LOCUS       sequence-466837-       13014 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Non Targeting Control.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13014)
  AUTHORS   Miao R, Jiang C, Chang WY, Zhang H, An J, Ho F, Chen P, Zhang H, 
            Junqueira C, Amgalan D, Liang FG, Zhang J, Evavold CL, 
            Hafner-Bratkovic I, Zhang Z, Fontana P, Xia S, Waldeck-Weiermair M, 
            Pan Y, Michel T, Bar-Peled L, Wu H, Kagan JC, Kitsis RN, Zhang P, 
            Liu X, Lieberman J
  TITLE     Gasdermin D permeabilization of mitochondrial inner and outer 
            membranes accelerates and enhances pyroptosis.
  JOURNAL   Immunity. 2023 Nov 14;56(11):2523-2541.e8. doi: 
            10.1016/j.immuni.2023.10.004. Epub 2023 Nov 3.
  PUBMED    37924812
REFERENCE   2  (bases 1 to 13014)
  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: 
            "10.1016/j.immuni.2023.10.004"; journalName: "Immunity"; date: 
            "2023-11-14- 14"; volume: "56"; issue: "11"; pages: "2523-2541.e8"
COMMENT     Sequence Label: lentiCRISPRv2-sgCTL
FEATURES             Location/Qualifiers
     source          1..13014
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     464..484
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    583..1171
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(636..656)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1054..1065)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             1243..1476
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(1502..1519)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1508..1732
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1778..2206
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1865..1884)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2075..2096
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2215..2235)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2220..2549
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2400..2535
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2462..2481
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        2597..2644
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2663..3037
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    3167..3288
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3204..3223)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3258..3277
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3337..3353)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3337..3353)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3350..3372)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3361..3377
                     /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(3385..3391,3392..3409,3410..3415))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3430..3451
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3568..3585)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3739..4327)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3819..3838)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4498..5289,5290..5358))
                     /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     5121..5140
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5359..5463)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5538..5557)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        5729..6108
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6110..6308
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6263..6283
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             6326..6506
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6553..6678
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7171..7404
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7589..7633
                     /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    7931..8048
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        8099..8347
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8099..8119
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8270..8289
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        8370..8445
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        8507..8718
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      8737..8746
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             8743..12846
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             8743..12846
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             12847..12894
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             12895..12918
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             12928..12984
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             join(12985..13014,1..567)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="TEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERV
                     TELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAA
                     QQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSA
                     PRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
ORIGIN
