LOCUS       sequence-301650-       13159 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The piggyBac plasmid harboring Hsp70Bb-Cas9-T2A-eGFP-p10 and 
            Opie2-dsRed-SV10 (marker gene).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13159)
  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 13159)
  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: 1137=Hsp70Bb-Cas9[dsRed]
FEATURES             Location/Qualifiers
     source          1..13159
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   305..367
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     protein_bind    1401..1470
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     promoter        1820..2058
                     /label=hsp70 promoter
                     /note="Drosophila melanogaster hsp70Bb promoter"
     primer_bind     2005..2026
                     /label=pCasper-hs
                     /note="Drosophila Hsp70 promoter, forward primer"
     CDS             2072..2137
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2144..2164
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2189..6289
                     /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             2189..6289
                     /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             6290..6337
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6338..6391
                     /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(6392..6394,6395..6397,6398..7111)
                     /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(6698..6717)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7045..7066
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             join(8346..8348,8349..8351,8352..9026)
                     /codon_start=1
                     /product="wild-type DsRed"
                     /label=DsRed1
                     /note="mammalian codon-optimized"
                     /translation="MVRSSKNVIKEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTV
                     KLKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     VVTVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     KALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERTEGRH
                     HLFL"
     primer_bind     complement(8529..8549)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     8747..8766
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     8941..8964
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    9140..9261
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9177..9196)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9231..9250
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   9929..9963
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(10144..10160)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10144..10160)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10157..10179)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10168..10184
                     /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(10192..10198,10199..10216,10217..10222))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10237..10258
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10375..10392)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10546..11134)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10626..10645)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11305..12096,12097..12165))
                     /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     11928..11947
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12166..12270)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     12338..12356
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(12394..12416)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     12516..12535
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     12729..12751
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     12743..12760
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     12744..12760
