LOCUS       sequence-373359-       10635 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses sgRNA targeting Dora gene and Cas9-Puro in Drosophila S2 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10635)
  AUTHORS   Kingston ER, Blodgett LW, Bartel DP
  TITLE     Endogenous transcripts direct microRNA degradation in Drosophila, 
            and this targeted degradation is required for proper embryonic 
            development.
  JOURNAL   Mol Cell. 2022 Sep 15. pii: S1097-2765(22)00849-8. doi: 
            10.1016/j.molcel.2022.08.029.
  PUBMED    36150386
REFERENCE   2  (bases 1 to 10635)
  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: "Mol Cell. 
            2022 Sep 15. pii: S1097-2765(22)00849-8. doi: 
            10.1016/j.molcel.2022.08.029."
COMMENT     Sequence Label: pAc-sgRNA-Cas9.dora_1
FEATURES             Location/Qualifiers
     source          1..10635
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        827..1226
                     /label=dU6-2 promoter
                     /note="RNA polymerase III promoter for Drosophila U6-2 
                     snRNA (Das et al., 1987)"
     misc_RNA        1248..1323
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(1475..1493)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1561..1665
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1666..1734,1735..2526)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1884..1903)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2697..3285
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3186..3205
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3886..4020
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3923..3942)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3977..3996
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(4028..4627)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4111..4131)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     4608..4627
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     CDS             complement(4652..4705)
                     /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             complement(4721..4768)
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             complement(4769..8869)
                     /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             complement(4769..8869)
                     /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             complement(8894..8914)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(8921..8986)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
ORIGIN
        1 ccgccccacc cttccttccg ctgccccgca cccctttcct tccactaccg ttttgccgca
       61 tttggtattg ccatctcgct tttttggctt tactttgctt tttgaataag ccaagaaact
      121 gaaatcaaag cggacagtcc gaaaattgaa tattccaaaa aacaacaatt gcacatgcac
      181 tgcgttcggc aaaagacgca gaaattcaag ccttttggtc tggcatcatc ttaaaatatc
      241 atcagtatca tcattgccat tataaataca taagaaaatg agaatacatt aaagatatac
      301 atacaatctt tgtcatagtc aacatatttt gcaacaaaat tatctataga taacatgttt
      361 ttgtagaaaa ttccactaac tgattttcgt ataaatacaa attattctca ttcttcaaaa
      421 tactcgccga cgtcatcgcg aaattcctca gcattcttcg acaaatgaag agagacagcg
      481 agtgacacaa tcttatcttg ccgacgagtc tttgaaaatg gccccgggta acggtgaaaa
      541 acatatagcc aagaatggag cgatcgccag gcgatggaaa ttggcttcaa attgctccga
      601 atatggccag ttggtcagtt ggcattggat tgtggagaga tactagatca gctcgttctc
      661 cttctcctcc gcattctcct actccgcccg aagacgcttc tttgatcgct tctttctcga
      721 atgtgtacat attcattatg tatttggtgt ttggcgacga tcagccagct ttaatggcca
      781 taaaacgtgt cttggtttgt gggccacata acaggcagcg agatctgttc gacttgcagc
      841 ctgaaatacg gcacgagtag gaaaagccga gtcaaatgcc gaatgcagag tctcattaca
      901 gcacaatcaa ctcaagaaaa actcgacact tttttaccat ttgcacttaa atcctttttt
      961 attcgttatg tatacttttt ttggtcccta accaaaacaa aaccaaactc tcttagtcgt
