LOCUS       sequence-346232-        9127 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR/Cas9 plasmid to create GFP fusion proteins.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9127)
  TITLE     ENCODE collection: CRISPR constructs
REFERENCE   2  (bases 1 to 9127)
  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"
COMMENT     Sequence Label: ZMAT4.1.2-gDNA
FEATURES             Location/Qualifiers
     source          1..9127
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          123..350
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      362..371
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             371..436
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             443..463
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(488..511,512..514,515..4588)
                     /codon_start=1
                     /product="nickase mutant of the Cas9 endonuclease from the 
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(D10A)
                     /note="generates RNA-guided single strand nicks in DNA due 
                     to the D10A mutation in the RuvC catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             488..4588
                     /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
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             4589..4636
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             4652..4705
                     /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             4706..5305
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4706..4725)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5202..5222
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(5333..5350)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5339..5546
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5555..5695
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5555..5684
                     /label=AAV2 ITR
     rep_origin      5770..6225
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5857..5876)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6067..6088
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6242..6261)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6361..6383
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6421..6439)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6507..6611
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6612..6680,6681..7472)
                     /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     complement(6830..6849)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7643..8231
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8132..8151
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        8293..8541
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8293..8313
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8464..8483
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        8563..8638
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        8735..9020
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 ggggcggggc gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt
       61 ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg
      121 cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc
      181 cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc
      241 ctccgggctg taattagctg agcaagaggt aagggtttaa gggatggttg gttggtgggg
      301 tattaatgtt taattacctg gagcacctgc ctgaaatcac tttttttcag gttggaccgg
      361 tgccaccatg gactataagg accacgacgg agactacaag gatcatgata ttgattacaa
      421 agacgatgac gataagatgg ccccaaagaa gaagcggaag gtcggtatcc acggagtccc
      481 agcagccgac aagaagtaca gcatcggcct ggccatcggc accaactctg tgggctgggc
      541 cgtgatcacc gacgagtaca aggtgcccag caagaaattc aaggtgctgg gcaacaccga
      601 ccggcacagc atcaagaaga acctgatcgg agccctgctg ttcgacagcg gcgaaacagc
      661 cgaggccacc cggctgaaga gaaccgccag aagaagatac accagacgga agaaccggat
      721 ctgctatctg caagagatct tcagcaacga gatggccaag gtggacgaca gcttcttcca
      781 cagactggaa gagtccttcc tggtggaaga ggataagaag cacgagcggc accccatctt
      841 cggcaacatc gtggacgagg tggcctacca cgagaagtac cccaccatct accacctgag
      901 aaagaaactg gtggacagca ccgacaaggc cgacctgcgg ctgatctatc tggccctggc
      961 ccacatgatc aagttccggg gccacttcct gatcgagggc gacctgaacc ccgacaacag
     1021 cgacgtggac aagctgttca tccagctggt gcagacctac aaccagctgt tcgaggaaaa
     1081 ccccatcaac gccagcggcg tggacgccaa ggccatcctg tctgccagac tgagcaagag
     1141 cagacggctg gaaaatctga tcgcccagct gcccggcgag aagaagaatg gcctgttcgg
     1201 caacctgatt gccctgagcc tgggcctgac ccccaacttc aagagcaact tcgacctggc