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 cgtctgtaca cgaacgcgta ttaacgcaga gtgcaaagta taagagggtt aaaaaatata
       61 ttttacgcac catatacgca tcgggttgat atcgttaata tggatcaatt tgaacagttg
      121 attaacgtgt ctctgctcaa gtctttgatc aaaacgcaaa tcgacgaaaa tgtgtcggac
      181 aatatcaagt cgatgagcga aaaactaaaa aggctagaat acgacaatct cacagacagc
      241 gttgagatat acggtattca cgacagcagg ctgaataata aaaaaattag aaactattat
      301 ttaaccctag aaagataatc atattgtgac gtacgttaaa gataatcatg cgtaaaattg
      361 acgcatgtgt tttatcggtc tgtatatcga ggtttattta ttaatttgaa tagatattaa
      421 gttttattat atttacactt acatactaat aataaattca acaaacaatt tatttatgtt
      481 tatttattta ttaaaaaaaa acaaaaactc aaaatttctt ctataaagta acaaaacttt
      541 taaacattct ctcttttaca aaaataaact tattttgtac tttaaaaaca gtcatgttgt
      601 attataaaat aagtaattag cttaacttat acataataga aacaaattat acttattagt
      661 cagtcagaaa caactttggc acatatcaat attatgctct cgacaaataa cttttttgca
      721 ttttttgcac gatgcatttg cctttcgcct tattttagag gggcagtaag tacagtaagt
      781 acgttttttc attactggct cttcagtact gtcatctgat gtaccaggca cttcatttgg
      841 caaaatatta gagatattat cgcgcaaata tctcttcaaa gtaggagctt ctaaacgctt
      901 acgcataaac gatgacgtca ggctcatgta aaggtttctc ataaattttt tgcgactttg
      961 aaccttttct cccttgctac tgacattatg gctgtatata ataaaagaat ttatgcaggc
     1021 aatgtttatc attccgtaca ataatgccat aggccaccta ttcgtcttcc tactgcaggt
     1081 catcacagaa cacatttggt ctagcgtgtc cactccgcct ttagtttgat tataatacat
     1141 aaccatttgc ggtttaccgg tactttcgtt gatagaagca tcctcatcac aagatgataa
     1201 taagtatacc atcttagctg gcttcggttt atatgagacg agagtaaggg gtccgtcaaa
     1261 acaaaacatc gatgttccca ctggcctgga gcgactgttt ttcagtactt ccggtatctc
     1321 gcgtttgttt gatcgcacgg ttcccacaat ggttaattcg agctcgcccg gggtcctagg
     1381 tcgacgatgt aggtcacggt ctcgaagccg cggtgcgggt gccagggcgt gcccttgggc
     1441 tccccgggcg cgtactccac ctcacccatc tggtccatca tgatgaacgg gtcgaggtgg
     1501 cggtagttga tcccggcgaa cgcgcggcgc accgggaagc cctcgccctc gaaaccgctg
     1561 ggcgcggtgg tcacggtgag cacgggacgt gcgacggcgt cggcgggtgc ggatacgcgg
     1621 ggcagcgtca gcgggttctc gacggtcacg gcgggcatgt cgactcgaga aatttctctg
     1681 gccgttattc gttattctct cttttctttt tgggtctctc cctctctgca ctaatgctct
     1741 ctcactctgt cacacagtaa acggcatact gctctcgttg gttcgagaga gcgcgcctcg
     1801 aatgttcgcg aaaagagcgc cggagtataa atagaggcgc ttcgtctacg gagcgacaat
     1861 tcaattcaaa caagcaaagt gaacacgtcg ctaagcgaaa gctaagcaaa taaacaagcg
     1921 cagctgaaca agctaaacaa tctgcagtaa agtgcaagtt aaagtgaatc aattaaaagt
     1981 aaccagcaac caagtaaatc aactgcaact actgaaatct gccaagaagt aattattgaa
     2041 tacaagaaga gaactctgtt taaacaaaat ggactataag gaccacgacg gagactacaa
     2101 ggatcatgat attgattaca aagacgatga cgataagatg gccccaaaga agaagcggaa
     2161 ggtcggtatc cacggagtcc cagcagccga caagaagtac agcatcggcc tggacatcgg
     2221 caccaactct gtgggctggg ccgtgatcac cgacgagtac aaggtgccca gcaagaaatt
     2281 caaggtgctg ggcaacaccg accggcacag catcaagaag aacctgatcg gagccctgct
     2341 gttcgacagc ggcgaaacag ccgaggccac ccggctgaag agaaccgcca gaagaagata
     2401 caccagacgg aagaaccgga tctgctatct gcaagagatc ttcagcaacg agatggccaa
     2461 ggtggacgac agcttcttcc acagactgga agagtccttc ctggtggaag aggataagaa
     2521 gcacgagcgg caccccatct tcggcaacat cgtggacgag gtggcctacc acgagaagta
     2581 ccccaccatc taccacctga gaaagaaact ggtggacagc accgacaagg ccgacctgcg
     2641 gctgatctat ctggccctgg cccacatgat caagttccgg ggccacttcc tgatcgaggg
     2701 cgacctgaac cccgacaaca gcgacgtgga caagctgttc atccagctgg tgcagaccta
     2761 caaccagctg ttcgaggaaa accccatcaa cgccagcggc gtggacgcca aggccatcct
     2821 gtctgccaga ctgagcaaga gcagacggct ggaaaatctg atcgcccagc tgcccggcga
     2881 gaagaagaat ggcctgttcg gaaacctgat tgccctgagc ctgggcctga cccccaactt
     2941 caagagcaac ttcgacctgg ccgaggatgc caaactgcag ctgagcaagg acacctacga
     3001 cgacgacctg gacaacctgc tggcccagat cggcgaccag tacgccgacc tgtttctggc
     3061 cgccaagaac ctgtccgacg ccatcctgct gagcgacatc ctgagagtga acaccgagat