     1021 gcctctatat ttaaaactat caatttatta tagtcaataa atcgaactgt gttttcaaca
     1081 aacgaacaat aggacacttt gattctaaag gaaattttga aaatcttaag cagagggttc
     1141 ttaagaccat ttgccaattc ttataattct caactgctct ttcctgatgt tgatcattta
     1201 tataggtatg ttttcctcaa tacttcgcgc tggaggctta ggagtgcgtt ttagagctag
     1261 aaatagcaag ttaaaataag gctagtccgt tatcaacttg aaaaagtggc accgagtcgg
     1321 tgctttttta gatctggaaa aatgatgtga cagtggaaat gagaagtagg tgcatctgca
     1381 aagatttgat tcagagttga tgccattcat gatttttttt tagaagtatc atttgtgttt
     1441 ttagaatata gaattaattc ttgaagacga aagggcctcg tgatacgcct atttttatag
     1501 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
     1561 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     1621 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     1681 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     1741 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     1801 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     1861 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg
     1921 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     1981 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     2041 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     2101 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     2161 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgc agcaatggca
     2221 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     2281 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     2341 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     2401 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     2461 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     2521 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     2581 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     2641 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     2701 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     2761 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     2821 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     2881 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     2941 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     3001 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     3061 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     3121 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     3181 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     3241 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     3301 gcctttttac ggttcctggc cttttgctgg ccttgaagct gtccctgatg gtcgtcatct
     3361 acctgcctgg acagcatggc ctgcaacgcg ggcatcccga tgccgccgga agcgagaaga
     3421 atcataatgg ggaaggccat ccagcctcgc gtcgcgaacg ccagcaagac gtagcccagc
     3481 gcgtcggccg ccatgccggc gataatggcc tgcttctcgc cgaaacgttt ggtggcggga
     3541 ccagtgacga aggcttgagc gagggcgtgc aagattccga ataccgcaag cgacaggccg
     3601 atcatcgtcg cgctccagcg aaagcggtcc tcgccgaaaa tgacccagag cgctgccggc
     3661 acctgtccta cgagttgcat gataaagaag acagtcataa gtgcggcgac gatagtcatg
     3721 ccccgcgccc accggaagga gctgactggg ttgaaggctc tcaagggcat cggtcgactg
     3781 atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
     3841 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
     3901 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     3961 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc
     4021 cctcgagtca ggcaccgggc ttgcgggtca tgcaccaggt gcgcggtcct tcgggcacct
     4081 cgacgtcggc ggtgacggtg aagccgagcc gctcgtagaa ggggaggttg cggggcgcgg
     4141 aggtctccag gaaggcgggc accccggcgc gctcggccgc ctccactccg gggagcacga
     4201 cggcgctgcc cagacccttg ccctggtggt cgggcgagac gccgacggtg gccaggaacc
     4261 acgcgggctc cttgggccgg tgcggcgcca ggaggccttc catctgttgc tgcgcggcca
     4321 gccgggaacc gctcaactcg gccatgcgcg ggccgatctc ggcgaacacc gcccccgctt
     4381 cgacgctctc cggcgtggtc cagaccgcca ccgcggcgcc gtcgtccgcg acccacacct
