LOCUS       sequence-306429-       16903 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of PE2 enzyme under control of ubiquitous Actin5c 
            promoter. Can be used to generate stable cell lines by Hygromycin 
            selection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 16903)
  AUTHORS   Bosch JA, Birchak G, Perrimon N
  TITLE     Precise genome engineering in Drosophila using prime editing.
  JOURNAL   Proc Natl Acad Sci U S A. 2021 Jan 5;118(1). pii: 2021996118. doi: 
            10.1073/pnas.2021996118.
  PUBMED    33443210
REFERENCE   2  (bases 1 to 16903)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2021-01-5"; volume: "118(1). pii: 
            2021996118. doi"; pages: " 10.1073/pnas.2021996118"
COMMENT     Sequence Label: pAct-PE2-HygroR
FEATURES             Location/Qualifiers
     source          1..16903
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(624..641)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(795..1383)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(875..894)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1554..2345,2346..2414))
                     /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     2177..2196
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2415..2519)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2587..2605
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4001..4280
                     /label=copia promoter
                     /note="strong promoter from the Drosophila transposable 
                     element copia (Sinclair et al., 1986)"
     CDS             4326..5351
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             4326..5351
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     intron          5714..5779
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             5909..5929
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    6354..6488
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6391..6410)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6445..6464
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        6539..9048
                     /label=Ac5 promoter
                     /note="Drosophila melanogaster actin 5C promoter"
     protein_bind    9164..9188
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             9245..9265
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(9266..11779,11780..11782,11783..13366)
                     /codon_start=1
                     /product="nickase mutant of the Cas9 endonuclease from the 
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(H840A)
                     /note="generates RNA-guided single strand nicks in DNA due 
                     to the H840A mutation in the HNH catalytic domain"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             9266..13366
                     /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
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             15539..15559
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    complement(15579..15603)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 cccgcgttgc aggccatgct gtccaggcag gtagatgacg accatcaggg acagcttcaa
       61 ggatcgctcg cggctcttac cagcctaact tcgatcattg gaccgctgat cgtcacggcg
      121 atttatgccg cctcggcgag cacatggaac gggttggcat ggattgtagg cgccgcccta
      181 taccttgtct gcctccccgc gttgcgtcgc ggtgcatgga gccgggccac ctcgacctga
