LOCUS       sequence-306424-       13414 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expression of PE2 enzyme under control of the germ cell specific 
            nanos promoter. Can be used to generate transgenic flies with 
            vermillion+ selection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13414)
  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 13414)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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: pNos-PE2-attB
FEATURES             Location/Qualifiers
     source          1..13414
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     17..37
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        19..37
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             complement(992..1012)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(join(3185..4768,4769..4771,4772..7285))
                     /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             complement(3185..7285)
                     /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             complement(7286..7306)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    8549..8618
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     CDS             join(8933..8958,9032..9192,9253..9386,9441..10047,
                     10139..10232,10309..10426)
                     /codon_start=1
                     /product="tryptophan oxygenase"
                     /label=vermilion
                     /note="selectable marker, required to synthesize the brown 
                     eye pigment in Drosophila"
                     /translation="MSCPYAGNGNDHDDSAVPLTTEVGKIYGEYLMLDKLLDAQCMLSE
                     EDKRPVHDEHLFIITHQAYELWFKQIIFEFDSIRDMLDAEVIDETKTLEIVKRLNRVVL
                     ILKLLVDQVPILETMTPLDFMDFRKYLAPASGFQSLQFRLIENKLGVLTEQRVRYNQKY
                     SDVFSDEEARNSIRNSEKDPSLLELVQRWLERTPGLEESGFNFWAKFQESVDRFLEAQV
                     QSAMEEPVEKAKNYRLMDIEKRREVYRSIFDPAVHDALVRRGDRRFSHRALQGAIMITF
                     YRDEPRFSQPHQLLTLLMDIDSLITKWRYNHVIMVQRMIGSQQLGTGGSSGYQYLRSTL
                     SDRYKVFLDLFNLSTFLIPREAIPPLDETIRKKLINKSV"
     primer_bind     complement(10617..10636)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(10618..10636)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(10646..10663)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(10646..10662)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10655..10677)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(10803..11259)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10941..10962)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     11153..11172
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        11286..11390
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(11391..11459,11460..12251)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(11609..11628)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      12422..13010
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     12911..12930
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    13298..13319
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(13334..13339,13340..13357,13358..13364)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    13372..13388
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     13377..13399
