LOCUS       sequence-305007-       14904 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The attB plasmid with the mini-white harboring two U6.3-gRNAs 
            targeting Dmel sxl and bTub genes, and Hsp70Bb-Cas9-T2A-eGFP-p10..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14904)
  AUTHORS   Kandul NP, Liu J, Akbari OS
  TITLE     Temperature-Inducible Precision-Guided Sterile Insect Technique.
  JOURNAL   CRISPR J. 2021 Dec;4(6):822-835. doi: 10.1089/crispr.2021.0077. Epub
            2021 Nov 3.
  PUBMED    34735288
REFERENCE   2  (bases 1 to 14904)
  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: 
            "10.1089/crispr.2021.0077"; journalName: "CRISPR J"; date: 
            "2021-12"; volume: "4"; issue: "6"; pages: "822-835"
COMMENT     Sequence Label: 1098A=TI-pgSIT[sxl,bTub,Hasp70Bb-Cas9]
FEATURES             Location/Qualifiers
     source          1..14904
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(156..174)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(195..211)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(195..211)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(208..230)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    219..235
                     /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(243..249,250..267,268..273))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    288..309
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(426..443)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(597..1185)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(677..696)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1356..2147,2148..2216))
                     /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     1979..1998
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2217..2321)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2347..2802)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2484..2505)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2696..2715
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2929..2951
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2943..2960
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2944..2960
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2967..2986
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2967..2985
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3019..3037)
                     /label=pCasper-F
                     /note="Drosophila mini-white gene, reverse primer"
     CDS             join(3489..3560,3962..4235,4310..4964,5026..5341,5545..5676,
                     5747..6361)
                     /codon_start=1
                     /gene="white"
                     /product="Drosophila white gene eye color pigment"
                     /label=mini-white
                     /note="This is a modified version of the white gene lacking
                     part of the first intron."
                     /translation="MGQEDQELLIRGGSKHPSAEHLNNGDSGAASQSCINQGFGQAKNY
                     GTLRPPSPPEDSGSGSGQLAENLTYAWHNMDIFGAVNQPGSGWRQLVNRTRGLFCNERH
                     IPAPRKHLLKNVCGVAYPGELLAVMGSSGAGKTTLLNALAFRSPQGIQVSPSGMRLLNG
                     QPVDAKEMQARCAYVQQDDLFIGSLTAREHLIFQAMVRMPRHLTYRQRVARVDQVIQEL
                     SLSKCQHTIIGVPGRVKGLSGGERKRLAFASEALTDPPLLICDEPTSGLDSFTAHSVVQ
                     VLKKLSQKGKTVILTIHQPSSELFELFDKILLMAEGRVAFLGTPSEAVDFFSYVGAQCP
