LOCUS       sequence-134647-        9289 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes gRNA for 3' target of human ZNF3 along with Cas9 with 2A 
            GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9289)
  TITLE     Mendenhall CRISPR tagging plasmids
REFERENCE   2  (bases 1 to 9289)
  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: PX458_ZNF3_2
FEATURES             Location/Qualifiers
     source          1..9289
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          22..250
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      262..271
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             271..336
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             343..363
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             388..4488
                     /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             388..4488
                     /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             4489..4536
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             4552..4605
                     /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             join(4606..4608,4609..5319)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(4606..4608,4609..5319)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(4909..4928)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5256..5277
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5347..5364)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5353..5560
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5569..5709
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5569..5698
                     /label=AAV2 ITR
     rep_origin      5784..6239
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5871..5890)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6081..6102
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6256..6275)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6375..6397
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6435..6453)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6521..6625
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6626..6694,6695..7486)
                     /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(6844..6863)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7657..8245
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8146..8165
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        8307..8555
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8307..8327
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8478..8497
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        8574..8649
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        8746..9031
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        join(9033..9289,1..21)
                     /label=chicken beta-actin promoter
ORIGIN
        1 aagcgaagcg cgcggcgggc gggagtcgct gcgacgctgc cttcgccccg tgccccgctc
       61 cgccgccgcc tcgcgccgcc cgccccggct ctgactgacc gcgttactcc cacaggtgag
      121 cgggcgggac ggcccttctc ctccgggctg taattagctg agcaagaggt aagggtttaa
      181 gggatggttg gttggtgggg tattaatgtt taattacctg gagcacctgc ctgaaatcac
      241 tttttttcag gttggaccgg tgccaccatg gactataagg accacgacgg agactacaag
      301 gatcatgata ttgattacaa agacgatgac gataagatgg ccccaaagaa gaagcggaag
      361 gtcggtatcc acggagtccc agcagccgac aagaagtaca gcatcggcct ggacatcggc
      421 accaactctg tgggctgggc cgtgatcacc gacgagtaca aggtgcccag caagaaattc
      481 aaggtgctgg gcaacaccga ccggcacagc atcaagaaga acctgatcgg agccctgctg
      541 ttcgacagcg gcgaaacagc cgaggccacc cggctgaaga gaaccgccag aagaagatac
      601 accagacgga agaaccggat ctgctatctg caagagatct tcagcaacga gatggccaag
      661 gtggacgaca gcttcttcca cagactggaa gagtccttcc tggtggaaga ggataagaag
      721 cacgagcggc accccatctt cggcaacatc gtggacgagg tggcctacca cgagaagtac
      781 cccaccatct accacctgag aaagaaactg gtggacagca ccgacaaggc cgacctgcgg
      841 ctgatctatc tggccctggc ccacatgatc aagttccggg gccacttcct gatcgagggc
      901 gacctgaacc ccgacaacag cgacgtggac aagctgttca tccagctggt gcagacctac
      961 aaccagctgt tcgaggaaaa ccccatcaac gccagcggcg tggacgccaa ggccatcctg
     1021 tctgccagac tgagcaagag cagacggctg gaaaatctga tcgcccagct gcccggcgag
     1081 aagaagaatg gcctgttcgg aaacctgatt gccctgagcc tgggcctgac ccccaacttc
     1141 aagagcaact tcgacctggc cgaggatgcc aaactgcagc tgagcaagga cacctacgac
     1201 gacgacctgg acaacctgct ggcccagatc ggcgaccagt acgccgacct gtttctggcc
     1261 gccaagaacc tgtccgacgc catcctgctg agcgacatcc tgagagtgaa caccgagatc
     1321 accaaggccc ccctgagcgc ctctatgatc aagagatacg acgagcacca ccaggacctg
