LOCUS       sequence-369303-       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        72..176
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(177..245,246..1037)
                     /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(395..414)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1208..1796
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1697..1716
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    2397..2531
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2434..2453)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2488..2507
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(2539..3138)
                     /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(2622..2642)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     3119..3138
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     CDS             complement(3163..3216)
                     /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(3232..3279)
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             complement(3280..7380)
                     /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(3280..7380)
                     /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(7405..7425)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(7432..7497)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     promoter        9973..10372
                     /label=dU6-2 promoter
                     /note="RNA polymerase III promoter for Drosophila U6-2 
                     snRNA (Das et al., 1987)"
     misc_RNA        10394..10469
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(join(10621..10635,1..4))
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
       61 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
      121 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
      181 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
      241 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
      301 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
      361 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg
      421 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
      481 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
      541 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
      601 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
      661 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
      721 cagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
      781 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
      841 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
      901 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
      961 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     1021 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     1081 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     1141 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     1201 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     1261 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     1321 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
     1381 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     1441 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     1501 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     1561 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     1621 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     1681 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     1741 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     1801 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttgaagc tgtccctgat
     1861 ggtcgtcatc tacctgcctg gacagcatgg cctgcaacgc gggcatcccg atgccgccgg
     1921 aagcgagaag aatcataatg gggaaggcca tccagcctcg cgtcgcgaac gccagcaaga
     1981 cgtagcccag cgcgtcggcc gccatgccgg cgataatggc ctgcttctcg ccgaaacgtt
     2041 tggtggcggg accagtgacg aaggcttgag cgagggcgtg caagattccg aataccgcaa
     2101 gcgacaggcc gatcatcgtc gcgctccagc gaaagcggtc ctcgccgaaa atgacccaga
     2161 gcgctgccgg cacctgtcct acgagttgca tgataaagaa gacagtcata agtgcggcga
     2221 cgatagtcat gccccgcgcc caccggaagg agctgactgg gttgaaggct ctcaagggca
     2281 tcggtcgact gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa
     2341 acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact
     2401 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     2461 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     2521 atgtctggat ccctcgagtc aggcaccggg cttgcgggtc atgcaccagg tgcgcggtcc
     2581 ttcgggcacc tcgacgtcgg cggtgacggt gaagccgagc cgctcgtaga aggggaggtt
     2641 gcggggcgcg gaggtctcca ggaaggcggg caccccggcg cgctcggccg cctccactcc
     2701 ggggagcacg acggcgctgc ccagaccctt gccctggtgg tcgggcgaga cgccgacggt
     2761 ggccaggaac cacgcgggct ccttgggccg gtgcggcgcc aggaggcctt ccatctgttg
     2821 ctgcgcggcc agccgggaac cgctcaactc ggccatgcgc gggccgatct cggcgaacac
     2881 cgcccccgct tcgacgctct ccggcgtggt ccagaccgcc accgcggcgc cgtcgtccgc
     2941 gacccacacc ttgccgatgt cgagcccgac gcgcgtgagg aagagttctt gcagctcggt
     3001 gacccgctcg atgtggcggt ccgggtcgac ggtgtggcgc gtggcggggt agtcggcgaa
     3061 cgcggcggcg agggtgcgta cggcccgggg gacgtcgtcg cgggtggcga ggcgcaccgt
     3121 gggcttgtac tcggtcatgc taccgctgcc gctaccgcta gcagggccgg gattctcctc
     3181 cacgtcaccg catgttagaa gacttcctct gccctcaagc ttgcctgatc cctttttctt
     3241 ttttgcctgg ccggcctttt tcgtggccgc cggccttttg tcgcctccca gctgagacag
     3301 gtcgatccgt gtctcgtaca ggccggtgat gctctggtgg atcagggtgg cgtccagcac
     3361 ctctttggtg ctggtgtacc tcttccggtc gatggtggtg tcaaagtact tgaaggcggc
     3421 aggggctccc agattggtca gggtaaacag gtggatgata ttctcggcct gctctctgat
     3481 gggcttatcc cggtgcttgt tgtaggcgga cagcactttg tccagattag cgtcggccag
     3541 gatcactctc ttggagaact cgctgatctg ctcgatgatc tcgtccaggt agtgcttgtg
     3601 ctgttccaca aacagctgtt tctgctcatt atcctcgggg gagcccttca gcttctcata
     3661 gtggctggcc aggtacagga agttcacata tttggagggc agggccagtt cgtttccctt
     3721 ctgcagttcg ccggcagagg ccagcattct cttccggccg ttttccagct cgaacaggga
     3781 gtacttaggc agcttgatga tcaggtcctt tttcacttct ttgtagccct tggcttccag
     3841 aaagtcgatg ggattcttct cgaagctgct tctttccatg atggtgatcc ccagcagctc
     3901 tttcacactc ttcagtttct tggacttgcc cttttccact ttggccacca ccagcacaga
     3961 ataggccacg gtggggctgt cgaagccgcc gtacttctta gggtcccagt ccttctttct
     4021 ggcgatcagc ttatcgctgt tcctcttggg caggatagac tctttgctga agccgcctgt
     4081 ctgcacctcg gtctttttca cgatattcac ttggggcatg ctcagcactt tccgcacggt
     4141 ggcaaaatcc cggcccttat cccacacgat ctccccggtt tcgccgtttg tctcgatcag
     4201 aggccgcttc cggatctcgc cgttggccag ggtaatctcg gtcttgaaaa agttcatgat
     4261 gttgctgtag aagaagtact tggcggtagc cttgccgatt tcctgctcgc tcttggcgat
     4321 catcttccgc acgtcgtaca ccttgtagtc gccgtacacg aactcgcttt ccagcttagg
     4381 gtactttttg atcagggcgg ttcccacgac ggcgttcagg taggcgtcgt gggcgtggtg
     4441 gtagttgttg atctcgcgca ctttgtaaaa ctggaaatcc ttccggaaat cggacaccag
     4501 cttggacttc agggtgatca ctttcacttc ccggatcagc ttgtcattct cgtcgtactt
     4561 agtgttcatc cgggagtcca ggatctgtgc cacgtgcttt gtgatctgcc gggtttccac
     4621 cagctgtctc ttgatgaagc cggccttatc cagttcgctc aggccgcctc tctcggcctt
     4681 ggtcagattg tcgaactttc tctgggtaat cagcttggcg ttcagcagct gccgccagta
     4741 gttcttcatc ttcttcacga cctcttcgga gggcacgttg tcgctcttgc cccggttctt
