LOCUS       sequence-244418-       10004 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  CRISPaint universal donor plasmid for gene exon insertion. Encodes 
            T2A-Cas9-T2A-GFP-SV40 and 3xP3-RFP visible eye marker..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 10004)
  AUTHORS   Bosch JA, Colbeth R, Zirin J, Perrimon N
  TITLE     Gene Knock-Ins in Drosophila Using Homology-Independent Insertion of
            Universal Donor Plasmids.
  JOURNAL   Genetics. 2019 Nov 4. pii: genetics.119.302819. doi: 
            10.1534/genetics.119.302819.
  PUBMED    31685521
REFERENCE   2  (bases 1 to 10004)
  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: "Genetics. 
            2019 Nov 4. pii: genetics.119.302819. doi: 
            10.1534/genetics.119.302819."
COMMENT     Sequence Label: pCRISPaint-T2A-Cas9-T2A-GFP-3xP3-RFP
FEATURES             Location/Qualifiers
     source          1..10004
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     rep_origin      complement(6..461)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(143..164)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     355..374
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     588..610
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     602..619
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     603..619
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             737..790
                     /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             800..865
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             872..892
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             917..5017
                     /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             917..5017
                     /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             5018..5065
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             5069..5122
                     /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(5138..5140,5141..5143,5144..5854)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELY"
     CDS             join(5138..5140,5141..5143,5144..5851)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELY"
     primer_bind     complement(5183..5204)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5444..5463)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5791..5812
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     intron          5944..6009
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             6139..6159
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    6431..6552
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    6553..6586
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      6812..6821
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6818..7495
                     /codon_start=1
                     /product="rapidly maturing tetrameric variant of DsRed 
                     fluorescent protein (Bevis and Glick, 2002)"
                     /label=DsRed-Express
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGSFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERAEGRHH
                     LFL"
     primer_bind     complement(6948..6966)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(6998..7018)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     7216..7235
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     7410..7433
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    7616..7737
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    7767..7800
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        complement(7819..7837)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7858..7874)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7858..7874)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7871..7893)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7882..7898
                     /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(7906..7912,7913..7930,7931..7936))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7951..7972
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8089..8106)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8260..8848)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8340..8359)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9019..9810,9811..9879))
                     /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     9642..9661
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9880..9984)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cacctaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
       61 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
      121 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
      181 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
      241 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
      301 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
      361 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
      421 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcccattcgc cattcaggct
      481 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
      541 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
      601 ttgtaaaacg acggccagtg aattgtaata cgactcacta tagggcgaat tgggtacaag
      661 cttcacgacc tgaggcgcgc cgggccagta cccaaaaagc ggggggtccg gtggaagcgg
      721 aggtagcgcg gccgccgagg gccgcggcag cctgctgacc tgcggcgatg tggaggagaa
      781 ccccgggccc gctagcatgg attacaagga tcacgatggc gactacaagg accatgacat
