LOCUS       sequence-153106-       11270 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cas9, mCherry expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11270)
  AUTHORS   Schmid-Burgk JL, Honing K, Ebert TS, Hornung V
  TITLE     CRISPaint allows modular base-specific gene tagging using a 
            ligase-4-dependent mechanism.
  JOURNAL   Nat Commun. 2016 Jul 28;7:12338. doi: 10.1038/ncomms12338.
  PUBMED    27465542
REFERENCE   2  (bases 1 to 11270)
  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 
            Commun."; date: "2016-07-28"; volume: "7:12338. doi"; pages: " 
            10.1038/ncomms12338"
COMMENT     Sequence Label: pRZ-CAS9-mCherry
FEATURES             Location/Qualifiers
     source          1..11270
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        215..594
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        595..798
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             799..974
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    1037..1236
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1437..1853
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1783..1805
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1821..1837
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1865..2239
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     enhancer        2375..2754
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2755..2958
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2955..2979
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      3067..3076
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3076..3099
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3100..3804
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(3242..3260)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3510..3529
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             3829..3882
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             3883..7986
                     /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             3883..7986
                     /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             7999..8019
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     LTR             8076..8669
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(8752..8772)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        8770..9067
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8937..9072
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     8999..9018
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     complement(9113..9129)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9113..9129)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9126..9148)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9137..9153
                     /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(9161..9167,9168..9185,9186..9191))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9206..9227
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(9344..9361)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9515..10103)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9595..9614)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10274..11065,11066..11134))
                     /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     10897..10916
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11135..11239)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg agttgacatt gattattgac tagttattaa
      241 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
      301 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
      361 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
      421 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
      481 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
      541 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg
      601 cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt
      661 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca
      721 aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag
      781 gtctatataa gcagagctca ataaaagagc ccacaacccc tcactcggcg cgccagtctt
      841 ccgatagact gcgtcgcccg ggtacccgta ttcccaataa agcctcttgc tgtttgcatc