     1381 accctgctga aagctctcgt gcggcagcag ctgcctgaga agtacaaaga gattttcttc
     1441 gaccagagca agaacggcta cgccggctac attgacggcg gagccagcca ggaagagttc
     1501 tacaagttca tcaagcccat cctggaaaag atggacggca ccgaggaact gctcgtgaag
     1561 ctgaacagag aggacctgct gcggaagcag cggaccttcg acaacggcag catcccccac
     1621 cagatccacc tgggagagct gcacgccatt ctgcggcggc aggaagattt ttacccattc
     1681 ctgaaggaca accgggaaaa gatcgagaag atcctgacct tccgcatccc ctactacgtg
     1741 ggccctctgg ccaggggaaa cagcagattc gcctggatga ccagaaagag cgaggaaacc
     1801 atcaccccct ggaacttcga ggaagtggtg gacaagggcg cttccgccca gagcttcatc
     1861 gagcggatga ccaacttcga taagaacctg cccaacgaga aggtgctgcc caagcacagc
     1921 ctgctgtacg agtacttcac cgtgtataac gagctgacca aagtgaaata cgtgaccgag
     1981 ggaatgagaa agcccgcctt cctgagcggc gagcagaaaa aggccatcgt ggacctgctg
     2041 ttcaagacca accggaaagt gaccgtgaag cagctgaaag aggactactt caagaaaatc
     2101 gagtgcttcg actccgtgga aatctccggc gtggaagatc ggttcaacgc ctccctgggc
     2161 acataccacg atctgctgaa aattatcaag gacaaggact tcctggacaa tgaggaaaac
     2221 gaggacattc tggaagatat cgtgctgacc ctgacactgt ttgaggacag agagatgatc
     2281 gaggaacggc tgaaaaccta tgcccacctg ttcgacgaca aagtgatgaa gcagctgaag
     2341 cggcggagat acaccggctg gggcaggctg agccggaagc tgatcaacgg catccgggac
     2401 aagcagtccg gcaagacaat cctggatttc ctgaagtccg acggcttcgc caacagaaac
     2461 ttcatgcagc tgatccacga cgacagcctg acctttaaag aggacatcca gaaagcccag
     2521 gtgtccggcc agggcgatag cctgcacgag cacattgcca atctggccgg cagccccgcc
     2581 attaagaagg gcatcctgca gacagtgaag gtggtggacg agctcgtgaa agtgatgggc
     2641 cggcacaagc ccgagaacat cgtgatcgaa atggccagag agaaccagac cacccagaag
     2701 ggacagaaga acagccgcga gagaatgaag cggatcgaag agggcatcaa agagctgggc
     2761 agccagatcc tgaaagaaca ccccgtggaa aacacccagc tgcagaacga gaagctgtac
     2821 ctgtactacc tgcagaatgg gcgggatatg tacgtggacc aggaactgga catcaaccgg
     2881 ctgtccgact acgatgtgga ccatatcgtg cctcagagct ttctgaagga cgactccatc
     2941 gacaacaagg tgctgaccag aagcgacaag aaccggggca agagcgacaa cgtgccctcc
     3001 gaagaggtcg tgaagaagat gaagaactac tggcggcagc tgctgaacgc caagctgatt
     3061 acccagagaa agttcgacaa tctgaccaag gccgagagag gcggcctgag cgaactggat
     3121 aaggccggct tcatcaagag acagctggtg gaaacccggc agatcacaaa gcacgtggca
     3181 cagatcctgg actcccggat gaacactaag tacgacgaga atgacaagct gatccgggaa
     3241 gtgaaagtga tcaccctgaa gtccaagctg gtgtccgatt tccggaagga tttccagttt
     3301 tacaaagtgc gcgagatcaa caactaccac cacgcccacg acgcctacct gaacgccgtc
     3361 gtgggaaccg ccctgatcaa aaagtaccct aagctggaaa gcgagttcgt gtacggcgac
     3421 tacaaggtgt acgacgtgcg gaagatgatc gccaagagcg agcaggaaat cggcaaggct
     3481 accgccaagt acttcttcta cagcaacatc atgaactttt tcaagaccga gattaccctg
     3541 gccaacggcg agatccggaa gcggcctctg atcgagacaa acggcgaaac cggggagatc
     3601 gtgtgggata agggccggga ttttgccacc gtgcggaaag tgctgagcat gccccaagtg
     3661 aatatcgtga aaaagaccga ggtgcagaca ggcggcttca gcaaagagtc tatcctgccc
     3721 aagaggaaca gcgataagct gatcgccaga aagaaggact gggaccctaa gaagtacggc
     3781 ggcttcgaca gccccaccgt ggcctattct gtgctggtgg tggccaaagt ggaaaagggc
     3841 aagtccaaga aactgaagag tgtgaaagag ctgctgggga tcaccatcat ggaaagaagc
     3901 agcttcgaga agaatcccat cgactttctg gaagccaagg gctacaaaga agtgaaaaag
     3961 gacctgatca tcaagctgcc taagtactcc ctgttcgagc tggaaaacgg ccggaagaga
     4021 atgctggcct ctgccggcga actgcagaag ggaaacgaac tggccctgcc ctccaaatat
     4081 gtgaacttcc tgtacctggc cagccactat gagaagctga agggctcccc cgaggataat
     4141 gagcagaaac agctgtttgt ggaacagcac aagcactacc tggacgagat catcgagcag
     4201 atcagcgagt tctccaagag agtgatcctg gccgacgcta atctggacaa agtgctgtcc
     4261 gcctacaaca agcaccggga taagcccatc agagagcagg ccgagaatat catccacctg
     4321 tttaccctga ccaatctggg agcccctgcc gccttcaagt actttgacac caccatcgac
     4381 cggaagaggt acaccagcac caaagaggtg ctggacgcca ccctgatcca ccagagcatc
     4441 accggcctgt acgagacacg gatcgacctg tctcagctgg gaggcgacaa aaggccggcg
     4501 gccacgaaaa aggccggcca ggcaaaaaag aaaaaggaat tcggcagtgg agagggcaga
     4561 ggaagtctgc taacatgcgg tgacgtcgag gagaatcctg gcccagtgag caagggcgag
     4621 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
     4681 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
     4741 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc
     4801 tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag
     4861 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
     4921 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
     4981 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
     5041 aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc
     5101 aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
