LOCUS       sequence-230497-        8554 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR plasmid for expression of Cas9 and gRNA targeting human 
            transcription factor ELF4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8554)
  TITLE     White lab ENCODE plasmids to tag human transcription factors with 
            GFP
REFERENCE   2  (bases 1 to 8554)
  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: pELF4.1.0-gDNA
FEATURES             Location/Qualifiers
     source          1..8554
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        169..454
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     intron          780..1007
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      1019..1028
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1028..1093
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1100..1120
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1145..5245
                     /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             1145..5245
                     /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             5246..5293
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(5321..5338)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5327..5534
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5543..5683
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5543..5672
                     /label=AAV2 ITR
     rep_origin      5758..6213
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5845..5864)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6055..6076
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6230..6249)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6349..6371
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6409..6427)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6495..6599
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6600..6668,6669..7460)
                     /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(6818..6837)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7631..8219
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8120..8139
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        8281..8529
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8281..8301
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8452..8471
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        join(8551..8554,1..72)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
ORIGIN
        1 tagagctaga aatagcaagt taaaataagg ctagtccgtt atcaacttga aaaagtggca
       61 ccgagtcggt gcttttttgt tttagagcta gaaatagcaa gttaaaataa ggctagtccg
      121 tttttagcgc gtgcgccaat tctgcagaca aatggctcta gaggtacccg ttacataact
      181 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaatagt
      241 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
      301 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
      361 taaatggccc gcctggcatt gtgcccagta catgacctta tgggactttc ctacttggca
      421 gtacatctac gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt
      481 cactctcccc atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt
      541 attttgtgca gcgatggggg cggggggggg gggggggcgc gccccagggs ggggggggsg
      601 gggggggggg gggggggggg ggsgggsggk sgkgggggnn ggggcggggc gaggggcggg
      661 gcggggcgag gcggagaggt gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc
      721 cttttatggc gaggcggcgg cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg
      781 gagtcgctgc gcgctgcctt cgccccgtgc cccgctccgc cgccgcctcg cgccgcccgc
      841 cccggctctg actgaccgcg ttactcccac aggtgagcgg gcgggacggc ccttctcctc
      901 cgggctgtaa ttagctgagc aagaggtaag ggtttaaggg atggttggtt ggtggggtat
      961 taatgtttaa ttacctggag cacctgcctg aaatcacttt ttttcaggtt ggaccggtgc
     1021 caccatggac tataaggacc acgacggaga ctacaaggat catgatattg attacaaaga
     1081 cgatgacgat aagatggccc caaagaagaa gcggaaggtc ggtatccacg gagtcccagc
     1141 agccgacaag aagtacagca tcggcctgga catcggcacc aactctgtgg gctgggccgt
     1201 gatcaccgac gagtacaagg tgcccagcaa gaaattcaag gtgctgggca acaccgaccg
     1261 gcacagcatc aagaagaacc tgatcggagc cctgctgttc gacagcggcg aaacagccga
     1321 ggccacccgg ctgaagagaa ccgccagaag aagatacacc agacggaaga accggatctg
