LOCUS       sequence-373334-       10635 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses scrambled sgRNA 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.scr_2
FEATURES             Location/Qualifiers
     source          1..10635
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(89..107)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        175..279
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(280..348,349..1140)
                     /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(498..517)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1311..1899
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1800..1819
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    2500..2634
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2537..2556)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2591..2610
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(2642..3241)
                     /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(2725..2745)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     3222..3241
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     CDS             complement(3266..3319)
                     /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(3335..3382)
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             complement(3383..7483)
                     /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(3383..7483)
                     /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(7508..7528)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(7535..7600)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     promoter        10076..10475
                     /label=dU6-2 promoter
                     /note="RNA polymerase III promoter for Drosophila U6-2 
                     snRNA (Das et al., 1987)"
     misc_RNA        10497..10572
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
ORIGIN
        1 tgattcagag ttgatgccat tcatgatttt tttttagaag tatcatttgt gtttttagaa
       61 tatagaatta attcttgaag acgaaagggc ctcgtgatac gcctattttt ataggttaat
      121 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
      181 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
      241 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
      301 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
      361 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
      421 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
      481 agcactttta aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag
      541 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
      601 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
      661 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
      721 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
      781 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg
      841 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
      901 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
      961 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     1021 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     1081 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     1141 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     1201 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     1261 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     1321 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     1381 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     1441 cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct