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     13396..13412
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     13396..13412
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 tgattacgcc aagcgcgcaa ttaaccctca ctaaagggaa caaaagctgg agctcactag
       61 tgcaagaagg acatttgttt ccttaaattg taaccatttc tttatttggc agtcgagcca
      121 ttgaattttt cattttcaga atatgtgtac acatttttta aaaaaataaa aaaattatat
      181 aatgctggcg gttgtttcat gtgtgaaaaa ttgatcaatg gtaaacaaaa ttgaataaat
      241 atataacata tatatatata tatatatatg ttatatattt atatagatat gtgtgttgaa
      301 atgaatactt gcgatacatg taataaaaat actcttcgct tatctatcaa aaagtgcgga
      361 atgtcaaaat ttaaaatttt acaatgaatg cgtagccgac gacgaaagtg ttccttgcta
      421 tttcctttag caagatttaa atttagatta aattctaatg atacgattga cagttcgaaa
      481 ttcaaagtgt tcctttttca aaatttagta aagattgtag atgaattgta gatatatcga
      541 aatttttcgg ccgcaagcga acattttacg aaatgaaggc gaccagttgc agaccaattc
      601 cattcatcaa ctttcggatt gtaagatatt tctatcggcc acgacgattg aacaagtatt
      661 acgatattgt aagtcttctt taacaaaatt agtttccctt tcacagaaac agacataaat
      721 tcttgaatta ttgacttgga tttgagtgat cgttcgttgt ctatactata tgatctatag
      781 gcacgggata acgatctaaa tctctttaaa atcgaacgcg ccaggcgcta tttaaacgtt
      841 actatctatc tatctggtta acccagcttt gatcggaatg cgtatatata tttcatgtta
      901 tataaacgct gcaaaagctg ccagagcctc tgctccagag ctggattcgc cctctctaaa
      961 cctcgagtgg atcctcctag gtgctagatt agactttcct cttcttcttg ggctcgaatt
     1021 cgctgccgtc ggcggttctt tttgagccgc cagagggtga tgaattttct atgaggaggg
     1081 tagaggtgtc tggagtctct gtgatggctg cctttcgggc cgcttggtca gccatccggt
     1141 tgcctctagc ctcggcgctg tgtccctttt gatgtcctgg acaatggatt atgctaagtc
     1201 ttttgggcag aaagagggct tttagtaggg ccaagatctc gtctttattt ttgatctctt
     1261 tgccttctga tgtgagccac ccacgccttc tgtatatttc tccatggata tgggcagtag
     1321 caaaagcata acggctatca gtataaacat ttagcttctt accttctgcc atctttaggg
     1381 cctgggtgag tgctatcagt tcagcccgct gagcggatgt cccggctggc agggctttag
     1441 cccagattac ctcggtctcg gtggtcaccg cagctcccgc cttacgctgt ccctcttgta
     1501 agagactgct tccatccgtg taccaggtgt ggtcggcgtc tgggagcggc tggtccgtta
     1561 ggtcgggtcg ggttccgtgg gcttcggcca ggatatcaag gcagttgtgt tgcagccctt
     1621 cctcaggcag tgggagcagc gtagccgggt tcagggctac caccggtccg aactggaccc
     1681 ggtccgtgtc caaaagcaag gcctgatagt gagtcatccg ggcgttggaa agccagcggt
     1741 cggggggttg tttgactagt gcctctactg catggggggc cagaatgact agtggctgtc
     1801 ccatggttag cttgcctgca tcctttgtca gtacggcaat ggctgctacc atccgtaggc
     1861 aagggggcca cccagctgct actgggtcta gctttttgga caggtaggcc accggccgac
     1921 gccaaggtcc cagtttttgc gttaggacac ctttggcgta gccctgcttc tcgtcgacaa
     1981 agagttcaaa gggcttagtc aaatctggca accccagggc tggggcagtt agaagagctt
     2041 gcttgatttc ttgataggcc ttttgttggt ctgggcccca attaaacaga gtccccggtt
     2101 tggtgagagg gtacaggggg gctgccattt ctgcaaaccc agggatgaag aggcgacaga
     2161 agcctgcctt ccctaggaac tcccttagtt gtcgaggggt cttcggagta ggctgcccca
     2221 tcacagtctc ttttctggcc tcagtcagcc atctctgacc ctcttttaga agatacccca
     2281 gatacttgac ctgtttctgg caaatttggg ctttcttggc cgaggcccga tacccgaggt
     2341 tccctagggt ttgtaacagg gcccgagtac cttgttggca gtctagctca gaagtggcgg
     2401 ccagcagtaa gtcatccacg tactgtagca ggatcaagtc tgggtgctgg atccggaagt