     1261 cgaggatgcc aaactgcagc tgagcaagga cacctacgac gacgacctgg acaacctgct
     1321 ggcccagatc ggcgaccagt acgccgacct gtttctggcc gccaagaacc tgtccgacgc
     1381 catcctgctg agcgacatcc tgagagtgaa caccgagatc accaaggccc ccctgagcgc
     1441 ctctatgatc aagagatacg acgagcacca ccaggacctg accctgctga aagctctcgt
     1501 gcggcagcag ctgcctgaga agtacaaaga gattttcttc gaccagagca agaacggcta
     1561 cgccggctac attgacggcg gagccagcca ggaagagttc tacaagttca tcaagcccat
     1621 cctggaaaag atggacggca ccgaggaact gctcgtgaag ctgaacagag aggacctgct
     1681 gcggaagcag cggaccttcg acaacggcag catcccccac cagatccacc tgggagagct
     1741 gcacgccatt ctgcggcggc aggaagattt ttacccattc ctgaaggaca accgggaaaa
     1801 gatcgagaag atcctgacct tccgcatccc ctactacgtg ggccctctgg ccaggggaaa
     1861 cagcagattc gcctggatga ccagaaagag cgaggaaacc atcaccccct ggaacttcga
     1921 ggaagtggtg gacaagggcg cttccgccca gagcttcatc gagcggatga ccaacttcga
     1981 taagaacctg cccaacgaga aggtgctgcc caagcacagc ctgctgtacg agtacttcac
     2041 cgtgtataac gagctgacca aagtgaaata cgtgaccgag ggaatgagaa agcccgcctt
     2101 cctgagcggc gagcagaaaa aggccatcgt ggacctgctg ttcaagacca accggaaagt
     2161 gaccgtgaag cagctgaaag aggactactt caagaaaatc gagtgcttcg actccgtgga
     2221 aatctccggc gtggaagatc ggttcaacgc ctccctgggc acataccacg atctgctgaa
     2281 aattatcaag gacaaggact tcctggacaa tgaggaaaac gaggacattc tggaagatat
     2341 cgtgctgacc ctgacactgt ttgaggacag agagatgatc gaggaacggc tgaaaaccta
     2401 tgcccacctg ttcgacgaca aagtgatgaa gcagctgaag cggcggagat acaccggctg
     2461 gggcaggctg agccggaagc tgatcaacgg catccgggac aagcagtccg gcaagacaat
     2521 cctggatttc ctgaagtccg acggcttcgc caacagaaac ttcatgcagc tgatccacga
     2581 cgacagcctg acctttaaag aggacatcca gaaagcccag gtgtccggcc agggcgatag
     2641 cctgcacgag cacattgcca atctggccgg cagccccgcc attaagaagg gcatcctgca
     2701 gacagtgaag gtggtggacg agctcgtgaa agtgatgggc cggcacaagc ccgagaacat
     2761 cgtgatcgaa atggccagag agaaccagac cacccagaag ggacagaaga acagccgcga
     2821 gagaatgaag cggatcgaag agggcatcaa agagctgggc agccagatcc tgaaagaaca
     2881 ccccgtggaa aacacccagc tgcagaacga gaagctgtac ctgtactacc tgcagaatgg
     2941 gcgggatatg tacgtggacc aggaactgga catcaaccgg ctgtccgact acgatgtgga
     3001 ccatatcgtg cctcagagct ttctgaagga cgactccatc gacaacaagg tgctgaccag
     3061 aagcgacaag aaccggggca agagcgacaa cgtgccctcc gaagaggtcg tgaagaagat
     3121 gaagaactac tggcggcagc tgctgaacgc caagctgatt acccagagaa agttcgacaa
     3181 tctgaccaag gccgagagag gcggcctgag cgaactggat aaggccggct tcatcaagag
     3241 acagctggtg gaaacccggc agatcacaaa gcacgtggca cagatcctgg actcccggat
     3301 gaacactaag tacgacgaga atgacaagct gatccgggaa gtgaaagtga tcaccctgaa
     3361 gtccaagctg gtgtccgatt tccggaagga tttccagttt tacaaagtgc gcgagatcaa
     3421 caactaccac cacgcccacg acgcctacct gaacgccgtc gtgggaaccg ccctgatcaa
     3481 aaagtaccct aagctggaaa gcgagttcgt gtacggcgac tacaaggtgt acgacgtgcg
     3541 gaagatgatc gccaagagcg agcaggaaat cggcaaggct accgccaagt acttcttcta
     3601 cagcaacatc atgaactttt tcaagaccga gattaccctg gccaacggcg agatccggaa
     3661 gcggcctctg atcgagacaa acggcgaaac cggggagatc gtgtgggata agggccggga
     3721 ttttgccacc gtgcggaaag tgctgagcat gccccaagtg aatatcgtga aaaagaccga
     3781 ggtgcagaca ggcggcttca gcaaagagtc tatcctgccc aagaggaaca gcgataagct
     3841 gatcgccaga aagaaggact gggaccctaa gaagtacggc ggcttcgaca gccccaccgt
     3901 ggcctattct gtgctggtgg tggccaaagt ggaaaagggc aagtccaaga aactgaagag
     3961 tgtgaaagag ctgctgggga tcaccatcat ggaaagaagc agcttcgaga agaatcccat
     4021 cgactttctg gaagccaagg gctacaaaga agtgaaaaag gacctgatca tcaagctgcc
     4081 taagtactcc ctgttcgagc tggaaaacgg ccggaagaga atgctggcct ctgccggcga
     4141 actgcagaag ggaaacgaac tggccctgcc ctccaaatat gtgaacttcc tgtacctggc
     4201 cagccactat gagaagctga agggctcccc cgaggataat gagcagaaac agctgtttgt
     4261 ggaacagcac aagcactacc tggacgagat catcgagcag atcagcgagt tctccaagag
     4321 agtgatcctg gccgacgcta atctggacaa agtgctgtcc gcctacaaca agcaccggga
     4381 taagcccatc agagagcagg ccgagaatat catccacctg tttaccctga ccaatctggg
     4441 agcccctgcc gccttcaagt actttgacac caccatcgac cggaagaggt acaccagcac
     4501 caaagaggtg ctggacgcca ccctgatcca ccagagcatc accggcctgt acgagacacg
     4561 gatcgacctg tctcagctgg gaggcgacaa aaggccggcg gccacgaaaa aggccggcca
     4621 ggcaaaaaag aaaaaggaat tcggcagtgg agagggcaga ggaagtctgc taacatgcgg
     4681 tgacgtcgag gagaatcctg gcccaatgac cgagtacaag cccacggtgc gcctcgccac
     4741 ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc
     4801 cacgcgccac accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact
     4861 cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc
     4921 ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg
