LOCUS       sequence-310410-       16663 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Drives expression of Cas9 and a TdTomato fluorescent marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 16663)
  AUTHORS   Gamez S, Chaverra-Rodriguez D, Buchman A, Kandul NP, Mendez-Sanchez 
            SC, Bennett JB, Sanchez C HM, Yang T, Antoshechkin I, Duque JE, 
            Papathanos PA, Marshall JM, Akbari OS
  TITLE     Exploiting a Y chromosome-linked Cas9 for sex selection and gene 
            drive.
  JOURNAL   Nat Commun. 2021 Dec 10;12(1):7202. doi: 10.1038/s41467-021-27333-1.
  PUBMED    34893590
REFERENCE   2  (bases 1 to 16663)
  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: "Nat 
            Commun."; date: "2021-12-10"; volume: "12(1):7202. doi"; pages: " 
            10.1038/s41467-021-27333-1"
COMMENT     Sequence Label: SGyA
FEATURES             Location/Qualifiers
     source          1..16663
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(95..683)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(175..194)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(854..1645,1646..1714))
                     /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     1477..1496
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1715..1819)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1887..1905
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1943..1965)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2065..2084
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2278..2300
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2292..2309
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2293..2309
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   2998..3060
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     protein_bind    4593..5023
                     /label=gypsy insulator
                     /bound_moiety="Su(Hw), CP190, andMod(mdg4)67.2"
                     /note="chromatin insulator from Drosophila"
     CDS             7280..7345
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             7352..7372
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7397..11497
                     /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             7397..11497
                     /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             11498..11545
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             11546..11599
                     /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             join(11600..11602,11603..11605,11606..12319)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTSKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(11906..11925)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     12253..12274
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     regulatory      13268..13277
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(13407..13425)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     13675..13694
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     13869..13892
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    14826..14947
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(14863..14882)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     14917..14936
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    15025..15124
                     /label=phage phi-C31 attP
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attachment site of phage phi-C31"