                     TNYNPADFYVQVLAVVPGREIESRDRIAKICDNFAISKVARDMEQLLATKNLEKPLEQP
                     ENGYTYKATWFMQFRAVLWRSWLSVLKEPLLVKVRLIQTTMVAILIGLIFLGQQLTQVG
                     VMNINGAIFLFLTNMTFQNVFATINVFTSELPVFMREARSRLYRCDTYFLGKTIAELPL
                     FLTVPLVFTAIAYPMIGLRAGVLHFFNCLALVTLVANVSTSFGYLISCASSSTSMALSV
                     GPPVIIPFLLFGGFFLNSGSVPVYLKWLSYLSWFRYANEGLLINQWADVEPGEISCTSS
                     NTTCPSSGKVILETLNFSAADLPLDYVGLAILIVSFRVLAYLALRLRARRKE"
     protein_bind    7136..7169
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        7188..7608
                     /label=dU6-3 promoter
                     /note="RNA polymerase III promoter for Drosophila U6-3 
                     snRNA (Port et al., 2014)"
     misc_RNA        7629..7704
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        7767..8187
                     /label=dU6-3 promoter
                     /note="RNA polymerase III promoter for Drosophila U6-3 
                     snRNA (Port et al., 2014)"
     misc_RNA        8208..8283
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        8757..8995
                     /label=hsp70 promoter
                     /note="Drosophila melanogaster hsp70Bb promoter"
     primer_bind     8942..8963
                     /label=pCasper-hs
                     /note="Drosophila Hsp70 promoter, forward primer"
     CDS             9009..9074
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             9081..9101
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             9126..13226
                     /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             9126..13226
                     /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             13227..13274
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             13275..13328
                     /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(13329..13331,13332..13334,13335..14048)
                     /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(13635..13654)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     13982..14003
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    14772..14841
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
ORIGIN
        1 gggaagccct cgccctcgaa accgctgggc gcggtggtca cggtgagcac gggacgtgcg
       61 acggcgtcgg cgggtgcgga tacgcggggc agcgtcagcg ggttctcgac ggtcacggcg
      121 ggcatgtcga cactagttct agccagcttt tgttcccttt agtgagggtt aatttcgagc
      181 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
      241 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
      301 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
      361 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
      421 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
      481 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
      541 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
      601 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
      661 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
      721 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
      781 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
      841 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
      901 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