      841 cgactataag gacgacgacg ataagatggc ccccaagaag aagcgcaaag tgggcatcca
      901 tggcgtgcca gccgccgata agaagtacag catcggcctg gatatcggca ccaacagcgt
      961 gggctgggcc gtgatcaccg atgagtacaa ggtgcccagc aagaagttca aggtgctggg
     1021 caacaccgat cgccacagca tcaagaagaa cctgatcggc gccctgctgt tcgatagcgg
     1081 agaaaccgcc gaggccaccc gcctgaagcg caccgcccgc cgccgctata cccgccgcaa
     1141 gaatcgcatc tgctatctgc aagagatctt cagcaacgag atggccaagg tggacgatag
     1201 cttcttccac cgcctggagg agtccttcct ggtggaggag gataagaagc acgagcgcca
     1261 ccccatcttc ggcaacatcg tggatgaggt ggcctaccac gagaagtacc ccaccatcta
     1321 ccatctgcgc aagaagctgg tggatagcac cgataaggcc gatctgcgcc tgatctacct
     1381 ggccctggcc cacatgatca agttccgcgg ccacttcctg atcgagggcg atctgaaccc
     1441 cgataactcc gatgtggata agctgttcat ccagctggtg cagacctaca accagctgtt
     1501 cgaggagaac cccatcaacg ccagcggcgt ggacgccaag gccattctga gtgcccgcct
     1561 gagcaagagt cgccgcctgg agaatctgat cgcccagctg ccaggcgaga agaagaacgg
     1621 cctgttcggc aacctgattg ccctgagcct gggcctgacc cccaacttca agagcaactt
     1681 cgatctggcc gaggatgcca agctgcagct gagcaaggat acctacgatg atgatctgga
     1741 taacctgctg gcccagatcg gcgatcagta cgccgacctg tttctggccg ccaagaacct
     1801 gtccgatgcc atcctgctga gcgacatcct gcgcgtgaac accgagatca ccaaggcccc
     1861 actgagcgcc tcgatgatca agcgctacga tgagcaccac caggatctga ccctgctgaa
     1921 ggccctcgtg cgccagcagc tgcccgagaa gtataaggag atcttcttcg accagagcaa
     1981 gaacggctac gccggctaca tcgatggcgg agccagccag gaggagttct acaagttcat
     2041 caagcccatc ctggagaaga tggatggcac cgaggagctg ctcgtgaagc tgaaccgcga
     2101 ggatctgctg cgcaagcagc gcaccttcga taacggcagc atcccccacc agatccacct
     2161 gggagagctg cacgccattc tgcgccgcca ggaggatttt tacccgttcc tgaaggataa
     2221 ccgcgagaag attgagaaga tcctgacctt ccggatcccc tactacgtgg gaccactggc
     2281 ccgcggcaat agccgctttg cctggatgac ccgcaagagc gaggaaacca tcaccccctg
     2341 gaacttcgag gaggtggtgg ataagggcgc cagcgcccag agcttcatcg agcgcatgac
     2401 caacttcgac aagaatctgc ccaacgagaa ggtgctgccc aagcacagcc tgctgtacga
     2461 gtacttcacc gtgtacaacg agctgacgaa agtgaaatac gtgaccgagg gaatgcgcaa
     2521 gcccgccttc ctgagcggag agcagaagaa ggccatcgtg gacctgctgt ttaagaccaa
     2581 ccgcaaagtg accgtgaagc agctgaagga ggactacttc aagaagatcg agtgcttcga
     2641 cagcgtggag atctcgggcg tggaggatcg cttcaatgcc agcctgggca cgtaccacga
     2701 tctgctgaag attatcaagg acaaggactt cctggacaac gaggagaatg aggacattct
     2761 ggaggacatc gtgctgaccc tgaccctgtt tgaggatcgc gagatgatcg aggagcgcct
     2821 caagacctac gcccatctgt ttgatgacaa agtgatgaag cagctgaagc gccgccgcta
     2881 caccggatgg ggacgcctga gtcgcaagct gatcaacggc atccgcgata agcagagcgg
     2941 caagacgatc ctggatttcc tgaagtccga tggcttcgcc aatcgcaact tcatgcagct
     3001 gatccacgac gattcgctga ccttcaagga ggatattcag aaggcccagg tgtccggcca
     3061 gggcgatagc ctgcatgagc acattgccaa cctggccggc agccccgcca ttaagaaggg
     3121 cattctgcag accgtgaagg tggtggacga gctcgtgaaa gtgatgggcc gccacaagcc
     3181 cgagaatatc gtgattgaga tggcccgcga gaaccagacc acccagaagg gccagaagaa
     3241 cagccgcgag cgcatgaagc gcatcgagga gggcattaag gagctgggca gccagattct
     3301 gaaggagcac cccgtggaga acacccagct gcagaacgag aagctgtacc tgtactacct
     3361 gcagaacggc cgcgatatgt acgtggacca ggagctggac atcaaccgcc tgtccgacta
     3421 cgatgtggac catatcgtgc cgcagagctt tctgaaggat gacagcatcg ataacaaggt
     3481 gctgacccgc agcgataaga accgcggcaa gtccgataac gtgccctcgg aggaggtcgt
     3541 gaagaagatg aagaactatt ggcgccagct gctgaacgcc aagctgatta cccagcgcaa
     3601 gttcgacaat ctgaccaagg ccgagcgcgg aggcctgagc gagctggata aggccggctt
     3661 catcaagcgc cagctggtgg aaacccgcca gattaccaag cacgtggccc agattctgga
     3721 tagccgcatg aacacgaagt acgacgagaa tgataagctg attcgcgaag tgaaagtgat
     3781 caccctgaag tccaagctgg tgtccgattt ccgcaaggat ttccagttct ataaagtgcg
     3841 cgagatcaac aactaccacc acgcccacga tgcctacctg aacgccgtcg tgggaaccgc
     3901 cctgattaag aagtatccca agctggagag cgagttcgtg tacggcgatt ataaggtgta
     3961 cgacgtgcgc aagatgatcg ccaagtccga gcaggagatc ggcaaggcca ccgccaagta
     4021 ctttttctac agcaacatca tgaatttctt caagaccgag attaccctgg ccaacggcga
     4081 gatccgcaag cgcccactga tcgaaacgaa tggcgaaacc ggcgagattg tgtgggacaa
     4141 gggccgcgat tttgccaccg tgcgcaaggt gctgtccatg ccccaagtga acattgtgaa
     4201 aaagaccgag gtgcagacgg gcggcttcag caaggagtcc attctgccga agcgcaacag
     4261 cgacaagctg atcgcccgca agaaggattg ggaccccaag aagtacggcg gcttcgatag
     4321 ccccaccgtg gcctatagtg tgctggtggt ggccaaggtg gagaagggca agtccaagaa
     4381 gctgaagtcc gtgaaggagc tgctgggcat caccatcatg gagcgcagca gctttgagaa
     4441 gaacccaatc gatttcctgg aggccaaggg ctacaaggaa gtgaagaagg atctgatcat
     4501 caagctgccg aagtactccc tgttcgagct ggagaacgga cgcaagcgca tgctggccag
     4561 tgccggcgag ctgcagaagg gaaacgagct ggccctgccc tcgaaatatg tgaacttcct
     4621 gtacctggcc agccactacg agaagctgaa gggcagcccc gaggataacg agcagaagca
     4681 gctgtttgtg gagcagcaca agcactacct ggacgagatc attgagcaga tcagcgagtt
     4741 cagcaagcgc gtgatcctgg ccgacgccaa tctggataag gtgctgagcg cctacaacaa
     4801 gcaccgcgac aagccaatcc gcgagcaggc cgagaacatc atccacctgt tcaccctgac
     4861 caacctggga gccccagccg ccttcaagta cttcgatacc accatcgatc gcaagcgcta
     4921 taccagcacc aaggaggtgc tggacgccac cctgatccac cagagcatca ccggcctgta
     4981 cgaaacccgc atcgatctgt cgcagctggg aggcgataag cgcccagccg ccaccaagaa
     5041 ggccggacag gccaaaaaga agaagcttga gggcagagga agtcttctaa catgcggtga
     5101 cgtggaggag aatcccggcc ctgctagcgg tactagcatg gtgagcaagg gcgaggagct