     primer_bind     complement(16356..16372)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(16356..16372)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(16369..16391)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    16380..16396
                     /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(16404..16410,16411..16428,16429..16434))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    16449..16470
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(16587..16604)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
       61 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
      121 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
      181 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
      241 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct
      301 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
      361 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
      421 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
      481 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
      541 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
      601 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
      661 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
      721 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
      781 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
      841 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
      901 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
      961 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     1021 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     1081 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     1141 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     1201 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     1261 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     1321 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     1381 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     1441 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     1501 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     1561 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     1621 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     1681 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     1741 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     1801 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     1861 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc
     1921 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
     1981 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     2041 ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
     2101 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc
     2161 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
     2221 agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc
     2281 agtcacgacg ttgtaaaacg acggccagtg ccaagctttg tttaaaatat aacaaaattg
     2341 tgatcccaca aaatgaagtg gggcaaaatc aaataattaa ctagtgtccg taaacttgtt
     2401 ggtcttcaac tttttgagga acacgttgga cggcaaatcg tgactataac acaagttgat
     2461 ttaataattt tagccaacac gtcgggctgc gtgttttttg cgctctgtgt acacgttgat
     2521 taactggtcg attaaataat ttaatttttg gttcttcttt aaatctgtga tgaaattttt
     2581 taaaataact ttaaattctt cattggtaaa aaatgccacg ttttgcaact tgtgagggtc
     2641 taatatgagg tcaaactcag taggagtttt atccaaaaaa gaaaacatga ttacgtctgt
     2701 acacgaacgc gtattaacgc agagtgcaaa gtataagagg gttaaaaaat atattttacg
