LOCUS       sequence-373455-       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_2
FEATURES             Location/Qualifiers
     source          1..10635
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        813..1212
                     /label=dU6-2 promoter
                     /note="RNA polymerase III promoter for Drosophila U6-2 
                     snRNA (Das et al., 1987)"
     misc_RNA        1234..1309
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(1461..1479)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1547..1651
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1652..1720,1721..2512)
                     /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(1870..1889)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2683..3271
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3172..3191
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3872..4006
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3909..3928)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3963..3982
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(4014..4613)
                     /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(4097..4117)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     4594..4613
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     CDS             complement(4638..4691)
                     /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(4707..4754)
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             complement(4755..8855)
                     /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(4755..8855)
                     /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(8880..8900)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(8907..8972)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
ORIGIN
        1 cttccgctgc cccgcacccc tttccttcca ctaccgtttt gccgcatttg gtattgccat
       61 ctcgcttttt tggctttact ttgctttttg aataagccaa gaaactgaaa tcaaagcgga
      121 cagtccgaaa attgaatatt ccaaaaaaca acaattgcac atgcactgcg ttcggcaaaa
      181 gacgcagaaa ttcaagcctt ttggtctggc atcatcttaa aatatcatca gtatcatcat
      241 tgccattata aatacataag aaaatgagaa tacattaaag atatacatac aatctttgtc
      301 atagtcaaca tattttgcaa caaaattatc tatagataac atgtttttgt agaaaattcc
      361 actaactgat tttcgtataa atacaaatta ttctcattct tcaaaatact cgccgacgtc
      421 atcgcgaaat tcctcagcat tcttcgacaa atgaagagag acagcgagtg acacaatctt
      481 atcttgccga cgagtctttg aaaatggccc cgggtaacgg tgaaaaacat atagccaaga
      541 atggagcgat cgccaggcga tggaaattgg cttcaaattg ctccgaatat ggccagttgg
      601 tcagttggca ttggattgtg gagagatact agatcagctc gttctccttc tcctccgcat
      661 tctcctactc cgcccgaaga cgcttctttg atcgcttctt tctcgaatgt gtacatattc
      721 attatgtatt tggtgtttgg cgacgatcag ccagctttaa tggccataaa acgtgtcttg
      781 gtttgtgggc cacataacag gcagcgagat ctgttcgact tgcagcctga aatacggcac
      841 gagtaggaaa agccgagtca aatgccgaat gcagagtctc attacagcac aatcaactca
      901 agaaaaactc gacacttttt taccatttgc acttaaatcc ttttttattc gttatgtata
      961 ctttttttgg tccctaacca aaacaaaacc aaactctctt agtcgtgcct ctatatttaa
     1021 aactatcaat ttattatagt caataaatcg aactgtgttt tcaacaaacg aacaatagga
     1081 cactttgatt ctaaaggaaa ttttgaaaat cttaagcaga gggttcttaa gaccatttgc
     1141 caattcttat aattctcaac tgctctttcc tgatgttgat catttatata ggtatgtttt
     1201 cctcaatact tcgcagctcg aacggtatgt agggttttag agctagaaat agcaagttaa
     1261 aataaggcta gtccgttatc aacttgaaaa agtggcaccg agtcggtgct tttttagatc
     1321 tggaaaaatg atgtgacagt ggaaatgaga agtaggtgca tctgcaaaga tttgattcag
     1381 agttgatgcc attcatgatt tttttttaga agtatcattt gtgtttttag aatatagaat
     1441 taattcttga agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat
