LOCUS       sequence-106781-        9281 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible; gRNA seq is ATGAGAATCAAGGCGGTCGA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9281)
  AUTHORS   Moore R, Spinhirne A, Lai MJ, Preisser S, Li Y, Kang T, Bleris L
  TITLE     CRISPR-based self-cleaving mechanism for controllable gene delivery 
            in human cells.
  JOURNAL   Nucleic Acids Res. 2015 Jan 30;43(2):1297-303. doi: 
            10.1093/nar/gku1326. Epub 2014 Dec 18.
  PUBMED    25527740
REFERENCE   2  (bases 1 to 9281)
  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: 
            "10.1093/nar/gku1326"; journalName: "Nucleic Acids Res"; date: 
            "2015-01-30- 30"; volume: "43"; issue: "2"; pages: "1297-303"
COMMENT     Sequence Label: ptreCas9-mKate2ps-T1gRNA
FEATURES             Location/Qualifiers
     source          1..9281
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      302..890
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     791..810
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    1032..1050
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1068..1086
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1104..1122
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1140..1158
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1176..1194
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1212..1230
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1248..1266
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1311..1335
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      1411..1420
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1417..5520
                     /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             1417..5520
                     /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             1417..5520
                     /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             5533..5553
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5557..5610
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             5623..6318
                     /codon_start=1
                     /product="monomeric far-red fluorescent protein (Shcherbo 
                     et al., 2009)"
                     /label=mKate2
                     /note="mammalian codon-optimized"
                     /translation="MVSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK
                     AVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRADMAL
                     KLVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAVARY
                     CDLPSKLGHR"
     CDS             6337..6456
                     /codon_start=1
                     /product="PEST degradation sequence from mouse ornithine 
                     decarboxylase"
                     /label=hPEST
                     /note="human codon-optimized"
                     /translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
     polyA_signal    6590..6711
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6627..6646)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6681..6700
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(6718..7173)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6855..6876)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7067..7086
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        7200..7304
                     /gene="bla"
                     /label=AmpR promoter
     promoter        7306..7663
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        7306..7497
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(7329..7349)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      7514..7649
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             7698..8492
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             7698..8492
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(7752..7771)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     8362..8381
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(8680..8699)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    8724..8771
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     promoter        8864..9112
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8864..8884
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     9035..9054
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        9133..9208
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(9220..9244)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 ctagtggatc cgccacccca ccgagacccc attggggcca atacgcccgc gtttcttcct
       61 tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg
      121 cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc
      181 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
      241 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
      301 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
      361 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
      421 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
      481 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
      541 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