      241 atggaagccg gcggcacctc gctaacggat tcaccactcc aagaattgga gccaatcaat
      301 tcttgcggag aactgtgaat gcgcaaacca acccttggca gaacatatcc atcgcgtccg
      361 ccatctccag cagccgcacg cggcgcatct cgggcagcgt tgggtcctgg ccacgggtgc
      421 gcatgatcgt gctcctgtcg ttgaggaccc ggctaggctg gcggggttgc cttactggtt
      481 agcagaatga atcaccgata cgcgagcgaa cgtgaagcga ctgctgctgc aaaacgtctg
      541 cgacctgagc aacaacatga atggtcttcg gtttccgtgt ttcgtaaagt ctggaaacgc
      601 ggaagtcagc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
      661 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
      721 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
      781 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
      841 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
      901 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
      961 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     1021 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     1081 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     1141 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     1201 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     1261 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     1321 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     1381 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     1441 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     1501 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     1561 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     1621 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     1681 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     1741 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     1801 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     1861 gccattgctg caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     1921 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     1981 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     2041 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     2101 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     2161 ccggcgtcaa cacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     2221 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     2281 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     2341 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     2401 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     2461 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     2521 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
     2581 tataaaaata ggcgtatcac gaggcccttt cgtcttcaag aattcacatt tgtacgaatt
     2641 ttttttttat caaaagttcg agtttttcac caatttcctc atcaaccgag caaggcaaac
     2701 ggctttgaat aatatggtgt tatatataca tatatcaaat cgctgctgac tgcgtgattg
     2761 atggccccaa gattacatat tatcgaatca ggattcagaa ggagatcaat gtcaaatgcg
     2821 gacaggaaca tgaaagacgc ctgttatgcg caattaaaaa tttgggttta attgctgtgg
     2881 aaactgttgt tggcggcatc ttaagttcct gtttaacaac atcaactact tatgtacgta
     2941 gaagcgttta agccatttgc atacagatga gaactggctt ttgtgctaat cagtcaagat
     3001 gactccgatg atgatgactc attacctgac cagttttcgc tgctttcttt tcaacaacta
     3061 cttgtatatg tattgtatcc aatagcaata cattgaattt ccatggtcta gtcacgtatt
     3121 atcatttaat tgacaccaag tcgtgttatt gttgagctat cgagttcagc tcaaacattt
     3181 cttattccca tgaataagcc ggcaaaaata tgcaatctat gaaagttaat ataagcaaac
     3241 cttactttga ctcaatacca atgcactttg tgtcgatagg ttcacgcaat tgaggcgatt