     1501 aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     1561 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     1621 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     1681 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     1741 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     1801 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     1861 taaagttctg ctatgtggcg cggtattatc ccgtgttgac gccgggcaag agcaactcgg
     1921 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     1981 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     2041 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     2101 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     2161 cataccaaac gacgagcgtg acaccacgat gcctgcagca atggcaacaa cgttgcgcaa
     2221 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     2281 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     2341 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     2401 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     2461 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     2521 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     2581 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     2641 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     2701 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     2761 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     2821 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     2881 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     2941 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     3001 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     3061 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     3121 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     3181 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     3241 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     3301 cctggccttt tgctggcctt gaagctgtcc ctgatggtcg tcatctacct gcctggacag
     3361 catggcctgc aacgcgggca tcccgatgcc gccggaagcg agaagaatca taatggggaa
     3421 ggccatccag cctcgcgtcg cgaacgccag caagacgtag cccagcgcgt cggccgccat
     3481 gccggcgata atggcctgct tctcgccgaa acgtttggtg gcgggaccag tgacgaaggc
     3541 ttgagcgagg gcgtgcaaga ttccgaatac cgcaagcgac aggccgatca tcgtcgcgct
     3601 ccagcgaaag cggtcctcgc cgaaaatgac ccagagcgct gccggcacct gtcctacgag
     3661 ttgcatgata aagaagacag tcataagtgc ggcgacgata gtcatgcccc gcgcccaccg
     3721 gaaggagctg actgggttga aggctctcaa gggcatcggt cgactgatca taatcagcca
     3781 taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct
     3841 gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta
     3901 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     3961 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatccctc gagtcaggca
     4021 ccgggcttgc gggtcatgca ccaggtgcgc ggtccttcgg gcacctcgac gtcggcggtg
     4081 acggtgaagc cgagccgctc gtagaagggg aggttgcggg gcgcggaggt ctccaggaag
     4141 gcgggcaccc cggcgcgctc ggccgcctcc actccgggga gcacgacggc gctgcccaga
     4201 cccttgccct ggtggtcggg cgagacgccg acggtggcca ggaaccacgc gggctccttg
     4261 ggccggtgcg gcgccaggag gccttccatc tgttgctgcg cggccagccg ggaaccgctc
     4321 aactcggcca tgcgcgggcc gatctcggcg aacaccgccc ccgcttcgac gctctccggc
     4381 gtggtccaga ccgccaccgc ggcgccgtcg tccgcgaccc acaccttgcc gatgtcgagc
     4441 ccgacgcgcg tgaggaagag ttcttgcagc tcggtgaccc gctcgatgtg gcggtccggg
     4501 tcgacggtgt ggcgcgtggc ggggtagtcg gcgaacgcgg cggcgagggt gcgtacggcc
     4561 cgggggacgt cgtcgcgggt ggcgaggcgc accgtgggct tgtactcggt catgctaccg
     4621 ctgccgctac cgctagcagg gccgggattc tcctccacgt caccgcatgt tagaagactt
     4681 cctctgccct caagcttgcc tgatcccttt ttcttttttg cctggccggc ctttttcgtg