     1501 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     1561 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     1621 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     1681 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     1741 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     1801 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     1861 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     1921 ttacggttcc tggccttttg ctggccttga agctgtccct gatggtcgtc atctacctgc
     1981 ctggacagca tggcctgcaa cgcgggcatc ccgatgccgc cggaagcgag aagaatcata
     2041 atggggaagg ccatccagcc tcgcgtcgcg aacgccagca agacgtagcc cagcgcgtcg
     2101 gccgccatgc cggcgataat ggcctgcttc tcgccgaaac gtttggtggc gggaccagtg
     2161 acgaaggctt gagcgagggc gtgcaagatt ccgaataccg caagcgacag gccgatcatc
     2221 gtcgcgctcc agcgaaagcg gtcctcgccg aaaatgaccc agagcgctgc cggcacctgt
     2281 cctacgagtt gcatgataaa gaagacagtc ataagtgcgg cgacgatagt catgccccgc
     2341 gcccaccgga aggagctgac tgggttgaag gctctcaagg gcatcggtcg actgatcata
     2401 atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc
     2461 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat
     2521 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     2581 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatccctcga
     2641 gtcaggcacc gggcttgcgg gtcatgcacc aggtgcgcgg tccttcgggc acctcgacgt
     2701 cggcggtgac ggtgaagccg agccgctcgt agaaggggag gttgcggggc gcggaggtct
     2761 ccaggaaggc gggcaccccg gcgcgctcgg ccgcctccac tccggggagc acgacggcgc
     2821 tgcccagacc cttgccctgg tggtcgggcg agacgccgac ggtggccagg aaccacgcgg
     2881 gctccttggg ccggtgcggc gccaggaggc cttccatctg ttgctgcgcg gccagccggg
     2941 aaccgctcaa ctcggccatg cgcgggccga tctcggcgaa caccgccccc gcttcgacgc
     3001 tctccggcgt ggtccagacc gccaccgcgg cgccgtcgtc cgcgacccac accttgccga
     3061 tgtcgagccc gacgcgcgtg aggaagagtt cttgcagctc ggtgacccgc tcgatgtggc
     3121 ggtccgggtc gacggtgtgg cgcgtggcgg ggtagtcggc gaacgcggcg gcgagggtgc
     3181 gtacggcccg ggggacgtcg tcgcgggtgg cgaggcgcac cgtgggcttg tactcggtca
     3241 tgctaccgct gccgctaccg ctagcagggc cgggattctc ctccacgtca ccgcatgtta
     3301 gaagacttcc tctgccctca agcttgcctg atcccttttt cttttttgcc tggccggcct
     3361 ttttcgtggc cgccggcctt ttgtcgcctc ccagctgaga caggtcgatc cgtgtctcgt
     3421 acaggccggt gatgctctgg tggatcaggg tggcgtccag cacctctttg gtgctggtgt
     3481 acctcttccg gtcgatggtg gtgtcaaagt acttgaaggc ggcaggggct cccagattgg
     3541 tcagggtaaa caggtggatg atattctcgg cctgctctct gatgggctta tcccggtgct
     3601 tgttgtaggc ggacagcact ttgtccagat tagcgtcggc caggatcact ctcttggaga
     3661 actcgctgat ctgctcgatg atctcgtcca ggtagtgctt gtgctgttcc acaaacagct
     3721 gtttctgctc attatcctcg ggggagccct tcagcttctc atagtggctg gccaggtaca
     3781 ggaagttcac atatttggag ggcagggcca gttcgtttcc cttctgcagt tcgccggcag
     3841 aggccagcat tctcttccgg ccgttttcca gctcgaacag ggagtactta ggcagcttga
     3901 tgatcaggtc ctttttcact tctttgtagc ccttggcttc cagaaagtcg atgggattct
     3961 tctcgaagct gcttctttcc atgatggtga tccccagcag ctctttcaca ctcttcagtt
     4021 tcttggactt gcccttttcc actttggcca ccaccagcac agaataggcc acggtggggc
     4081 tgtcgaagcc gccgtacttc ttagggtccc agtccttctt tctggcgatc agcttatcgc
     4141 tgttcctctt gggcaggata gactctttgc tgaagccgcc tgtctgcacc tcggtctttt
     4201 tcacgatatt cacttggggc atgctcagca ctttccgcac ggtggcaaaa tcccggccct
     4261 tatcccacac gatctccccg gtttcgccgt ttgtctcgat cagaggccgc ttccggatct
     4321 cgccgttggc cagggtaatc tcggtcttga aaaagttcat gatgttgctg tagaagaagt
     4381 acttggcggt agccttgccg atttcctgct cgctcttggc gatcatcttc cgcacgtcgt
     4441 acaccttgta gtcgccgtac acgaactcgc tttccagctt agggtacttt ttgatcaggg
     4501 cggttcccac gacggcgttc aggtaggcgt cgtgggcgtg gtggtagttg ttgatctcgc
     4561 gcactttgta aaactggaaa tccttccgga aatcggacac cagcttggac ttcagggtga
     4621 tcactttcac ttcccggatc agcttgtcat tctcgtcgta cttagtgttc atccgggagt
