LOCUS       sequence-417966-        8149 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Coding for Sp Cas9 alongside Sp sgRNA targeting GFP Target 4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8149)
  AUTHORS   Mangin A, Dion V, Menzies G
  TITLE     Developing small Cas9 hybrids using molecular modeling
  JOURNAL   bioRxiv 2023.10.24.563270
REFERENCE   2  (bases 1 to 8149)
  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: "bioRxiv 
            2023.10.24.563270"
COMMENT     Sequence Label: pUC57-ITR2-H1-GFP sgRNA Target 4- eCMV-
            S.pyoe...
FEATURES             Location/Qualifiers
     source          1..8149
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   432..572
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   443..572
                     /label=AAV2 ITR
     promoter        605..695
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     primer_bind     616..635
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     misc_RNA        724..799
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        895..1198
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1199..1402
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1353..1373
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1441..1450
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1447..5550
                     /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             1447..5550
                     /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             1447..5550
                     /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             5551..5622
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags"
                     /label=3xFLAG
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
     CDS             5626..5646
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    5665..5713
                     /note="synthetic polyadenylation signal"
     repeat_region   5730..5870
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5730..5859
                     /label=AAV2 ITR
     primer_bind     complement(5928..5944)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5928..5944)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5941..5963)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5952..5968
                     /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(5976..5982,5983..6000,6001..6006))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6021..6042
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(6330..6918)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6410..6429)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7089..7880,7881..7949))
                     /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     7712..7731
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7950..8054)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8122..8140
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cgagctcggt acctcgcgaa
      421 tgcatctaga tcctgcaggc agctgcgcgc tcgctcgctc actgaggccg cccgggcaaa
      481 gcccgggcgt cgggcgacct ttggtcgccc ggcctcagtg agcgagcgag cgcgcagaga
      541 gggagtggcc aactccatca ctaggggttc ctgcggcccc ccctaggtag gctaggctag
      601 gcgtatattt gcatgtcgct atgtgttctg ggaaatcacc ataaacgtga aatgtctttg
      661 gatttgggaa tcttataagt tctgtatgag accactcttt cccgggcgag gagctgttca
      721 ccggttttag agctagaaat agcaagttaa aataaggcta gtccgttatc aacttgaaaa
      781 agtggcaccg agtcggtgct ttttttttct agagggtacc ggggcccgcc taggtagtta
      841 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
      901 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
      961 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
     1021 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
     1081 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
     1141 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
     1201 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
     1261 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
     1321 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
     1381 ggaggtctat ataagcagag ctggtttagt gaaccgtcag atccgctagc gctaccggtc
     1441 gccaccatgg acaagaagta ctccattggg ctcgacatcg gcacaaacag cgtcggctgg
     1501 gccgtcatta cggacgagta caaggtgccg agcaaaaaat tcaaagttct gggcaatacc
     1561 gatcgccaca gcataaagaa gaacctcatt ggcgccctcc tgttcgactc cggggagacg
     1621 gccgaagcca cgcggctcaa aagaacagca cggcgcagat atacccgcag aaagaatcgg
     1681 atctgctacc tgcaggagat ctttagtaat gagatggcta aggtggatga ctctttcttc
     1741 cataggctgg aggagtcctt tttggtggag gaggataaaa agcacgagcg ccacccaatc
     1801 tttggcaata tcgtggacga ggtggcgtac catgaaaagt acccaaccat atatcatctg
     1861 aggaagaagc ttgtagacag tactgataag gctgacttgc ggttgatcta tctcgcgctg
     1921 gcgcatatga tcaaatttcg gggacacttc ctcatcgagg gggacctgaa cccagacaac
     1981 agcgatgtcg acaaactctt tatccaactg gttcagactt acaatcagct tttcgaagag
     2041 aacccgatca acgcatccgg agttgacgcc aaagcaatcc tgagcgctag gctgtccaaa
     2101 tcccggcggc tcgaaaacct catcgcacag ctccctgggg agaagaagaa cggcctgttt
     2161 ggtaatctta tcgccctgtc actcgggctg acccccaact ttaaatctaa cttcgacctg
     2221 gccgaagatg ccaagcttca actgagcaaa gacacctacg atgatgatct cgacaatctg
     2281 ctggcccaga tcggcgacca gtacgcagac ctttttttgg cggcaaagaa cctgtcagac
     2341 gccattctgc tgagtgatat tctgcgagtg aacacggaga tcaccaaagc tccgctgagc
     2401 gctagtatga tcaagcgcta tgatgagcac caccaagact tgactttgct gaaggccctt
     2461 gtcagacagc aactgcctga gaagtacaag gaaattttct tcgatcagtc taaaaatggc
     2521 tacgccggat acattgacgg cggagcaagc caggaggaat tttacaaatt tattaagccc
     2581 atcttggaaa aaatggacgg caccgaggag ctgctggtaa agcttaacag agaagatctg
     2641 ttgcgcaaac agcgcacttt cgacaatgga agcatccccc accagattca cctgggcgaa
     2701 ctgcacgcta tcctcaggcg gcaagaggat ttctacccct ttttgaaaga taacagggaa
     2761 aagattgaga aaatcctcac atttcggata ccctactatg taggccccct cgcccgggga
     2821 aattccagat tcgcgtggat gactcgcaaa tcagaagaga ccatcactcc ctggaacttc
     2881 gaggaagtcg tggataaggg ggcctctgcc cagtccttca tcgaaaggat gactaacttt