     3301 attccgataa cccaagcgat tgactgttcc cgtttcgatt ccaattgaaa tttggaaatg
     3361 tacaatagtt ttgctatatg ctgtcaagta cgctcttatc ttctctgggt tttcttcaga
     3421 gtttcgaaac gcttcttctt ttttttgttt tttttttttt ggaatctcgt attttggaag
     3481 gggctcccct ctggaatttg ttacactgtc gttatcattg cgaacaagcg gcccgaagct
     3541 atcagcgact ttaacattta caatgcactt ttttacgacc aattaaatgt acattttcct
     3601 ttcttcgccc gttgataagc gaacgcgatg tggcgcaggc aatgtgttgc tcttgcgaca
     3661 caaacgcaat caaaatggat tcaatttcgc tttttcccag tgaaacgaag aacgaaccga
     3721 ccatcatgat atgctcctct gcatgttgcg tattgaatca atgacaattt caattaagcc
     3781 gcccgttcgt catgcgtttt cgtgcgcttc gaaatgctga taacgctgct gtcctccaac
     3841 tgctttgcat gtggacacaa ttccatttat ttaattcttt tatttggatc ggttaaatta
     3901 aaaagcgcct tgttacgcat ttaacgttgt ttccggtgcg tggtggtttc atgcttctgg
     3961 gaacggcaaa tgggtttagg attgggaacc cctcatcatc tgttggaata tactattcaa
     4021 cctacaaaag taacgttaaa caacactact ttatatttga tatgaatggc cacacctttt
     4081 atgccataaa acatattgta agagaatacc actcttttta ttccttcttt ccttcttgta
     4141 cgttttttgc tgtaagtagg tcgtggtgct ggtgttgcag ttgaaataac ttaaaatata
     4201 aatcataaaa ctcaaacata aacttgacta tttatttatt tattaagaaa ggaaatataa
     4261 attataaatt acaacaggtt atggacctgc agccaagctt ggcgctcgtc cgggggcaat
     4321 gagatatgaa aaagcctgaa ctcaccgcga cgtctgtcga gaagtttctg atcgaaaagt
     4381 tcgacagcgt ctccgacctg atgcagctct cggagggcga agaatctcgt gctttcagct
     4441 tcgatgtagg agggcgtgga tatgtcctgc gggtaaatag ctgcgccgat ggtttctaca
     4501 aagatcgtta tgtttatcgg cactttgcat cggccgcgct cccgattccg gaagtgcttg
     4561 acattgggga attcagcgag agcctgacct attgcatctc ccgccgtgca cagggtgtca
     4621 cgttgcaaga cctgcctgaa accgaactgc ccgctgttct gcagccggtc gcggaggcca
     4681 tggatgcgat cgctgcggcc gatcttagcc agacgagcgg gttcggccca ttcggaccgc
     4741 aaggaatcgg tcaatacact acatggcgtg atttcatatg cgcgattgct gatccccatg
     4801 tgtatcactg gcaaactgtg atggacgaca ccgtcagtgc gtccgtcgcg caggctctcg
     4861 atgagctgat gctttgggcc gaggactgcc ccgaagtccg gcacctcgtg cacgcggatt
     4921 tcggctccaa caatgtcctg acggacaatg gccgcataac agcggtcatt gactggagcg
     4981 aggcgatgtt cggggattcc caatacgagg tcgccaacat cttcttctgg aggccgtggt
     5041 tggcttgtat ggagcagcag acgcgctact tcgagcggag gcatccggag cttgcaggat
     5101 cgccgcggct ccgggcgtat atgctccgca ttggtcttga ccaactctat cagagcttgg
     5161 ttgacggcaa tttcgatgat gcagcttggg cgcagggtcg atgcgacgca atcgtccgat
     5221 ccggagccgg gactgtcggg cgtacacaaa tcgcccgcag aagcgcggcc gtctggaccg
     5281 atggctgtgt agaagtactc gccgatagtg gaaaccgacg ccccagcact cgtccgaggg
     5341 caaaggaata gagtagatgc cgaccgaaca agagctgatt tcgagaacgc ctcagccagc
     5401 aactcgcgcg agcctagcaa ggcaaatgcg agagaacggc cttacgcttg gtggcacagt
     5461 tctcgtccac agttcgctaa gctcgctcgg ctgggtcgcg ggagggccgg tcgcagtgat
     5521 tcaggccctt ctggattgtg ttggtcccca gggcacgatt gtcatgccca cgcactcggg
     5581 tgatctgact gatcccgcag attggagatc gccgcccgtg cctgccgatt gggtgcagat
     5641 ctttgtgaag gaaccttact tctgtggtgt gacataattg gacaaactac ctacagagat
     5701 ttaaagctct aaggtaaata taaaattttt aagtgtataa tgtgttaaac tactgattct
     5761 aattgtttgt gtattttaga ttccaaccta tggaactgat gaatgggagc agtggtggaa
     5821 tgcctttaat gaggaaaacc tgttttgctc agaagaaatg ccatctagtg atgatgaggc
     5881 tactgctgac tctcaacatt ctactcctcc aaaaaagaag agaaaggtag aagaccccaa
     5941 ggactttcct tcagaattgc taagtttttt gagtcatgct gtgtttagta atagaactct