     2761 caccatatac gcatcgggtt gatatcgtta atatggatca atttgaacag ttgattaacg
     2821 tgtctctgct caagtctttg atcaaaacgc aaatcgacga aaatgtgtcg gacaatatca
     2881 agtcgatgag cgaaaaacta aaaaggctag aatacgacaa tctcacagac agcgttgaga
     2941 tatacggtat tcacgacagc aggctgaata ataaaaaaat tagaaactat tatttaaccc
     3001 tagaaagata atcatattgt gacgtacgtt aaagataatc atgcgtaaaa ttgacgcatg
     3061 tgttttatcg gtctgtatat cgaggtttat ttattaattt gaatagatat taagttttat
     3121 tatatttaca cttacatact aataataaat tcaacaaaca atttatttat gtttatttat
     3181 ttattaaaaa aaaacaaaaa ctcaaaattt cttctataaa gtaacaaaac ttttaaacat
     3241 tctctctttt acaaaaataa acttattttg tactttaaaa acagtcatgt tgtattataa
     3301 aataagtaat tagcttaact tatacataat agaaacaaat tatacttatt agtcagtcag
     3361 aaacaacttt ggcacatatc aatattatgc tctcgacaaa taactttttt gcattttttg
     3421 cacgatgcat ttgcctttcg ccttatttta gaggggcagt aagtacagta agtacgtttt
     3481 ttcattactg gctcttcagt actgtcatct gatgtaccag gcacttcatt tggcaaaata
     3541 ttagagatat tatcgcgcaa atatctcttc aaagtaggag cttctaaacg cttacgcata
     3601 aacgatgacg tcaggctcat gtaaaggttt ctcataaatt ttttgcgact ttgaaccttt
     3661 tctcccttgc tactgacatt atggctgtat ataataaaag aatttatgca ggcaatgttt
     3721 atcattccgt acaataatgc cataggccac ctattcgtct tcctactgca ggtgtacctc
     3781 gcccagttgg tcctattggt tgagggccta cacggtcgct cgatgtgttg tagttggtgc
     3841 aaaagatgaa agattaccgg acagtgacca gatgagagtt ctggaagtgc ctcagctgta
     3901 atcattgatt gggatatcct ttttgtgatg gcaaagtcaa tcaaatctgg aagcttcatg
     3961 ggatctgctg gccattatgt aggcgctccc ggggaaacat agttaagttt gtgttgcggt
     4021 ttgatgattg cgttgtacag caacaaacct ttcaccaaac gagtgaaaag tgttgtgaat
     4081 ttttttctcc aacgccaacg cttaactata ctctataact actcattcac caaaatctat
     4141 ttgctctatc cataaactag ttctttccaa gtaacttaat ctcccgaatc gtaatttctc
     4201 actcgaacta tggcgcgcat agttgtgctt aaggttggag gtgaggtgtt ttgtagatgc
     4261 ccgaagtcca cctgtggctt gttcggtgac aaaaactgat ctgattctat attctagctc
     4321 tggtctcaat tagacctccg ctcagagaag taaataggaa atagtgtctt aaatgctaca
     4381 tttgtaagtg ttaattcata agtgctgcgt cgcacttatt ccgaattgct gtgttgccta
     4441 ctcttgagtg gatcgattct ttgctgagtc gttctcctat tacatagtcc ggaggagtag
     4501 gggtttcctg tccaaatgtc atgttaacgg ccgttcgctg aacggtcgtt gtgtggccta
     4561 attaaatccg ttgtttacat tttgtttaaa cttggccacg taataagtgt gcgttgaatt
     4621 tattcgcaaa aacattgcat attttcggca aagtaaaatt ttgttgcata ccttatcaaa
     4681 aaataagtgc tgcatacttt ttagagaaac caaataattt tttattgcat acccgttttt
     4741 aataaaatac attgcatacc ctcttttaat aaaaaatatt gcatactttg acgaaacaaa
     4801 ttttcgttgc atacccaata aaagattatt atattgcata cccgttttta ataaaataca
     4861 ttgcataccc tcttttaata aaaaatattg catacgttga cgaaacaaat tttcgttgca
     4921 tacccaataa aagattatta tattgcatac cttttcttgc cataccattt agccgatcaa
     4981 ttgtgctcgg caacagtata tttgtggtgt gccaaccaac aacggcgcgc ctgcagctgg
     5041 ttgtaggtgc agttgcgctt cttgcacttg gcgcacttga ggagatcggt cttggtgccc
     5101 tgcacggtgg ccagctgggc atcgttgatg gcctccttga cgaacttctc gcgcagcttc
     5161 ttcatctcgt cgctggccat ctcctccggc gtcattttgg ccagctgctt ggcagtgacg
     5221 gcgccgcaca taaagttgcc gcgcaatcca ggattcttgg ggtccttcag attggccacg
     5281 cgtgacctaa tgcgattctt gtacttcata tccgtgttat tgaactcgga gtaaatggca
     5341 tcctccagtt cggcggccat ttcctccggc tcaccgcatc cctcgggcac ttcgccaatc
     5401 ttcagtgcgg tggccagcat ttcgcggcat ttgatgcgga ccgcatctgt catgccaccg
     5461 gagggaaacg aggtctgtga cgaggatgtc gagcctttct tctctttatc cttggatgag