      961 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     1021 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     1081 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     1141 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     1201 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     1261 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     1321 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     1381 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     1441 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     1501 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     1561 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     1621 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     1681 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     1741 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     1801 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     1861 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     1921 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     1981 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     2041 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     2101 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     2161 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     2221 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     2281 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     2341 ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     2401 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     2461 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     2521 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     2581 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     2641 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     2701 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     2761 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg ccattcaggc
     2821 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga
     2881 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac
     2941 gttgtaaaac gacggccagt gaattgtaat acgactcact atagggcgaa ttgggtacgt
     3001 accgggcccc tagtatgtat gtaagttaat aaaacccatt tttgcggaaa gtagataaaa
     3061 aaaacatttt ttttttttac tgcactggat atcattgaac ttatctgatc agttttaaat
     3121 ttacttcgat ccaagggtat ttgatgtacc aggttctttc gattacctct cactcaaaat
     3181 gacattccac tcaaagtcag cgctgtttgc ctccttctct gtccacagaa atatcgccgt
     3241 ctctttcgcc gctgcgtccg ctatctcttt cgccaccgtt tgtagcgtta cgtagcgtca
     3301 atgtccgcct tcagttgcat tttgtcagcg gtttcgtgac gaagctccaa gcggtttacg
     3361 ccatcaatta aacacaaagt gctgtgccaa aactcctctc gcttcttatt tttgtttgtt
     3421 ttttgagtga ttggggtggt gattggtttt gggtgggtaa gcaggggaaa gtgtgaaaaa
     3481 tcccggcaat gggccaagag gatcaggagc tattaattcg cggaggcagc aaacacccat
     3541 ctgccgagca tctgaacaat gtgagtagta catgtgcata catcttaagt tcacttgatc
     3601 tataggaact gcgattgcaa catcaaattg tctgcggcgt gagaactgcg acccacaaaa
     3661 atcccaaacc gcaattgcac aaacaaatag tgacacgaaa cagattattc tggtagctgt
     3721 tctcgctata taagacaatt tttgagatca tatcatgatc aagacatcta aaggcattca
     3781 ttttcgacta tattcttttt tacaaaaaat ataacaacca gatattttaa gctgatccta
     3841 gatgcacaaa aaataaataa aagtataaac ctacttcgta ggatacttcg gggtactttt
     3901 tgttcggggt tagatgagca taacgcttgt agttgatatt tgagatcccc tatcattgca
     3961 gggtgacagc ggagcggctt cgcagagctg cattaaccag ggcttcgggc aggccaaaaa