     1381 ctatctgcaa gagatcttca gcaacgagat ggccaaggtg gacgacagct tcttccacag
     1441 actggaagag tccttcctgg tggaagagga taagaagcac gagcggcacc ccatcttcgg
     1501 caacatcgtg gacgaggtgg cctaccacga gaagtacccc accatctacc acctgagaaa
     1561 gaaactggtg gacagcaccg acaaggccga cctgcggctg atctatctgg ccctggccca
     1621 catgatcaag ttccggggcc acttcctgat cgagggcgac ctgaaccccg acaacagcga
     1681 cgtggacaag ctgttcatcc agctggtgca gacctacaac cagctgttcg aggaaaaccc
     1741 catcaacgcc agcggcgtgg acgccaaggc catcctgtct gccagactga gcaagagcag
     1801 acggctggaa aatctgatcg cccagctgcc cggcgagaag aagaatggcc tgttcggaaa
     1861 cctgattgcc ctgagcctgg gcctgacccc caacttcaag agcaacttcg acctggccga
     1921 ggatgccaaa ctgcagctga gcaaggacac ctacgacgac gacctggaca acctgctggc
     1981 ccagatcggc gaccagtacg ccgacctgtt tctggccgcc aagaacctgt ccgacgccat
     2041 cctgctgagc gacatcctga gagtgaacac cgagatcacc aaggcccccc tgagcgcctc
     2101 tatgatcaag agatacgacg agcaccacca ggacctgacc ctgctgaaag ctctcgtgcg
     2161 gcagcagctg cctgagaagt acaaagagat tttcttcgac cagagcaaga acggctacgc
     2221 cggctacatt gacggcggag ccagccagga agagttctac aagttcatca agcccatcct
     2281 ggaaaagatg gacggcaccg aggaactgct cgtgaagctg aacagagagg acctgctgcg
     2341 gaagcagcgg accttcgaca acggcagcat cccccaccag atccacctgg gagagctgca
     2401 cgccattctg cggcggcagg aagattttta cccattcctg aaggacaacc gggaaaagat
     2461 cgagaagatc ctgaccttcc gcatccccta ctacgtgggc cctctggcca ggggaaacag
     2521 cagattcgcc tggatgacca gaaagagcga ggaaaccatc accccctgga acttcgagga
     2581 agtggtggac aagggcgctt ccgcccagag cttcatcgag cggatgacca acttcgataa
     2641 gaacctgccc aacgagaagg tgctgcccaa gcacagcctg ctgtacgagt acttcaccgt
     2701 gtataacgag ctgaccaaag tgaaatacgt gaccgaggga atgagaaagc ccgccttcct
     2761 gagcggcgag cagaaaaagg ccatcgtgga cctgctgttc aagaccaacc ggaaagtgac
     2821 cgtgaagcag ctgaaagagg actacttcaa gaaaatcgag tgcttcgact ccgtggaaat
     2881 ctccggcgtg gaagatcggt tcaacgcctc cctgggcaca taccacgatc tgctgaaaat
     2941 tatcaaggac aaggacttcc tggacaatga ggaaaacgag gacattctgg aagatatcgt
     3001 gctgaccctg acactgtttg aggacagaga gatgatcgag gaacggctga aaacctatgc
     3061 ccacctgttc gacgacaaag tgatgaagca gctgaagcgg cggagataca ccggctgggg
     3121 caggctgagc cggaagctga tcaacggcat ccgggacaag cagtccggca agacaatcct
     3181 ggatttcctg aagtccgacg gcttcgccaa cagaaacttc atgcagctga tccacgacga
     3241 cagcctgacc tttaaagagg acatccagaa agcccaggtg tccggccagg gcgatagcct
     3301 gcacgagcac attgccaatc tggccggcag ccccgccatt aagaagggca tcctgcagac
     3361 agtgaaggtg gtggacgagc tcgtgaaagt gatgggccgg cacaagcccg agaacatcgt
     3421 gatcgaaatg gccagagaga accagaccac ccagaaggga cagaagaaca gccgcgagag
     3481 aatgaagcgg atcgaagagg gcatcaaaga gctgggcagc cagatcctga aagaacaccc
     3541 cgtggaaaac acccagctgc agaacgagaa gctgtacctg tactacctgc agaatgggcg
     3601 ggatatgtac gtggaccagg aactggacat caaccggctg tccgactacg atgtggacca
     3661 tatcgtgcct cagagctttc tgaaggacga ctccatcgac aacaaggtgc tgaccagaag
     3721 cgacaagaac cggggcaaga gcgacaacgt gccctccgaa gaggtcgtga agaagatgaa
     3781 gaactactgg cggcagctgc tgaacgccaa gctgattacc cagagaaagt tcgacaatct
     3841 gaccaaggcc gagagaggcg gcctgagcga actggataag gccggcttca tcaagagaca
     3901 gctggtggaa acccggcaga tcacaaagca cgtggcacag atcctggact cccggatgaa
     3961 cactaagtac gacgagaatg acaagctgat ccgggaagtg aaagtgatca ccctgaagtc
     4021 caagctggtg tccgatttcc ggaaggattt ccagttttac aaagtgcgcg agatcaacaa
     4081 ctaccaccac gcccacgacg cctacctgaa cgccgtcgtg ggaaccgccc tgatcaaaaa
     4141 gtaccctaag ctggaaagcg agttcgtgta cggcgactac aaggtgtacg acgtgcggaa
     4201 gatgatcgcc aagagcgagc aggaaatcgg caaggctacc gccaagtact tcttctacag
     4261 caacatcatg aactttttca agaccgagat taccctggcc aacggcgaga tccggaagcg
     4321 gcctctgatc gagacaaacg gcgaaaccgg ggagatcgtg tgggataagg gccgggattt
     4381 tgccaccgtg cggaaagtgc tgagcatgcc ccaagtgaat atcgtgaaaa agaccgaggt
     4441 gcagacaggc ggcttcagca aagagtctat cctgcccaag aggaacagcg ataagctgat
     4501 cgccagaaag aaggactggg accctaagaa gtacggcggc ttcgacagcc ccaccgtggc
     4561 ctattctgtg ctggtggtgg ccaaagtgga aaagggcaag tccaagaaac tgaagagtgt
     4621 gaaagagctg ctggggatca ccatcatgga aagaagcagc ttcgagaaga atcccatcga
     4681 ctttctggaa gccaagggct acaaagaagt gaaaaaggac ctgatcatca agctgcctaa