     5521 ctggatgaac tggacttatc cttgccggag attgacgacg acgacttggc ggcactggtg
     5581 gacttcgagg cactgctgtt gttggaggag ccctccttag cggagctgtt gttcggtgtg
     5641 gttggcgccg ggctggcgag gaatcgtttc cagttcttga tcagcgtctt ggccagagcg
     5701 atcacctcgt cgtccttgct actcttgcgc agctcgttta cggtcatgcc gatgcgcgtt
     5761 ttggtcagaa tgtcgagatt gatgttaagc gtttgcaggg ccttcagcag gtccagagcc
     5821 tgatcctgtc cctgtacggt ggatatatgt atatttgtga catgatgcgc ggcacgttag
     5881 ctaatcccga tacttactgt gccgtcgctg gccatcttgc tcatcttctt ttggattcga
     5941 aacacttcct cctccacgct cattttggcc cacttaattt gatcagaggt taggttaact
     6001 ggcgtcggct gtcggcttta atcggtgctg cgggtctgcg acagttgttt atgcggaagc
     6061 cgggctattt ctttttttcc ctccaataaa tgttgtttac tatttttttc gtttctgtat
     6121 tcgcagaaca agttgacaga acaccctaac gcgttttagt gttaccgggc ggtggcttgt
     6181 caagaaatac cttcccatgg ctaatacaat tcccaactgg aagtagtttg ggtttcagaa
     6241 gtcaacagtg ttaccgtatt ttaatgtaca tctggctttt aagcgagtac acacacatag
     6301 cattgcgatg ttttttaaac ttgcactgct tgcaagcagg ggtccactgt atttcaatga
     6361 aagaacagcg tgttgggcgt ttttgaatta tactaattac cttcaataac aattccctat
     6421 atatcactta gttttaataa ataaatcgtt tgagtgtggt ctagagatga tttattatga
     6481 tgaagtgcgg ttgttgggga caagcgggat acgtctgcgg gtcaaatcgg cggtgggaca
     6541 tagtgagagg gcaacggctc caaaaccaag cgttgcgaac aatcaccaac gcgccctggt
     6601 acgcgagaaa tgttgatatc gcacatgacc tcagcatcca ccttcagttt gagaacaaat
     6661 taaatcaggc acataccaga caaccaggag aacactgagg aggcgtcatc aaagatatct
     6721 ttggacacgt ggcataaaca agccaaacta ttatctaatc aatattttta tacaaatttt
     6781 gtatgtttcc atgttatctt cgttgctcct gttagttact acgtttcata gcccttaaat
     6841 tggttgctta acgtaaaata aaattatatt gtataaaaat aaagaggaat tcacctataa
     6901 caaacgaaaa agtaaataaa taagaacttc cccatgttaa aaatgccacc accatcttta
     6961 catataagta tgtacatatg aatgaatcac ttaggttgct tgaatattac aatttcatta
     7021 atagctaaat cacatttgat gtgttagtgg aaaacggcta tatatataaa ttatcgaaat
     7081 tgtgaatatc gaattgcgat agcacaatgg gaaattccac cactagattt ttggtacttt
     7141 taacagatcc ttttcggttt tgcgttgcgc gaagtgatct gaacttatca aagtttgtaa
     7201 ggtaatacat aaagtgaaaa agaattaatt tgctcttgaa aggcaggcca aattaaaaaa
     7261 aaaatatcaa ttcgagatgg actataagga ccacgacgga gactacaagg atcatgatat
     7321 tgattacaaa gacgatgacg ataagatggc cccaaagaag aagcggaagg tcggtatcca
     7381 cggagtccca gcagccgaca agaagtacag catcggcctg gacatcggca ccaactctgt
     7441 gggctgggcc gtgatcaccg acgagtacaa ggtgcccagc aagaaattca aggtgctggg
     7501 caacaccgac cggcacagca tcaagaagaa cctgatcgga gccctgctgt tcgacagcgg
     7561 cgaaacagcc gaggccaccc ggctgaagag aaccgccaga agaagataca ccagacggaa
     7621 gaaccggatc tgctatctgc aagagatctt cagcaacgag atggccaagg tggacgacag
     7681 cttcttccac agactggaag agtccttcct ggtggaagag gataagaagc acgagcggca
     7741 ccccatcttc ggcaacatcg tggacgaggt ggcctaccac gagaagtacc ccaccatcta
     7801 ccacctgaga aagaaactgg tggacagcac cgacaaggcc gacctgcggc tgatctatct
     7861 ggccctggcc cacatgatca agttccgggg ccacttcctg atcgagggcg acctgaaccc
     7921 cgacaacagc gacgtggaca agctgttcat ccagctggtg cagacctaca accagctgtt
     7981 cgaggaaaac cccatcaacg ccagcggcgt ggacgccaag gccatcctgt ctgccagact
     8041 gagcaagagc agacggctgg aaaatctgat cgcccagctg cccggcgaga agaagaatgg
     8101 cctgttcgga aacctgattg ccctgagcct gggcctgacc cccaacttca agagcaactt
     8161 cgacctggcc gaggatgcca aactgcagct gagcaaggac acctacgacg acgacctgga
     8221 caacctgctg gcccagatcg gcgaccagta cgccgacctg tttctggccg ccaagaacct
