LOCUS       sequence-248026-       10008 bp    DNA     circular SYN 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      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10008)
  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 10008)
  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..10008
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     78..100
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     92..109
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     93..109
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     119..138
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        119..137
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             230..283
                     /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             293..358
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             365..385
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             410..4510
                     /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             410..4510
                     /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             4511..4558
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             4562..4615
                     /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(4631..4633,4634..4636,4637..5347)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKGD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELY"
     primer_bind     complement(4676..4697)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     5284..5305
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     intron          5437..5502
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             5632..5652
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    5924..6045
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    6046..6079
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      6305..6314
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6311..6988
                     /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(6441..6459)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(6491..6511)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     6709..6728
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     6903..6926
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    7109..7230
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    7260..7293
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(7312..7332)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(7312..7330)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7351..7367)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7351..7367)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7364..7386)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7375..7391
                     /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(7399..7405,7406..7423,7424..7429))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7444..7465
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7582..7599)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7753..8341)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7833..7852)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8512..9303,9304..9372))
                     /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     9135..9154
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9373..9477)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      9504..9960
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9591..9610)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9801..9822
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg
       61 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg agcgcgcgta
      121 atacgactca ctatagggcg aattgggtac aagcttcacg acctgaggcg cgccgggcca
      181 gtacccaaaa agcggggggt ccggtggaag cggaggtagc gcggccgccg agggccgcgg
      241 cagcctgctg acctgcggcg atgtggagga gaaccccggg cccgctagca tggattacaa
      301 ggatcacgat ggcgactaca aggaccatga catcgactat aaggacgacg acgataagat
      361 ggcccccaag aagaagcgca aagtgggcat ccatggcgtg ccagccgccg ataagaagta
      421 cagcatcggc ctggatatcg gcaccaacag cgtgggctgg gccgtgatca ccgatgagta
      481 caaggtgccc agcaagaagt tcaaggtgct gggcaacacc gatcgccaca gcatcaagaa
      541 gaacctgatc ggcgccctgc tgttcgatag cggagaaacc gccgaggcca cccgcctgaa
      601 gcgcaccgcc cgccgccgct atacccgccg caagaatcgc atctgctatc tgcaagagat
      661 cttcagcaac gagatggcca aggtggacga tagcttcttc caccgcctgg aggagtcctt
      721 cctggtggag gaggataaga agcacgagcg ccaccccatc ttcggcaaca tcgtggatga
      781 ggtggcctac cacgagaagt accccaccat ctaccatctg cgcaagaagc tggtggatag
      841 caccgataag gccgatctgc gcctgatcta cctggccctg gcccacatga tcaagttccg
      901 cggccacttc ctgatcgagg gcgatctgaa ccccgataac tccgatgtgg ataagctgtt
      961 catccagctg gtgcagacct acaaccagct gttcgaggag aaccccatca acgccagcgg
     1021 cgtggacgcc aaggccattc tgagtgcccg cctgagcaag agtcgccgcc tggagaatct
     1081 gatcgcccag ctgccaggcg agaagaagaa cggcctgttc ggcaacctga ttgccctgag
     1141 cctgggcctg acccccaact tcaagagcaa cttcgatctg gccgaggatg ccaagctgca
     1201 gctgagcaag gatacctacg atgatgatct ggataacctg ctggcccaga tcggcgatca