     2461 ctgctaggtc tctgtgcagt gcctcattaa acagggtggg actgtttttg aaaccctgtg
     2521 ggagtctggt ccaggtcaat tgtcctgaga ttcccatctc tggatctctc cactcaaagg
     2581 cgaagagagg ctgactggtg gggtggagtc tcaggcagaa aaaggcatcc tttaaatcaa
     2641 gcacagtgta ccactggtgg gacggtggga gcccgctcaa gaggttgtaa gggttgggca
     2701 cggtggggtg gatgtcttcc acccgcttgt tgacttctct cagatcctgg acaggcctat
     2761 aatcattagt ccctggtttc ttaacgggta gcaggggcgt gttccagggg gactggcagg
     2821 gtaccagtat tccctggtcc aacagtctct gtatgtgggg cttgatcccc agtctggctt
     2881 cttgtgacat ggggtattgt tttatggaca cgggggtaga ggttgctttc agaggtatga
     2941 tcagaggagc ttggcgaact gccagtccca tgcccccggt ttccgcccag gcctgaggaa
     3001 aatcagacag ccatgtggac cctagagaaa catctggctc ttttgaggtc tcatgtagcc
     3061 gatactcatc ttctatattt agggtgctgc tgccgccgct gctgccgcca ctgctctctg
     3121 gtgttgctga ctcgcttgtt cctggtgtct cgctgccaga ggatcctccg ctagatcctc
     3181 cagagtcacc tcccagctga gacaggtcga tccgtgtctc gtacaggccg gtgatgctct
     3241 ggtggatcag ggtggcgtcc agcacctctt tggtgctggt gtacctcttc cggtcgatgg
     3301 tggtgtcaaa gtacttgaag gcggcagggg ctcccagatt ggtcagggta aacaggtgga
     3361 tgatattctc ggcctgctct ctgatgggct tatcccggtg cttgttgtag gcggacagca
     3421 ctttgtccag attagcgtcg gccaggatca ctctcttgga gaactcgctg atctgctcga
     3481 tgatctcgtc caggtagtgc ttgtgctgtt ccacaaacag ctgtttctgc tcattatcct
     3541 cgggggagcc cttcagcttc tcatagtggc tggccaggta caggaagttc acatatttgg
     3601 agggcagggc cagttcgttt cccttctgca gttcgccggc agaggccagc attctcttcc
     3661 ggccgttttc cagctcgaac agggagtact taggcagctt gatgatcagg tcctttttca
     3721 cttctttgta gcccttggct tccagaaagt cgatgggatt cttctcgaag ctgcttcttt
     3781 ccatgatggt gatccccagc agctctttca cactcttcag tttcttggac ttgccctttt
     3841 ccactttggc caccaccagc acagaatagg ccacggtggg gctgtcgaag ccgccgtact
     3901 tcttagggtc ccagtccttc tttctggcga tcagcttatc gctgttcctc ttgggcagga
     3961 tagactcttt gctgaagccg cctgtctgca cctcggtctt tttcacgata ttcacttggg
     4021 gcatgctcag cactttccgc acggtggcaa aatcccggcc cttatcccac acgatctccc
     4081 cggtttcgcc gtttgtctcg atcagaggcc gcttccggat ctcgccgttg gccagggtaa
     4141 tctcggtctt gaaaaagttc atgatgttgc tgtagaagaa gtacttggcg gtagccttgc
     4201 cgatttcctg ctcgctcttg gcgatcatct tccgcacgtc gtacaccttg tagtcgccgt
     4261 acacgaactc gctttccagc ttagggtact ttttgatcag ggcggttccc acgacggcgt
     4321 tcaggtaggc gtcgtgggcg tggtggtagt tgttgatctc gcgcactttg taaaactgga
     4381 aatccttccg gaaatcggac accagcttgg acttcagggt gatcactttc acttcccgga
     4441 tcagcttgtc attctcgtcg tacttagtgt tcatccggga gtccaggatc tgtgccacgt
     4501 gctttgtgat ctgccgggtt tccaccagct gtctcttgat gaagccggcc ttatccagtt
     4561 cgctcaggcc gcctctctcg gccttggtca gattgtcgaa ctttctctgg gtaatcagct
     4621 tggcgttcag cagctgccgc cagtagttct tcatcttctt cacgacctct tcggagggca
     4681 cgttgtcgct cttgccccgg ttcttgtcgc ttctggtcag caccttgttg tcgatggagt
     4741 cgtccttcag aaagctctga ggcacgatag cgtccacatc gtagtcggac agccggttga
     4801 tgtccagttc ctggtccacg tacatatccc gcccattctg caggtagtac aggtacagct
     4861 tctcgttctg cagctgggtg ttttccacgg ggtgttcttt caggatctgg ctgcccagct
     4921 ctttgatgcc ctcttcgatc cgcttcattc tctcgcggct gttcttctgt cccttctggg
     4981 tggtctggtt ctctctggcc atttcgatca cgatgttctc gggcttgtgc cggcccatca
     5041 ctttcacgag ctcgtccacc accttcactg tctgcaggat gcccttctta atggcggggc
