LOCUS       sequence-255032-       11372 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Cas9 C-terminal gateway fusion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11372)
  TITLE     Gateway compatible mammalian Cas9 vectors
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 11372)
  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: 
            "Unpublished"
COMMENT     Sequence Label: Cas9 gateway compatible
FEATURES             Location/Qualifiers
     source          1..11372
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(4..23)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        195..574
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        575..778
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     729..749
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     775..799
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      887..896
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             893..4996
                     /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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             893..4996
                     /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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             893..4996
                     /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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             5009..5029
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    5066..5190
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(5215..5220,5221..5238,5239..5245)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             5299..5958
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(5365..5384)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             6300..6605
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     6484..6503
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(6646..6770)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(6903..6922)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6947..6995
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7197..7625
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7284..7303)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7494..7515
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7634..7654)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7639..7968
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7819..7954
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7881..7900
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             8035..8829
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(8089..8108)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     8699..8718
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    9005..9126
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9042..9061)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9096..9115
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(9175..9191)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9175..9191)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9188..9210)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9199..9215
                     /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)."
     promoter        complement(join(9223..9229,9230..9247,9248..9253))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9268..9289
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(9406..9423)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9577..10165)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9657..9676)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10336..11127,11128..11196))
                     /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     10959..10978
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11197..11301)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt
       61 ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga
      121 caattgcatg aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc
      181 cagatatacg cgttgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc
      241 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
      301 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
      361 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
      421 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
      481 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
      541 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa
      601 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
      661 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc
      721 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg
      781 tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag
      841 acaccgggac cgatccagcc tccggactct agaggatcga acccttgcca ccatggacaa
      901 gaagtactcc attgggctcg atatcggcac aaacagcgtc ggctgggccg tcattacgga
      961 cgagtacaag gtgccgagca aaaaattcaa agttctgggc aataccgatc gccacagcat
     1021 aaagaagaac ctcattggcg ccctcctgtt cgactccggg gagacggccg aagccacgcg
     1081 gctcaaaaga acagcacggc gcagatatac ccgcagaaag aatcggatct gctacctgca
     1141 ggagatcttt agtaatgaga tggctaaggt ggatgactct ttcttccata ggctggagga
     1201 gtcctttttg gtggaggagg ataaaaagca cgagcgccac ccaatctttg gcaatatcgt
     1261 ggacgaggtg gcgtaccatg aaaagtaccc aaccatatat catctgagga agaagcttgt
     1321 agacagtact gataaggctg acttgcggtt gatctatctc gcgctggcgc atatgatcaa
     1381 atttcgggga cacttcctca tcgaggggga cctgaaccca gacaacagcg atgtcgacaa
     1441 actctttatc caactggttc agacttacaa tcagcttttc gaagagaacc cgatcaacgc
     1501 atccggagtt gacgccaaag caatcctgag cgctaggctg tccaaatccc ggcggctcga
     1561 aaacctcatc gcacagctcc ctggggagaa gaagaacggc ctgtttggta atcttatcgc