     5161 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc
     5221 gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     5281 gccgccggga tcactctcgg catggacgag ctgtacaagg aattctaact agagctcgct
     5341 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     5401 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     5461 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     5521 agggggagga ttgggaagag aatagcaggc atgctgggga gcggccgcag gaacccctag
     5581 tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa
     5641 aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct
     5701 gcctgcaggg gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     5761 gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     5821 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     5881 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     5941 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt
     6001 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     6061 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     6121 ctcgggctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     6181 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt
     6241 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
     6301 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     6361 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     6421 cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     6481 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     6541 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     6601 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     6661 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     6721 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     6781 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     6841 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     6901 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     6961 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     7021 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     7081 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     7141 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     7201 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     7261 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     7321 atctggagcc ggtgagcgtg gaagccgcgg tatcattgca gcactggggc cagatggtaa
     7381 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     7441 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     7501 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     7561 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     7621 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     7681 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     7741 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     7801 tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     7861 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     7921 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     7981 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     8041 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     8101 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     8161 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     8221 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     8281 cttttgctgg ccttttgctc acatgtgagg gcctatttcc catgattcct tcatatttgc
     8341 atatacgata caaggctgtt agagagataa ttggaattaa tttgactgta aacacaaaga
     8401 tattagtaca aaatacgtga cgtagaaagt aataatttct tgggtagttt gcagttttaa
     8461 aattatgttt taaaatggac tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct
     8521 tggctttata tatcttgtgg aaaggacgaa acaccgtgag gaaaatgggt gtggttttag
     8581 agctagaaat agcaagttaa aataaggcta gtccgttatc aacttgaaaa agtggcaccg
     8641 agtcggtgct tttttgtttt agagctagaa atagcaagtt aaaataaggc tagtccgttt
     8701 ttagcgcgtg cgccaattct gcagacaaat ggctctagag gtacccgtta cataacttac
     8761 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caatagtaac
     8821 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
     8881 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
     8941 atggcccgcc tggcattgtg cccagtacat gaccttatgg gactttccta cttggcagta
     9001 catctacgta ttagtcatcg ctattaccat ggtcgaggtg agccccacgt tctgcttcac
     9061 tctccccatc tcccccccct ccccaccccc aattttgtat ttatttattt tttaattatt
     9121 ttgtgcagcg atgggggcgg gggggggggg ggggcgcgcg ccaggcgggg cggggcgggg
     9181 cgaggggcgg ggcggggcga ggcggagagg tgcggcggca gccaatcaga gcggcgcgct
     9241 ccgaaagttt ccttttatgg cgaggcggcg gcggcggcgg ccctataaa
//