     2941 gataaaaatc tgcctaacga aaaggtgctt cctaaacact ctctgctgta cgagtacttc
     3001 acagtttata acgagctcac caaggtcaaa tacgtcacag aagggatgag aaagccagca
     3061 ttcctgtctg gagagcagaa gaaagctatc gtggacctcc tcttcaagac gaaccggaaa
     3121 gttaccgtga aacagctcaa agaagactat ttcaaaaaga ttgaatgttt cgactctgtt
     3181 gaaatcagcg gagtggagga tcgcttcaac gcatccctgg gaacgtatca cgatctcctg
     3241 aaaatcatta aagacaagga cttcctggac aatgaggaga acgaggacat tcttgaggac
     3301 attgtcctca cccttacgtt gtttgaagat agggagatga ttgaagaacg cttgaaaact
     3361 tacgctcatc tcttcgacga caaagtcatg aaacagctca agaggcgccg atatacagga
     3421 tgggggcggc tgtcaagaaa actgatcaat gggatccgag acaagcagag tggaaagaca
     3481 atcctggatt ttcttaagtc cgatggattt gccaacagga acttcatgca gttgatccat
     3541 gatgactctc tcacctttaa ggaggacatc cagaaagcac aagtttctgg ccagggggac
     3601 agtcttcacg agcacatcgc taatcttgca ggtagcccag ctatcaaaaa gggaatactg
     3661 cagaccgtta aggtcgtgga tgaactcgtc aaagtaatgg gaaggcataa gcccgagaat
     3721 atcgttatcg agatggcccg agagaaccaa actacccaga agggacagaa gaacagtagg
     3781 gaaaggatga agaggattga agagggtata aaagaactgg ggtcccaaat ccttaaggaa
     3841 cacccagttg aaaacaccca gcttcagaat gagaagctct acctgtacta cctgcagaac
     3901 ggcagggaca tgtacgtgga tcaggaactg gacatcaatc ggctctccga ctacgacgtg
     3961 gatcatatcg tgccccagtc ttttctcaaa gatgattcta ttgataataa agtgttgaca
     4021 agatccgata aaaatagagg gaagagtgat aacgtcccct cagaagaagt tgtcaagaaa
     4081 atgaaaaatt attggcggca gctgctgaac gccaaactga tcacacaacg gaagttcgat
     4141 aatctgacta aggctgaacg aggtggcctg tctgagttgg ataaagccgg cttcatcaaa
     4201 aggcagcttg ttgagacacg ccagatcacc aagcacgtgg cccaaattct cgattcacgc
     4261 atgaacacca agtacgatga aaatgacaaa ctgattcgag aggtgaaagt tattactctg
     4321 aagtctaagc tggtctcaga tttcagaaag gactttcagt tttataaggt gagagagatc
     4381 aacaattacc accatgcgca tgatgcctac ctgaatgcag tggtaggcac tgcacttatc
     4441 aaaaaatatc ccaagcttga atctgaattt gtttacggag actataaagt gtacgatgtt
     4501 aggaaaatga tcgcaaagtc tgagcaggaa ataggcaagg ccaccgctaa gtacttcttt
     4561 tacagcaata ttatgaattt tttcaagacc gagattacac tggccaatgg agagattcgg
     4621 aagcgaccac ttatcgaaac aaacggagaa acaggagaaa tcgtgtggga caagggtagg
     4681 gatttcgcga cagtccggaa ggtcctgtcc atgccgcagg tgaacatcgt taaaaagacc
     4741 gaagtacaga ccggaggctt ctccaaggaa agtatcctcc cgaaaaggaa cagcgacaag
     4801 ctgatcgcac gcaaaaaaga ttgggacccc aagaaatacg gcggattcga ttctcctaca
     4861 gtcgcttaca gtgtactggt tgtggccaaa gtggagaaag ggaagtctaa aaaactcaaa
     4921 agcgtcaagg aactgctggg catcacaatc atggagcgat caagcttcga aaaaaacccc
     4981 atcgactttc tcgaggcgaa aggatataaa gaggtcaaaa aagacctcat cattaagctt
     5041 cccaagtact ctctctttga gcttgaaaac ggccggaaac gaatgctcgc tagtgcgggc
     5101 gagctgcaga aaggtaacga gctggcactg ccctctaaat acgttaattt cttgtatctg
     5161 gccagccact atgaaaagct caaagggtct cccgaagata atgagcagaa gcagctgttc
     5221 gtggaacaac acaaacacta ccttgatgag atcatcgagc aaataagcga attctccaaa
     5281 agagtgatcc tcgccgacgc taacctcgat aaggtgcttt ctgcttacaa taagcacagg
     5341 gataagccca tcagggagca ggcagaaaac attatccact tgtttactct gaccaacttg
     5401 ggcgcgcctg cagccttcaa gtacttcgac accaccatag acagaaagcg gtacacctct
     5461 acaaaggagg tcctggacgc cacactgatt catcagtcaa ttacggggct ctatgaaaca
     5521 agaatcgacc tctctcagct cggtggagac gactacaagg acgacgacga caaggactat
     5581 aaggacgatg atgataagga ttacaaagat gacgacgata agccccccaa gaagaagagg
     5641 aaggtggagg ccagctaaga attcaataaa agatctttat tttcattaga tctgtgtgtt
     5701 ggttttttgt gtgtaagaat tccggccgca ggaaccccta gtgatggagt tggccactcc
     5761 ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg
     5821 ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg atcggatccc
     5881 gggcccgtcg actgcagagg cctgcatgca agcttggcgt aatcatggtc atagctgttt
     5941 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     6001 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     6061 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     6121 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     6181 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     6241 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     6301 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     6361 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     6421 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     6481 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     6541 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     6601 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     6661 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     6721 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     6781 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     6841 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     6901 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     6961 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     7021 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     7081 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     7141 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     7201 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     7261 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     7321 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     7381 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     7441 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     7501 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     7561 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     7621 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     7681 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     7741 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     7801 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     7861 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     7921 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     7981 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     8041 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt
     8101 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtc
//