     4021 ctacggcacg ctccggccac ccagtccgcc ggaggactcc ggttcaggga gcggccaact
     4081 agccgagaac ctcacctatg cctggcacaa tatggacatc tttggggcgg tcaatcagcc
     4141 gggctccgga tggcggcagc tggtcaaccg gacacgcgga ctattctgca acgagcgaca
     4201 cataccggcg cccaggaaac atttgctcaa gaacggtgag tttctattcg cagtcggctg
     4261 atctgtgtga aatcttaata aagggtccaa ttaccaattt gaaactcagt ttgcggcgtg
     4321 gcctatccgg gcgaactttt ggccgtgatg ggcagttccg gtgccggaaa gacgaccctg
     4381 ctgaatgccc ttgcctttcg atcgccgcag ggcatccaag tatcgccatc cgggatgcga
     4441 ctgctcaatg gccaacctgt ggacgccaag gagatgcagg ccaggtgcgc ctatgtccag
     4501 caggatgacc tctttatcgg ctccctaacg gccagggaac acctgatttt ccaagccatg
     4561 gtgcggatgc cacgacatct gacctatcgg cagcgagtgg cccgcgtgga tcaggtgatc
     4621 caggagcttt cgctcagcaa atgtcagcac acgatcatcg gtgtgcccgg cagggtgaaa
     4681 ggtctgtccg gcggagaaag gaagcgtctg gcattcgcct ccgaggctct aaccgatccg
     4741 ccgcttctga tctgcgatga gcccacctcc ggactggact cctttaccgc ccacagcgtc
     4801 gtccaggtgc tgaagaagct gtcgcagaag ggcaagaccg tcatcctgac cattcatcag
     4861 ccgtcttccg agctgtttga gctctttgac aagatccttc tgatggccga gggcagggta
     4921 gctttcttgg gcactcccag cgaagccgtc gacttctttt cctagtgagt tcgatgtgtt
     4981 tattaagggt atctagtatt acataacatc tcaactccta tccagcgtgg gtgcccagtg
     5041 tcctaccaac tacaatccgg cggactttta cgtacaggtg ttggccgttg tgcccggacg
     5101 ggagatcgag tcccgtgatc ggatcgccaa gatatgcgac aattttgcca ttagcaaagt
     5161 agcccgggat atggagcagt tgttggccac caaaaatctg gagaagccac tggagcagcc
     5221 ggagaatggg tacacctaca aggccacctg gttcatgcag ttccgggcgg tcctgtggcg
     5281 atcctggctg tcggtgctca aggaaccact cctcgtaaaa gtgcgactta ttcagacaac
     5341 ggtgagtggt tccagtggaa acaaatgata taacgcttac aattcttgga aacaaattcg
     5401 ctagatttta gatagaattg cctgattcca cacccttctt agtttttttc aatgagatgt
     5461 atagtttata gttttgcaga agataaataa atttcattta actcgcgaat attaatgaga
     5521 tgcgagtaac attttaattt gcagatggtt gccatcttga ttggcctcat ctttttgggc
     5581 caacaactca cgcaagtggg tgtgatgaat atcaacggag ccatcttcct cttcctgacc
     5641 aacatgacct ttcaaaacgt ctttgccacg ataaatgtaa gtcatgttta gaatacattt
     5701 gcatttcaat aatttactaa ctttctaatg aatcgattcg atttaggtgt tcacctcaga
     5761 gctgccagtt tttatgaggg aggcccgaag tcgactttat cgctgtgaca catactttct
     5821 gggcaaaacg attgccgaat tgccgctttt tctcacagtg ccactggtct tcacggcgat
     5881 tgcctatccg atgatcggac tgcgggccgg agtgctgcac ttcttcaact gcctggcgct
     5941 ggtcactctg gtggccaatg tgtcaacgtc cttcggatat ctaatatcct gcgccagctc
     6001 ctcgacctcg atggcgctgt ctgtgggtcc gccggttatc ataccattcc tgctctttgg
     6061 cggcttcttc ttgaactcgg gctcggtgcc agtatacctc aaatggttgt cgtacctctc
     6121 atggttccgt tacgccaacg agggtctgct gattaaccaa tgggcggacg tggagccggg
     6181 cgaaattagc tgcacatcgt cgaacaccac gtgccccagt tcgggcaagg tcatcctgga
     6241 gacgcttaac ttctccgccg ccgatctgcc gctggactac gtgggtctgg ccattctcat
     6301 cgtgagcttc cgggtgctcg catatctggc tctaagactt cgggcccgac gcaaggagta
     6361 gccgacatat atccgaaata actgcttgtt tttttttttt accattatta ccatcgtgtt
     6421 tactgtttat tgccccctca aaaagctaat gtaattatat ttgtgccaat aaaaacaaga
     6481 tatgacctat agaatacaag tatttcccct tcgaacatcc ccacaagtag actttggatt
     6541 tgtcttctaa ccaaaagact tacacacctg cataccttac atcaaaaact cgtttatcgc
     6601 tacataaaac accgggatat attttttata tacatacttt tcaaatcgcg cgccctcttc
     6661 ataattcacc tccaccacac cacgtttcgt agttgctctt tcgctgtctc ccacccgctc