     4441 tgccgatgtc gagcccgacg cgcgtgagga agagttcttg cagctcggtg acccgctcga
     4501 tgtggcggtc cgggtcgacg gtgtggcgcg tggcggggta gtcggcgaac gcggcggcga
     4561 gggtgcgtac ggcccggggg acgtcgtcgc gggtggcgag gcgcaccgtg ggcttgtact
     4621 cggtcatgct accgctgccg ctaccgctag cagggccggg attctcctcc acgtcaccgc
     4681 atgttagaag acttcctctg ccctcaagct tgcctgatcc ctttttcttt tttgcctggc
     4741 cggccttttt cgtggccgcc ggccttttgt cgcctcccag ctgagacagg tcgatccgtg
     4801 tctcgtacag gccggtgatg ctctggtgga tcagggtggc gtccagcacc tctttggtgc
     4861 tggtgtacct cttccggtcg atggtggtgt caaagtactt gaaggcggca ggggctccca
     4921 gattggtcag ggtaaacagg tggatgatat tctcggcctg ctctctgatg ggcttatccc
     4981 ggtgcttgtt gtaggcggac agcactttgt ccagattagc gtcggccagg atcactctct
     5041 tggagaactc gctgatctgc tcgatgatct cgtccaggta gtgcttgtgc tgttccacaa
     5101 acagctgttt ctgctcatta tcctcggggg agcccttcag cttctcatag tggctggcca
     5161 ggtacaggaa gttcacatat ttggagggca gggccagttc gtttcccttc tgcagttcgc
     5221 cggcagaggc cagcattctc ttccggccgt tttccagctc gaacagggag tacttaggca
     5281 gcttgatgat caggtccttt ttcacttctt tgtagccctt ggcttccaga aagtcgatgg
     5341 gattcttctc gaagctgctt ctttccatga tggtgatccc cagcagctct ttcacactct
     5401 tcagtttctt ggacttgccc ttttccactt tggccaccac cagcacagaa taggccacgg
     5461 tggggctgtc gaagccgccg tacttcttag ggtcccagtc cttctttctg gcgatcagct
     5521 tatcgctgtt cctcttgggc aggatagact ctttgctgaa gccgcctgtc tgcacctcgg
     5581 tctttttcac gatattcact tggggcatgc tcagcacttt ccgcacggtg gcaaaatccc
     5641 ggcccttatc ccacacgatc tccccggttt cgccgtttgt ctcgatcaga ggccgcttcc
     5701 ggatctcgcc gttggccagg gtaatctcgg tcttgaaaaa gttcatgatg ttgctgtaga
     5761 agaagtactt ggcggtagcc ttgccgattt cctgctcgct cttggcgatc atcttccgca
     5821 cgtcgtacac cttgtagtcg ccgtacacga actcgctttc cagcttaggg tactttttga
     5881 tcagggcggt tcccacgacg gcgttcaggt aggcgtcgtg ggcgtggtgg tagttgttga
     5941 tctcgcgcac tttgtaaaac tggaaatcct tccggaaatc ggacaccagc ttggacttca
     6001 gggtgatcac tttcacttcc cggatcagct tgtcattctc gtcgtactta gtgttcatcc
     6061 gggagtccag gatctgtgcc acgtgctttg tgatctgccg ggtttccacc agctgtctct
     6121 tgatgaagcc ggccttatcc agttcgctca ggccgcctct ctcggccttg gtcagattgt
     6181 cgaactttct ctgggtaatc agcttggcgt tcagcagctg ccgccagtag ttcttcatct
     6241 tcttcacgac ctcttcggag ggcacgttgt cgctcttgcc ccggttcttg tcgcttctgg
     6301 tcagcacctt gttgtcgatg gagtcgtcct tcagaaagct ctgaggcacg atatggtcca
     6361 catcgtagtc ggacagccgg ttgatgtcca gttcctggtc cacgtacata tcccgcccat
     6421 tctgcaggta gtacaggtac agcttctcgt tctgcagctg ggtgttttcc acggggtgtt
     6481 ctttcaggat ctggctgccc agctctttga tgccctcttc gatccgcttc attctctcgc
     6541 ggctgttctt ctgtcccttc tgggtggtct ggttctctct ggccatttcg atcacgatgt
     6601 tctcgggctt gtgccggccc atcactttca cgagctcgtc caccaccttc actgtctgca
     6661 ggatgccctt cttaatggcg gggctgccgg ccagattggc aatgtgctcg tgcaggctat
     6721 cgccctggcc ggacacctgg gctttctgga tgtcctcttt aaaggtcagg ctgtcgtcgt
     6781 ggatcagctg catgaagttt ctgttggcga agccgtcgga cttcaggaaa tccaggattg
     6841 tcttgccgga ctgcttgtcc cggatgccgt tgatcagctt ccggctcagc ctgccccagc
     6901 cggtgtatct ccgccgcttc agctgcttca tcactttgtc gtcgaacagg tgggcatagg
     6961 ttttcagccg ttcctcgatc atctctctgt cctcaaacag tgtcagggtc agcacgatat
     7021 cttccagaat gtcctcgttt tcctcattgt ccaggaagtc cttgtccttg ataattttca
     7081 gcagatcgtg gtatgtgccc agggaggcgt tgaaccgatc ttccacgccg gagatttcca
     7141 cggagtcgaa gcactcgatt ttcttgaagt agtcctcttt cagctgcttc acggtcactt
     7201 tccggttggt cttgaacagc aggtccacga tggccttttt ctgctcgccg ctcaggaagg
     7261 cgggctttct cattccctcg gtcacgtatt tcactttggt cagctcgtta tacacggtga
     7321 agtactcgta cagcaggctg tgcttgggca gcaccttctc gttgggcagg ttcttatcga
     7381 agttggtcat ccgctcgatg aagctctggg cggaagcgcc cttgtccacc acttcctcga
     7441 agttccaggg ggtgatggtt tcctcgctct ttctggtcat ccaggcgaat ctgctgtttc
     7501 ccctggccag agggcccacg tagtagggga tgcggaaggt caggatcttc tcgatctttt