     6001 tgcttgcttt gctatttaca ccacaaagga aaaagctgca ctgctataca agaaaattat
     6061 ggaaaaatat tctgtaacct ttataagtag gcataacagt tataatcata acatactgtt
     6121 ttttcttact ccacacaggc atagagtgtc tgctattaat aactatgctc aaaaattgtg
     6181 tacctttagc tttttaattt gtaaaggggt taataaggaa tatttgatgt atagtgcctt
     6241 gactagagat cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc
     6301 tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt
     6361 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     6421 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     6481 tctggatcgc atgcaattct atattctaaa aacacaaatg atacttctaa aaaaaaatca
     6541 tgaatggcat caactctgaa tcaaatcttt gcagatgcac ctacttctca tttccactgt
     6601 cacatcattt ttccagatct cgctgcctgt tatgtggccc acaaaccaag acacgtttta
     6661 tggccattaa agctggctga tcgtcgccaa acaccaaata cataatgaat atgtacacat
     6721 tcgagaaaga agcgatcaaa gaagcgtctt cgggcggagt aggagaatgc ggaggagaag
     6781 gagaacgagc tgatctagta tctctccaca atccaatgcc aactgaccaa ctggccatat
     6841 tcggagcaat ttgaagccaa tttccatcgc ctggcgatcg ctccattctt ggctatatgt
     6901 ttttcaccgt tacccggggc cattttcaaa gactcgtcgg caagataaga ttgtgtcact
     6961 cgctgtctct cttcatttgt cgaagaatgc tgaggaattt cgcgatgacg tcggcgagta
     7021 ttttgaagaa tgagaataat ttgtatttat acgaaaatca gttagtggaa ttttctacaa
     7081 aaacatgtta tctatagata attttgttgc aaaatatgtt gactatgaca aagattgtat
     7141 gtatatacct ttaatgtatt ctcattttct tatgtattta taatggcaat gatgatactg
     7201 atgatatttt aagatgatgc cagaccaaaa ggcttgaatt tctgcgtctt ttgccgaacg
     7261 cagtgcatgt gcaattgttg ttttttggaa tattcaattt tcggactgtc cgctttgatt
     7321 tcagtttctt ggcttattca aaaagcaaag taaagccaaa aaagcgagat ggcaatacca
     7381 aatgcggcaa aacggtagtg gaaggaaagg ggtgcggggc agcggaagga agggtggggc
     7441 ggggcgtggc ggggtctgtg gctgggcgcg acgtcaccga cgttggagcc actcctttga
     7501 ccatgtgtgc gtgtgtgtat tattcgtgtc tcgccactcg ccggttgttt ttttcttttt
     7561 atgctgcgct ctctctagcg ccatctcgct tacgcatgct caacgcaccg catgttgccg
     7621 tttcctttta tgcgtcattt tggctcgaaa taggcaatta tttaaacaaa gattagtcaa
     7681 cgaaaacgct aaaataaata agtctacaat atggttactt attgccatgt gtgtgcagcc
     7741 aacgatagca acaaaagcaa caacacaggt ggctttccct ctttcacttt ttgtttgcaa
     7801 gccgcgtgcg agcaagacgg cacgaccggc aaacgcaatt acgctgacaa agagcagacg
     7861 aagttttggc gaaaaacatc aaggcgcctg atacgaatgc atttgcaata acaattgcga
     7921 tatttaatat tgtttatgaa gctgtttgac ttcaaaacac acaaaaaaaa aaataaaaca
     7981 aattatttga aagagaatta ggaatcggac gcttatcgtt agggtaacaa caagaaatgc
     8041 ttactgagtc acagcctctg gaaaactgcc gcaagccaga gagagagaga aaaagaggga
     8101 gagcagctta gaccgcatgt gcttgtgtgt gaggcgtctc tctcttcgtc tctgttgcgc
     8161 aaacgcatag actgcactga aaaaatcgat tacctatttt ttatgaatga atatttgcac
     8221 tattactatt caaaactatt aagatagcaa tcacattcaa tagccaaata ctataccacc
     8281 tgagcgatgc aacgaaatga tcaatttgag caaaaatgct gcatatttag gacggcatca
     8341 ttatagaaat gcttcttgct gtgtactttt ctctcgtctg gcagctgttt cgccgttatt
     8401 gttaaaaccg gcttaagtta ggtgtgtttt ctacgactag tgaatgccct actagaagat
     8461 gtgtgttgca caaaatgtcc ctggaataac caatttgaag tgcagatagc agtaaacgta
     8521 agctaatatg aatattattt aactgtaatg ttttaatatc gctggacatt actaataaac
     8581 ccactataaa cacatgtaca tatgtatgtt ttggcataca atgagtagtt ggggaaaaaa
     8641 tgtgtaaaag caccgtgacc atcacagcat aaagataacc agctgaagta tcgaatatga
     8701 gtaaccccca aattgaatca catgccgcaa ctgataggac ccatggaagt acactcttca