      601 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
      661 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
      721 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
      781 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
      841 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
      901 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
      961 cgttatcccc tgattctgtg gataaccgta ttaccgcccc gtacacgcca cctcgacata
     1021 ctcgagttta ctccctatca gtgatagaga acgtatgaag agtttactcc ctatcagtga
     1081 tagagaacgt atgcagactt tactccctat cagtgataga gaacgtataa ggagtttact
     1141 ccctatcagt gatagagaac gtatgaccag tttactccct atcagtgata gagaacgtat
     1201 ctacagttta ctccctatca gtgatagaga acgtatatcc agtttactcc ctatcagtga
     1261 tagagaacgt ataagcttta ggcgtgtacg gtgggcgcct ataaaagcag agctcgttta
     1321 gtgaaccgtc agatcgcctg gagcaattcc acaacacttt tgtcttatac caactttccg
     1381 gctagtggat ccgcggccgt tactagcctt gccaccatgg acaagaagta ctccattggg
     1441 ctcgatatcg gcacaaacag cgtcggctgg gccgtcatta cggacgagta caaggtgccg
     1501 agcaaaaaat tcaaagttct gggcaatacc gatcgccaca gcataaagaa gaacctcatt
     1561 ggcgccctcc tgttcgactc cggggagacg gccgaagcca cgcggctcaa aagaacagca
     1621 cggcgcagat atacccgcag aaagaatcgg atctgctacc tgcaggagat ctttagtaat
     1681 gagatggcta aggtggatga ctctttcttc cataggctgg aggagtcctt tttggtggag
     1741 gaggataaaa agcacgagcg ccacccaatc tttggcaata tcgtggacga ggtggcgtac
     1801 catgaaaagt acccaaccat atatcatctg aggaagaagc ttgtagacag tactgataag
     1861 gctgacttgc ggttgatcta tctcgcgctg gcgcatatga tcaaatttcg gggacacttc
     1921 ctcatcgagg gggacctgaa cccagacaac agcgatgtcg acaaactctt tatccaactg
     1981 gttcagactt acaatcagct tttcgaagag aacccgatca acgcatccgg agttgacgcc
     2041 aaagcaatcc tgagcgctag gctgtccaaa tcccggcggc tcgaaaacct catcgcacag
     2101 ctccctgggg agaagaagaa cggcctgttt ggtaatctta tcgccctgtc actcgggctg
     2161 acccccaact ttaaatctaa cttcgacctg gccgaagatg ccaagcttca actgagcaaa
     2221 gacacctacg atgatgatct cgacaatctg ctggcccaga tcggcgacca gtacgcagac
     2281 ctttttttgg cggcaaagaa cctgtcagac gccattctgc tgagtgatat tctgcgagtg
     2341 aacacggaga tcaccaaagc tccgctgagc gctagtatga tcaagcgcta tgatgagcac
     2401 caccaagact tgactttgct gaaggccctt gtcagacagc aactgcctga gaagtacaag
     2461 gaaattttct tcgatcagtc taaaaatggc tacgccggat acattgacgg cggagcaagc
     2521 caggaggaat tttacaaatt tattaagccc atcttggaaa aaatggacgg caccgaggag
     2581 ctgctggtaa agcttaacag agaagatctg ttgcgcaaac agcgcacttt cgacaatgga
     2641 agcatccccc accagattca cctgggcgaa ctgcacgcta tcctcaggcg gcaagaggat
     2701 ttctacccct ttttgaaaga taacagggaa aagattgaga aaatcctcac atttcggata
     2761 ccctactatg taggccccct cgcccgggga aattccagat tcgcgtggat gactcgcaaa
     2821 tcagaagaga ccatcactcc ctggaacttc gaggaagtcg tggataaggg ggcctctgcc
     2881 cagtccttca tcgaaaggat gactaacttt gataaaaatc tgcctaacga aaaggtgctt
     2941 cctaaacact ctctgctgta cgagtacttc acagtttata acgagctcac caaggtcaaa
     3001 tacgtcacag aagggatgag aaagccagca ttcctgtctg gagagcagaa gaaagctatc
     3061 gtggacctcc tcttcaagac gaaccggaaa gttaccgtga aacagctcaa agaagactat
     3121 ttcaaaaaga ttgaatgttt cgactctgtt gaaatcagcg gagtggagga tcgcttcaac
     3181 gcatccctgg gaacgtatca cgatctcctg aaaatcatta aagacaagga cttcctggac
     3241 aatgaggaga acgaggacat tcttgaggac attgtcctca cccttacgtt gtttgaagat
     3301 agggagatga ttgaagaacg cttgaaaact tacgctcatc tcttcgacga caaagtcatg
     3361 aaacagctca agaggcgccg atatacagga tgggggcggc tgtcaagaaa actgatcaat
     3421 gggatccgag acaagcagag tggaaagaca atcctggatt ttcttaagtc cgatggattt
     3481 gccaaccgga acttcatgca gttgatccat gatgactctc tcacctttaa ggaggacatc
     3541 cagaaagcac aagtttctgg ccagggggac agtcttcacg agcacatcgc taatcttgca
     3601 ggtagcccag ctatcaaaaa gggaatactg cagaccgtta aggtcgtgga tgaactcgtc
     3661 aaagtaatgg gaaggcataa gcccgagaat atcgttatcg agatggcccg agagaaccaa
     3721 actacccaga agggacagaa gaacagtagg gaaaggatga agaggattga agagggtata
     3781 aaagaactgg ggtcccaaat ccttaaggaa cacccagttg aaaacaccca gcttcagaat
     3841 gagaagctct acctgtacta cctgcagaac ggcagggaca tgtacgtgga tcaggaactg
     3901 gacatcaatc ggctctccga ctacgacgtg gatcatatcg tgccccagtc ttttctcaaa
     3961 gatgattcta ttgataataa agtgttgaca agatccgata aaaatagagg gaagagtgat
     4021 aacgtcccct cagaagaagt tgtcaagaaa atgaaaaatt attggcggca gctgctgaac
     4081 gccaaactga tcacacaacg gaagttcgat aatctgacta aggctgaacg aggtggcctg
     4141 tctgagttgg ataaagccgg cttcatcaaa aggcagcttg ttgagacacg ccagatcacc
     4201 aagcacgtgg cccaaattct cgattcacgc atgaacacca agtacgatga aaatgacaaa
     4261 ctgattcgag aggtgaaagt tattactctg aagtctaagc tggtctcaga tttcagaaag
     4321 gactttcagt tttataaggt gagagagatc aacaattacc accatgcgca tgatgcctac
     4381 ctgaatgcag tggtaggcac tgcacttatc aaaaaatatc ccaagcttga atctgaattt
     4441 gtttacggag actataaagt gtacgatgtt aggaaaatga tcgcaaagtc tgagcaggaa
     4501 ataggcaagg ccaccgctaa gtacttcttt tacagcaata ttatgaattt tttcaagacc
     4561 gagattacac tggccaatgg agagattcgg aagcgaccac ttatcgaaac aaacggagaa
     4621 acaggagaaa tcgtgtggga caagggtagg gatttcgcga cagtccggaa ggtcctgtcc
     4681 atgccgcagg tgaacatcgt taaaaagacc gaagtacaga ccggaggctt ctccaaggaa
     4741 agtatcctcc cgaaaaggaa cagcgacaag ctgatcgcac gcaaaaaaga ttgggacccc