     7561 cccggttgtc cttcaggaat gggtaaaaat cttcctgccg ccgcagaatg gcgtgcagct
     7621 ctcccaggtg gatctggtgg gggatgctgc cgttgtcgaa ggtccgctgc ttccgcagca
     7681 ggtcctctct gttcagcttc acgagcagtt cctcggtgcc gtccatcttt tccaggatgg
     7741 gcttgatgaa cttgtagaac tcttcctggc tggctccgcc gtcaatgtag ccggcgtagc
     7801 cgttcttgct ctggtcgaag aaaatctctt tgtacttctc aggcagctgc tgccgcacga
     7861 gagctttcag cagggtcagg tcctggtggt gctcgtcgta tctcttgatc atagaggcgc
     7921 tcaggggggc cttggtgatc tcggtgttca ctctcaggat gtcgctcagc aggatggcgt
     7981 cggacaggtt cttggcggcc agaaacaggt cggcgtactg gtcgccgatc tgggccagca
     8041 ggttgtccag gtcgtcgtcg taggtgtcct tgctcagctg cagtttggca tcctcggcca
     8101 ggtcgaagtt gctcttgaag ttgggggtca ggcccaggct cagggcaatc aggtttccga
     8161 acaggccatt cttcttctcg ccgggcagct gggcgatcag attttccagc cgtctgctct
     8221 tgctcagtct ggcagacagg atggccttgg cgtccacgcc gctggcgttg atggggtttt
     8281 cctcgaacag ctggttgtag gtctgcacca gctggatgaa cagcttgtcc acgtcgctgt
     8341 tgtcggggtt caggtcgccc tcgatcagga agtggccccg gaacttgatc atgtgggcca
     8401 gggccagata gatcagccgc aggtcggcct tgtcggtgct gtccaccagt ttctttctca
     8461 ggtggtagat ggtggggtac ttctcgtggt aggccacctc gtccacgatg ttgccgaaga
     8521 tggggtgccg ctcgtgcttc ttatcctctt ccaccaggaa ggactcttcc agtctgtgga
     8581 agaagctgtc gtccaccttg gccatctcgt tgctgaagat ctcttgcaga tagcagatcc
     8641 ggttcttccg tctggtgtat cttcttctgg cggttctctt cagccgggtg gcctcggctg
     8701 tttcgccgct gtcgaacagc agggctccga tcaggttctt cttgatgctg tgccggtcgg
     8761 tgttgcccag caccttgaat ttcttgctgg gcaccttgta ctcgtcggtg atcacggccc
     8821 agcccacaga gttggtgccg atgtccaggc cgatgctgta cttcttgtcg gctgctggga
     8881 ctccgtggat accgaccttc cgcttcttct ttggggccat cttatcgtca tcgtctttgt
     8941 aatcaatatc atgatccttg tagtctccgt cgtggtcctt atagtccatg aattccacca
     9001 cactggacta gtaggtaccc cgatccgggg tctctggatt agacgactgc tggctgatgg
     9061 agcggctttg tgtcgggagg agtatccaca cagcacaaga actcaaacgg tagtgatatg
     9121 aaaactggtc ccgcagcccg ttttataaag cccgcttagc gctaaaacct tatcagccgc
     9181 aatgctctcc gctctcaagt cgcgttcaaa acttttacca tatatgggta tgaattgttc
     9241 gccgattggg ttttcattta cggagaattt cctccgcaac tgggtgttcg tgcttgtgtg
     9301 tgtcttgtat atcgccatga agagtgtact tccatgggtc ctatcagttg cggcatgtga
     9361 ttcaatttgg gggttactca tattcgatac ttcagctggt tatctttatg ctgtgatggt
     9421 cacggtgctt ttacacattt tttccccaac tactcattgt atgccaaaac atacatatgt
     9481 acatgtgttt atagtgggtt tattagtaat gtccagcgat attaaaacat tacagttaaa
     9541 taatattcat attagcttac gtttactgct atctgcactt caaattggtt attccaggga
     9601 cattttgtgc aacacacatc ttctagtagg gcattcacta gtcgtagaaa acacacctaa
     9661 cttaagccgg ttttaacaat aacggcgaaa cagctgccag acgagagaaa agtacacagc
     9721 aagaagcatt tctataatga tgccgtccta aatatgcagc atttttgctc aaattgatca
     9781 tttcgttgca tcgctcaggt ggtatagtat ttggctattg aatgtgattg ctatcttaat
     9841 agttttgaat agtaatagtg caaatattca ttcataaaaa ataggtaatc gattttttca
     9901 gtgcagtcta tgcgtttgcg caacagagac gaagagagag acgcctcaca cacaagcaca
     9961 tgcggtctaa gctgctctcc ctctttttct ctctctctct ggcttgcggc agttttccag
    10021 aggctgtgac tcagtaagca tttcttgttg ttaccctaac gataagcgtc cgattcctaa
    10081 ttctctttca aataatttgt tttatttttt tttttgtgtg ttttgaagtc aaacagcttc
    10141 ataaacaata ttaaatatcg caattgttat tgcaaatgca ttcgtatcag gcgccttgat
    10201 gtttttcgcc aaaacttcgt ctgctctttg tcagcgtaat tgcgtttgcc ggtcgtgccg
    10261 tcttgctcgc acgcggcttg caaacaaaaa gtgaaagagg gaaagccacc tgtgttgttg
    10321 cttttgttgc tatcgttggc tgcacacaca tggcaataag taaccatatt gtagacttat
    10381 ttattttagc gttttcgttg actaatcttt gtttaaataa ttgcctattt cgagccaaaa
    10441 tgacgcataa aaggaaacgg caacatgcgg tgcgttgagc atgcgtaagc gagatggcgc
    10501 tagagagagc gcagcataaa aagaaaaaaa caaccggcga gtggcgagac acgaataata
    10561 cacacacgca cacatggtca aaggagtggc tccaacgtcg gtgacgtcgc gcccagccac
    10621 agaccccgcc acgcc
//