     8761 tggcgatata caagacacac acaagcacga acacccagtt gcggaggaaa ttctccgtaa
     8821 atgaaaaccc aatcggcgaa caattcatac ccatatatgg taaaagtttt gaacgcgact
     8881 tgagagcgga gagcattgcg gctgataagg ttttagcgct aagcgggctt tataaaacgg
     8941 gctgcgggac cagttttcat atcactaccg tttgagttct tgtgctgtgt ggatactcct
     9001 cccgacacaa agccgctcca tcagccagca gtcgtctaat ccagagaccc cggatccaga
     9061 taccgagagc atctggccaa tgtgcatcag ttgtggtcag cagcaaaatc aagtgaatca
     9121 tctcagtgca actaaagggg ggatctcgag ggatccgata tcaacaagtt tgtacaaaaa
     9181 agcaggctcc gcggccgccc ccttcaccat gaaacggaca gccgacggaa gcgagttcga
     9241 gtcaccaaag aagaagcgga aagtcgacaa gaagtacagc atcggcctgg acatcggcac
     9301 caactctgtg ggctgggccg tgatcaccga cgagtacaag gtgcccagca agaaattcaa
     9361 ggtgctgggc aacaccgacc ggcacagcat caagaagaac ctgatcggag ccctgctgtt
     9421 cgacagcggc gaaacagccg aggccacccg gctgaagaga accgccagaa gaagatacac
     9481 cagacggaag aaccggatct gctatctgca agagatcttc agcaacgaga tggccaaggt
     9541 ggacgacagc ttcttccaca gactggaaga gtccttcctg gtggaagagg ataagaagca
     9601 cgagcggcac cccatcttcg gcaacatcgt ggacgaggtg gcctaccacg agaagtaccc
     9661 caccatctac cacctgagaa agaaactggt ggacagcacc gacaaggccg acctgcggct
     9721 gatctatctg gccctggccc acatgatcaa gttccggggc cacttcctga tcgagggcga
     9781 cctgaacccc gacaacagcg acgtggacaa gctgttcatc cagctggtgc agacctacaa
     9841 ccagctgttc gaggaaaacc ccatcaacgc cagcggcgtg gacgccaagg ccatcctgtc
     9901 tgccagactg agcaagagca gacggctgga aaatctgatc gcccagctgc ccggcgagaa
     9961 gaagaatggc ctgttcggaa acctgattgc cctgagcctg ggcctgaccc ccaacttcaa
    10021 gagcaacttc gacctggccg aggatgccaa actgcagctg agcaaggaca cctacgacga
    10081 cgacctggac aacctgctgg cccagatcgg cgaccagtac gccgacctgt ttctggccgc
    10141 caagaacctg tccgacgcca tcctgctgag cgacatcctg agagtgaaca ccgagatcac
    10201 caaggccccc ctgagcgcct ctatgatcaa gagatacgac gagcaccacc aggacctgac
    10261 cctgctgaaa gctctcgtgc ggcagcagct gcctgagaag tacaaagaga ttttcttcga
    10321 ccagagcaag aacggctacg ccggctacat tgacggcgga gccagccagg aagagttcta
    10381 caagttcatc aagcccatcc tggaaaagat ggacggcacc gaggaactgc tcgtgaagct
    10441 gaacagagag gacctgctgc ggaagcagcg gaccttcgac aacggcagca tcccccacca
    10501 gatccacctg ggagagctgc acgccattct gcggcggcag gaagattttt acccattcct
    10561 gaaggacaac cgggaaaaga tcgagaagat cctgaccttc cgcatcccct actacgtggg
    10621 ccctctggcc aggggaaaca gcagattcgc ctggatgacc agaaagagcg aggaaaccat
    10681 caccccctgg aacttcgagg aagtggtgga caagggcgct tccgcccaga gcttcatcga
    10741 gcggatgacc aacttcgata agaacctgcc caacgagaag gtgctgccca agcacagcct
    10801 gctgtacgag tacttcaccg tgtataacga gctgaccaaa gtgaaatacg tgaccgaggg
    10861 aatgagaaag cccgccttcc tgagcggcga gcagaaaaag gccatcgtgg acctgctgtt
    10921 caagaccaac cggaaagtga ccgtgaagca gctgaaagag gactacttca agaaaatcga
    10981 gtgcttcgac tccgtggaaa tctccggcgt ggaagatcgg ttcaacgcct ccctgggcac
    11041 ataccacgat ctgctgaaaa ttatcaagga caaggacttc ctggacaatg aggaaaacga
    11101 ggacattctg gaagatatcg tgctgaccct gacactgttt gaggacagag agatgatcga
    11161 ggaacggctg aaaacctatg cccacctgtt cgacgacaaa gtgatgaagc agctgaagcg
    11221 gcggagatac accggctggg gcaggctgag ccggaagctg atcaacggca tccgggacaa
    11281 gcagtccggc aagacaatcc tggatttcct gaagtccgac ggcttcgcca acagaaactt
    11341 catgcagctg atccacgacg acagcctgac ctttaaagag gacatccaga aagcccaggt
    11401 gtccggccag ggcgatagcc tgcacgagca cattgccaat ctggccggca gccccgccat