     4741 gccgccggcc ttttgtcgcc tcccagctga gacaggtcga tccgtgtctc gtacaggccg
     4801 gtgatgctct ggtggatcag ggtggcgtcc agcacctctt tggtgctggt gtacctcttc
     4861 cggtcgatgg tggtgtcaaa gtacttgaag gcggcagggg ctcccagatt ggtcagggta
     4921 aacaggtgga tgatattctc ggcctgctct ctgatgggct tatcccggtg cttgttgtag
     4981 gcggacagca ctttgtccag attagcgtcg gccaggatca ctctcttgga gaactcgctg
     5041 atctgctcga tgatctcgtc caggtagtgc ttgtgctgtt ccacaaacag ctgtttctgc
     5101 tcattatcct cgggggagcc cttcagcttc tcatagtggc tggccaggta caggaagttc
     5161 acatatttgg agggcagggc cagttcgttt cccttctgca gttcgccggc agaggccagc
     5221 attctcttcc ggccgttttc cagctcgaac agggagtact taggcagctt gatgatcagg
     5281 tcctttttca cttctttgta gcccttggct tccagaaagt cgatgggatt cttctcgaag
     5341 ctgcttcttt ccatgatggt gatccccagc agctctttca cactcttcag tttcttggac
     5401 ttgccctttt ccactttggc caccaccagc acagaatagg ccacggtggg gctgtcgaag
     5461 ccgccgtact tcttagggtc ccagtccttc tttctggcga tcagcttatc gctgttcctc
     5521 ttgggcagga tagactcttt gctgaagccg cctgtctgca cctcggtctt tttcacgata
     5581 ttcacttggg gcatgctcag cactttccgc acggtggcaa aatcccggcc cttatcccac
     5641 acgatctccc cggtttcgcc gtttgtctcg atcagaggcc gcttccggat ctcgccgttg
     5701 gccagggtaa tctcggtctt gaaaaagttc atgatgttgc tgtagaagaa gtacttggcg
     5761 gtagccttgc cgatttcctg ctcgctcttg gcgatcatct tccgcacgtc gtacaccttg
     5821 tagtcgccgt acacgaactc gctttccagc ttagggtact ttttgatcag ggcggttccc
     5881 acgacggcgt tcaggtaggc gtcgtgggcg tggtggtagt tgttgatctc gcgcactttg
     5941 taaaactgga aatccttccg gaaatcggac accagcttgg acttcagggt gatcactttc
     6001 acttcccgga tcagcttgtc attctcgtcg tacttagtgt tcatccggga gtccaggatc
     6061 tgtgccacgt gctttgtgat ctgccgggtt tccaccagct gtctcttgat gaagccggcc
     6121 ttatccagtt cgctcaggcc gcctctctcg gccttggtca gattgtcgaa ctttctctgg
     6181 gtaatcagct tggcgttcag cagctgccgc cagtagttct tcatcttctt cacgacctct
     6241 tcggagggca cgttgtcgct cttgccccgg ttcttgtcgc ttctggtcag caccttgttg
     6301 tcgatggagt cgtccttcag aaagctctga ggcacgatat ggtccacatc gtagtcggac
     6361 agccggttga tgtccagttc ctggtccacg tacatatccc gcccattctg caggtagtac
     6421 aggtacagct tctcgttctg cagctgggtg ttttccacgg ggtgttcttt caggatctgg
     6481 ctgcccagct ctttgatgcc ctcttcgatc cgcttcattc tctcgcggct gttcttctgt
     6541 cccttctggg tggtctggtt ctctctggcc atttcgatca cgatgttctc gggcttgtgc
     6601 cggcccatca ctttcacgag ctcgtccacc accttcactg tctgcaggat gcccttctta
     6661 atggcggggc tgccggccag attggcaatg tgctcgtgca ggctatcgcc ctggccggac
     6721 acctgggctt tctggatgtc ctctttaaag gtcaggctgt cgtcgtggat cagctgcatg
     6781 aagtttctgt tggcgaagcc gtcggacttc aggaaatcca ggattgtctt gccggactgc
     6841 ttgtcccgga tgccgttgat cagcttccgg ctcagcctgc cccagccggt gtatctccgc
     6901 cgcttcagct gcttcatcac tttgtcgtcg aacaggtggg cataggtttt cagccgttcc
     6961 tcgatcatct ctctgtcctc aaacagtgtc agggtcagca cgatatcttc cagaatgtcc
     7021 tcgttttcct cattgtccag gaagtccttg tccttgataa ttttcagcag atcgtggtat
     7081 gtgcccaggg aggcgttgaa ccgatcttcc acgccggaga tttccacgga gtcgaagcac
     7141 tcgattttct tgaagtagtc ctctttcagc tgcttcacgg tcactttccg gttggtcttg
     7201 aacagcaggt ccacgatggc ctttttctgc tcgccgctca ggaaggcggg ctttctcatt
     7261 ccctcggtca cgtatttcac tttggtcagc tcgttataca cggtgaagta ctcgtacagc
     7321 aggctgtgct tgggcagcac cttctcgttg ggcaggttct tatcgaagtt ggtcatccgc
     7381 tcgatgaagc tctgggcgga agcgcccttg tccaccactt cctcgaagtt ccagggggtg
     7441 atggtttcct cgctctttct ggtcatccag gcgaatctgc tgtttcccct ggccagaggg
     7501 cccacgtagt aggggatgcg gaaggtcagg atcttctcga tcttttcccg gttgtccttc
     7561 aggaatgggt aaaaatcttc ctgccgccgc agaatggcgt gcagctctcc caggtggatc
     7621 tggtggggga tgctgccgtt gtcgaaggtc cgctgcttcc gcagcaggtc ctctctgttc