     1261 gtacgccgac ctgtttctgg ccgccaagaa cctgtccgat gccatcctgc tgagcgacat
     1321 cctgcgcgtg aacaccgaga tcaccaaggc cccactgagc gcctcgatga tcaagcgcta
     1381 cgatgagcac caccaggatc tgaccctgct gaaggccctc gtgcgccagc agctgcccga
     1441 gaagtataag gagatcttct tcgaccagag caagaacggc tacgccggct acatcgatgg
     1501 cggagccagc caggaggagt tctacaagtt catcaagccc atcctggaga agatggatgg
     1561 caccgaggag ctgctcgtga agctgaaccg cgaggatctg ctgcgcaagc agcgcacctt
     1621 cgataacggc agcatccccc accagatcca cctgggagag ctgcacgcca ttctgcgccg
     1681 ccaggaggat ttttacccgt tcctgaagga taaccgcgag aagattgaga agatcctgac
     1741 cttccggatc ccctactacg tgggaccact ggcccgcggc aatagccgct ttgcctggat
     1801 gacccgcaag agcgaggaaa ccatcacccc ctggaacttc gaggaggtgg tggataaggg
     1861 cgccagcgcc cagagcttca tcgagcgcat gaccaacttc gacaagaatc tgcccaacga
     1921 gaaggtgctg cccaagcaca gcctgctgta cgagtacttc accgtgtaca acgagctgac
     1981 gaaagtgaaa tacgtgaccg agggaatgcg caagcccgcc ttcctgagcg gagagcagaa
     2041 gaaggccatc gtggacctgc tgtttaagac caaccgcaaa gtgaccgtga agcagctgaa
     2101 ggaggactac ttcaagaaga tcgagtgctt cgacagcgtg gagatctcgg gcgtggagga
     2161 tcgcttcaat gccagcctgg gcacgtacca cgatctgctg aagattatca aggacaagga
     2221 cttcctggac aacgaggaga atgaggacat tctggaggac atcgtgctga ccctgaccct
     2281 gtttgaggat cgcgagatga tcgaggagcg cctcaagacc tacgcccatc tgtttgatga
     2341 caaagtgatg aagcagctga agcgccgccg ctacaccgga tggggacgcc tgagtcgcaa
     2401 gctgatcaac ggcatccgcg ataagcagag cggcaagacg atcctggatt tcctgaagtc
     2461 cgatggcttc gccaatcgca acttcatgca gctgatccac gacgattcgc tgaccttcaa
     2521 ggaggatatt cagaaggccc aggtgtccgg ccagggcgat agcctgcatg agcacattgc
     2581 caacctggcc ggcagccccg ccattaagaa gggcattctg cagaccgtga aggtggtgga
     2641 cgagctcgtg aaagtgatgg gccgccacaa gcccgagaat atcgtgattg agatggcccg
     2701 cgagaaccag accacccaga agggccagaa gaacagccgc gagcgcatga agcgcatcga
     2761 ggagggcatt aaggagctgg gcagccagat tctgaaggag caccccgtgg agaacaccca
     2821 gctgcagaac gagaagctgt acctgtacta cctgcagaac ggccgcgata tgtacgtgga
     2881 ccaggagctg gacatcaacc gcctgtccga ctacgatgtg gaccatatcg tgccgcagag
     2941 ctttctgaag gatgacagca tcgataacaa ggtgctgacc cgcagcgata agaaccgcgg
     3001 caagtccgat aacgtgccct cggaggaggt cgtgaagaag atgaagaact attggcgcca
     3061 gctgctgaac gccaagctga ttacccagcg caagttcgac aatctgacca aggccgagcg
     3121 cggaggcctg agcgagctgg ataaggccgg cttcatcaag cgccagctgg tggaaacccg
     3181 ccagattacc aagcacgtgg cccagattct ggatagccgc atgaacacga agtacgacga
     3241 gaatgataag ctgattcgcg aagtgaaagt gatcaccctg aagtccaagc tggtgtccga
     3301 tttccgcaag gatttccagt tctataaagt gcgcgagatc aacaactacc accacgccca
     3361 cgatgcctac ctgaacgccg tcgtgggaac cgccctgatt aagaagtatc ccaagctgga
     3421 gagcgagttc gtgtacggcg attataaggt gtacgacgtg cgcaagatga tcgccaagtc
     3481 cgagcaggag atcggcaagg ccaccgccaa gtactttttc tacagcaaca tcatgaattt
     3541 cttcaagacc gagattaccc tggccaacgg cgagatccgc aagcgcccac tgatcgaaac
     3601 gaatggcgaa accggcgaga ttgtgtggga caagggccgc gattttgcca ccgtgcgcaa
     3661 ggtgctgtcc atgccccaag tgaacattgt gaaaaagacc gaggtgcaga cgggcggctt
     3721 cagcaaggag tccattctgc cgaagcgcaa cagcgacaag ctgatcgccc gcaagaagga
     3781 ttgggacccc aagaagtacg gcggcttcga tagccccacc gtggcctata gtgtgctggt
     3841 ggtggccaag gtggagaagg gcaagtccaa gaagctgaag tccgtgaagg agctgctggg
     3901 catcaccatc atggagcgca gcagctttga gaagaaccca atcgatttcc tggaggccaa
     3961 gggctacaag gaagtgaaga aggatctgat catcaagctg ccgaagtact ccctgttcga
     4021 gctggagaac ggacgcaagc gcatgctggc cagtgccggc gagctgcaga agggaaacga
     4081 gctggccctg ccctcgaaat atgtgaactt cctgtacctg gccagccact acgagaagct
     4141 gaagggcagc cccgaggata acgagcagaa gcagctgttt gtggagcagc acaagcacta
     4201 cctggacgag atcattgagc agatcagcga gttcagcaag cgcgtgatcc tggccgacgc
     4261 caatctggat aaggtgctga gcgcctacaa caagcaccgc gacaagccaa tccgcgagca
     4321 ggccgagaac atcatccacc tgttcaccct gaccaacctg ggagccccag ccgccttcaa
     4381 gtacttcgat accaccatcg atcgcaagcg ctataccagc accaaggagg tgctggacgc
     4441 caccctgatc caccagagca tcaccggcct gtacgaaacc cgcatcgatc tgtcgcagct
     4501 gggaggcgat aagcgcccag ccgccaccaa gaaggccgga caggccaaaa agaagaagct
     4561 tgagggcaga ggaagtcttc taacatgcgg tgacgtggag gagaatcccg gccctgctag
     4621 cggtactagc atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
     4681 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga
     4741 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
     4801 ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga
     4861 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
     4921 caccatcttc ttcaagggcg acggcaacta caagacccgc gccgaggtga agttcgaggg
     4981 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
     5041 cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa
     5101 gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt
     5161 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     5221 cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga
     5281 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
     5341 gtactaaggt acctctagag gatctttgtg aaggaacctt acttctgtgg tgtgacataa
     5401 ttggacaaac tacctacaga gatttaaagc tctaaggtaa atataaaatt tttaagtgta