     5101 tgccggccag attggcaatg tgctcgtgca ggctatcgcc ctggccggac acctgggctt
     5161 tctggatgtc ctctttaaag gtcaggctgt cgtcgtggat cagctgcatg aagtttctgt
     5221 tggcgaagcc gtcggacttc aggaaatcca ggattgtctt gccggactgc ttgtcccgga
     5281 tgccgttgat cagcttccgg ctcagcctgc cccagccggt gtatctccgc cgcttcagct
     5341 gcttcatcac tttgtcgtcg aacaggtggg cataggtttt cagccgttcc tcgatcatct
     5401 ctctgtcctc aaacagtgtc agggtcagca cgatatcttc cagaatgtcc tcgttttcct
     5461 cattgtccag gaagtccttg tccttgataa ttttcagcag atcgtggtat gtgcccaggg
     5521 aggcgttgaa ccgatcttcc acgccggaga tttccacgga gtcgaagcac tcgattttct
     5581 tgaagtagtc ctctttcagc tgcttcacgg tcactttccg gttggtcttg aacagcaggt
     5641 ccacgatggc ctttttctgc tcgccgctca ggaaggcggg ctttctcatt ccctcggtca
     5701 cgtatttcac tttggtcagc tcgttataca cggtgaagta ctcgtacagc aggctgtgct
     5761 tgggcagcac cttctcgttg ggcaggttct tatcgaagtt ggtcatccgc tcgatgaagc
     5821 tctgggcgga agcgcccttg tccaccactt cctcgaagtt ccagggggtg atggtttcct
     5881 cgctctttct ggtcatccag gcgaatctgc tgtttcccct ggccagaggg cccacgtagt
     5941 aggggatgcg gaaggtcagg atcttctcga tcttttcccg gttgtccttc aggaatgggt
     6001 aaaaatcttc ctgccgccgc agaatggcgt gcagctctcc caggtggatc tggtggggga
     6061 tgctgccgtt gtcgaaggtc cgctgcttcc gcagcaggtc ctctctgttc agcttcacga
     6121 gcagttcctc ggtgccgtcc atcttttcca ggatgggctt gatgaacttg tagaactctt
     6181 cctggctggc tccgccgtca atgtagccgg cgtagccgtt cttgctctgg tcgaagaaaa
     6241 tctctttgta cttctcaggc agctgctgcc gcacgagagc tttcagcagg gtcaggtcct
     6301 ggtggtgctc gtcgtatctc ttgatcatag aggcgctcag gggggccttg gtgatctcgg
     6361 tgttcactct caggatgtcg ctcagcagga tggcgtcgga caggttcttg gcggccagaa
     6421 acaggtcggc gtactggtcg ccgatctggg ccagcaggtt gtccaggtcg tcgtcgtagg
     6481 tgtccttgct cagctgcagt ttggcatcct cggccaggtc gaagttgctc ttgaagttgg
     6541 gggtcaggcc caggctcagg gcaatcaggt ttccgaacag gccattcttc ttctcgccgg
     6601 gcagctgggc gatcagattt tccagccgtc tgctcttgct cagtctggca gacaggatgg
     6661 ccttggcgtc cacgccgctg gcgttgatgg ggttttcctc gaacagctgg ttgtaggtct
     6721 gcaccagctg gatgaacagc ttgtccacgt cgctgttgtc ggggttcagg tcgccctcga
     6781 tcaggaagtg gccccggaac ttgatcatgt gggccagggc cagatagatc agccgcaggt
     6841 cggccttgtc ggtgctgtcc accagtttct ttctcaggtg gtagatggtg gggtacttct
     6901 cgtggtaggc cacctcgtcc acgatgttgc cgaagatggg gtgccgctcg tgcttcttat
     6961 cctcttccac caggaaggac tcttccagtc tgtggaagaa gctgtcgtcc accttggcca
     7021 tctcgttgct gaagatctct tgcagatagc agatccggtt cttccgtctg gtgtatcttc
     7081 ttctggcggt tctcttcagc cgggtggcct cggctgtttc gccgctgtcg aacagcaggg
     7141 ctccgatcag gttcttcttg atgctgtgcc ggtcggtgtt gcccagcacc ttgaatttct
     7201 tgctgggcac cttgtactcg tcggtgatca cggcccagcc cacagagttg gtgccgatgt
     7261 ccaggccgat gctgtacttc ttgtcgactt tccgcttctt ctttggtgac tcgaactcgc
     7321 ttccgtcggc tgtccgtttc atggtggcgg tacctctaga gagatctcaa ttcaggcgaa
     7381 aatccgggtc gaaagttacg gttatcgcgc actctacttt ccacaaatcc tcacccaaaa
     7441 accaagcaca gtttattcaa ctgaagtatt cgcgatactt ctttatctaa taataatgta
     7501 catgtaacta aactcgcttt tgggttaaaa acgtgacgca gaggcaaaaa aaatcgtatg
     7561 tcccttagac aacttgaaac aactgcgaag cgtacggcaa ttccaggaat tttgtggtaa
     7621 agctacgcgc caactaacgg ttcttgctta gaggtggaat aatgtagttt tccagcgata
     7681 ataaatatat cgatattttt agtaaaattg aaaaggtaaa cttaatttta gaaaataatt