     4801 gtcgcttctg gtcagcacct tgttgtcgat ggagtcgtcc ttcagaaagc tctgaggcac
     4861 gatatggtcc acatcgtagt cggacagccg gttgatgtcc agttcctggt ccacgtacat
     4921 atcccgccca ttctgcaggt agtacaggta cagcttctcg ttctgcagct gggtgttttc
     4981 cacggggtgt tctttcagga tctggctgcc cagctctttg atgccctctt cgatccgctt
     5041 cattctctcg cggctgttct tctgtccctt ctgggtggtc tggttctctc tggccatttc
     5101 gatcacgatg ttctcgggct tgtgccggcc catcactttc acgagctcgt ccaccacctt
     5161 cactgtctgc aggatgccct tcttaatggc ggggctgccg gccagattgg caatgtgctc
     5221 gtgcaggcta tcgccctggc cggacacctg ggctttctgg atgtcctctt taaaggtcag
     5281 gctgtcgtcg tggatcagct gcatgaagtt tctgttggcg aagccgtcgg acttcaggaa
     5341 atccaggatt gtcttgccgg actgcttgtc ccggatgccg ttgatcagct tccggctcag
     5401 cctgccccag ccggtgtatc tccgccgctt cagctgcttc atcactttgt cgtcgaacag
     5461 gtgggcatag gttttcagcc gttcctcgat catctctctg tcctcaaaca gtgtcagggt
     5521 cagcacgata tcttccagaa tgtcctcgtt ttcctcattg tccaggaagt ccttgtcctt
     5581 gataattttc agcagatcgt ggtatgtgcc cagggaggcg ttgaaccgat cttccacgcc
     5641 ggagatttcc acggagtcga agcactcgat tttcttgaag tagtcctctt tcagctgctt
     5701 cacggtcact ttccggttgg tcttgaacag caggtccacg atggcctttt tctgctcgcc
     5761 gctcaggaag gcgggctttc tcattccctc ggtcacgtat ttcactttgg tcagctcgtt
     5821 atacacggtg aagtactcgt acagcaggct gtgcttgggc agcaccttct cgttgggcag
     5881 gttcttatcg aagttggtca tccgctcgat gaagctctgg gcggaagcgc ccttgtccac
     5941 cacttcctcg aagttccagg gggtgatggt ttcctcgctc tttctggtca tccaggcgaa
     6001 tctgctgttt cccctggcca gagggcccac gtagtagggg atgcggaagg tcaggatctt
     6061 ctcgatcttt tcccggttgt ccttcaggaa tgggtaaaaa tcttcctgcc gccgcagaat
     6121 ggcgtgcagc tctcccaggt ggatctggtg ggggatgctg ccgttgtcga aggtccgctg
     6181 cttccgcagc aggtcctctc tgttcagctt cacgagcagt tcctcggtgc cgtccatctt
     6241 ttccaggatg ggcttgatga acttgtagaa ctcttcctgg ctggctccgc cgtcaatgta
     6301 gccggcgtag ccgttcttgc tctggtcgaa gaaaatctct ttgtacttct caggcagctg
     6361 ctgccgcacg agagctttca gcagggtcag gtcctggtgg tgctcgtcgt atctcttgat
     6421 catagaggcg ctcagggggg ccttggtgat ctcggtgttc actctcagga tgtcgctcag
     6481 caggatggcg tcggacaggt tcttggcggc cagaaacagg tcggcgtact ggtcgccgat
     6541 ctgggccagc aggttgtcca ggtcgtcgtc gtaggtgtcc ttgctcagct gcagtttggc
     6601 atcctcggcc aggtcgaagt tgctcttgaa gttgggggtc aggcccaggc tcagggcaat
     6661 caggtttccg aacaggccat tcttcttctc gccgggcagc tgggcgatca gattttccag
     6721 ccgtctgctc ttgctcagtc tggcagacag gatggccttg gcgtccacgc cgctggcgtt
     6781 gatggggttt tcctcgaaca gctggttgta ggtctgcacc agctggatga acagcttgtc
     6841 cacgtcgctg ttgtcggggt tcaggtcgcc ctcgatcagg aagtggcccc ggaacttgat
     6901 catgtgggcc agggccagat agatcagccg caggtcggcc ttgtcggtgc tgtccaccag
     6961 tttctttctc aggtggtaga tggtggggta cttctcgtgg taggccacct cgtccacgat
     7021 gttgccgaag atggggtgcc gctcgtgctt cttatcctct tccaccagga aggactcttc
     7081 cagtctgtgg aagaagctgt cgtccacctt ggccatctcg ttgctgaaga tctcttgcag
     7141 atagcagatc cggttcttcc gtctggtgta tcttcttctg gcggttctct tcagccgggt
     7201 ggcctcggct gtttcgccgc tgtcgaacag cagggctccg atcaggttct tcttgatgct
     7261 gtgccggtcg gtgttgccca gcaccttgaa tttcttgctg ggcaccttgt actcgtcggt
     7321 gatcacggcc cagcccacag agttggtgcc gatgtccagg ccgatgctgt acttcttgtc
     7381 ggctgctggg actccgtgga taccgacctt ccgcttcttc tttggggcca tcttatcgtc
     7441 atcgtctttg taatcaatat catgatcctt gtagtctccg tcgtggtcct tatagtccat
     7501 gaattccacc acactggact agtaggtacc ccgatccggg gtctctggat tagacgactg
     7561 ctggctgatg gagcggcttt gtgtcgggag gagtatccac acagcacaag aactcaaacg
     7621 gtagtgatat gaaaactggt cccgcagccc gttttataaa gcccgcttag cgctaaaacc
     7681 ttatcagccg caatgctctc cgctctcaag tcgcgttcaa aacttttacc atatatgggt