        1 acccgcgacg acgtccccag ggccgtacgc accctcgccg ccgcgttcgc cgactacccc
       61 gccacgcgcc acaccgtcga tccggaccgc cacatcgagc gggtcaccga gctgcaagaa
      121 ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc
      181 gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc
      241 ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggaaggc
      301 ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggagtctcg
      361 cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc
      421 gagcgcgccg gggtgcccgc cttcctggag acctccgcgc cccgcaacct ccccttctac
      481 gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg
      541 tgcatgaccc gcaagcccgg tgcctgaacg cgttaagtcg acaatcaacc tctggattac
      601 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga
      661 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc
      721 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa
      781 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc
      841 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc
      901 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc
      961 gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg
     1021 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct
     1081 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg
     1141 agtcggatct ccctttgggc cgcctccccg cgtcgacttt aagaccaatg acttacaagg
     1201 cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact
     1261 cccaacgaag acaagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct
     1321 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
     1381 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
     1441 tcagaccctt ttagtcagtg tggaaaatct ctagcagggc ccgtttaaac ccgctgatca
     1501 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     1561 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     1621 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     1681 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag
     1741 gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta
     1801 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     1861 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     1921 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     1981 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     2041 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     2101 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     2161 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg
     2221 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     2281 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     2341 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     2401 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     2461 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
     2521 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg
     2581 gatctgatca gcacgtgttg acaattaatc atcggcatag tatatcggca tagtataata
     2641 cgacaaggtg aggaactaaa ccatggccaa gttgaccagt gccgttccgg tgctcaccgc
     2701 gcgcgacgtc gccggagcgg tcgagttctg gaccgaccgg ctcgggttct cccgggactt
     2761 cgtggaggac gacttcgccg gtgtggtccg ggacgacgtg accctgttca tcagcgcggt
     2821 ccaggaccag gtggtgccgg acaacaccct ggcctgggtg tgggtgcgcg gcctggacga
     2881 gctgtacgcc gagtggtcgg aggtcgtgtc cacgaacttc cgggacgcct ccgggccggc
     2941 catgaccgag atcggcgagc agccgtgggg gcgggagttc gccctgcgcg acccggccgg
     3001 caactgcgtg cacttcgtgg ccgaggagca ggactgacac gtgctacgag atttcgattc
     3061 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     3121 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     3181 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     3241 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     3301 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     3361 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     3421 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     3481 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     3541 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     3601 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     3661 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     3721 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     3781 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     3841 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     3901 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     3961 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     4021 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     4081 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     4141 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     4201 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     4261 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     4321 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     4381 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     4441 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     4501 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     4561 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     4621 