     7741 tataagaaat ttaatagtat gcaaaataat ttttacttgc taagaatatg tgccactaat
     7801 taaaagctgg acaccgcgca atggaaaata gtactacaac acagcaacaa agcctgagtt
     7861 atcaacaaaa aaatacgaaa acatctccca aaactaagca cccacacgcg ccactcgccg
     7921 tcacaacaca atcactgcac accaccattc gaatttcgcg cactgtgaca acatcacatg
     7981 atatcggcgc ggcaacatcg gattaccgac aaaacgaact atcgcacgag ccaccgccgg
     8041 cgaagagcgc tcgttttgca acaccggcgc gcgctgaacg aagagaacag ctgactgctt
     8101 gatacgtgcg tgtttcgcgg caggaattac ataaagttta gagcctctga cgccagaccc
     8161 cccgaacatt cgctccgatc aaactacctg cgaacggtca cctaatcccc accatggtag
     8221 gttacctctg atcccggtca tcactggcgt tcgctcacat ccgtccttac atgtgcatat
     8281 ttcgaggtta aaacggtcga agccctagag atcctgctag acgaattagg ccttctagtg
     8341 gatcaattcg gcttgtcgac atgcccgccg tgaccgtcga gaacccgctg acgctgcccc
     8401 gcgtatccgc accagccgac gccgtcgcac gtcccgtgct caccgtgacc accgcgccca
     8461 gcggtttcga gggcgagggc ttcccggtgc gccgcgcgtt cgccgggatc aactaccgcc
     8521 acctcgaccc gttcatcatg atggaccaga tgggtgaggt ggagtacgcg cccggggagc
     8581 ccaagggcac gccctggcac ccgcaccgcg gcttcgagac cgtgacctac atcgtcgacg
     8641 gtacgtgaag ccgaattgat ccggagagct cccaacgcgt tggatgcagc gtcgagatcg
     8701 atgatatcaa gcttggattt gtgtgcgccg cactttcacc tcaagtgatt gataattccc
     8761 agcctatctg gcagtgccca tcgcccagat caccgactgt gcaatcagtc ggaactggag
     8821 ctctctcgct ctgttatcgg ttcgctgggg tctcatctcc ggtccgctgg cggagatcag
     8881 ttcgccagca tccgccggtc gaggagtcac gatctgatct gagctgtgca ccatgagctg
     8941 tccctatgca ggaaacgggt gagcaccagc acgtgctgtc caggaatgcc aatcgatctt
     9001 cagttctgcg attcaattca aacccataca gaaacgatca cgatgattcg gcggtgccat
     9061 taaccacgga agtgggcaaa atctatggag agtatctgat gctggacaaa ctgctggatg
     9121 cccagtgtat gctgtccgag gaggacaagc gacccgtgca cgatgagcat ctgttcatca
     9181 tcacgcacca gggtgagtag gtttacaact ttgatgacaa cactcaatgg catttaagta
     9241 ccttcgccac agcctacgag ctttggttca agcagatcat ctttgagttc gactccatac
     9301 gagacatgtt ggatgcagag gtcatcgatg aaaccaagac gctggagatt gtcaagcgac
     9361 tgaaccgagt ggttctgatt ctaaaagtga gtgctttctg aatctcttac caaaatccgt
     9421 ttataacttc ctttgtacag ctcctggtgg accaagtgcc cattctggag accatgaccc
     9481 cgctagactt catggacttc cgcaagtacc tggcacccgc atctggtttt cagtcgctgc
     9541 agttccgttt gatcgagaac aagctgggag ttctgacaga gcagcgggtg agatacaacc
     9601 agaagtactc ggatgtcttt agcgacgagg aggcgcggaa ctcgattcgc aactcggaga
     9661 aagatccctc gctactggag ctagtgcagc gatggctgga gaggacgccc ggactggagg
     9721 agagtggctt caacttctgg gccaagtttc aggagagcgt cgatcgattc ctggaggcgc
     9781 aggtacagag cgccatggag gagcccgtgg agaaggcgaa aaactaccgc ctcatggaca
     9841 ttgagaagcg acgcgaggtg tatcgctcca tctttgatcc ggcagtgcac gatgcactgg
     9901 tgcgtcgtgg ggatcgccgg tttagccatc gtgcccttca gggagccatc atgatcacct
     9961 tctataggga tgaacccagg ttcagccaac cacaccagtt gctcaccctg ctcatggaca
    10021 tcgactcgtt aataaccaag tggagatgta agtattgcat tctttgatac tcttttataa
    10081 atatatctta tgtttaagac tggttttcct aaccaaatac tttctattcc cgccgcagac
    10141 aatcacgtga tcatggtgca acgcatgatt ggatcgcaac agttgggcac tggtggctcg
    10201 tctggttatc aatatctgcg ctccactctc aggtgatcat cgcagatgtg attatatcgg
    10261 ggatcaatga actcaaactg ttctcccttt gttttttttt ggtttcagtg atcggtacaa
    10321 ggtgtttctg gatctgttca atctgtccac ttttctgatt ccccgcgagg cgattccacc