     8281 gtccgacgcc atcctgctga gcgacatcct gagagtgaac accgagatca ccaaggcccc
     8341 cctgagcgcc tctatgatca agagatacga cgagcaccac caggacctga ccctgctgaa
     8401 agctctcgtg cggcagcagc tgcctgagaa gtacaaagag attttcttcg accagagcaa
     8461 gaacggctac gccggctaca ttgacggcgg agccagccag gaagagttct acaagttcat
     8521 caagcccatc ctggaaaaga tggacggcac cgaggaactg ctcgtgaagc tgaacagaga
     8581 ggacctgctg cggaagcagc ggaccttcga caacggcagc atcccccacc agatccacct
     8641 gggagagctg cacgccattc tgcggcggca ggaagatttt tacccattcc tgaaggacaa
     8701 ccgggaaaag atcgagaaga tcctgacctt ccgcatcccc tactacgtgg gccctctggc
     8761 caggggaaac agcagattcg cctggatgac cagaaagagc gaggaaacca tcaccccctg
     8821 gaacttcgag gaagtggtgg acaagggcgc ttccgcccag agcttcatcg agcggatgac
     8881 caacttcgat aagaacctgc ccaacgagaa ggtgctgccc aagcacagcc tgctgtacga
     8941 gtacttcacc gtgtataacg agctgaccaa agtgaaatac gtgaccgagg gaatgagaaa
     9001 gcccgccttc ctgagcggcg agcagaaaaa ggccatcgtg gacctgctgt tcaagaccaa
     9061 ccggaaagtg accgtgaagc agctgaaaga ggactacttc aagaaaatcg agtgcttcga
     9121 ctccgtggaa atctccggcg tggaagatcg gttcaacgcc tccctgggca cataccacga
     9181 tctgctgaaa attatcaagg acaaggactt cctggacaat gaggaaaacg aggacattct
     9241 ggaagatatc gtgctgaccc tgacactgtt tgaggacaga gagatgatcg aggaacggct
     9301 gaaaacctat gcccacctgt tcgacgacaa agtgatgaag cagctgaagc ggcggagata
     9361 caccggctgg ggcaggctga gccggaagct gatcaacggc atccgggaca agcagtccgg
     9421 caagacaatc ctggatttcc tgaagtccga cggcttcgcc aacagaaact tcatgcagct
     9481 gatccacgac gacagcctga cctttaaaga ggacatccag aaagcccagg tgtccggcca
     9541 gggcgatagc ctgcacgagc acattgccaa tctggccggc agccccgcca ttaagaaggg
     9601 catcctgcag acagtgaagg tggtggacga gctcgtgaaa gtgatgggcc ggcacaagcc
     9661 cgagaacatc gtgatcgaaa tggccagaga gaaccagacc acccagaagg gacagaagaa
     9721 cagccgcgag agaatgaagc ggatcgaaga gggcatcaaa gagctgggca gccagatcct
     9781 gaaagaacac cccgtggaaa acacccagct gcagaacgag aagctgtacc tgtactacct
     9841 gcagaatggg cgggatatgt acgtggacca ggaactggac atcaaccggc tgtccgacta
     9901 cgatgtggac catatcgtgc ctcagagctt tctgaaggac gactccatcg acaacaaggt
     9961 gctgaccaga agcgacaaga accggggcaa gagcgacaac gtgccctccg aagaggtcgt
    10021 gaagaagatg aagaactact ggcggcagct gctgaacgcc aagctgatta cccagagaaa
    10081 gttcgacaat ctgaccaagg ccgagagagg cggcctgagc gaactggata aggccggctt
    10141 catcaagaga cagctggtgg aaacccggca gatcacaaag cacgtggcac agatcctgga
    10201 ctcccggatg aacactaagt acgacgagaa tgacaagctg atccgggaag tgaaagtgat
    10261 caccctgaag tccaagctgg tgtccgattt ccggaaggat ttccagtttt acaaagtgcg
    10321 cgagatcaac aactaccacc acgcccacga cgcctacctg aacgccgtcg tgggaaccgc
    10381 cctgatcaaa aagtacccta agctggaaag cgagttcgtg tacggcgact acaaggtgta
    10441 cgacgtgcgg aagatgatcg ccaagagcga gcaggaaatc ggcaaggcta ccgccaagta
    10501 cttcttctac agcaacatca tgaacttttt caagaccgag attaccctgg ccaacggcga
    10561 gatccggaag cggcctctga tcgagacaaa cggcgaaacc ggggagatcg tgtgggataa
    10621 gggccgggat tttgccaccg tgcggaaagt gctgagcatg ccccaagtga atatcgtgaa
    10681 aaagaccgag gtgcagacag gcggcttcag caaagagtct atcctgccca agaggaacag
    10741 cgataagctg atcgccagaa agaaggactg ggaccctaag aagtacggcg gcttcgacag
    10801 ccccaccgtg gcctattctg tgctggtggt ggccaaagtg gaaaagggca agtccaagaa
    10861 actgaagagt gtgaaagagc tgctggggat caccatcatg gaaagaagca gcttcgagaa
    10921 gaatcccatc gactttctgg aagccaaggg ctacaaagaa gtgaaaaagg acctgatcat
    10981 caagctgcct aagtactccc tgttcgagct ggaaaacggc cggaagagaa tgctggcctc