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     4681 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     4741 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     4801 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     4861 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     4921 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     4981 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     5041 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     5101 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     5161 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     5221 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     5281 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     5341 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     5401 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     5461 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc
     5521 ctatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc
     5581 cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg
     5641 caaggcttga ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt
     5701 cgcgatgtac gggccagata tacgcgttga cattgattat tgactagtta ttaatagtaa
     5761 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
     5821 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
     5881 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
     5941 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
     6001 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
     6061 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt
     6121 tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
     6181 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
     6241 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
     6301 ataagcagcg cgttttgcct gtactgggtc tctctggtta gaccagatct gagcctggga
     6361 gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct
     6421 tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt
     6481 ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa agcgaaaggg
     6541 aaaccagagg agctctctcg acgcaggact cggcttgctg aagcgcgcac ggcaagaggc
     6601 gaggggcggc gactggtgag tacgccaaaa attttgacta gcggaggcta gaaggagaga
     6661 gatgggtgcg agagcgtcag tattaagcgg gggagaatta gatcgcgatg ggaaaaaatt
     6721 cggttaaggc cagggggaaa gaaaaaatat aaattaaaac atatagtatg ggcaagcagg
     6781 gagctagaac gattcgcagt taatcctggc ctgttagaaa catcagaagg ctgtagacaa
     6841 atactgggac agctacaacc atcccttcag acaggatcag aagaacttag atcattatat
     6901 aatacagtag caaccctcta ttgtgtgcat caaaggatag agataaaaga caccaaggaa
     6961 gctttagaca agatagagga agagcaaaac aaaagtaaga ccaccgcaca gcaagcggcc
     7021 gctgatcttc agacctggag gaggagatat gagggacaat tggagaagtg aattatataa
     7081 atataaagta gtaaaaattg aaccattagg agtagcaccc accaaggcaa agagaagagt
     7141 ggtgcagaga gaaaaaagag cagtgggaat aggagctttg ttccttgggt tcttgggagc
     7201 agcaggaagc actatgggcg cagcgtcaat gacgctgacg gtacaggcca gacaattatt
     7261 gtctggtata gtgcagcagc agaacaattt gctgagggct attgaggcgc aacagcatct
     7321 gttgcaactc acagtctggg gcatcaagca gctccaggca agaatcctgg ctgtggaaag
     7381 atacctaaag gatcaacagc tcctggggat ttggggttgc tctggaaaac tcatttgcac
     7441 cactgctgtg ccttggaatg ctagttggag taataaatct ctggaacaga tttggaatca
     7501 cacgacctgg atggagtggg acagagaaat taacaattac acaagcttaa tacactcctt
     7561 aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa gaattattgg aattagataa
     7621 atgggcaagt ttgtggaatt ggtttaacat aacaaattgg ctgtggtata taaaattatt
     7681 cataatgata gtaggaggct tggtaggttt aagaatagtt tttgctgtac tttctatagt
     7741 gaatagagtt aggcagggat attcaccatt atcgtttcag acccacctcc caaccccgag
     7801 gggacccgac aggcccgaag gaatagaaga agaaggtgga gagagagaca gagacagatc
     7861 cattcgatta gtgaacggat cggcactgcg tgcgccaatt ctgcagacaa atggcagtat
     7921 tcatccacaa ttttaaaaga aaagggggga ttggggggta cagtgcaggg gaaagaatag
     7981 tagacataat agcaacagac atacaaacta aagaattaca aaaacaaatt acaaaaattc
     8041 aaaattttcg ggtttattac agggacagca gagatccagt ttggttaatt aaggtaccga
     8101 gggcctattt cccatgattc cttcatattt gcatatacga tacaaggctg ttagagagat
     8161 aattagaatt aatttgactg taaacacaaa gatattagta caaaatacgt gacgtagaaa
     8221 gtaataattt cttgggtagt ttgcagtttt aaaattatgt tttaaaatgg actatcatat
     8281 gcttaccgta acttgaaagt atttcgattt cttggcttta tatatcttgt ggaaaggacg
     8341 aaacaccgca acaagatgaa gagcaccaag ttttagagct agaaatagca agttaaaata
     8401 aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt tgaattcgct
     8461 agctaggtct tgaaaggagt gggaattggc tccggtgccc gtcagtgggc agagcgcaca
     8521 tcgcccacag tccccgagaa gttgggggga ggggtcggca attgatccgg tgcctagaga
     8581 aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag
     8641 ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgcaacggg
     8701 tttgccgcca gaacacagga ccggttctag agcgctgcca ccatggacaa gaagtacagc
     8761 atcggcctgg acatcggcac caactctgtg ggctgggccg tgatcaccga cgagtacaag
     8821 gtgcccagca agaaattcaa ggtgctgggc aacaccgacc ggcacagcat caagaagaac
     8881 ctgatcggag ccctgctgtt cgacagcggc gaaacagccg aggccacccg gctgaagaga