     1621 cctgtcactc gggctgaccc ccaactttaa atctaacttc gacctggccg aagatgccaa
     1681 gcttcaactg agcaaagaca cctacgatga tgatctcgac aatctgctgg cccagatcgg
     1741 cgaccagtac gcagaccttt ttttggcggc aaagaacctg tcagacgcca ttctgctgag
     1801 tgatattctg cgagtgaaca cggagatcac caaagctccg ctgagcgcta gtatgatcaa
     1861 gcgctatgat gagcaccacc aagacttgac tttgctgaag gcccttgtca gacagcaact
     1921 gcctgagaag tacaaggaaa ttttcttcga tcagtctaaa aatggctacg ccggatacat
     1981 tgacggcgga gcaagccagg aggaatttta caaatttatt aagcccatct tggaaaaaat
     2041 ggacggcacc gaggagctgc tggtaaagct taacagagaa gatctgttgc gcaaacagcg
     2101 cactttcgac aatggaagca tcccccacca gattcacctg ggcgaactgc acgctatcct
     2161 caggcggcaa gaggatttct accccttttt gaaagataac agggaaaaga ttgagaaaat
     2221 cctcacattt cggataccct actatgtagg ccccctcgcc cggggaaatt ccagattcgc
     2281 gtggatgact cgcaaatcag aagagaccat cactccctgg aacttcgagg aagtcgtgga
     2341 taagggggcc tctgcccagt ccttcatcga aaggatgact aactttgata aaaatctgcc
     2401 taacgaaaag gtgcttccta aacactctct gctgtacgag tacttcacag tttataacga
     2461 gctcaccaag gtcaaatacg tcacagaagg gatgagaaag ccagcattcc tgtctggaga
     2521 gcagaagaaa gctatcgtgg acctcctctt caagacgaac cggaaagtta ccgtgaaaca
     2581 gctcaaagaa gactatttca aaaagattga atgtttcgac tctgttgaaa tcagcggagt
     2641 ggaggatcgc ttcaacgcat ccctgggaac gtatcacgat ctcctgaaaa tcattaaaga
     2701 caaggacttc ctggacaatg aggagaacga ggacattctt gaggacattg tcctcaccct
     2761 tacgttgttt gaagataggg agatgattga agaacgcttg aaaacttacg ctcatctctt
     2821 cgacgacaaa gtcatgaaac agctcaagag gcgccgatat acaggatggg ggcggctgtc
     2881 aagaaaactg atcaatggga tccgagacaa gcagagtgga aagacaatcc tggattttct
     2941 taagtccgat ggatttgcca accggaactt catgcagttg atccatgatg actctctcac
     3001 ctttaaggag gacatccaga aagcacaagt ttctggccag ggggacagtc ttcacgagca
     3061 catcgctaat cttgcaggta gcccagctat caaaaaggga atactgcaga ccgttaaggt
     3121 cgtggatgaa ctcgtcaaag taatgggaag gcataagccc gagaatatcg ttatcgagat
     3181 ggcccgagag aaccaaacta cccagaaggg acagaagaac agtagggaaa ggatgaagag
     3241 gattgaagag ggtataaaag aactggggtc ccaaatcctt aaggaacacc cagttgaaaa
     3301 cacccagctt cagaatgaga agctctacct gtactacctg cagaacggca gggacatgta
     3361 cgtggatcag gaactggaca tcaatcggct ctccgactac gacgtggatc atatcgtgcc
     3421 ccagtctttt ctcaaagatg attctattga taataaagtg ttgacaagat ccgataaaaa
     3481 tagagggaag agtgataacg tcccctcaga agaagttgtc aagaaaatga aaaattattg
     3541 gcggcagctg ctgaacgcca aactgatcac acaacggaag ttcgataatc tgactaaggc
     3601 tgaacgaggt ggcctgtctg agttggataa agccggcttc atcaaaaggc agcttgttga
     3661 gacacgccag atcaccaagc acgtggccca aattctcgat tcacgcatga acaccaagta
     3721 cgatgaaaat gacaaactga ttcgagaggt gaaagttatt actctgaagt ctaagctggt
     3781 ctcagatttc agaaaggact ttcagtttta taaggtgaga gagatcaaca attaccacca
     3841 tgcgcatgat gcctacctga atgcagtggt aggcactgca cttatcaaaa aatatcccaa
     3901 gcttgaatct gaatttgttt acggagacta taaagtgtac gatgttagga aaatgatcgc
     3961 aaagtctgag caggaaatag gcaaggccac cgctaagtac ttcttttaca gcaatattat
     4021 gaattttttc aagaccgaga ttacactggc caatggagag attcggaagc gaccacttat
     4081 cgaaacaaac ggagaaacag gagaaatcgt gtgggacaag ggtagggatt tcgcgacagt
     4141 ccggaaggtc ctgtccatgc cgcaggtgaa catcgttaaa aagaccgaag tacagaccgg
     4201 aggcttctcc aaggaaagta tcctcccgaa aaggaacagc gacaagctga tcgcacgcaa
     4261 aaaagattgg gaccccaaga aatacggcgg attcgattct cctacagtcg cttacagtgt
     4321 actggttgtg gccaaagtgg agaaagggaa gtctaaaaaa ctcaaaagcg tcaaggaact
     4381 gctgggcatc acaatcatgg agcgatcaag cttcgaaaaa aaccccatcg actttctcga
     4441 ggcgaaagga tataaagagg tcaaaaaaga cctcatcatt aagcttccca agtactctct
     4501 ctttgagctt gaaaacggcc ggaaacgaat gctcgctagt gcgggcgagc tgcagaaagg
     4561 taacgagctg gcactgccct ctaaatacgt taatttcttg tatctggcca gccactatga
     4621 aaagctcaaa gggtctcccg aagataatga gcagaagcag ctgttcgtgg aacaacacaa
     4681 acactacctt gatgagatca tcgagcaaat aagcgaattc tccaaaagag tgatcctcgc
     4741 cgacgctaac ctcgataagg tgctttctgc ttacaataag cacagggata agcccatcag
     4801 ggagcaggca gaaaacatta tccacttgtt tactctgacc aacttgggcg cgcctgcagc
     4861 cttcaagtac ttcgacacca ccatagacag aaagcggtac acctctacaa aggaggtcct
     4921 ggacgccaca ctgattcatc agtcaattac ggggctctat gaaacaagaa tcgacctctc
     4981 tcagctcggt ggagacagca gggctgaccc caagaagaag aggaaggtgt cgccagggat
     5041 ccgtcgactt gacgcgttaa tatcaacaag tttgtacaaa aaagctgaac gagaaacgta
     5101 aaatgatata aatatcaata tattaaatta gattttgcat aaaaaacaga ctacataata
     5161 ctgtaaaaca caacatatcc agtcactatg gcggccgcat taggcacccc aggctttaca
     5221 ctttatgctt ccggctcgta taatgtgtgg attttgagtt aggatccgtc gagattttca
     5281 ggagctaagg aagctaaaat ggagaaaaaa atcactggat ataccaccgt tgatatatcc
     5341 caatggcatc gtaaagaaca ttttgaggca tttcagtcag ttgctcaatg tacctataac
     5401 cagaccgttc agctggatat tacggccttt ttaaagaccg taaagaaaaa taagcacaag
     5461 ttttatccgg cctttattca cattcttgcc cgcctgatga atgctcatcc ggaattccgt
     5521 atggcaatga aagacggtga gctggtgata tgggatagtg ttcacccttg ttacaccgtt
     5581 ttccatgagc aaactgaaac gttttcatcg ctctggagtg aataccacga cgatttccgg
     5641 cagtttctac acatatattc gcaagatgtg gcgtgttacg gtgaaaacct ggcctatttc
     5701 cctaaagggt ttattgagaa tatgtttttc gtctcagcca atccctgggt gagtttcacc
     5761 agttttgatt taaacgtggc caatatggac aacttcttcg cccccgtttt caccatgggc
     5821 aaatattata cgcaaggcga caaggtgctg atgccgctgg cgattcaggt tcatcatgcc
     5881 gtttgtgatg gcttccatgt cggcagaatg cttaatgaat tacaacagta ctgcgatgag
     5941 tggcagggcg gggcgtaaag atctggatcc ggcttactaa aagccagata acagtatgcg
     6001 tatttgcgcg ctgatttttg cggtataaga atatatactg atatgtatac ccgaagtatg
     6061 tcaaaaagag gtatgctatg aagcagcgta ttacagtgac agttgacagc gacagctatc
     6121 agttgctcaa ggcatatatg atgtcaatat ctccggtctg gtaagcacaa ccatgcagaa
