LOCUS       sequence-374594-        9630 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid ensures constitutive expression of the C-terminal fragment 
            of split DHFR (aa 1-173), fused to the catalytically inactive Cas9 
            (dCas9) protein, a flexible GS linker and the SV40 NLS signal at the
            N-terminus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9630)
  AUTHORS   Rihtar E, Lebar T, Lainscek D, Kores K, Lesnik S, Bren U, Jerala R
  TITLE     Chemically inducible split protein regulators for mammalian cells.
  JOURNAL   Nat Chem Biol. 2022 Sep 26. doi: 10.1038/s41589-022-01136-x.
  PUBMED    36163385
REFERENCE   2  (bases 1 to 9630)
  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: "Nat Chem 
            Biol. 2022 Sep 26. doi: 10.1038/s41589-022-01136-x."
COMMENT     Sequence Label: dCas9-NLS-gs10-cDHFR174
FEATURES             Location/Qualifiers
     source          1..9630
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(88..107)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        279..658
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        659..862
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     813..833
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     907..926
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        907..925
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             997..1014
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             join(1015..1038,1039..1041,1042..3528,3529..3531,
                     3532..5112)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the H840A mutation in the HNH 
                     catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGG"
     CDS             1015..5112
                     /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="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGG"
     CDS             5119..5139
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        complement(5227..5245)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(5228..5245)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(5265..5282)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5271..5495
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5541..5969
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5628..5647)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5838..5859
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5978..5998)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5983..6312
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6163..6298
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6225..6244
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             6379..7173
                     /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(6433..6452)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7043..7062
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7347..7468
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7384..7403)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7438..7457
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7517..7533)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7517..7533)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7530..7552)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7541..7557
                     /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(7565..7571,7572..7589,7590..7595))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7610..7631
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7748..7765)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7919..8507)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7999..8018)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8678..9469,9470..9538))
                     /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     9301..9320
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(9539..9630,1..13))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc
       61 tcccgatccc ctatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
      121 gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc
      181 tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttaggg ttaggcgttt
      241 tgcgctgctt cgcgatgtac gggccagata tacgcgttga cattgattat tgactagtta
      301 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
      361 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
      421 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
      481 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
      541 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
      601 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
      661 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
      721 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
      781 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
      841 ggaggtctat ataagcagag ctctctggct aactagagaa cccactgctt actggcttat
      901 cgaaattaat acgactcact atagggagac ccaagcttgg taccgagctc ggatccacta