     8941 accgccagaa gaagatacac cagacggaag aaccggatct gctatctgca agagatcttc
     9001 agcaacgaga tggccaaggt ggacgacagc ttcttccaca gactggaaga gtccttcctg
     9061 gtggaagagg ataagaagca cgagcggcac cccatcttcg gcaacatcgt ggacgaggtg
     9121 gcctaccacg agaagtaccc caccatctac cacctgagaa agaaactggt ggacagcacc
     9181 gacaaggccg acctgcggct gatctatctg gccctggccc acatgatcaa gttccggggc
     9241 cacttcctga tcgagggcga cctgaacccc gacaacagcg acgtggacaa gctgttcatc
     9301 cagctggtgc agacctacaa ccagctgttc gaggaaaacc ccatcaacgc cagcggcgtg
     9361 gacgccaagg ccatcctgtc tgccagactg agcaagagca gacggctgga aaatctgatc
     9421 gcccagctgc ccggcgagaa gaagaatggc ctgttcggaa acctgattgc cctgagcctg
     9481 ggcctgaccc ccaacttcaa gagcaacttc gacctggccg aggatgccaa actgcagctg
     9541 agcaaggaca cctacgacga cgacctggac aacctgctgg cccagatcgg cgaccagtac
     9601 gccgacctgt ttctggccgc caagaacctg tccgacgcca tcctgctgag cgacatcctg
     9661 agagtgaaca ccgagatcac caaggccccc ctgagcgcct ctatgatcaa gagatacgac
     9721 gagcaccacc aggacctgac cctgctgaaa gctctcgtgc ggcagcagct gcctgagaag
     9781 tacaaagaga ttttcttcga ccagagcaag aacggctacg ccggctacat tgacggcgga
     9841 gccagccagg aagagttcta caagttcatc aagcccatcc tggaaaagat ggacggcacc
     9901 gaggaactgc tcgtgaagct gaacagagag gacctgctgc ggaagcagcg gaccttcgac
     9961 aacggcagca tcccccacca gatccacctg ggagagctgc acgccattct gcggcggcag
    10021 gaagattttt acccattcct gaaggacaac cgggaaaaga tcgagaagat cctgaccttc
    10081 cgcatcccct actacgtggg ccctctggcc aggggaaaca gcagattcgc ctggatgacc
    10141 agaaagagcg aggaaaccat caccccctgg aacttcgagg aagtggtgga caagggcgct
    10201 tccgcccaga gcttcatcga gcggatgacc aacttcgata agaacctgcc caacgagaag
    10261 gtgctgccca agcacagcct gctgtacgag tacttcaccg tgtataacga gctgaccaaa
    10321 gtgaaatacg tgaccgaggg aatgagaaag cccgccttcc tgagcggcga gcagaaaaag
    10381 gccatcgtgg acctgctgtt caagaccaac cggaaagtga ccgtgaagca gctgaaagag
    10441 gactacttca agaaaatcga gtgcttcgac tccgtggaaa tctccggcgt ggaagatcgg
    10501 ttcaacgcct ccctgggcac ataccacgat ctgctgaaaa ttatcaagga caaggacttc
    10561 ctggacaatg aggaaaacga ggacattctg gaagatatcg tgctgaccct gacactgttt
    10621 gaggacagag agatgatcga ggaacggctg aaaacctatg cccacctgtt cgacgacaaa
    10681 gtgatgaagc agctgaagcg gcggagatac accggctggg gcaggctgag ccggaagctg
    10741 atcaacggca tccgggacaa gcagtccggc aagacaatcc tggatttcct gaagtccgac
    10801 ggcttcgcca acagaaactt catgcagctg atccacgacg acagcctgac ctttaaagag
    10861 gacatccaga aagcccaggt gtccggccag ggcgatagcc tgcacgagca cattgccaat
    10921 ctggccggca gccccgccat taagaagggc atcctgcaga cagtgaaggt ggtggacgag
    10981 ctcgtgaaag tgatgggccg gcacaagccc gagaacatcg tgatcgaaat ggccagagag
    11041 aaccagacca cccagaaggg acagaagaac agccgcgaga gaatgaagcg gatcgaagag
    11101 ggcatcaaag agctgggcag ccagatcctg aaagaacacc ccgtggaaaa cacccagctg
    11161 cagaacgaga agctgtacct gtactacctg cagaatgggc gggatatgta cgtggaccag
    11221 gaactggaca tcaaccggct gtccgactac gatgtggacc atatcgtgcc tcagagcttt
    11281 ctgaaggacg actccatcga caacaaggtg ctgaccagaa gcgacaagaa ccggggcaag
    11341 agcgacaacg tgccctccga agaggtcgtg aagaagatga agaactactg gcggcagctg
    11401 ctgaacgcca agctgattac ccagagaaag ttcgacaatc tgaccaaggc cgagagaggc
    11461 ggcctgagcg aactggataa ggccggcttc atcaagagac agctggtgga aacccggcag
    11521 atcacaaagc acgtggcaca gatcctggac tcccggatga acactaagta cgacgagaat
    11581 gacaagctga tccgggaagt gaaagtgatc accctgaagt ccaagctggt gtccgatttc
    11641 cggaaggatt tccagtttta caaagtgcgc gagatcaaca actaccacca cgcccacgac
    11701 gcctacctga acgccgtcgt gggaaccgcc ctgatcaaaa agtaccctaa gctggaaagc
    11761 gagttcgtgt acggcgacta caaggtgtac gacgtgcgga agatgatcgc caagagcgag
    11821 caggaaatcg gcaaggctac cgccaagtac ttcttctaca gcaacatcat gaactttttc
    11881 aagaccgaga ttaccctggc caacggcgag atccggaagc ggcctctgat cgagacaaac
    11941 ggcgaaaccg gggagatcgt gtgggataag ggccgggatt ttgccaccgt gcggaaagtg
    12001 ctgagcatgc cccaagtgaa tatcgtgaaa aagaccgagg tgcagacagg cggcttcagc
    12061 aaagagtcta tcctgcccaa gaggaacagc gataagctga tcgccagaaa gaaggactgg
    12121 gaccctaaga agtacggcgg cttcgacagc cccaccgtgg cctattctgt gctggtggtg
    12181 gccaaagtgg aaaagggcaa gtccaagaaa ctgaagagtg tgaaagagct gctggggatc
    12241 accatcatgg aaagaagcag cttcgagaag aatcccatcg actttctgga agccaagggc
    12301 tacaaagaag tgaaaaagga cctgatcatc aagctgccta agtactccct gttcgagctg
    12361 gaaaacggcc ggaagagaat gctggcctct gccggcgaac tgcagaaggg aaacgaactg
    12421 gccctgccct ccaaatatgt gaacttcctg tacctggcca gccactatga gaagctgaag
    12481 ggctcccccg aggataatga gcagaaacag ctgtttgtgg aacagcacaa gcactacctg
    12541 gacgagatca tcgagcagat cagcgagttc tccaagagag tgatcctggc cgacgctaat
    12601 ctggacaaag tgctgtccgc ctacaacaag caccgggata agcccatcag agagcaggcc
    12661 gagaatatca tccacctgtt taccctgacc aatctgggag cccctgccgc cttcaagtac
    12721 tttgacacca ccatcgaccg gaagaggtac accagcacca aagaggtgct ggacgccacc
    12781 ctgatccacc agagcatcac cggcctgtac gagacacgga tcgacctgtc tcagctggga
    12841 ggcgacaagc gacctgccgc cacaaagaag gctggacagg ctaagaagaa gaaagattac
    12901 aaagacgatg acgataaggg atccggcgca acaaacttct ctctgctgaa acaagccgga
    12961 gatgtcgaag agaatcctgg accgaccgag tacaagccca cggtgcgcct cgcc
//