     4681 ccaggatctg tgccacgtgc tttgtgatct gccgggtttc caccagctgt ctcttgatga
     4741 agccggcctt atccagttcg ctcaggccgc ctctctcggc cttggtcaga ttgtcgaact
     4801 ttctctgggt aatcagcttg gcgttcagca gctgccgcca gtagttcttc atcttcttca
     4861 cgacctcttc ggagggcacg ttgtcgctct tgccccggtt cttgtcgctt ctggtcagca
     4921 ccttgttgtc gatggagtcg tccttcagaa agctctgagg cacgatatgg tccacatcgt
     4981 agtcggacag ccggttgatg tccagttcct ggtccacgta catatcccgc ccattctgca
     5041 ggtagtacag gtacagcttc tcgttctgca gctgggtgtt ttccacgggg tgttctttca
     5101 ggatctggct gcccagctct ttgatgccct cttcgatccg cttcattctc tcgcggctgt
     5161 tcttctgtcc cttctgggtg gtctggttct ctctggccat ttcgatcacg atgttctcgg
     5221 gcttgtgccg gcccatcact ttcacgagct cgtccaccac cttcactgtc tgcaggatgc
     5281 ccttcttaat ggcggggctg ccggccagat tggcaatgtg ctcgtgcagg ctatcgccct
     5341 ggccggacac ctgggctttc tggatgtcct ctttaaaggt caggctgtcg tcgtggatca
     5401 gctgcatgaa gtttctgttg gcgaagccgt cggacttcag gaaatccagg attgtcttgc
     5461 cggactgctt gtcccggatg ccgttgatca gcttccggct cagcctgccc cagccggtgt
     5521 atctccgccg cttcagctgc ttcatcactt tgtcgtcgaa caggtgggca taggttttca
     5581 gccgttcctc gatcatctct ctgtcctcaa acagtgtcag ggtcagcacg atatcttcca
     5641 gaatgtcctc gttttcctca ttgtccagga agtccttgtc cttgataatt ttcagcagat
     5701 cgtggtatgt gcccagggag gcgttgaacc gatcttccac gccggagatt tccacggagt
     5761 cgaagcactc gattttcttg aagtagtcct ctttcagctg cttcacggtc actttccggt
     5821 tggtcttgaa cagcaggtcc acgatggcct ttttctgctc gccgctcagg aaggcgggct
     5881 ttctcattcc ctcggtcacg tatttcactt tggtcagctc gttatacacg gtgaagtact
     5941 cgtacagcag gctgtgcttg ggcagcacct tctcgttggg caggttctta tcgaagttgg
     6001 tcatccgctc gatgaagctc tgggcggaag cgcccttgtc caccacttcc tcgaagttcc
     6061 agggggtgat ggtttcctcg ctctttctgg tcatccaggc gaatctgctg tttcccctgg
     6121 ccagagggcc cacgtagtag gggatgcgga aggtcaggat cttctcgatc ttttcccggt
     6181 tgtccttcag gaatgggtaa aaatcttcct gccgccgcag aatggcgtgc agctctccca
     6241 ggtggatctg gtgggggatg ctgccgttgt cgaaggtccg ctgcttccgc agcaggtcct
     6301 ctctgttcag cttcacgagc agttcctcgg tgccgtccat cttttccagg atgggcttga
     6361 tgaacttgta gaactcttcc tggctggctc cgccgtcaat gtagccggcg tagccgttct
     6421 tgctctggtc gaagaaaatc tctttgtact tctcaggcag ctgctgccgc acgagagctt
     6481 tcagcagggt caggtcctgg tggtgctcgt cgtatctctt gatcatagag gcgctcaggg
     6541 gggccttggt gatctcggtg ttcactctca ggatgtcgct cagcaggatg gcgtcggaca
     6601 ggttcttggc ggccagaaac aggtcggcgt actggtcgcc gatctgggcc agcaggttgt
     6661 ccaggtcgtc gtcgtaggtg tccttgctca gctgcagttt ggcatcctcg gccaggtcga
     6721 agttgctctt gaagttgggg gtcaggccca ggctcagggc aatcaggttt ccgaacaggc
     6781 cattcttctt ctcgccgggc agctgggcga tcagattttc cagccgtctg ctcttgctca
     6841 gtctggcaga caggatggcc ttggcgtcca cgccgctggc gttgatgggg ttttcctcga
     6901 acagctggtt gtaggtctgc accagctgga tgaacagctt gtccacgtcg ctgttgtcgg
     6961 ggttcaggtc gccctcgatc aggaagtggc cccggaactt gatcatgtgg gccagggcca
     7021 gatagatcag ccgcaggtcg gccttgtcgg tgctgtccac cagtttcttt ctcaggtggt
     7081 agatggtggg gtacttctcg tggtaggcca cctcgtccac gatgttgccg aagatggggt
     7141 gccgctcgtg cttcttatcc tcttccacca ggaaggactc ttccagtctg tggaagaagc
     7201 tgtcgtccac cttggccatc tcgttgctga agatctcttg cagatagcag atccggttct
     7261 tccgtctggt gtatcttctt ctggcggttc tcttcagccg ggtggcctcg gctgtttcgc
     7321 cgctgtcgaa cagcagggct ccgatcaggt tcttcttgat gctgtgccgg tcggtgttgc
     7381 ccagcacctt gaatttcttg ctgggcacct tgtactcgtc ggtgatcacg gcccagccca
     7441 cagagttggt gccgatgtcc aggccgatgc tgtacttctt gtcggctgct gggactccgt
     7501 ggataccgac cttccgcttc ttctttgggg ccatcttatc gtcatcgtct ttgtaatcaa
     7561 tatcatgatc cttgtagtct ccgtcgtggt ccttatagtc catgaattcc accacactgg
     7621 actagtaggt accccgatcc ggggtctctg gattagacga ctgctggctg atggagcggc