      961 gtaacggccg ccagtgtgct ggaattcgcc accatgcacc accaccacca ccacgacaag
     1021 aagtacagca tcggcctggc catcggcacc aactctgtgg gctgggccgt gatcaccgac
     1081 gagtacaagg tgcccagcaa gaaattcaag gtgctgggca acaccgaccg gcacagcatc
     1141 aagaagaacc tgatcggcgc cctgctgttc gacagcggag aaacagccga ggccacccgg
     1201 ctgaagagaa ccgccagaag aagatacacc agacggaaga accggatctg ctatctgcaa
     1261 gagatcttca gcaacgagat ggccaaggtg gacgacagct tcttccacag actggaagag
     1321 tccttcctgg tggaagagga taagaagcac gagcggcacc ccatcttcgg caacatcgtg
     1381 gacgaggtgg cctaccacga gaagtacccc accatctacc acctgagaaa gaaactggtg
     1441 gacagcaccg acaaggccga cctgcggctg atctatctgg ccctggccca catgatcaag
     1501 ttccggggcc acttcctgat cgagggcgac ctgaaccccg acaacagcga cgtggacaag
     1561 ctgttcatcc agctggtgca gacctacaac cagctgttcg aggaaaaccc catcaacgcc
     1621 agcggcgtgg acgccaaggc catcctgtct gccagactga gcaagagcag acggctggaa
     1681 aatctgatcg cccagctgcc cggcgagaag aagaatggcc tgttcggcaa cctgattgcc
     1741 ctgagcctgg gcctgacccc caacttcaag agcaacttcg acctggccga ggatgccaaa
     1801 ctgcagctga gcaaggacac ctacgacgac gacctggaca acctgctggc ccagatcggc
     1861 gaccagtacg ccgacctgtt tctggccgcc aagaacctgt ccgacgccat cctgctgagc
     1921 gacatcctga gagtgaacac cgagatcacc aaggcccccc tgagcgcctc tatgatcaag
     1981 agatacgacg agcaccacca ggacctgacc ctgctgaaag ctctcgtgcg gcagcagctg
     2041 cctgagaagt acaaagagat tttcttcgac cagagcaaga acggctacgc cggctacatc
     2101 gatggcggag ccagccagga agagttctac aagttcatca agcccatcct ggaaaagatg
     2161 gacggcaccg aggaactgct cgtgaagctg aacagagagg acctgctgcg gaagcagcgg
     2221 accttcgaca acggcagcat cccccaccag atccacctgg gagagctgca cgccattctg
     2281 cggcggcagg aagattttta cccattcctg aaggacaacc gggaaaagat cgagaagatc
     2341 ctgaccttcc gcatccccta ctacgtgggc cctctggcca ggggaaacag cagattcgcc
     2401 tggatgacca gaaagagcga ggaaaccatc accccctgga acttcgagga agtggtggac
     2461 aagggcgcca gcgcccagag cttcatcgag cggatgacca acttcgataa gaacctgccc
     2521 aacgagaagg tgctgcccaa gcacagcctg ctgtacgagt acttcaccgt gtacaacgag
     2581 ctgaccaaag tgaaatacgt gaccgaggga atgagaaagc ccgccttcct gagcggcgag
     2641 cagaaaaaag ccatcgtgga cctgctgttc aagaccaacc ggaaagtgac cgtgaagcag
     2701 ctgaaagagg actacttcaa gaaaatcgag tgcttcgact ccgtggaaat ctccggcgtg
     2761 gaagatcggt tcaacgcctc cctgggcaca taccacgatc tgctgaaaat tatcaaggac
     2821 aaggacttcc tggacaatga ggaaaacgag gacattctgg aagatatcgt gctgaccctg
     2881 acactgtttg aggacagaga gatgatcgag gaacggctga aaacctatgc ccacctgttc
     2941 gacgacaaag tgatgaagca gctgaagcgg cggagataca ccggctgggg caggctgagc
     3001 cggaagctga tcaacggcat ccgggacaag cagtccggca agacaatcct ggatttcctg
     3061 aagtccgacg gcttcgccaa cagaaacttc atgcagctga tccacgacga cagcctgacc
     3121 tttaaagagg acatccagaa agcccaggtg tccggccagg gcgatagcct gcacgagcac
     3181 attgccaatc tggccggcag ccccgccatt aagaagggca tcctgcagac agtgaaggtg
     3241 gtggacgagc tcgtgaaagt gatgggccgg cacaagcccg agaacatcgt gatcgaaatg
     3301 gccagagaga accagaccac ccagaaggga cagaagaaca gccgcgagag aatgaagcgg
     3361 atcgaagagg gcatcaaaga gctgggcagc cagatcctga aagaacaccc cgtggaaaac
     3421 acccagctgc agaacgagaa gctgtacctg tactacctgc agaatgggcg ggatatgtac
     3481 gtggaccagg aactggacat caaccggctg tccgactacg atgtggacgc tatcgtgcct
     3541 cagagctttc tgaaggacga ctccatcgat aacaaagtgc tgactcggag cgacaagaac
     3601 cggggcaaga gcgacaacgt gccctccgaa gaggtcgtga agaagatgaa gaactactgg
     3661 cgccagctgc tgaatgccaa gctgattacc cagaggaagt tcgacaatct gaccaaggcc
     3721 gagagaggcg gcctgagcga actggataag gccggcttca tcaagagaca gctggtggaa
     3781 acccggcaga tcacaaagca cgtggcacag atcctggact cccggatgaa cactaagtac
     3841 gacgagaacg acaaactgat ccgggaagtg aaagtgatca ccctgaagtc caagctggtg
     3901 tccgatttcc ggaaggattt ccagttttac aaagtgcgcg agatcaacaa ctaccaccac
     3961 gcccacgacg cctacctgaa cgccgtcgtg ggaaccgccc tgatcaaaaa gtaccctaag
     4021 ctggaaagcg agttcgtgta cggcgactac aaggtgtacg acgtgcggaa gatgatcgcc
     4081 aagagcgagc aggaaatcgg caaggctacc gccaagtact tcttctacag caacatcatg
     4141 aactttttca agaccgagat taccctggcc aacggcgaga tccggaagcg gcctctgatc
     4201 gagacaaacg gcgaaacagg cgagatcgtg tgggataagg gccgggactt tgccaccgtg
     4261 cggaaagtgc tgtctatgcc ccaagtgaat atcgtgaaaa agaccgaggt gcagacaggc
     4321 ggcttcagca aagagtctat cctgcccaag aggaacagcg acaagctgat cgccagaaag
     4381 aaggactggg accctaagaa gtacggcggc ttcgacagcc ccaccgtggc ctattctgtg
     4441 ctggtggtgg ccaaagtgga aaagggcaag tccaagaaac tgaagagtgt gaaagagctg
     4501 ctggggatca ccatcatgga aagaagcagc ttcgagaaga atcccatcga ctttctggaa
     4561 gccaagggct acaaagaagt gaaaaaggac ctgatcatca agctgcctaa gtactccctg
     4621 ttcgagctgg aaaacggccg gaagagaatg ctggcctctg ccggcgaact gcagaaggga
     4681 aacgaactgg ccctgccctc caaatatgtg aacttcctgt acctggccag ccactatgag
     4741 aagctgaagg gctcccccga ggataatgag cagaaacagc tgtttgtgga acagcacaaa
     4801 cactacctgg acgagatcat cgagcagatc agcgagttct ccaagagagt gatcctggcc
     4861 gacgctaatc tggacaaggt gctgagcgcc tacaacaagc acagagacaa gcctatcaga
