LOCUS       sequence-363020-        8859 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of spCas9, gRNA targeting intron 3 and gRNA targeting 
            3'UTR of the mouse Nefl gene. Can be used for the tagging of 
            endogenous NFL via Targeted Knock-In with Two guides (TKIT) 
            approach..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8859)
  AUTHORS   Arsic A, Hagemann C, Stajkovic N, Schubert T, Nikic-Spiegel I
  TITLE     Minimal genetically encoded tags for fluorescent protein labeling in
            living neurons.
  JOURNAL   Nat Commun. 2022 Jan 14;13(1):314. doi: 10.1038/s41467-022-27956-y.
  PUBMED    35031604
REFERENCE   2  (bases 1 to 8859)
  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: "2022-01-14"; volume: "13(1):314. doi"; pages: " 
            10.1038/s41467-022-27956-y"
COMMENT     Sequence Label: TKIT NFL gRNA1 gRNA2
FEATURES             Location/Qualifiers
     source          1..8859
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        119..404
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     misc_RNA        complement(524..599)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(620..868)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        complement(961..1036)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(1058..1306)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     rep_origin      complement(1368..1956)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1448..1467)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2127..2918,2919..2987))
                     /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     2750..2769
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2988..3092)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3160..3178
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3216..3238)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3338..3357
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(3374..3829)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3511..3532)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3723..3742
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   3904..4044
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3915..4044
                     /label=AAV2 ITR
     polyA_signal    4053..4260
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     4249..4266
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(4306..4326)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(4330..8430)
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             complement(4330..8430)
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             complement(8440..8460)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(8464..8490)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     intron          8511..8738
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
ORIGIN
        1 cncmsccccc cgcccccatc gctgcacaaa ataattaaaa aataaataaa tacaaaattg
       61 ggggtgggga ggggggggag atggggagag tgaagcagaa cgtggggctc acctcgacca
      121 tggtaatagc gatgactaat acgtagatgt actgccaagt aggaaagtcc cataaggtca
      181 tgtactgggc acaatgccag gcgggccatt taccgtcatt gacgtcaata gggggcgtac
      241 ttggcatatg atacacttga tgtactgcca agtgggcagt ttaccgtaaa tactccaccc
      301 attgacgtca atggaaagtc cctattggcg ttactattga cgtcaatggg cgggggtcgt
      361 tgggcggtca gccaggcggg ccatttaccg taagttatgt aacgggtacc tctggacgcg
      421 tcctaggacg cgtcctagga tcctcgagtc gacgaattgg cgcacgcgct aaaaacggac
      481 tagccttatt ttaacttgct atttctagct ctaaaacaaa aaagcaccga ctcggtgcca
      541 ctttttcaag ttgataacgg actagcctta ttttaacttg ctatttctag ctctaaaaca
      601 agtcatctca gaattatacg gtgtttcgtc ctttccacaa gatatataaa gccaagaaat
      661 cgaaatactt tcaagttacg gtaagcatat gatagtccat tttaaaacat aattttaaaa
      721 ctgcaaacta cccaagaaat tattactttc tacgtcacgt attttgtact aatatctttg
      781 tgtttacagt caaattaatt ccaattatct ctctaacagc cttgtatcgt atatgcaaat
      841 atgaaggaat catgggaaat aggccctcac tctagagcca tttgtctgca gaattggcgc
      901 acgcgctaaa aacggactag ccttatttta acttgctatt tctagctcta aaacaaaaaa
      961 gcaccgactc ggtgccactt tttcaagttg ataacggact agccttattt taacttgcta
     1021 tttctagctc taaaaccact ctcctaaagg atatgtcggt gtttcgtcct ttccacaaga
     1081 tatataaagc caagaaatcg aaatactttc aagttacggt aagcatatga tagtccattt
     1141 taaaacataa ttttaaaact gcaaactacc caagaaatta ttactttcta cgtcacgtat
     1201 tttgtactaa tatctttgtg tttacagtca aattaattcc aattatctct ctaacagcct
     1261 tgtatcgtat atgcaaatat gaaggaatca tgggaaatag gccctcacat gtgagcaaaa