    10381 gctggacgag accattcgca agaaactgat caacaaaagt gtctgacaat cggcagggta
    10441 tccaattcgt caatgtttgg ctatgcgttg tttgttctgc ctactgtttt gtcgttttgg
    10501 tgtaataaaa ttacttgttt agtctttgtt atcacatttg atgtgttcct tttctttatg
    10561 tctgacatat aatacatata acataacaaa ataaatccaa gcttgtaccc aattcgccct
    10621 atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa
    10681 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta
    10741 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
    10801 ggaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
    10861 atttttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
    10921 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
    10981 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
    11041 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
    11101 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
    11161 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
    11221 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa
    11281 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
    11341 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
    11401 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
    11461 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
    11521 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
    11581 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
    11641 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
    11701 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
    11761 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
    11821 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
    11881 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
    11941 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
    12001 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
    12061 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
    12121 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
    12181 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
    12241 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
    12301 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
    12361 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
    12421 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
    12481 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
    12541 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
    12601 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
    12661 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
    12721 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
    12781 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
    12841 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
    12901 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
    12961 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
    13021 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
    13081 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
    13141 gccgcagccg aacgaccgag cgcagcgagt cagtgaacga ggaagcggaa gagcgcccaa
    13201 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
    13261 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt
    13321 aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg
    13381 gataacaatt tcacacagga aacagctatg acca
//