     6721 tccgcaacac attcaccttt tgttcgacga ccttggagcg actgtcgtta gttccgcgcg
     6781 attcggttcg ctcaaatggt tccgagtggt tcatttcgtc tcaatagaaa ttagtaataa
     6841 atatttgtat gtacaattta tttgctccaa tatatttgta tatatttccc tcacagctat
     6901 atttattcta atttaatatt atgacttttt aaggtaattt tttgtgacct gttcggagtg
     6961 attagcgtta caatttgaac tgaaagtgac atccagtgtt tgttccttgt gtagatgcat
     7021 ctcaaaaaaa tggtgggcat aatagtgttg tttatatata tcaaaaataa caactataat
     7081 aataagaata catttaattt agaaaatgct tggatttcac tggaactagg ctagcataac
     7141 ttcgtataat gtatgctata cgaagttatg ctagcggatc cgaattcttt tttgctcacc
     7201 tgtgattgct cctactcaaa tacaaaaaca tcaaattttc tgtcaataaa gcatatttat
     7261 ttatatttat tttacaggaa agaattcctt ttaaagtgta ttttaaccta taatgaaaaa
     7321 cgattaaaaa aaatacataa aataattcga aaatttttga atagcccagg ttgataaaaa
     7381 ttcatttcat acgttttata acttatgccc ctaagtattt tttgaccata gtgtttcaat
     7441 tctacattaa ttttacagag tagaatgaaa cgccacctac tcagccaaga ggcgaaaagg
     7501 ttagctcgcc aagcagagag ggcgccagtg ctcactactt tttataattc tcaacttctt
     7561 tttccagact cagttcgtat atatagacct attttcaatt taacgtcgga ttgtcaacta
     7621 cttgccccgt tttagagcta gaaatagcaa gttaaaataa ggctagtccg ttatcaactt
     7681 gaaaaagtgg caccgagtcg gtgctttttt ttttggtacc gggaattggg aattgggcaa
     7741 tatttaaatg gcggcgcgcc gaattctttt ttgctcacct gtgattgctc ctactcaaat
     7801 acaaaaacat caaattttct gtcaataaag catatttatt tatatttatt ttacaggaaa
     7861 gaattccttt taaagtgtat tttaacctat aatgaaaaac gattaaaaaa aatacataaa
     7921 ataattcgaa aatttttgaa tagcccaggt tgataaaaat tcatttcata cgttttataa
     7981 cttatgcccc taagtatttt ttgaccatag tgtttcaatt ctacattaat tttacagagt
     8041 agaatgaaac gccacctact cagccaagag gcgaaaaggt tagctcgcca agcagagagg
     8101 gcgccagtgc tcactacttt ttataattct caacttcttt ttccagactc agttcgtata
     8161 tatagaccta ttttcaattt aacgtcgcct gagtgtgcat cagctgggtt ttagagctag
     8221 aaatagcaag ttaaaataag gctagtccgt tatcaacttg aaaaagtggc accgagtcgg
     8281 tgcttttttt tttgcctacc tggagcctga gagttgttca ataaaataaa aatgtttcgt
     8341 ttttttgctt tcgccagtat ttattatttt tcatcaatat gtattcaatt tggtatgtat
     8401 ttagtaattg taatatatag acaatggttt tccgttgacg tacatacatc tgacgtgtgt
     8461 ttatttagac ataatagtta tgttttcaca tctttttaat gttcgcttaa tgcgtatgca
     8521 ttctagacaa ttgtgctcgg caacagtata tttgtggtgt gccaaccaac aacctgcagg
     8581 agctccagct tttgtctcga gtcgagaaat ttctctggcc gttattcgtt attctctctt
     8641 ttctttttgg gtctctccct ctctgcacta atgctctctc actctgtcac acagtaaacg
     8701 gcatactgct ctcgttggtt cgagagagcg cgcctcgaat gttcgcgaaa agagcgccgg
     8761 agtataaata gaggcgcttc gtctacggag cgacaattca attcaaacaa gcaaagtgaa
     8821 cacgtcgcta agcgaaagct aagcaaataa acaagcgcag ctgaacaagc taaacaatct
     8881 gcagtaaagt gcaagttaaa gtgaatcaat taaaagtaac cagcaaccaa gtaaatcaac
     8941 tgcaactact gaaatctgcc aagaagtaat tattgaatac aagaagagaa ctctgtttaa
     9001 acaaaatgga ctataaggac cacgacggag actacaagga tcatgatatt gattacaaag
     9061 acgatgacga taagatggcc ccaaagaaga agcggaaggt cggtatccac ggagtcccag
     9121 cagccgacaa gaagtacagc atcggcctgg acatcggcac caactctgtg ggctgggccg
     9181 tgatcaccga cgagtacaag gtgcccagca agaaattcaa ggtgctgggc aacaccgacc
     9241 ggcacagcat caagaagaac ctgatcggag ccctgctgtt cgacagcggc gaaacagccg
     9301 aggccacccg gctgaagaga accgccagaa gaagatacac cagacggaag aaccggatct
     9361 gctatctgca agagatcttc agcaacgaga tggccaaggt ggacgacagc ttcttccaca