     1321 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     1381 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     1441 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     1501 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     1561 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     1621 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     1681 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     1741 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     1801 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     1861 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     1921 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     1981 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     2041 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     2101 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     2161 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     2221 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcggct tccacgctca
     2281 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     2341 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     2401 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     2461 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     2521 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     2581 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     2641 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     2701 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     2761 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     2821 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     2881 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     2941 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     3001 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     3061 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     3121 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     3181 tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
     3241 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
     3301 gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg
     3361 agagtgcacc ataaaattgt aaacgttaat attttgttaa aattcgcgtt aaatttttgt
     3421 taaatcagct cattttttaa ccaatagacc gaaatcggca aaatccctta taaatcaaaa
     3481 gaatagcccg agatagagtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     3541 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     3601 gaaccatcac ccaaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     3661 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     3721 gaagggaaga aagcgaaagg agcgggcgct aaggcgctgg caagtgtagc ggtcacgctg
     3781 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgta ctatggttgc
     3841 tttgacgtat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc
     3901 gcccctgcag gcagctgcgc gctcgctcgc tcactgaggc cgcccgggca aagcccgggc
     3961 gtcgggcgac ctttggtcgc ccggcctcag tgagcgagcg agcgcgcaga gagggagtgg
     4021 ccaactccat cactaggggt tcctgcggcc gctccccagc atgcctgcta ttctcttccc
     4081 aatcctcccc cttgctgtcc tgccccaccc caccccccag aatagaatga cacctactca
     4141 gacaatgcga tgcaatttcc tcattttatt aggaaaggac agtgggagtg gcaccttcca
     4201 gggtcaagga aggcacgggg gaggggcaaa caacagatgg ctggcaacta gaaggcacag
     4261 tcgaggctga tcagcgagct ctagttagaa ttcttagctg gcctccacct ttctcttctt
     4321 cttggggctg tcgcctccca gctgagacag gtcgatccgt gtctcgtaca ggccggtgat
     4381 gctctggtgg atcagggtgg cgtccagcac ctctttggtg ctggtgtacc tcttccggtc
     4441 gatggtggtg tcaaagtact tgaaggcggc aggggctccc agattggtca gggtaaacag
     4501 gtggatgata ttctcggcct gctctctgat gggcttatcc cggtgcttgt tgtaggcgga
     4561 cagcactttg tccagattag cgtcggccag gatcactctc ttggagaact cgctgatctg
     4621 ctcgatgatc tcgtccaggt agtgcttgtg ctgttccaca aacagctgtt tctgctcatt
     4681 atcctcgggg gagcccttca gcttctcata gtggctggcc aggtacagga agttcacata
     4741 tttggagggc agggccagtt cgtttccctt ctgcagttcg ccggcagagg ccagcattct
     4801 cttccggccg ttttccagct cgaacaggga gtacttaggc agcttgatga tcaggtcctt
     4861 tttcacttct ttgtagccct tggcttccag aaagtcgatg ggattcttct cgaagctgct
     4921 tctttccatg atggtgatcc ccagcagctc tttcacactc ttcagtttct tggacttgcc
     4981 cttttccact ttggccacca ccagcacaga ataggccacg gtggggctgt cgaagccgcc
     5041 gtacttctta gggtcccagt ccttctttct ggcgatcagc ttatcgctgt tcctcttggg
     5101 caggatagac tctttgctga agccgcctgt ctgcacctcg gtctttttca cgatattcac
     5161 ttggggcatg ctcagcactt tccgcacggt ggcaaaatcc cggcccttat cccacacgat
     5221 ctccccggtt tcgccgtttg tctcgatcag aggccgcttc cggatctcgc cgttggccag