     4741 gtactccctg ttcgagctgg aaaacggccg gaagagaatg ctggcctctg ccggcgaact
     4801 gcagaaggga aacgaactgg ccctgccctc caaatatgtg aacttcctgt acctggccag
     4861 ccactatgag aagctgaagg gctcccccga ggataatgag cagaaacagc tgtttgtgga
     4921 acagcacaag cactacctgg acgagatcat cgagcagatc agcgagttct ccaagagagt
     4981 gatcctggcc gacgctaatc tggacaaagt gctgtccgcc tacaacaagc accgggataa
     5041 gcccatcaga gagcaggccg agaatatcat ccacctgttt accctgacca atctgggagc
     5101 ccctgccgcc ttcaagtact ttgacaccac catcgaccgg aagaggtaca ccagcaccaa
     5161 agaggtgctg gacgccaccc tgatccacca gagcatcacc ggcctgtacg agacacggat
     5221 cgacctgtct cagctgggag gcgacaaaag gccggcggcc acgaaaaagg ccggccaggc
     5281 aaaaaagaaa aagtaagaat tcctagagct cgctgatcag cctcgactgt gccttctagt
     5341 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     5401 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     5461 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agagaatagc
     5521 aggcatgctg gggagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg
     5581 cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc
     5641 cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat
     5701 tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg
     5761 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     5821 cacttgccag cgccttagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     5881 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     5941 ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat
     6001 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     6061 tcttgttcca aactggaaca acactcaact ctatctcggg ctattctttt gatttataag
     6121 ggattttgcc gatttcggtc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     6181 cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct
     6241 ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac
     6301 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     6361 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac
     6421 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     6481 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     6541 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     6601 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     6661 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     6721 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     6781 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     6841 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     6901 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     6961 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     7021 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     7081 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     7141 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     7201 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     7261 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtggaagcc
     7321 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     7381 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     7441 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     7501 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     7561 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     7621 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     7681 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     7741 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     7801 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     7861 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     7921 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     7981 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     8041 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     8101 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     8161 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     8221 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     8281 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag
     8341 ataattggaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga
     8401 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat
     8461 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga
     8521 cgaaacaccg cccctttgct tatatgtcat gttt
//