     9421 gactggaaga gtccttcctg gtggaagagg ataagaagca cgagcggcac cccatcttcg
     9481 gcaacatcgt ggacgaggtg gcctaccacg agaagtaccc caccatctac cacctgagaa
     9541 agaaactggt ggacagcacc gacaaggccg acctgcggct gatctatctg gccctggccc
     9601 acatgatcaa gttccggggc cacttcctga tcgagggcga cctgaacccc gacaacagcg
     9661 acgtggacaa gctgttcatc cagctggtgc agacctacaa ccagctgttc gaggaaaacc
     9721 ccatcaacgc cagcggcgtg gacgccaagg ccatcctgtc tgccagactg agcaagagca
     9781 gacggctgga aaatctgatc gcccagctgc ccggcgagaa gaagaatggc ctgttcggaa
     9841 acctgattgc cctgagcctg ggcctgaccc ccaacttcaa gagcaacttc gacctggccg
     9901 aggatgccaa actgcagctg agcaaggaca cctacgacga cgacctggac aacctgctgg
     9961 cccagatcgg cgaccagtac gccgacctgt ttctggccgc caagaacctg tccgacgcca
    10021 tcctgctgag cgacatcctg agagtgaaca ccgagatcac caaggccccc ctgagcgcct
    10081 ctatgatcaa gagatacgac gagcaccacc aggacctgac cctgctgaaa gctctcgtgc
    10141 ggcagcagct gcctgagaag tacaaagaga ttttcttcga ccagagcaag aacggctacg
    10201 ccggctacat tgacggcgga gccagccagg aagagttcta caagttcatc aagcccatcc
    10261 tggaaaagat ggacggcacc gaggaactgc tcgtgaagct gaacagagag gacctgctgc
    10321 ggaagcagcg gaccttcgac aacggcagca tcccccacca gatccacctg ggagagctgc
    10381 acgccattct gcggcggcag gaagattttt acccattcct gaaggacaac cgggaaaaga
    10441 tcgagaagat cctgaccttc cgcatcccct actacgtggg ccctctggcc aggggaaaca
    10501 gcagattcgc ctggatgacc agaaagagcg aggaaaccat caccccctgg aacttcgagg
    10561 aagtggtgga caagggcgct tccgcccaga gcttcatcga gcggatgacc aacttcgata
    10621 agaacctgcc caacgagaag gtgctgccca agcacagcct gctgtacgag tacttcaccg
    10681 tgtataacga gctgaccaaa gtgaaatacg tgaccgaggg aatgagaaag cccgccttcc
    10741 tgagcggcga gcagaaaaag gccatcgtgg acctgctgtt caagaccaac cggaaagtga
    10801 ccgtgaagca gctgaaagag gactacttca agaaaatcga gtgcttcgac tccgtggaaa
    10861 tctccggcgt ggaagatcgg ttcaacgcct ccctgggcac ataccacgat ctgctgaaaa
    10921 ttatcaagga caaggacttc ctggacaatg aggaaaacga ggacattctg gaagatatcg
    10981 tgctgaccct gacactgttt gaggacagag agatgatcga ggaacggctg aaaacctatg
    11041 cccacctgtt cgacgacaaa gtgatgaagc agctgaagcg gcggagatac accggctggg
    11101 gcaggctgag ccggaagctg atcaacggca tccgggacaa gcagtccggc aagacaatcc
    11161 tggatttcct gaagtccgac ggcttcgcca acagaaactt catgcagctg atccacgacg
    11221 acagcctgac ctttaaagag gacatccaga aagcccaggt gtccggccag ggcgatagcc
    11281 tgcacgagca cattgccaat ctggccggca gccccgccat taagaagggc atcctgcaga
    11341 cagtgaaggt ggtggacgag ctcgtgaaag tgatgggccg gcacaagccc gagaacatcg
    11401 tgatcgaaat ggccagagag aaccagacca cccagaaggg acagaagaac agccgcgaga
    11461 gaatgaagcg gatcgaagag ggcatcaaag agctgggcag ccagatcctg aaagaacacc
    11521 ccgtggaaaa cacccagctg cagaacgaga agctgtacct gtactacctg cagaatgggc
    11581 gggatatgta cgtggaccag gaactggaca tcaaccggct gtccgactac gatgtggacc
    11641 atatcgtgcc tcagagcttt ctgaaggacg actccatcga caacaaggtg ctgaccagaa
    11701 gcgacaagaa ccggggcaag agcgacaacg tgccctccga agaggtcgtg aagaagatga
    11761 agaactactg gcggcagctg ctgaacgcca agctgattac ccagagaaag ttcgacaatc
    11821 tgaccaaggc cgagagaggc ggcctgagcg aactggataa ggccggcttc atcaagagac
    11881 agctggtgga aacccggcag atcacaaagc acgtggcaca gatcctggac tcccggatga
    11941 acactaagta cgacgagaat gacaagctga tccgggaagt gaaagtgatc accctgaagt
    12001 ccaagctggt gtccgatttc cggaaggatt tccagtttta caaagtgcgc gagatcaaca
    12061 actaccacca cgcccacgac gcctacctga acgccgtcgt gggaaccgcc ctgatcaaaa