     5281 ggtaatctcg gtcttgaaaa agttcatgat gttgctgtag aagaagtact tggcggtagc
     5341 cttgccgatt tcctgctcgc tcttggcgat catcttccgc acgtcgtaca ccttgtagtc
     5401 gccgtacacg aactcgcttt ccagcttagg gtactttttg atcagggcgg ttcccacgac
     5461 ggcgttcagg taggcgtcgt gggcgtggtg gtagttgttg atctcgcgca ctttgtaaaa
     5521 ctggaaatcc ttccggaaat cggacaccag cttggacttc agggtgatca ctttcacttc
     5581 ccggatcagc ttgtcattct cgtcgtactt agtgttcatc cgggagtcca ggatctgtgc
     5641 cacgtgcttt gtgatctgcc gggtttccac cagctgtctc ttgatgaagc cggccttatc
     5701 cagttcgctc aggccgcctc tctcggcctt ggtcagattg tcgaactttc tctgggtaat
     5761 cagcttggcg ttcagcagct gccgccagta gttcttcatc ttcttcacga cctcttcgga
     5821 gggcacgttg tcgctcttgc cccggttctt gtcgcttctg gtcagcacct tgttgtcgat
     5881 ggagtcgtcc ttcagaaagc tctgaggcac gatatggtcc acatcgtagt cggacagccg
     5941 gttgatgtcc agttcctggt ccacgtacat atcccgccca ttctgcaggt agtacaggta
     6001 cagcttctcg ttctgcagct gggtgttttc cacggggtgt tctttcagga tctggctgcc
     6061 cagctctttg atgccctctt cgatccgctt cattctctcg cggctgttct tctgtccctt
     6121 ctgggtggtc tggttctctc tggccatttc gatcacgatg ttctcgggct tgtgccggcc
     6181 catcactttc acgagctcgt ccaccacctt cactgtctgc aggatgccct tcttaatggc
     6241 ggggctgccg gccagattgg caatgtgctc gtgcaggcta tcgccctggc cggacacctg
     6301 ggctttctgg atgtcctctt taaaggtcag gctgtcgtcg tggatcagct gcatgaagtt
     6361 tctgttggcg aagccgtcgg acttcaggaa atccaggatt gtcttgccgg actgcttgtc
     6421 ccggatgccg ttgatcagct tccggctcag cctgccccag ccggtgtatc tccgccgctt
     6481 cagctgcttc atcactttgt cgtcgaacag gtgggcatag gttttcagcc gttcctcgat
     6541 catctctctg tcctcaaaca gtgtcagggt cagcacgata tcttccagaa tgtcctcgtt
     6601 ttcctcattg tccaggaagt ccttgtcctt gataattttc agcagatcgt ggtatgtgcc
     6661 cagggaggcg ttgaaccgat cttccacgcc ggagatttcc acggagtcga agcactcgat
     6721 tttcttgaag tagtcctctt tcagctgctt cacggtcact ttccggttgg tcttgaacag
     6781 caggtccacg atggcctttt tctgctcgcc gctcaggaag gcgggctttc tcattccctc
     6841 ggtcacgtat ttcactttgg tcagctcgtt atacacggtg aagtactcgt acagcaggct
     6901 gtgcttgggc agcaccttct cgttgggcag gttcttatcg aagttggtca tccgctcgat
     6961 gaagctctgg gcggaagcgc ccttgtccac cacttcctcg aagttccagg gggtgatggt
     7021 ttcctcgctc tttctggtca tccaggcgaa tctgctgttt cccctggcca gagggcccac
     7081 gtagtagggg atgcggaagg tcaggatctt ctcgatcttt tcccggttgt ccttcaggaa
     7141 tgggtaaaaa tcttcctgcc gccgcagaat ggcgtgcagc tctcccaggt ggatctggtg
     7201 ggggatgctg ccgttgtcga aggtccgctg cttccgcagc aggtcctctc tgttcagctt
     7261 cacgagcagt tcctcggtgc cgtccatctt ttccaggatg ggcttgatga acttgtagaa
     7321 ctcttcctgg ctggctccgc cgtcaatgta gccggcgtag ccgttcttgc tctggtcgaa
     7381 gaaaatctct ttgtacttct caggcagctg ctgccgcacg agagctttca gcagggtcag
     7441 gtcctggtgg tgctcgtcgt atctcttgat catagaggcg ctcagggggg ccttggtgat
     7501 ctcggtgttc actctcagga tgtcgctcag caggatggcg tcggacaggt tcttggcggc
     7561 cagaaacagg tcggcgtact ggtcgccgat ctgggccagc aggttgtcca ggtcgtcgtc
     7621 gtaggtgtcc ttgctcagct gcagtttggc atcctcggcc aggtcgaagt tgctcttgaa
     7681 gttgggggtc aggcccaggc tcagggcaat caggtttccg aacaggccat tcttcttctc
     7741 gccgggcagc tgggcgatca gattttccag ccgtctgctc ttgctcagtc tggcagacag
     7801 gatggccttg gcgtccacgc cgctggcgtt gatggggttt tcctcgaaca gctggttgta
     7861 ggtctgcacc agctggatga acagcttgtc cacgtcgctg ttgtcggggt tcaggtcgcc
     7921 ctcgatcagg aagtggcccc ggaacttgat catgtgggcc agggccagat agatcagccg
     7981 caggtcggcc ttgtcggtgc tgtccaccag tttctttctc aggtggtaga tggtggggta
     8041 cttctcgtgg taggccacct cgtccacgat gttgccgaag atggggtgcc gctcgtgctt
     8101 cttatcctct tccaccagga aggactcttc cagtctgtgg aagaagctgt cgtccacctt
     8161 ggccatctcg ttgctgaaga tctcttgcag atagcagatc cggttcttcc gtctggtgta
     8221 tcttcttctg gcggttctct tcagccgggt ggcctcggct gtttcgccgc tgtcgaacag
     8281 cagggctccg atcaggttct tcttgatgct gtgccggtcg gtgttgccca gcaccttgaa
     8341 tttcttgctg ggcaccttgt actcgtcggt gatcacggcc cagcccacag agttggtgcc
     8401 gatgtccagg ccgatgctgt acttcttgtc ggacgcttcg accttgcgct ttttcttcgg
     8461 cgaagcgtaa tctggaacat cgtatgggta catggtggca ccggtccaac ctgaaaaaaa
     8521 gtgatttcag gcaggtgctc caggtaatta aacattaata ccccaccaac caaccatccc
     8581 ttaaaccctt acctcttgct cagctaatta cagcccggag gagaagggcc gtcccgcccg
     8641 ctcacctgtg ggagtaacgc ggtcagtcag agccggggcg ggcggcgcga ggcggcggcg
     8701 gagcggggca cggggcgaag gcagcgcgca gcgactcccg cccgccgcgc gcttcgcttt
     8761 ttatagggcc gccgccgccg ccgcctcgcc ataaaaggaa actttcggag cgcgccgctc
     8821 tgattggctg ccgccgcacc tctccgcctc gccccgccc
//