    11041 tgccggcgaa ctgcagaagg gaaacgaact ggccctgccc tccaaatatg tgaacttcct
    11101 gtacctggcc agccactatg agaagctgaa gggctccccc gaggataatg agcagaaaca
    11161 gctgtttgtg gaacagcaca agcactacct ggacgagatc atcgagcaga tcagcgagtt
    11221 ctccaagaga gtgatcctgg ccgacgctaa tctggacaaa gtgctgtccg cctacaacaa
    11281 gcaccgggat aagcccatca gagagcaggc cgagaatatc atccacctgt ttaccctgac
    11341 caatctggga gcccctgccg ccttcaagta ctttgacacc accatcgacc ggaagaggta
    11401 caccagcacc aaagaggtgc tggacgccac cctgatccac cagagcatca ccggcctgta
    11461 cgagacacgg atcgacctgt ctcagctggg aggcgacaaa aggccggcgg ccacgaaaaa
    11521 ggccggccag gcaaaaaaga aaaaggaggg cagaggaagt cttctaacat gcggtgacgt
    11581 ggaggagaat cccggcccta tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
    11641 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
    11701 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccagcaagct
    11761 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg
    11821 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
    11881 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
    11941 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
    12001 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat
    12061 ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga
    12121 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
    12181 gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga
    12241 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
    12301 ggacgagctg tacaagtaat taattaacta gaatgaatcg tttttaaaat aacaaatcaa
    12361 ttgttttata atattcgtac gattctttga ttatgtaata aaatgtgatc attaggaaga
    12421 ttacgaaaaa tataaaaaat atgagttctg tgtgtataac aaatgctgta aacgccacaa
    12481 ttgtgtttgt tgcaaataaa cccatgatta tttgattaaa attgttgttt tctttgttca
    12541 tagacaatag tgtgttttgc ctaaacgtgt actgcataaa ctccatgcga gtgtatagcg
    12601 agctagtggc taacgcttgc cccaccaaag tagattcgtc aaaatcctca atttcatcac
    12661 cctcctccaa gtttaacatt tggccgtcgg aattaacttc taaagatgcc acataatcta
    12721 ataaatgaaa tagagattca aacgtggcgt catcgtccgt ttcgaccatt tccgaaaaga
    12781 actcgggcat aaactctatg atttctctgg acgtggtgtt gtcgaaactc tcaaagtacg
    12841 cagtcaggaa cgtgcgcgac atgtcgtcgg gaaactcgcg cggaaacatg ttgttgtaac
    12901 cgaacgggtc ccatagcgcc aaaaccaaat ctgccagcgt caatagaatg agcacgatgc
    12961 cgacaatgga gctggcttgg atagcgattc gagttaacgg ccggctagca attcgagctc
    13021 gcccggggat ctaattcaat tagagactaa ttcaattaga gctaattcaa ttaggatcca
    13081 agcttatcga tttcgaaccc tcgaccgccg gagtataaat agaggcgctt cgtctacgga
    13141 gcgacaattc aattcaaaca agcaaagtga acacgtcgct aagcgaaagc taagcaaata
    13201 aacaagcgca gctgaacaag ctaaacaatc ggggtaccgc tagagtcgac ggtacgatcc
    13261 accggtcgcc accatggtga gcaagggcga ggaggtcatc aaagagttca tgcgcttcaa
    13321 ggtgcgcatg gagggctcca tgaacggcca cgagttcgag atcgagggcg agggcgaggg
    13381 ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggcg gccccctgcc
    13441 cttcgcctgg gacatcctgt ccccccagtt catgtacggc tccaaggcgt acgtgaagca
    13501 ccccgccgac atccccgatt acaagaagct gtccttcccc gagggcttca agtgggagcg
    13561 cgtgatgaac ttcgaggacg gcggtctggt gaccgtgacc caggactcct ccctgcagga
    13621 cggcacgctg atctacaagg tgaagatgcg cggcaccaac ttcccccccg acggccccgt
    13681 aatgcagaag aagaccatgg gctgggaggc ctccaccgag cgcctgtacc cccgcgacgg
    13741 cgtgctgaag ggcgagatcc accaggccct gaagctgaag gacggcggcc actacctggt
    13801 ggagttcaag accatctaca tggccaagaa gcccgtgcaa ctgcccggct actactacgt