     7681 agcttcacga gcagttcctc ggtgccgtcc atcttttcca ggatgggctt gatgaacttg
     7741 tagaactctt cctggctggc tccgccgtca atgtagccgg cgtagccgtt cttgctctgg
     7801 tcgaagaaaa tctctttgta cttctcaggc agctgctgcc gcacgagagc tttcagcagg
     7861 gtcaggtcct ggtggtgctc gtcgtatctc ttgatcatag aggcgctcag gggggccttg
     7921 gtgatctcgg tgttcactct caggatgtcg ctcagcagga tggcgtcgga caggttcttg
     7981 gcggccagaa acaggtcggc gtactggtcg ccgatctggg ccagcaggtt gtccaggtcg
     8041 tcgtcgtagg tgtccttgct cagctgcagt ttggcatcct cggccaggtc gaagttgctc
     8101 ttgaagttgg gggtcaggcc caggctcagg gcaatcaggt ttccgaacag gccattcttc
     8161 ttctcgccgg gcagctgggc gatcagattt tccagccgtc tgctcttgct cagtctggca
     8221 gacaggatgg ccttggcgtc cacgccgctg gcgttgatgg ggttttcctc gaacagctgg
     8281 ttgtaggtct gcaccagctg gatgaacagc ttgtccacgt cgctgttgtc ggggttcagg
     8341 tcgccctcga tcaggaagtg gccccggaac ttgatcatgt gggccagggc cagatagatc
     8401 agccgcaggt cggccttgtc ggtgctgtcc accagtttct ttctcaggtg gtagatggtg
     8461 gggtacttct cgtggtaggc cacctcgtcc acgatgttgc cgaagatggg gtgccgctcg
     8521 tgcttcttat cctcttccac caggaaggac tcttccagtc tgtggaagaa gctgtcgtcc
     8581 accttggcca tctcgttgct gaagatctct tgcagatagc agatccggtt cttccgtctg
     8641 gtgtatcttc ttctggcggt tctcttcagc cgggtggcct cggctgtttc gccgctgtcg
     8701 aacagcaggg ctccgatcag gttcttcttg atgctgtgcc ggtcggtgtt gcccagcacc
     8761 ttgaatttct tgctgggcac cttgtactcg tcggtgatca cggcccagcc cacagagttg
     8821 gtgccgatgt ccaggccgat gctgtacttc ttgtcggctg ctgggactcc gtggataccg
     8881 accttccgct tcttctttgg ggccatctta tcgtcatcgt ctttgtaatc aatatcatga
     8941 tccttgtagt ctccgtcgtg gtccttatag tccatgaatt ccaccacact ggactagtag
     9001 gtaccccgat ccggggtctc tggattagac gactgctggc tgatggagcg gctttgtgtc
     9061 gggaggagta tccacacagc acaagaactc aaacggtagt gatatgaaaa ctggtcccgc
     9121 agcccgtttt ataaagcccg cttagcgcta aaaccttatc agccgcaatg ctctccgctc
     9181 tcaagtcgcg ttcaaaactt ttaccatata tgggtatgaa ttgttcgccg attgggtttt
     9241 catttacgga gaatttcctc cgcaactggg tgttcgtgct tgtgtgtgtc ttgtatatcg
     9301 ccatgaagag tgtacttcca tgggtcctat cagttgcggc atgtgattca atttgggggt
     9361 tactcatatt cgatacttca gctggttatc tttatgctgt gatggtcacg gtgcttttac
     9421 acattttttc cccaactact cattgtatgc caaaacatac atatgtacat gtgtttatag
     9481 tgggtttatt agtaatgtcc agcgatatta aaacattaca gttaaataat attcatatta
     9541 gcttacgttt actgctatct gcacttcaaa ttggttattc cagggacatt ttgtgcaaca
     9601 cacatcttct agtagggcat tcactagtcg tagaaaacac acctaactta agccggtttt
     9661 aacaataacg gcgaaacagc tgccagacga gagaaaagta cacagcaaga agcatttcta
     9721 taatgatgcc gtcctaaata tgcagcattt ttgctcaaat tgatcatttc gttgcatcgc
     9781 tcaggtggta tagtatttgg ctattgaatg tgattgctat cttaatagtt ttgaatagta
     9841 atagtgcaaa tattcattca taaaaaatag gtaatcgatt ttttcagtgc agtctatgcg
     9901 tttgcgcaac agagacgaag agagagacgc ctcacacaca agcacatgcg gtctaagctg
     9961 ctctccctct ttttctctct ctctctggct tgcggcagtt ttccagaggc tgtgactcag
    10021 taagcatttc ttgttgttac cctaacgata agcgtccgat tcctaattct ctttcaaata
    10081 atttgtttta tttttttttt tgtgtgtttt gaagtcaaac agcttcataa acaatattaa
    10141 atatcgcaat tgttattgca aatgcattcg tatcaggcgc cttgatgttt ttcgccaaaa
    10201 cttcgtctgc tctttgtcag cgtaattgcg tttgccggtc gtgccgtctt gctcgcacgc
    10261 ggcttgcaaa caaaaagtga aagagggaaa gccacctgtg ttgttgcttt tgttgctatc
    10321 gttggctgca cacacatggc aataagtaac catattgtag acttatttat tttagcgttt
    10381 tcgttgacta atctttgttt aaataattgc ctatttcgag ccaaaatgac gcataaaagg
    10441 aaacggcaac atgcggtgcg ttgagcatgc gtaagcgaga tggcgctaga gagagcgcag
    10501 cataaaaaga aaaaaacaac cggcgagtgg cgagacacga ataatacaca cacgcacaca
    10561 tggtcaaagg agtggctcca acgtcggtga cgtcgcgccc agccacagac cccgccacgc
    10621 cccgccccac ccttc
//