      901 cgaatcgtgg tctcgctgtt ccttgggagg gtctcctctg agtgattgac tacccacgac
      961 gggggtcttt catttggggg ctcgtccggg atttggagac ccctgcccag ggaccaccga
     1021 cccaccaccg ggaggtaagc tggccagcaa cttatctgtg tctgtccgat tgtctagtgt
     1081 ctatgtttga tgttatgcgc ctgcgtctgt actagttagc taactagctc tgtatctggc
     1141 ggacccgtgg tggaactgac gagttctgaa cacccggccg caaccctggg agacgtccca
     1201 gggactttgg gggccgtttt tgtggcccga cctgaggaag ggagtcgatg tggaatccga
     1261 ccccgtcagg atatgtggtt ctggtaggag acgagaacct aaaacagttc ccgcctccgt
     1321 ctgaattttt gctttcggtt tggaaccgaa gccgcgcgtc ttgtctgctg cagcgctgca
     1381 gcatcgttct gtgttgtctc tgtctgactg tgtttctgta tttgtctgaa aattagggcc
     1441 agactgttac cactccctta agtttgacct taggtcactg gaaagatgtc gagcggatcg
     1501 ctcacaacca gtcggtagat gtcaagaaga gacgttgggt taccttctgc tctgcagaat
     1561 ggccaacctt taacgtcgga tggccgcgag acggcacctt taaccgagac ctcatcaccc
     1621 aggttaagat caaggtcttt tcacctggcc cgcatggaca cccagaccag gtcccctaca
     1681 tcgtgacctg ggaagccttg gcttttgacc cccctccctg ggtcaagccc tttgtacacc
     1741 ctaagcctcc gcctcctctt cctccatccg ccccgtctct cccccttgaa cctcctcgtt
     1801 cgaccccgcc tcgatcctcc ctttatccag ccctcactcc ttctctaggc gcctccggac
     1861 caccatggcc aagttgacca gtgccgttcc ggtgctcacc gcgcgcgacg tcgccggagc
     1921 ggtcgagttc tggaccgacc ggctcgggtt ctcccgggac ttcgtggagg acgacttcgc
     1981 cggtgtggtc cgggacgacg tgaccctgtt catcagcgcg gtccaggacc aggtggtgcc
     2041 ggacaacacc ctggcctggg tgtgggtgcg cggcctggac gagctgtacg ccgagtggtc
     2101 ggaggtcgtg tccacgaact tccgggacgc ctccgggccg gccatgaccg agatcggcga
     2161 gcagccgtgg gggcgggagt tcgccctgcg cgacccggcc ggcaactgcg tgcacttcgt
     2221 ggccgaggag caggactgaa gatccggcca ttagccatat tattcattgg ttatatagca
     2281 taaatcaata ttggctattg gccattgcat acgttgtatc catatcataa tatgtacatt
     2341 tatattggct catgtccaac attaccgcca tgttgacatt gattattgac tagttattaa
     2401 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
     2461 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
     2521 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
     2581 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
     2641 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
     2701 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg
     2761 cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt
     2821 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca
     2881 aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag
     2941 gtctatataa gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc
     3001 tgttttgacc tccatagaag acaccgggac cgatccagcc tccgcggccc caagcttccg
     3061 cgtacggcca ccatggatta taaggatgac gacgataagg tgagcaaggg cgaggaggat
     3121 aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg ctccgtgaac
     3181 ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg cacccagacc
     3241 gccaagctga aggtgaccaa gggtggcccc ctgcccttcg cctgggacat cctgtcccct
     3301 cagttcatgt acggctccaa ggcctacgtg aagcaccccg ccgacatccc cgactacttg
     3361 aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc
     3421 gtggtgaccg tgacccagga ctcctccctg caggacggcg agttcatcta caaggtgaag
     3481 ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc agaagaagac catgggctgg
     3541 gaggcctcct ccgagcggat gtaccccgag gacggcgccc tgaagggcga gatcaagcag
     3601 aggctgaagc tgaaggacgg cggccactac gacgctgagg tcaagaccac ctacaaggcc
     3661 aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca tcaagttgga catcacctcc
     3721 cacaacgagg actacaccat cgtggaacag tacgaacgcg ccgagggccg ccactccacc
     3781 ggcggcatgg acgagctgta caaggtcgct accatcgagg gatccggaga gggcagagga
     3841 agtctgctaa catgcggtga cgtcgaggag aatcctggcc caatggacaa gaagtacagc
     3901 atcggcctgg acatcggcac caacagcgtg ggctgggccg tgatcaccga cgagtacaag
     3961 gtgcccagca agaagttcaa ggtgctgggc aacaccgacc gccacagcat caagaagaac
     4021 ctgatcggcg ccctgctgtt cgacagcggc gagaccgccg aggccacccg cctgaagcgc
     4081 accgcccgcc gccgctacac ccgccgcaag aaccgcatct gctacctgca ggagatcttc
     4141 agcaacgaga tggccaaggt ggacgacagc ttcttccacc gcctggagga gagcttcctg
     4201 gtggaggagg acaagaagca cgagcgccac cccatcttcg gcaacatcgt ggacgaggtg