     7741 atgaattgtt cgccgattgg gttttcattt acggagaatt tcctccgcaa ctgggtgttc
     7801 gtgcttgtgt gtgtcttgta tatcgccatg aagagtgtac ttccatgggt cctatcagtt
     7861 gcggcatgtg attcaatttg ggggttactc atattcgata cttcagctgg ttatctttat
     7921 gctgtgatgg tcacggtgct tttacacatt ttttccccaa ctactcattg tatgccaaaa
     7981 catacatatg tacatgtgtt tatagtgggt ttattagtaa tgtccagcga tattaaaaca
     8041 ttacagttaa ataatattca tattagctta cgtttactgc tatctgcact tcaaattggt
     8101 tattccaggg acattttgtg caacacacat cttctagtag ggcattcact agtcgtagaa
     8161 aacacaccta acttaagccg gttttaacaa taacggcgaa acagctgcca gacgagagaa
     8221 aagtacacag caagaagcat ttctataatg atgccgtcct aaatatgcag catttttgct
     8281 caaattgatc atttcgttgc atcgctcagg tggtatagta tttggctatt gaatgtgatt
     8341 gctatcttaa tagttttgaa tagtaatagt gcaaatattc attcataaaa aataggtaat
     8401 cgattttttc agtgcagtct atgcgtttgc gcaacagaga cgaagagaga gacgcctcac
     8461 acacaagcac atgcggtcta agctgctctc cctctttttc tctctctctc tggcttgcgg
     8521 cagttttcca gaggctgtga ctcagtaagc atttcttgtt gttaccctaa cgataagcgt
     8581 ccgattccta attctctttc aaataatttg ttttattttt ttttttgtgt gttttgaagt
     8641 caaacagctt cataaacaat attaaatatc gcaattgtta ttgcaaatgc attcgtatca
     8701 ggcgccttga tgtttttcgc caaaacttcg tctgctcttt gtcagcgtaa ttgcgtttgc
     8761 cggtcgtgcc gtcttgctcg cacgcggctt gcaaacaaaa agtgaaagag ggaaagccac
     8821 ctgtgttgtt gcttttgttg ctatcgttgg ctgcacacac atggcaataa gtaaccatat
     8881 tgtagactta tttattttag cgttttcgtt gactaatctt tgtttaaata attgcctatt
     8941 tcgagccaaa atgacgcata aaaggaaacg gcaacatgcg gtgcgttgag catgcgtaag
     9001 cgagatggcg ctagagagag cgcagcataa aaagaaaaaa acaaccggcg agtggcgaga
     9061 cacgaataat acacacacgc acacatggtc aaaggagtgg ctccaacgtc ggtgacgtcg
     9121 cgcccagcca cagaccccgc cacgccccgc cccacccttc cttccgctgc cccgcacccc
     9181 tttccttcca ctaccgtttt gccgcatttg gtattgccat ctcgcttttt tggctttact
     9241 ttgctttttg aataagccaa gaaactgaaa tcaaagcgga cagtccgaaa attgaatatt
     9301 ccaaaaaaca acaattgcac atgcactgcg ttcggcaaaa gacgcagaaa ttcaagcctt
     9361 ttggtctggc atcatcttaa aatatcatca gtatcatcat tgccattata aatacataag
     9421 aaaatgagaa tacattaaag atatacatac aatctttgtc atagtcaaca tattttgcaa
     9481 caaaattatc tatagataac atgtttttgt agaaaattcc actaactgat tttcgtataa
     9541 atacaaatta ttctcattct tcaaaatact cgccgacgtc atcgcgaaat tcctcagcat
     9601 tcttcgacaa atgaagagag acagcgagtg acacaatctt atcttgccga cgagtctttg
     9661 aaaatggccc cgggtaacgg tgaaaaacat atagccaaga atggagcgat cgccaggcga
     9721 tggaaattgg cttcaaattg ctccgaatat ggccagttgg tcagttggca ttggattgtg
     9781 gagagatact agatcagctc gttctccttc tcctccgcat tctcctactc cgcccgaaga
     9841 cgcttctttg atcgcttctt tctcgaatgt gtacatattc attatgtatt tggtgtttgg
     9901 cgacgatcag ccagctttaa tggccataaa acgtgtcttg gtttgtgggc cacataacag
     9961 gcagcgagat ctgttcgact tgcagcctga aatacggcac gagtaggaaa agccgagtca
    10021 aatgccgaat gcagagtctc attacagcac aatcaactca agaaaaactc gacacttttt
    10081 taccatttgc acttaaatcc ttttttattc gttatgtata ctttttttgg tccctaacca
    10141 aaacaaaacc aaactctctt agtcgtgcct ctatatttaa aactatcaat ttattatagt
    10201 caataaatcg aactgtgttt tcaacaaacg aacaatagga cactttgatt ctaaaggaaa
    10261 ttttgaaaat cttaagcaga gggttcttaa gaccatttgc caattcttat aattctcaac
    10321 tgctctttcc tgatgttgat catttatata ggtatgtttt cctcaatact tcgcgctgga
    10381 ggcttaggag tgcgttttag agctagaaat agcaagttaa aataaggcta gtccgttatc
    10441 aacttgaaaa agtggcaccg agtcggtgct tttttagatc tggaaaaatg atgtgacagt
    10501 ggaaatgaga agtaggtgca tctgcaaaga tttgattcag agttgatgcc attcatgatt
    10561 tttttttaga agtatcattt gtgtttttag aatatagaat taattcttga agacgaaagg
    10621 gcctcgtgat acgcc
//