     4921 gagcaggccg agaatatcat ccacctgttt accctgacca atctgggagc ccctgccgcc
     4981 ttcaagtact ttgacaccac catcgaccgg aagaggtaca ccagcaccaa agaggtgctg
     5041 gacgccaccc tgatccacca gagcatcacc ggcctgtacg agacacggat cgacctgtct
     5101 cagctgggag gctctgatcc aaaaaagaag agaaaggtag gcggctctgg cggcggctcc
     5161 ggaggctctg gtattaagta caagtttgag gtctatgaaa agaatgacta atctagaggg
     5221 ccctattcta tagtgtcacc taaatgctag agctcgctga tcagcctcga ctgtgccttc
     5281 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     5341 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     5401 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     5461 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
     5521 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     5581 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     5641 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     5701 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     5761 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccttt tgacgttgga
     5821 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     5881 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     5941 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
     6001 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     6061 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     6121 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     6181 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     6241 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     6301 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca
     6361 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     6421 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     6481 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     6541 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc
     6601 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     6661 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     6721 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     6781 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     6841 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     6901 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     6961 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     7021 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     7081 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     7141 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     7201 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     7261 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     7321 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
     7381 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     7441 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
     7501 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     7561 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     7621 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     7681 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     7741 ccacttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     7801 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     7861 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     7921 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     7981 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     8041 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     8101 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     8161 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     8221 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     8281 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     8341 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     8401 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     8461 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     8521 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     8581 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     8641 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     8701 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     8761 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     8821 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     8881 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     8941 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     9001 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     9061 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     9121 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     9181 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     9241 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     9301 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     9361 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     9421 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     9481 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     9541 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     9601 tatttgaatg tatttagaaa aataaacaaa
//
