
    11881 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
    11941 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
    12001 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
    12061 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
    12121 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
    12181 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
    12241 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
    12301 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
    12361 aaaagatcaa aggatcttc
//