    13861 ggacaccaag ctggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga
    13921 gcgctccgag ggccgccacc acctgttcct ggggcatggc accggcagca ccggcagcgg
    13981 cagctccggc accgcctcct ccgaggacaa caacatggcc gttatcaagg aatttatgcg
    14041 cttcaaagtt aggatggagg gatccatgaa cggacatgag ttcgagatcg agggagaggg
    14101 cgagggacgc ccgtatgaag gcacacaaac agccaaactc aaggtcacca agggcggacc
    14161 actgcccttc gcctgggata tcctgagtcc ccagtttatg tacggcagca aggcctacgt
    14221 taagcacccc gctgacatac cggactacaa aaagctgtcc tttccggaag gcttcaagtg
    14281 ggagcgcgtg atgaatttcg aagacggagg actggtcact gtgacccaag atagcagttt
    14341 gcaggacggt acactgatct ataaggttaa aatgcgcggc actaactttc cgccagatgg
    14401 cccagtgatg cagaagaaga ccatgggttg ggaggcatcc accgaacgtc tgtaccctcg
    14461 agacggagtg ctcaagggcg agatccatca ggccctcaaa ctgaaagatg gtggtcacta
    14521 cctggtcgaa tttaagacca tttacatggc caagaagccg gttcagctgc ccggatatta
    14581 ttatgtggat acgaaactgg atataacttc gcataacgaa gactacacca ttgtcgagca
    14641 gtatgagcgc agcgaaggcc gacatcacct gttcctctac ggcatggacg agctgtacaa
    14701 gtaggcggcc gcgactctag atcataatca gccataccac atttgtagag gttttacttg
    14761 ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg
    14821 ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
    14881 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
    14941 tatcttaaag cttcctaggg tactgacgga cacaccgaag ccccggcggc aaccctcagc
    15001 ggatgccccg gggcttcacg ttttcccagg tcagaagcgg ttttcgggag tagtgcccca
    15061 actggggtaa cctttgagtt ctctcagttg ggggcgtagg gccgccgaca tgacacaagg
    15121 ggttgtgacc ggggtggaca cgtacgcggg tgcttacgac cgtcagtcgc gcgagcgcga
    15181 gggcccgggt tttttaaatg gtaccccttg cagcgccacc tggccgcgaa gagttccatt
    15241 ctctgcttcg aagtacgaag agagaaagag tactttaaat ttccacattc ccgccttgca
    15301 gcgccacctg gccttggtaa tgtagaacta ggaaggaaag caccaacaca agatgtcgct
    15361 ctccgacgat ggacatgtcg cgtaaaaaat gttcgataac tttcaatggt tcgattgaac
    15421 agacaataag tgtatttaag acaccagttc ttatattcaa aaatcctaac aactcacacc
    15481 ttgcagcgcc acctggccct agagatccac caactctaga gagatctgta agccggcgtg
    15541 gcttagctag agggtatgtg tgccgtttgg aaccggtaac tcctccctcc ttggattgaa
    15601 gtgacagcca caacgaacta gtagtcttca ctaagctcgg ctctggtacc gttttaaatg
    15661 ccactcttct tcttctattg atgaggtata ttgtcgtcac tactccaact aatagtaata
    15721 cgtaaagtgc tccgaaattt gtagtcgtaa attttttcca atatatgatg gaattagaag
    15781 catggttgga tcttatgttc agcttatgca ggttaatgtc ttctgtgttc tcggtgaaat
    15841 atatattttt tataattggg gattgtgcgg atcttcttat ggaaattttg gtacaatttg
    15901 attcaacggt agtgttttgt gtgtttaata ttaaaatgtg agctggttct acttctttta
    15961 taatttcttg attccctatg tctttagttt taaaattggc agtttccatc ttatgtatgt
    16021 tattgagaca ttcagtttta atattacctc tgattgccat tgcattgtcg cagatatggt
    16081 atttatatat cttgttatat ttgttggaaa agtagtaaaa aacatgccat atagttaggt
    16141 atatttatat atttatattt atatttcatt tatgggtaat gggacaatgt gtgtgaactt
    16201 ttagtaattt tgtttgaatg cagggattct tattctaaag tttatcttaa ttccaatcat
    16261 gaaagtatct aattctaaca tttgaaatat gtgtttgtct gacatttccc gtaactccgg
    16321 gtcgtttaat tcttgtctga gaattcgtaa tcatggtcat agctgtttcc tgtgtgaaat
    16381 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
    16441 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
    16501 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
    16561 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
    16621 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggt
//