     3121 caccaaggcc cccctgagcg cctctatgat caagagatac gacgagcacc accaggacct
     3181 gaccctgctg aaagctctcg tgcggcagca gctgcctgag aagtacaaag agattttctt
     3241 cgaccagagc aagaacggct acgccggcta cattgacggc ggagccagcc aggaagagtt
     3301 ctacaagttc atcaagccca tcctggaaaa gatggacggc accgaggaac tgctcgtgaa
     3361 gctgaacaga gaggacctgc tgcggaagca gcggaccttc gacaacggca gcatccccca
     3421 ccagatccac ctgggagagc tgcacgccat tctgcggcgg caggaagatt tttacccatt
     3481 cctgaaggac aaccgggaaa agatcgagaa gatcctgacc ttccgcatcc cctactacgt
     3541 gggccctctg gccaggggaa acagcagatt cgcctggatg accagaaaga gcgaggaaac
     3601 catcaccccc tggaacttcg aggaagtggt ggacaagggc gcttccgccc agagcttcat
     3661 cgagcggatg accaacttcg ataagaacct gcccaacgag aaggtgctgc ccaagcacag
     3721 cctgctgtac gagtacttca ccgtgtataa cgagctgacc aaagtgaaat acgtgaccga
     3781 gggaatgaga aagcccgcct tcctgagcgg cgagcagaaa aaggccatcg tggacctgct
     3841 gttcaagacc aaccggaaag tgaccgtgaa gcagctgaaa gaggactact tcaagaaaat
     3901 cgagtgcttc gactccgtgg aaatctccgg cgtggaagat cggttcaacg cctccctggg
     3961 cacataccac gatctgctga aaattatcaa ggacaaggac ttcctggaca atgaggaaaa
     4021 cgaggacatt ctggaagata tcgtgctgac cctgacactg tttgaggaca gagagatgat
     4081 cgaggaacgg ctgaaaacct atgcccacct gttcgacgac aaagtgatga agcagctgaa
     4141 gcggcggaga tacaccggct ggggcaggct gagccggaag ctgatcaacg gcatccggga
     4201 caagcagtcc ggcaagacaa tcctggattt cctgaagtcc gacggcttcg ccaacagaaa
     4261 cttcatgcag ctgatccacg acgacagcct gacctttaaa gaggacatcc agaaagccca
     4321 ggtgtccggc cagggcgata gcctgcacga gcacattgcc aatctggccg gcagccccgc
     4381 cattaagaag ggcatcctgc agacagtgaa ggtggtggac gagctcgtga aagtgatggg
     4441 ccggcacaag cccgagaaca tcgtgatcga aatggccaga gagaaccaga ccacccagaa
     4501 gggacagaag aacagccgcg agagaatgaa gcggatcgaa gagggcatca aagagctggg
     4561 cagccagatc ctgaaagaac accccgtgga aaacacccag ctgcagaacg agaagctgta
     4621 cctgtactac ctgcagaatg ggcgggatat gtacgtggac caggaactgg acatcaaccg
     4681 gctgtccgac tacgatgtgg accatatcgt gcctcagagc tttctgaagg acgactccat
     4741 cgacaacaag gtgctgacca gaagcgacaa gaaccggggc aagagcgaca acgtgccctc
     4801 cgaagaggtc gtgaagaaga tgaagaacta ctggcggcag ctgctgaacg ccaagctgat
     4861 tacccagaga aagttcgaca atctgaccaa ggccgagaga ggcggcctga gcgaactgga
     4921 taaggccggc ttcatcaaga gacagctggt ggaaacccgg cagatcacaa agcacgtggc
     4981 acagatcctg gactcccgga tgaacactaa gtacgacgag aatgacaagc tgatccggga
     5041 agtgaaagtg atcaccctga agtccaagct ggtgtccgat ttccggaagg atttccagtt
     5101 ttacaaagtg cgcgagatca acaactacca ccacgcccac gacgcctacc tgaacgccgt
     5161 cgtgggaacc gccctgatca aaaagtaccc taagctggaa agcgagttcg tgtacggcga
     5221 ctacaaggtg tacgacgtgc ggaagatgat cgccaagagc gagcaggaaa tcggcaaggc
     5281 taccgccaag tacttcttct acagcaacat catgaacttt ttcaagaccg agattaccct
     5341 ggccaacggc gagatccgga agcggcctct gatcgagaca aacggcgaaa ccggggagat
     5401 cgtgtgggat aagggccggg attttgccac cgtgcggaaa gtgctgagca tgccccaagt
     5461 gaatatcgtg aaaaagaccg aggtgcagac aggcggcttc agcaaagagt ctatcctgcc
     5521 caagaggaac agcgataagc tgatcgccag aaagaaggac tgggacccta agaagtacgg
     5581 cggcttcgac agccccaccg tggcctattc tgtgctggtg gtggccaaag tggaaaaggg
     5641 caagtccaag aaactgaaga gtgtgaaaga gctgctgggg atcaccatca tggaaagaag
     5701 cagcttcgag aagaatccca tcgactttct ggaagccaag ggctacaaag aagtgaaaaa
     5761 ggacctgatc atcaagctgc ctaagtactc cctgttcgag ctggaaaacg gccggaagag
     5821 aatgctggcc tctgccggcg aactgcagaa gggaaacgaa ctggccctgc cctccaaata
     5881 tgtgaacttc ctgtacctgg ccagccacta tgagaagctg aagggctccc ccgaggataa
     5941 tgagcagaaa cagctgtttg tggaacagca caagcactac ctggacgaga tcatcgagca
     6001 gatcagcgag ttctccaaga gagtgatcct ggccgacgct aatctggaca aagtgctgtc
     6061 cgcctacaac aagcaccggg ataagcccat cagagagcag gccgagaata tcatccacct
     6121 gtttaccctg accaatctgg gagcccctgc cgccttcaag tactttgaca ccaccatcga
     6181 ccggaagagg tacaccagca ccaaagaggt gctggacgcc accctgatcc accagagcat
     6241 caccggcctg tacgagacac ggatcgacct gtctcagctg ggaggcgaca aaaggccggc