     4261 gcctaccacg agaagtaccc caccatctac cacctgcgca agaagctggt ggacagcacc
     4321 gacaaggccg acctgcgcct gatctacctg gccctggccc acatgatcaa gttccgcggc
     4381 cacttcctga tcgagggcga cctgaacccc gacaacagcg acgtggacaa gctgttcatc
     4441 cagctggtgc agacctacaa ccagctgttc gaggagaacc ccatcaacgc cagcggcgtg
     4501 gacgccaagg ccatcctgag cgcccgcctg agcaagagcc gccgcctgga gaacctgatc
     4561 gcccagctgc ccggcgagaa gaagaacggc ctgttcggca acctgatcgc cctgagcctg
     4621 ggcctgaccc ccaacttcaa gagcaacttc gacctggccg aggacgccaa gctgcagctg
     4681 agcaaggaca cctacgacga cgacctggac aacctgctgg cccagatcgg cgaccagtac
     4741 gccgacctgt tcctggccgc caagaacctg agcgacgcca tcctgctgag cgacatcctg
     4801 cgcgtgaaca ccgagatcac caaggccccc ctgagcgcca gcatgatcaa gcgctacgac
     4861 gagcaccacc aggacctgac cctgctgaag gccctggtgc gccagcagct gcccgagaag
     4921 tacaaggaga tcttcttcga ccagagcaag aacggctacg ccggctacat cgacggcggc
     4981 gccagccagg aggagttcta caagttcatc aagcccatcc tggagaagat ggacggcacc
     5041 gaggagctgc tggtgaagct gaaccgcgag gacctgctgc gcaagcagcg caccttcgac
     5101 aacggcagca tcccccacca gatccacctg ggcgagctgc acgccatcct gcgccgccag
     5161 gaggacttct accccttcct gaaggacaac cgcgagaaga tcgagaagat cctgaccttc
     5221 cgcatcccct actacgtggg ccccctggcc cgcggcaaca gccgcttcgc ctggatgacc
     5281 cgcaagagcg aggagaccat caccccctgg aacttcgagg aggtggtgga caagggcgcc
     5341 agcgcccaga gcttcatcga gcgcatgacc aacttcgaca agaacctgcc caacgagaag
     5401 gtgctgccca agcacagcct gctgtacgag tacttcaccg tgtacaacga gctgaccaag
     5461 gtgaagtacg tgaccgaggg catgcgcaag cccgccttcc tgagcggcga gcagaagaag
     5521 gccatcgtgg acctgctgtt caagaccaac cgcaaggtga ccgtgaagca gctgaaggag
     5581 gactacttca agaagatcga gtgcttcgac agcgtggaga tcagcggcgt ggaggaccgc
     5641 ttcaacgcca gcctgggcac ctaccacgac ctgctgaaga tcatcaagga caaggacttc
     5701 ctggacaacg aggagaacga ggacatcctg gaggacatcg tgctgaccct gaccctgttc
     5761 gaggaccgcg agatgatcga ggagcgcctg aagacctacg cccacctgtt cgacgacaag
     5821 gtgatgaagc agctgaagcg ccgccgctac accggctggg gccgcctgag ccgcaagctg
     5881 atcaacggca tccgcgacaa gcagagcggc aagaccatcc tggacttcct gaagagcgac
     5941 ggcttcgcca accgcaactt catgcagctg atccacgacg acagcctgac cttcaaggag
     6001 gacatccaga aggcccaggt gagcggccag ggcgacagcc tgcacgagca catcgccaac
     6061 ctggccggca gccccgccat caagaagggc atcctgcaga ccgtgaaggt ggtggacgag
     6121 ctggtgaagg tgatgggccg ccacaagccc gagaacatcg tgatcgagat ggcccgcgag
     6181 aaccagacca cccagaaggg ccagaagaac agccgcgagc gcatgaagcg catcgaggag
     6241 ggcatcaagg agctgggcag ccagatcctg aaggagcacc ccgtggagaa cacccagctg
     6301 cagaacgaga agctgtacct gtactacctg cagaacggcc gcgacatgta cgtggaccag
     6361 gagctggaca tcaaccgcct gagcgactac gacgtggacc acatcgtgcc ccagagcttc
     6421 ctgaaggacg acagcatcga caacaaggtg ctgacccgca gcgacaagaa ccgcggcaag
     6481 agcgacaacg tgcccagcga ggaggtggtg aagaagatga agaactactg gcgccagctg
     6541 ctgaacgcca agctgatcac ccagcgcaag ttcgacaacc tgaccaaggc cgagcgcggc
     6601 ggcctgagcg agctggacaa ggccggcttc atcaagcgcc agctggtgga gacccgccag
     6661 atcaccaagc acgtggccca gatcctggac agccgcatga acaccaagta cgacgagaac
     6721 gacaagctga tccgcgaggt gaaggtgatc accctgaaga gcaagctggt gagcgacttc
     6781 cgcaaggact tccagttcta caaggtgcgc gagatcaaca actaccacca cgcccacgac
     6841 gcctacctga acgccgtggt gggcaccgcc ctgatcaaga agtaccccaa gctggagagc
     6901 gagttcgtgt acggcgacta caaggtgtac gacgtgcgca agatgatcgc caagagcgag
     6961 caggagatcg gcaaggccac cgccaagtac ttcttctaca gcaacatcat gaacttcttc
     7021 aagaccgaga tcaccctggc caacggcgag atccgcaagc gccccctgat cgagaccaac
     7081 ggcgagaccg gcgagatcgt gtgggacaag ggccgcgact tcgccaccgt gcgcaaggtg
     7141 ctgagcatgc cccaggtgaa catcgtgaag aagaccgagg tgcagaccgg cggcttcagc
     7201 aaggagagca tcctgcccaa gcgcaacagc gacaagctga tcgcccgcaa gaaggactgg
     7261 gaccccaaga agtacggcgg cttcgacagc cccaccgtgg cctacagcgt gctggtggtg
     7321 gccaaggtgg agaagggcaa gagcaagaag ctgaagagcg tgaaggagct gctgggcatc
     7381 accatcatgg agcgcagcag cttcgagaag aaccccatcg acttcctgga ggccaagggc
     7441 tacaaggagg tgaagaagga cctgatcatc aagctgccca agtacagcct gttcgagctg
     7501 gagaacggcc gcaagcgcat gctggccagc gccggcgagc tgcagaaggg caacgagctg