     5161 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
     5221 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat
     5281 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg
     5341 cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc
     5401 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa
     5461 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
     5521 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag
     5581 ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat
     5641 ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc
     5701 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct
     5761 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
     5821 cgggatcact ctcggcatgg acgagctgta ctaaggtacc tctagaggat ctttgtgaag
     5881 gaaccttact tctgtggtgt gacataattg gacaaactac ctacagagat ttaaagctct
     5941 aaggtaaata taaaattttt aagtgtataa tgtgttaaac tactgattct aattgtttgt
     6001 gtattttaga ttccaaccta tggaactgat gaatgggagc agtggtggaa tgcctttaat
     6061 gaggaaaacc tgttttgctc agaagaaatg ccatctagtg atgatgaggc tactgctgac
     6121 tctcaacatt ctactcctcc aaaaaagaag agaaaggtag aagaccccaa ggactttcct
     6181 tcagaattgc taagtttttt gagtcatgct gtgtttagta atagaactct tgcttgcttt
     6241 gctatttaca ccacaaagga aaaagctgca ctgctataca agaaaattat ggaaaaatat
     6301 ttgatgtata gtgccttgac tagagatcat aatcagccat accacatttg tagaggtttt
     6361 acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat
     6421 tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     6481 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     6541 caatgtatct taataacttc gtataatgta tgctatacga agttatcgta cgggatctaa
     6601 ttcaattaga gactaattca attagagcta attcaattag gatccaagct tatcgatttc
     6661 gaaccctcga ccgccggagt ataaatagag gcgcttcgtc tacggagcga caattcaatt
     6721 caaacaagca aagtgaacac gtcgctaagc gaaagctaag caaataaaca agcgcagctg
     6781 aacaagctaa acaatcggct cgaagccggt cgccaccatg gcctcctccg aggacgtcat
     6841 caaggagttc atgcgcttca aggtgcgcat ggagggctcc gtgaacggcc acgagttcga
     6901 gatcgagggc gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt
     6961 gaccaagggc ggccccctgc ccttcgcctg ggacatcctg tccccccagt tccagtacgg
     7021 ctccaaggtg tacgtgaagc accccgccga catccccgac tacaagaagc tgtccttccc
     7081 cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac
     7141 ccaggactcc tccctccagg acggctcctt catctacaag gtgaagttca tcggcgtgaa
     7201 cttcccctcc gacggccccg taatgcagaa gaagactatg ggctgggagg cgtccaccga
     7261 gcgcctgtac ccccgcgacg gcgtgctgaa gggcgagatc cacaaggccc tgaagctgaa
     7321 ggacggcggc cactacctgg tggagttcaa gtccatctac atggccaaga agcccgtgca
     7381 gctgcccggc tactactacg tggactccaa gctggacatc acctcccaca acgaggacta
     7441 caccatcgtg gagcagtacg agcgcgccga gggccgccac cacctgttcc tgtaggggcc
     7501 gcgactctag atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa
     7561 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
     7621 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     7681 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaacc
     7741 gtctgtaagc agcgaaggcc catacgataa cttcgtataa tgtatgctat acgaagttat
     7801 gagctccagc ttttgttccc tttagtgagg gttaatttcg agcttggcgt aatcatggtc
     7861 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     7921 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     7981 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     8041 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     8101 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     8161 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     8221 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     8281 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     8341 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     8401 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     8461 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     8521 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     8581 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     8641 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     8701 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     8761 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     8821 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     8881 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     8941 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     9001 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     9061 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     9121 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     9181 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     9241 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     9301 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     9361 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     9421 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     9481 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     9541 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     9601 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     9661 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     9721 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     9781 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     9841 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     9901 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     9961 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgc
//