     6301 ggccacgaaa aaggccggcc aggcaaaaaa gaaaaaggag ggcagaggaa gtcttctaac
     6361 atgcggtgac gtggaggaga atcccggccc tatggtgagc aagggcgagg agctgttcac
     6421 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
     6481 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac
     6541 caccagcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca
     6601 gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
     6661 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
     6721 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
     6781 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa
     6841 cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca
     6901 caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
     6961 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa
     7021 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
     7081 cactctcggc atggacgagc tgtacaagta attaattaac tagaatgaat cgtttttaaa
     7141 ataacaaatc aattgtttta taatattcgt acgattcttt gattatgtaa taaaatgtga
     7201 tcattaggaa gattacgaaa aatataaaaa atatgagttc tgtgtgtata acaaatgctg
     7261 taaacgccac aattgtgttt gttgcaaata aacccatgat tatttgatta aaattgttgt
     7321 tttctttgtt catagacaat agtgtgtttt gcctaaacgt gtactgcata aactccatgc
     7381 gagtgtatag cgagctagtg gctaacgctt gccccaccaa agtagattcg tcaaaatcct
     7441 caatttcatc accctcctcc aagtttaaca tttggccgtc ggaattaact tctaaagatg
     7501 ccacataatc taataaatga aatagagatt caaacgtggc gtcatcgtcc gtttcgacca
     7561 tttccgaaaa gaactcgggc ataaactcta tgatttctct ggacgtggtg ttgtcgaaac
     7621 tctcaaagta cgcagtcagg aacgtgcgcg acatgtcgtc gggaaactcg cgcggaaaca
     7681 tgttgttgta accgaacggg tcccatagcg ccaaaaccaa atctgccagc gtcaatagaa
     7741 tgagcacgat gccgacaatg gagctggctt ggatagcgat tcgagttaac ggccggggcg
     7801 cgcccatact cggtggcctc cccaccacca acttttttgc actgcaaaaa aacacgcttt
     7861 tgcacgcggg cccatacata gtacaaactc tacgtttcgt agactatttt acataaatag
     7921 tctacaccgt tgtatacgct ccaaatacac taccacacat tgaacctttt tgcagtgcaa
     7981 aaaagtacgt gtcggcagtc acgtaggccg gccttatcgg gtcgcgtcct gtcacgtacg
     8041 aatcacatta tcggaccgga cgagtgttgt cttatcgtga caggacgcca gcttcctgtg
     8101 ttgctaaccg cagccggacg caactcctta tcggaacagg acgcgcctcc atatcagccg
     8161 cgcgttatct catgcgcgtg accggacacg aggcgcccgt cccgcttatc gcgcctataa
     8221 atacagcccg caacgatctg gtaaacacag ttgaacagca tctgttacag cgacacaaca
     8281 tgagccggtc caacaacgcc aacgcgccca cgccatccaa ccgccgccgc aacctgtctc
     8341 tggtgatggt gcgctcctcc aagaacgtca tcaaggagtt catgcgcttc aaggtgcgca
     8401 tggagggcac cgtgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct
     8461 acgagggcca caacaccgtg aagctgaagg tgaccaaggg cggccccctg cccttcgcct
     8521 gggacatcct gtccccccag ttccagtacg gctccaaggt gtacgtgaag caccccgccg
     8581 acatccccga ctacaagaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga
     8641 acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag gacggctgct
     8701 tcatctacaa ggtgaagttc atcggcgtga acttcccctc cgacggcccc gtaatgcaga
     8761 agaagaccat gggctgggag gcctccaccg agcgcctgta cccccgcgac ggcgtgctga
     8821 agggcgagat ccacaaggcc ctgaagctga aggacggcgg ccactacctg gtggagttca
     8881 agtccatcta catggccaag aagcccgtgc agctgcccgg ctactactac gtggactcca
     8941 agctggacat cacctcccac aacgaggact acaccatcgt ggagcagtac gagcgcaccg
     9001 agggccgcca ccacctgttc ctgtaggact ctagatcata atcagccata ccacatttgt
     9061 agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat
     9121 gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa
     9181 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     9241 caaactcatc aatgtatctt aaagcttcta ggccggccga tctcggatct gacaatgttc
     9301 agtgcagaga ctcggctacg cctcgtggac tttgaagttg accaacaatg tttattctta
     9361 cctctaatag tcctctgtgg caaggtcaag attctgttag aagccaatga agaacctggt
     9421 tgttcaataa cattttgttc gtctaatatt tcactaccgc ttgacgttgg ctgcacttca
     9481 tgtacctcat ctataaacgc ttcttctgta tcgctctgga cgtcatcttc acttacgtga
     9541 tctgatattt cactgtcaga atcctcacca acaagctcgt catcgctttg cagaagagca