     4981 cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct
     5041 cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg gagtctcgcc
     5101 cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga
     5161 gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga
     5221 gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg
     5281 catgacccgc aagcccggtg cctgagaatt ctaactagag ctcgctgatc agcctcgact
     5341 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     5401 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     5461 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     5521 gaagagaata gcaggcatgc tggggagcgg ccgcaggaac ccctagtgat ggagttggcc
     5581 actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc
     5641 ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc
     5701 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc
     5761 aaccatagta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     5821 gcgtgaccgc tacacttgcc agcgccttag cgcccgctcc tttcgctttc ttcccttcct
     5881 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     5941 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac
     6001 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     6061 ttaatagtgg actcttgttc caaactggaa caacactcaa ctctatctcg ggctattctt
     6121 ttgatttata agggattttg ccgatttcgg tctattggtt aaaaaatgag ctgatttaac
     6181 aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca
     6241 gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg
     6301 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     6361 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg
     6421 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     6481 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     6541 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     6601 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     6661 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     6721 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     6781 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     6841 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     6901 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     6961 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     7021 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     7081 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     7141 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     7201 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     7261 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     7321 agcgtggaag ccgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     7381 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     7441 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     7501 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     7561 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     7621 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     7681 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     7741 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     7801 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     7861 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     7921 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     7981 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     8041 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     8101 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     8161 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     8221 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     8281 ttgctcacat gtgagggcct atttcccatg attccttcat atttgcatat acgatacaag
     8341 gctgttagag agataattgg aattaatttg actgtaaaca caaagatatt agtacaaaat
     8401 acgtgacgta gaaagtaata atttcttggg tagtttgcag ttttaaaatt atgttttaaa
     8461 atggactatc atatgcttac cgtaacttga aagtatttcg atttcttggc tttatatatc
     8521 ttgtggaaag gacgaaacac cgtcttcagg ctataagaca gagttttaga gctagaaata
     8581 gcaagttaaa ataaggctag tccgttatca acttgaaaaa gtggcaccga gtcggtgctt
     8641 ttttgtttta gagctagaaa tagcaagtta aaataaggct agtccgtttt tagcgcgtgc
     8701 gccaattctg cagacaaatg gctctagagg tacccgttac ataacttacg gtaaatggcc
     8761 cgcctggctg accgcccaac gacccccgcc cattgacgtc aatagtaacg ccaataggga
     8821 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
     8881 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
     8941 ggcattgtgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
     9001 tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct
     9061 cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga
     9121 tgggggc
//