     7561 gccctgccca gcaagtacgt gaacttcctg tacctggcca gccactacga gaagctgaag
     7621 ggcagccccg aggacaacga gcagaagcag ctgttcgtgg agcagcacaa gcactacctg
     7681 gacgagatca tcgagcagat cagcgagttc agcaagcgcg tgatcctggc cgacgccaac
     7741 ctggacaagg tgctgagcgc ctacaacaag caccgcgaca agcccatccg cgagcaggcc
     7801 gagaacatca tccacctgtt caccctgacc aacctgggcg cccccgccgc cttcaagtac
     7861 ttcgacacca ccatcgaccg caagcgctac accagcacca aggaggtgct ggacgccacc
     7921 ctgatccacc agagcatcac cggcctgtac gagacccgca tcgacctgag ccagctgggc
     7981 ggcgacagcc gcgccgaccc caagaagaag cgcaaggtgt gagcggccgc atcgataaaa
     8041 taaaagattt tatttagtct ccagaaaaag gggggaatga aagaccccac ctgtaggttt
     8101 ggcaagctag cttaagtaac gccattttgc aaggcatgga aaaatacata actgagaata
     8161 gagaagttca gatcaaggtc aggaacagat ggaacagctg aatatgggcc aaacaggata
     8221 tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggaac agctgaatat
     8281 gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc aagaacagat
     8341 ggtccccaga tgcggtccag ccctcagcag tttctagaga accatcagat gtttccaggg
     8401 tgccccaagg acctgaaatg accctgtgcc ttatttgaac taaccaatca gttcgcttct
     8461 cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa agagcccaca acccctcact
     8521 cggggcgcca gtcctccgat tgactgagtc gcccgggtac ccgtgtatcc aataaaccct
     8581 cttgcagttg catccgactt gtggtctcgc tgttccttgg gagggtctcc tctgagtgat
     8641 tgactacccg tcagcggggg tctttcattt gggggctcgt ccgggatcgg gagacccctg
     8701 cccagggacc accgacccac caccgggagg taagctggga actaattctg tggaatgtgt
     8761 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     8821 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     8881 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     8941 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     9001 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct
     9061 tttttggagg cctagtaccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
     9121 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     9181 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     9241 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     9301 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     9361 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     9421 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     9481 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     9541 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     9601 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     9661 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct
     9721 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     9781 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     9841 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     9901 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
     9961 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
    10021 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
    10081 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
    10141 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
    10201 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
    10261 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
    10321 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
    10381 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
    10441 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
    10501 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
    10561 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
    10621 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
    10681 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
    10741 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
    10801 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
    10861 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
    10921 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
    10981 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
    11041 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
    11101 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
    11161 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
    11221 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
//