     4801 aagaaatacg gcggattcga ttctcctaca gtcgcttaca gtgtactggt tgtggccaaa
     4861 gtggagaaag ggaagtctaa aaaactcaaa agcgtcaagg aactgctggg catcacaatc
     4921 atggagcgat caagcttcga aaaaaacccc atcgactttc tcgaggcgaa aggatataaa
     4981 gaggtcaaaa aagacctcat cattaagctt cccaagtact ctctctttga gcttgaaaac
     5041 ggccggaaac gaatgctcgc tagtgcgggc gagctgcaga aaggtaacga gctggcactg
     5101 ccctctaaat acgttaattt cttgtatctg gccagccact atgaaaagct caaagggtct
     5161 cccgaagata atgagcagaa gcagctgttc gtggaacaac acaaacacta ccttgatgag
     5221 atcatcgagc aaataagcga attctccaaa agagtgatcc tcgccgacgc taacctcgat
     5281 aaggtgcttt ctgcttacaa taagcacagg gataagccca tcagggagca ggcagaaaac
     5341 attatccact tgtttactct gaccaacttg ggcgcgcctg cagccttcaa gtacttcgac
     5401 accaccatag acagaaagcg gtacacctct acaaaggagg tcctggacgc cacactgatt
     5461 catcagtcaa ttacggggct ctatgaaaca agaatcgacc tctctcagct cggtggagac
     5521 agcagggctg accccaagaa gaagaggaag gtggctgagg gcagaggaag tcttctaaca
     5581 tgcggtgacg tggaggagaa tcccggccct tccggagggc ccatggtgag cgagctgatt
     5641 aaggagaaca tgcacatgaa gctgtacatg gagggcaccg tgaacaacca ccacttcaag
     5701 tgcacatccg agggcgaagg caagccctac gagggcaccc agaccatgag aatcaaggcg
     5761 gtcgagggcg gccctctccc cttcgccttc gacatcctgg ctaccagctt catgtacggc
     5821 agcaaaacct tcatcaacca cacccagggc atccccgact tctttaagca gtccttcccc
     5881 gagggcttca catgggagag agtcaccaca tacgaagacg ggggcgtgct gaccgctacc
     5941 caggacacca gcctccagga cggctgcctc atctacaacg tcaagatcag aggggtgaac
     6001 ttcccatcca acggccctgt gatgcagaag aaaacactcg gctgggaggc ctccaccgag
     6061 accctgtacc ccgctgacgg cggcctggaa ggcagagccg acatggccct gaagctcgtg
     6121 ggcgggggcc acctgatctg caacttgaag accacataca gatccaagaa acccgctaag
     6181 aacctcaaga tgcccggcgt ctactatgtg gacagaagac tggaaagaat caaggaggcc
     6241 gacaaagaga cctacgtcga gcagcacgag gtggctgtgg ccagatactg cgacctccct
     6301 agcaaactgg ggcacagagg tggaggaggt tccggatctc acggcttccc tcccgaggtg
     6361 gaggagcagg ccgccggcac cctgcccatg agctgcgccc aggagagcgg catggataga
     6421 caccctgctg cttgcgccag cgccaggatc aacgtctaag gccggccagc ggccgcgact
     6481 ctagatcata atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc
     6541 acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat
     6601 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     6661 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa
     6721 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     6781 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
     6841 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     6901 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
     6961 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     7021 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     7081 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     7141 ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc
     7201 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     7261 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa
     7321 ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     7381 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     7441 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     7501 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     7561 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     7621 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag
     7681 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     7741 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     7801 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     7861 gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     7921 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     7981 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     8041 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     8101 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     8161 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     8221 aggcgagcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     8281 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     8341 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     8401 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     8461 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
     8521 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     8581 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     8641 catgctggag ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat
     8701 accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg
     8761 gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc
     8821 gagacctcag tcgactggat ccggtaccaa ggtcgggcag gaagagggcc tatttcccat
     8881 gattccttca tatttgcata tacgatacaa ggctgttaga gagataatta gaattaattt
     8941 gactgtaaac acaaagatat tagtacaaaa tacgtgacgt agaaagtaat aatttcttgg
     9001 gtagtttgca gttttaaaat tatgttttaa aatggactat catatgctta ccgtaacttg
     9061 aaagtatttc gatttcttgg ctttatatat cttgtggaaa ggacgaaaca ccgtgagaat
     9121 caaggcggtc gagttttaga gctagaaata gcaagttaaa ataaggctag tccgttatca
     9181 acttgaaaaa gtggcaccga gtcggtgctt tttttctaga cccagctttc ttgtacaaag
     9241 ttggcattaa agggcgaatt ccagcacact ggcggccgtt a
//