     6181 tgaagcccgt cgtctgcgtg ccgaacgctg gaaagcggaa aatcaggaag ggatggctga
     6241 ggtcgcccgg tttattgaaa tgaacggctc ttttgctgac gagaacaggg gctggtgaaa
     6301 tgcagtttaa ggtttacacc tataaaagag agagccgtta tcgtctgttt gtggatgtac
     6361 agagtgatat tattgacacg cccgggcgac ggatggtgat ccccctggcc agtgcacgtc
     6421 tgctgtcaga taaagtctcc cgtgaacttt acccggtggt gcatatcggg gatgaaagct
     6481 ggcgcatgat gaccaccgat atggccagtg tgccggtctc cgttatcggg gaagaagtgg
     6541 ctgatctcag ccaccgcgaa aatgacatca aaaacgccat taacctgatg ttctggggaa
     6601 tataaatgtc aggctccctt atacacagcc agtctgcagg tcgaccatag tgactggata
     6661 tgttgtgttt tacagtatta tgtagtctgt tttttatgca aaatctaatt taatatattg
     6721 atatttatat cattttacgt ttctcgttca gctttcttgt acaaagtggt tgatatcatc
     6781 tagagcggcc gcaggtacct gaataactaa ggccgcttcc ctttagtgag ggttaatgct
     6841 tcgagcagac aaagggttcg atccctaccg gttagtaatg agtttaaacg ggggaggcta
     6901 actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga
     6961 cagaataaaa cgcacgggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg
     7021 ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc
     7081 cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg
     7141 ggcggcaggc cctgccatag cagatctgcg cagctggggc tctagggggt atccccacgc
     7201 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     7261 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     7321 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     7381 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     7441 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     7501 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     7561 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     7621 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     7681 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     7741 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     7801 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     7861 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
     7921 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     7981 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     8041 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     8101 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     8161 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     8221 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     8281 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     8341 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     8401 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     8461 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     8521 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     8581 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     8641 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     8701 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     8761 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     8821 ttcttctgag cgggactctg gggttcgcga aatgaccgac caagcgacgc ccaacctgcc
     8881 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     8941 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     9001 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     9061 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     9121 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     9181 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     9241 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     9301 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     9361 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     9421 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     9481 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     9541 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     9601 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     9661 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     9721 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     9781 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     9841 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     9901 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     9961 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
    10021 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
    10081 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
    10141 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
    10201 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
    10261 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
    10321 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
    10381 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
    10441 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
    10501 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
    10561 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
    10621 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
    10681 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
    10741 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
    10801 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
    10861 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
    10921 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
    10981 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
    11041 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
    11101 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
    11161 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
    11221 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
    11281 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc
    11341 gggagatctc ccgatcccct atggtgcact ct
//