     9601 gagaggatat gctcatcgtc taaagaacta cccattttat tatatattag tcacgatatc
     9661 tataacaaga aaatatatat ataataagtt atcacgtaag tagaacatga aataacaata
     9721 taattatcgt atgagttaaa tcttaaaagt cacgtaaaag ataatcatgc gtcattttga
     9781 ctcacgcggt cgttatagtt caaaatcagt gacacttacc gcattgacaa gcacgcctca
     9841 cgggagctcc aagcggcgac tgagatgtcc taaatgcaca gcgacggatt cgcgctattt
     9901 agaaagagag agcaatattt caagaatgca tgcgtcaatt ttacgcagac tatctttcta
     9961 gggttaaaaa agatttgcgc tttactcgac ctaaacttta aacacgtcat agaatcttcg
    10021 tttgacaaaa accacattgt ggccaagctg tgtgacgcga cgcgcgctaa agaatggcaa
    10081 accaagtcgc gcgagcgtcg actctagagg atccccgggt accgagctcg aattcgtaat
    10141 catgtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
    10201 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
    10261 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
    10321 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
    10381 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
    10441 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
    10501 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
    10561 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
    10621 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
    10681 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
    10741 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
    10801 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
    10861 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
    10921 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
    10981 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
    11041 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
    11101 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
    11161 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
    11221 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
    11281 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
    11341 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
    11401 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
    11461 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
    11521 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
    11581 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
    11641 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
    11701 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
    11761 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
    11821 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
    11881 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
    11941 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
    12001 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
    12061 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
    12121 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
    12181 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
    12241 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
    12301 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
    12361 tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac
    12421 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc
    12481 gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag
    12541 agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
    12601 gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
    12661 gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc
    12721 agggttttcc cagtcacgac gttgtaaaac gacggccagt gccaagcttt gtttaaaaat
    12781 ataacaaaat tgtgatccca caaaatgaag tggggcaaaa tcaaataatt aatagtgtcc
    12841 gtaaacttgt tggtcttcaa ctttttgagg aacacgttgg acggcaaatc cgtgactata
    12901 acacaagttg atttaataat tttagccaac acgtcgggct gcgtgttttt tgccgacgcg
    12961 tctgtgtaca cgttgattaa ctggtcgatt aaactgttga aataatttaa tttttggttc
    13021 ttctttaaat ctgtgatgaa attttttaaa ataactttaa attcttcatt ggtaaaaaat
    13081 gccacgtttt gcaacttgtg agggtctaat atgaggtcaa actcagtagg agttttatcc
    13141 aaaaaagaaa acatgatta
//