     5461 taatgtgtta aactactgat tctaattgtt tgtgtatttt agattccaac ctatggaact
     5521 gatgaatggg agcagtggtg gaatgccttt aatgaggaaa acctgttttg ctcagaagaa
     5581 atgccatcta gtgatgatga ggctactgct gactctcaac attctactcc tccaaaaaag
     5641 aagagaaagg tagaagaccc caaggacttt ccttcagaat tgctaagttt tttgagtcat
     5701 gctgtgttta gtaatagaac tcttgcttgc tttgctattt acaccacaaa ggaaaaagct
     5761 gcactgctat acaagaaaat tatggaaaaa tatttgatgt atagtgcctt gactagagat
     5821 cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct
     5881 ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc
     5941 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     6001 actgcattct agttgtggtt tgtccaaact catcaatgta tcttaataac ttcgtataat
     6061 gtatgctata cgaagttatc gtacgggatc taattcaatt agagactaat tcaattagag
     6121 ctaattcaat taggatccaa gcttatcgat ttcgaaccct cgaccgccgg agtataaata
     6181 gaggcgcttc gtctacggag cgacaattca attcaaacaa gcaaagtgaa cacgtcgcta
     6241 agcgaaagct aagcaaataa acaagcgcag ctgaacaagc taaacaatcg gctcgaagcc
     6301 ggtcgccacc atggcctcct ccgaggacgt catcaaggag ttcatgcgct tcaaggtgcg
     6361 catggagggc tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc
     6421 ctacgagggc acccagaccg ccaagctgaa ggtgaccaag ggcggccccc tgcccttcgc
     6481 ctgggacatc ctgtcccccc agttccagta cggctccaag gtgtacgtga agcaccccgc
     6541 cgacatcccc gactacaaga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat
     6601 gaacttcgag gacggcggcg tggtgaccgt gacccaggac tcctccctcc aggacggctc
     6661 cttcatctac aaggtgaagt tcatcggcgt gaacttcccc tccgacggcc ccgtaatgca
     6721 gaagaagact atgggctggg aggcgtccac cgagcgcctg tacccccgcg acggcgtgct
     6781 gaagggcgag atccacaagg ccctgaagct gaaggacggc ggccactacc tggtggagtt
     6841 caagtccatc tacatggcca agaagcccgt gcagctgccc ggctactact acgtggactc
     6901 caagctggac atcacctccc acaacgagga ctacaccatc gtggagcagt acgagcgcgc
     6961 cgagggccgc caccacctgt tcctgtaggg gccgcgactc tagatcataa tcagccatac
     7021 cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa
     7081 acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa
     7141 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     7201 tggtttgtcc aaactcatca atgtatctta accgtctgta agcagcgaag gcccatacga
     7261 taacttcgta taatgtatgc tatacgaagt tatgagctcc agcttttgtt ccctttagtg
     7321 agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     7381 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     7441 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     7501 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     7561 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     7621 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     7681 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     7741 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     7801 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     7861 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     7921 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     7981 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     8041 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     8101 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     8161 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     8221 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     8281 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     8341 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     8401 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     8461 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     8521 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     8581 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     8641 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     8701 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     8761 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     8821 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     8881 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     8941 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     9001 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     9061 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     9121 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     9181 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     9241 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     9301 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     9361 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     9421 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     9481 atttccccga aaagtgccac ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     9541 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     9601 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     9661 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     9721 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     9781 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     9841 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     9901 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt
     9961 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcg
//