     7681 tttgtgtcgg gaggagtatc cacacagcac aagaactcaa acggtagtga tatgaaaact
     7741 ggtcccgcag cccgttttat aaagcccgct tagcgctaaa accttatcag ccgcaatgct
     7801 ctccgctctc aagtcgcgtt caaaactttt accatatatg ggtatgaatt gttcgccgat
     7861 tgggttttca tttacggaga atttcctccg caactgggtg ttcgtgcttg tgtgtgtctt
     7921 gtatatcgcc atgaagagtg tacttccatg ggtcctatca gttgcggcat gtgattcaat
     7981 ttgggggtta ctcatattcg atacttcagc tggttatctt tatgctgtga tggtcacggt
     8041 gcttttacac attttttccc caactactca ttgtatgcca aaacatacat atgtacatgt
     8101 gtttatagtg ggtttattag taatgtccag cgatattaaa acattacagt taaataatat
     8161 tcatattagc ttacgtttac tgctatctgc acttcaaatt ggttattcca gggacatttt
     8221 gtgcaacaca catcttctag tagggcattc actagtcgta gaaaacacac ctaacttaag
     8281 ccggttttaa caataacggc gaaacagctg ccagacgaga gaaaagtaca cagcaagaag
     8341 catttctata atgatgccgt cctaaatatg cagcattttt gctcaaattg atcatttcgt
     8401 tgcatcgctc aggtggtata gtatttggct attgaatgtg attgctatct taatagtttt
     8461 gaatagtaat agtgcaaata ttcattcata aaaaataggt aatcgatttt ttcagtgcag
     8521 tctatgcgtt tgcgcaacag agacgaagag agagacgcct cacacacaag cacatgcggt
     8581 ctaagctgct ctccctcttt ttctctctct ctctggcttg cggcagtttt ccagaggctg
     8641 tgactcagta agcatttctt gttgttaccc taacgataag cgtccgattc ctaattctct
     8701 ttcaaataat ttgttttatt tttttttttg tgtgttttga agtcaaacag cttcataaac
     8761 aatattaaat atcgcaattg ttattgcaaa tgcattcgta tcaggcgcct tgatgttttt
     8821 cgccaaaact tcgtctgctc tttgtcagcg taattgcgtt tgccggtcgt gccgtcttgc
     8881 tcgcacgcgg cttgcaaaca aaaagtgaaa gagggaaagc cacctgtgtt gttgcttttg
     8941 ttgctatcgt tggctgcaca cacatggcaa taagtaacca tattgtagac ttatttattt
     9001 tagcgttttc gttgactaat ctttgtttaa ataattgcct atttcgagcc aaaatgacgc
     9061 ataaaaggaa acggcaacat gcggtgcgtt gagcatgcgt aagcgagatg gcgctagaga
     9121 gagcgcagca taaaaagaaa aaaacaaccg gcgagtggcg agacacgaat aatacacaca
     9181 cgcacacatg gtcaaaggag tggctccaac gtcggtgacg tcgcgcccag ccacagaccc
     9241 cgccacgccc cgccccaccc ttccttccgc tgccccgcac ccctttcctt ccactaccgt
     9301 tttgccgcat ttggtattgc catctcgctt ttttggcttt actttgcttt ttgaataagc
     9361 caagaaactg aaatcaaagc ggacagtccg aaaattgaat attccaaaaa acaacaattg
     9421 cacatgcact gcgttcggca aaagacgcag aaattcaagc cttttggtct ggcatcatct
     9481 taaaatatca tcagtatcat cattgccatt ataaatacat aagaaaatga gaatacatta
     9541 aagatataca tacaatcttt gtcatagtca acatattttg caacaaaatt atctatagat
     9601 aacatgtttt tgtagaaaat tccactaact gattttcgta taaatacaaa ttattctcat
     9661 tcttcaaaat actcgccgac gtcatcgcga aattcctcag cattcttcga caaatgaaga
     9721 gagacagcga gtgacacaat cttatcttgc cgacgagtct ttgaaaatgg ccccgggtaa
     9781 cggtgaaaaa catatagcca agaatggagc gatcgccagg cgatggaaat tggcttcaaa
     9841 ttgctccgaa tatggccagt tggtcagttg gcattggatt gtggagagat actagatcag
     9901 ctcgttctcc ttctcctccg cattctccta ctccgcccga agacgcttct ttgatcgctt
     9961 ctttctcgaa tgtgtacata ttcattatgt atttggtgtt tggcgacgat cagccagctt
    10021 taatggccat aaaacgtgtc ttggtttgtg ggccacataa caggcagcga gatctgttcg
    10081 acttgcagcc tgaaatacgg cacgagtagg aaaagccgag tcaaatgccg aatgcagagt
    10141 ctcattacag cacaatcaac tcaagaaaaa ctcgacactt ttttaccatt tgcacttaaa
    10201 tcctttttta ttcgttatgt atactttttt tggtccctaa ccaaaacaaa accaaactct
    10261 cttagtcgtg cctctatatt taaaactatc aatttattat agtcaataaa tcgaactgtg
    10321 ttttcaacaa acgaacaata ggacactttg attctaaagg aaattttgaa aatcttaagc
    10381 agagggttct taagaccatt tgccaattct tataattctc aactgctctt tcctgatgtt
    10441 gatcatttat ataggtatgt tttcctcaat acttcgcaag cctcaggtag atacaggttt
    10501 tagagctaga aatagcaagt taaaataagg ctagtccgtt atcaacttga aaaagtggca
    10561 ccgagtcggt gcttttttag atctggaaaa atgatgtgac agtggaaatg agaagtaggt
    10621 gcatctgcaa agatt
//