    11461 taagaagggc atcctgcaga cagtgaaggt ggtggacgag ctcgtgaaag tgatgggccg
    11521 gcacaagccc gagaacatcg tgatcgaaat ggccagagag aaccagacca cccagaaggg
    11581 acagaagaac agccgcgaga gaatgaagcg gatcgaagag ggcatcaaag agctgggcag
    11641 ccagatcctg aaagaacacc ccgtggaaaa cacccagctg cagaacgaga agctgtacct
    11701 gtactacctg cagaatgggc gggatatgta cgtggaccag gaactggaca tcaaccggct
    11761 gtccgactac gatgtggacg ctatcgtgcc tcagagcttt ctgaaggacg actccatcga
    11821 caacaaggtg ctgaccagaa gcgacaagaa ccggggcaag agcgacaacg tgccctccga
    11881 agaggtcgtg aagaagatga agaactactg gcggcagctg ctgaacgcca agctgattac
    11941 ccagagaaag ttcgacaatc tgaccaaggc cgagagaggc ggcctgagcg aactggataa
    12001 ggccggcttc atcaagagac agctggtgga aacccggcag atcacaaagc acgtggcaca
    12061 gatcctggac tcccggatga acactaagta cgacgagaat gacaagctga tccgggaagt
    12121 gaaagtgatc accctgaagt ccaagctggt gtccgatttc cggaaggatt tccagtttta
    12181 caaagtgcgc gagatcaaca actaccacca cgcccacgac gcctacctga acgccgtcgt
    12241 gggaaccgcc ctgatcaaaa agtaccctaa gctggaaagc gagttcgtgt acggcgacta
    12301 caaggtgtac gacgtgcgga agatgatcgc caagagcgag caggaaatcg gcaaggctac
    12361 cgccaagtac ttcttctaca gcaacatcat gaactttttc aagaccgaga ttaccctggc
    12421 caacggcgag atccggaagc ggcctctgat cgagacaaac ggcgaaaccg gggagatcgt
    12481 gtgggataag ggccgggatt ttgccaccgt gcggaaagtg ctgagcatgc cccaagtgaa
    12541 tatcgtgaaa aagaccgagg tgcagacagg cggcttcagc aaagagtcta tcctgcccaa
    12601 gaggaacagc gataagctga tcgccagaaa gaaggactgg gaccctaaga agtacggcgg
    12661 cttcgacagc cccaccgtgg cctattctgt gctggtggtg gccaaagtgg aaaagggcaa
    12721 gtccaagaaa ctgaagagtg tgaaagagct gctggggatc accatcatgg aaagaagcag
    12781 cttcgagaag aatcccatcg actttctgga agccaagggc tacaaagaag tgaaaaagga
    12841 cctgatcatc aagctgccta agtactccct gttcgagctg gaaaacggcc ggaagagaat
    12901 gctggcctct gccggcgaac tgcagaaggg aaacgaactg gccctgccct ccaaatatgt
    12961 gaacttcctg tacctggcca gccactatga gaagctgaag ggctcccccg aggataatga
    13021 gcagaaacag ctgtttgtgg aacagcacaa gcactacctg gacgagatca tcgagcagat
    13081 cagcgagttc tccaagagag tgatcctggc cgacgctaat ctggacaaag tgctgtccgc
    13141 ctacaacaag caccgggata agcccatcag agagcaggcc gagaatatca tccacctgtt
    13201 taccctgacc aatctgggag cccctgccgc cttcaagtac tttgacacca ccatcgaccg
    13261 gaagaggtac accagcacca aagaggtgct ggacgccacc ctgatccacc agagcatcac
    13321 cggcctgtac gagacacgga tcgacctgtc tcagctggga ggtgactctg gaggatctag
    13381 cggaggatcc tctggcagcg agacaccagg aacaagcgag tcagcaacac cagagagcag
    13441 tggcggcagc agcggcggca gcagcaccct aaatatagaa gatgagtatc ggctacatga
    13501 gacctcaaaa gagccagatg tttctctagg gtccacatgg ctgtctgatt ttcctcaggc
    13561 ctgggcggaa accgggggca tgggactggc agttcgccaa gctcctctga tcatacctct
    13621 gaaagcaacc tctacccccg tgtccataaa acaatacccc atgtcacaag aagccagact
    13681 ggggatcaag ccccacatac agagactgtt ggaccaggga atactggtac cctgccagtc
    13741 cccctggaac acgcccctgc tacccgttaa gaaaccaggg actaatgatt ataggcctgt
    13801 ccaggatctg agagaagtca acaagcgggt ggaagacatc caccccaccg tgcccaaccc
    13861 ttacaacctc ttgagcgggc tcccaccgtc ccaccagtgg tacactgtgc ttgatttaaa
    13921 ggatgccttt ttctgcctga gactccaccc caccagtcag cctctcttcg cctttgagtg
    13981 gagagatcca gagatgggaa tctcaggaca attgacctgg accagactcc cacagggttt
    14041 caaaaacagt cccaccctgt ttaatgaggc actgcacaga gacctagcag acttccggat
    14101 ccagcaccca gacttgatcc tgctacagta cgtggatgac ttactgctgg ccgccacttc