    12121 agtaccctaa gctggaaagc gagttcgtgt acggcgacta caaggtgtac gacgtgcgga
    12181 agatgatcgc caagagcgag caggaaatcg gcaaggctac cgccaagtac ttcttctaca
    12241 gcaacatcat gaactttttc aagaccgaga ttaccctggc caacggcgag atccggaagc
    12301 ggcctctgat cgagacaaac ggcgaaaccg gggagatcgt gtgggataag ggccgggatt
    12361 ttgccaccgt gcggaaagtg ctgagcatgc cccaagtgaa tatcgtgaaa aagaccgagg
    12421 tgcagacagg cggcttcagc aaagagtcta tcctgcccaa gaggaacagc gataagctga
    12481 tcgccagaaa gaaggactgg gaccctaaga agtacggcgg cttcgacagc cccaccgtgg
    12541 cctattctgt gctggtggtg gccaaagtgg aaaagggcaa gtccaagaaa ctgaagagtg
    12601 tgaaagagct gctggggatc accatcatgg aaagaagcag cttcgagaag aatcccatcg
    12661 actttctgga agccaagggc tacaaagaag tgaaaaagga cctgatcatc aagctgccta
    12721 agtactccct gttcgagctg gaaaacggcc ggaagagaat gctggcctct gccggcgaac
    12781 tgcagaaggg aaacgaactg gccctgccct ccaaatatgt gaacttcctg tacctggcca
    12841 gccactatga gaagctgaag ggctcccccg aggataatga gcagaaacag ctgtttgtgg
    12901 aacagcacaa gcactacctg gacgagatca tcgagcagat cagcgagttc tccaagagag
    12961 tgatcctggc cgacgctaat ctggacaaag tgctgtccgc ctacaacaag caccgggata
    13021 agcccatcag agagcaggcc gagaatatca tccacctgtt taccctgacc aatctgggag
    13081 cccctgccgc cttcaagtac tttgacacca ccatcgaccg gaagaggtac accagcacca
    13141 aagaggtgct ggacgccacc ctgatccacc agagcatcac cggcctgtac gagacacgga
    13201 tcgacctgtc tcagctggga ggcgacaaaa ggccggcggc cacgaaaaag gccggccagg
    13261 caaaaaagaa aaaggagggc agaggaagtc ttctaacatg cggtgacgtg gaggagaatc
    13321 ccggccctat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
    13381 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
    13441 ccacctacgg caagctgacc ctgaagttca tctgcaccac cagcaagctg cccgtgccct
    13501 ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc
    13561 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
    13621 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
    13681 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
    13741 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc
    13801 agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc
    13861 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
    13921 acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc
    13981 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
    14041 acaagtaatt aattaactag aatgaatcgt ttttaaaata acaaatcaat tgttttataa
    14101 tattcgtacg attctttgat tatgtaataa aatgtgatca ttaggaagat tacgaaaaat
    14161 ataaaaaata tgagttctgt gtgtataaca aatgctgtaa acgccacaat tgtgtttgtt
    14221 gcaaataaac ccatgattat ttgattaaaa ttgttgtttt ctttgttcat agacaatagt
    14281 gtgttttgcc taaacgtgta ctgcataaac tccatgcgag tgtatagcga gctagtggct
    14341 aacgcttgcc ccaccaaagt agattcgtca aaatcctcaa tttcatcacc ctcctccaag
    14401 tttaacattt ggccgtcgga attaacttct aaagatgcca cataatctaa taaatgaaat
    14461 agagattcaa acgtggcgtc atcgtccgtt tcgaccattt ccgaaaagaa ctcgggcata
    14521 aactctatga tttctctgga cgtggtgttg tcgaaactct caaagtacgc agtcaggaac
    14581 gtgcgcgaca tgtcgtcggg aaactcgcgc ggaaacatgt tgttgtaacc gaacgggtcc
    14641 catagcgcca aaaccaaatc tgccagcgtc aatagaatga gcacgatgcc gacaatggag
    14701 ctggcttgga tagcgattcg agttaacggc cggaggccag atccactagt gtcgacgatg
    14761 taggtcacgg tctcgaagcc gcggtgcggg tgccagggcg tgcccttggg ctccccgggc
    14821 gcgtactcca cctcacccat ctggtccatc atgatgaacg ggtcgaggtg gcggtagttg
    14881 atcccggcga acgcgcggcg cacc
//