    14161 tgagctagac tgccaacaag gtactcgggc cctgttacaa accctaggga acctcgggta
    14221 tcgggcctcg gccaagaaag cccaaatttg ccagaaacag gtcaagtatc tggggtatct
    14281 tctaaaagag ggtcagagat ggctgactga ggccagaaaa gagactgtga tggggcagcc
    14341 tactccgaag acccctcgac aactaaggga gttcctaggg aaggcaggct tctgtcgcct
    14401 cttcatccct gggtttgcag aaatggcagc ccccctgtac cctctcacca aaccggggac
    14461 tctgtttaat tggggcccag accaacaaaa ggcctatcaa gaaatcaagc aagctcttct
    14521 aactgcccca gccctggggt tgccagattt gactaagccc tttgaactct ttgtcgacga
    14581 gaagcagggc tacgccaaag gtgtcctaac gcaaaaactg ggaccttggc gtcggccggt
    14641 ggcctacctg tccaaaaagc tagacccagt agcagctggg tggccccctt gcctacggat
    14701 ggtagcagcc attgccgtac tgacaaagga tgcaggcaag ctaaccatgg gacagccact
    14761 agtcattctg gccccccatg cagtagaggc actagtcaaa caaccccccg accgctggct
    14821 ttccaacgcc cggatgactc actatcaggc cttgcttttg gacacggacc gggtccagtt
    14881 cggaccggtg gtagccctga acccggctac gctgctccca ctgcctgagg aagggctgca
    14941 acacaactgc cttgatatcc tggccgaagc ccacggaacc cgacccgacc taacggacca
    15001 gccgctccca gacgccgacc acacctggta cacggatgga agcagtctct tacaagaggg
    15061 acagcgtaag gcgggagctg cggtgaccac cgagaccgag gtaatctggg ctaaagccct
    15121 gccagccggg acatccgctc agcgggctga actgatagca ctcacccagg ccctaaagat
    15181 ggcagaaggt aagaagctaa atgtttatac tgatagccgt tatgcttttg ctactgccca
    15241 tatccatgga gaaatataca gaaggcgtgg gtggctcaca tcagaaggca aagagatcaa
    15301 aaataaagac gagatcttgg ccctactaaa agccctcttt ctgcccaaaa gacttagcat
    15361 aatccattgt ccaggacatc aaaagggaca cagcgccgag gctagaggca accggatggc
    15421 tgaccaagcg gcccgaaagg cagccatcac agagactcca gacacctcta ccctcctcat
    15481 agaaaattca tcaccctctg gcggctcaaa aagaaccgcc gacggcagcg aattcgagcc
    15541 caagaagaag aggaaagtct aaaagggtgg gcgcgccgac ccagctttct tgtacaaagt
    15601 ggttgatatc actagttcta gtcgaccatg aagatcaaga tcattgcccc gccagagcgc
    15661 aagtactctg tctggatcgg tggctccatc ctggcttcgc tgtccacctt ccagcagatg
    15721 tggatctcca agcaggagta cgacgagtcc ggcccctcca ttgtgcaccg caagtgcttc
    15781 taagaaggat cgcttgtctg ggcaagagga tcaggatcgg gatggtcttg attctgctgg
    15841 aggaggagga ggagaagtcg aggaagcagc agcgaaagtg caagtgcgag tggtggaagt
    15901 ttggagtgca gcacaacaaa atcaacaaca acaccaacta caagatgaaa agagcggaac
    15961 cacctgcaca ccatcatcac tatcatcatc gttttgggcg catgttgtgt ggttccagcg
    16021 tattaatata attaatttat tccacatgag atatgatatg atatactatg tattttttgt
    16081 ttttttttta tttgtaaacc tttaatataa caagaactac aaaaaatgaa aatgagcgaa
    16141 aatgcatatt ctgccattcc acacacacac caacaacacc caacacacgc acacccacaa
    16201 gcttacacac acacattcgc ggcatgacaa ggacatcaag ataaagaaga acttaaagaa
    16261 gatatttccc aaagcgcaaa aagaacacac acacattgca aaacacaaac aacacactag
    16321 cgttttgtac aattcgtcag caaccttatg tattattttt taattatgat gtaattataa
    16381 acaaagtgaa acaaaaatat gaaaacaaaa aggaaaatca aatctgtctt ctctttctcc
    16441 cgctctcctc gctctctgct gctaacctcg ccctctcctc tctcatcttt ttgtctgtct
    16501 ctcttccaca tttttgccgg ccggcaaaat aataacccac acacactcac acttggctgc
    16561 agtttcgcgt gcgatattca cacacattca agcatacaag catacataca tatgtatttt
    16621 ttttttattt gtacactttt ctaattgcat gcgtatcgat tgataagttt acgcctgaaa
    16681 atgttaatta aaatgtgaaa atgcaactga aaaactgatg aaatgaaaca acaacaagcg
    16741 aacaatttgc tacatgtgta ttgtctaaca accgttactg caacggttgc ttcggaaaag
    16801 ggtgaagagg aagagggcac gaggtcgact agagcggccg ccgacgcgag gctggatggc
    16861 cttccccatt atgattcttc tcgcttccgg cggcatcggg atg
//
