LOCUS       sequence-214737-        9176 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR plasmid for expression of Cas9 nickase (D10A) and gRNA 
            targeting human transcription factor E2F3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9176)
  TITLE     White lab ENCODE plasmids to tag human transcription factors with 
            GFP
REFERENCE   2  (bases 1 to 9176)
  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: pE2F3.3.1-gDNA
FEATURES             Location/Qualifiers
     source          1..9176
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        168..453
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        455..732
                     /label=chicken beta-actin promoter
     intron          733..961
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      973..982
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             982..1047
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1054..1074
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(1099..1122,1123..1125,1126..5199)
                     /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             1099..5199
                     /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             5200..5247
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             5263..5316
                     /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             5317..5916
                     /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(5317..5336)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5813..5833
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(5944..5961)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5950..6157
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   6166..6306
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   6166..6295
                     /label=AAV2 ITR
     rep_origin      6381..6836
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6468..6487)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6678..6699
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6853..6872)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6972..6994
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7032..7050)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7118..7222
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7223..7291,7292..8083)
                     /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(7441..7460)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8254..8842
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8743..8762
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        8904..9152
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8904..8924
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     9075..9094
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        join(9172..9176,1..71)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
ORIGIN
        1 agagctagaa atagcaagtt aaaataaggc tagtccgtta tcaacttgaa aaagtggcac
       61 cgagtcggtg cttttttgtt ttagagctag aaatagcaag ttaaaataag gctagtccgt
      121 ttttagcgcg tgcgccaatt ctgcagacaa atggctctag aggtacccgt tacataactt
      181 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaatagta
      241 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
      301 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
      361 aaatggcccg cctggcattg tgcccagtac atgaccttat gggactttcc tacttggcag
      421 tacatctacg tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc
      481 actctcccca tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta
      541 ttttgtgcag cgatgggggc gggggggggg ggggggcgcg cgccaggcgg ggcggggcgg
      601 ggcgaggggc ggggcggggc gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg
      661 ctccgaaagt ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc
      721 gcgcggcggg cgggagtcgc tgcgacgctg ccttcgcccc gtgccccgct ccgccgccgc
      781 ctcgcgccgc ccgccccggc tctgactgac cgcgttactc ccacaggtga gcgggcggga
      841 cggcccttct cctccgggct gtaattagct gagcaagagg taagggttta agggatggtt
      901 ggttggtggg gtattaatgt ttaattacct ggagcacctg cctgaaatca ctttttttca
      961 ggttggaccg gtgccaccat ggactataag gaccacgacg gagactacaa ggatcatgat
     1021 attgattaca aagacgatga cgataagatg gccccaaaga agaagcggaa ggtcggtatc
     1081 cacggagtcc cagcagccga caagaagtac agcatcggcc tggccatcgg caccaactct
     1141 gtgggctggg ccgtgatcac cgacgagtac aaggtgccca gcaagaaatt caaggtgctg
     1201 ggcaacaccg accggcacag catcaagaag aacctgatcg gagccctgct gttcgacagc
     1261 ggcgaaacag ccgaggccac ccggctgaag agaaccgcca gaagaagata caccagacgg
     1321 aagaaccgga tctgctatct gcaagagatc ttcagcaacg agatggccaa ggtggacgac
     1381 agcttcttcc acagactgga agagtccttc ctggtggaag aggataagaa gcacgagcgg
     1441 caccccatct tcggcaacat cgtggacgag gtggcctacc acgagaagta ccccaccatc
     1501 taccacctga gaaagaaact ggtggacagc accgacaagg ccgacctgcg gctgatctat
     1561 ctggccctgg cccacatgat caagttccgg ggccacttcc tgatcgaggg cgacctgaac
     1621 cccgacaaca gcgacgtgga caagctgttc atccagctgg tgcagaccta caaccagctg
     1681 ttcgaggaaa accccatcaa cgccagcggc gtggacgcca aggccatcct gtctgccaga
     1741 ctgagcaaga gcagacggct ggaaaatctg atcgcccagc tgcccggcga gaagaagaat
     1801 ggcctgttcg gaaacctgat tgccctgagc ctgggcctga cccccaactt caagagcaac
     1861 ttcgacctgg ccgaggatgc caaactgcag ctgagcaagg acacctacga cgacgacctg
     1921 gacaacctgc tggcccagat cggcgaccag tacgccgacc tgtttctggc cgccaagaac
     1981 ctgtccgacg ccatcctgct gagcgacatc ctgagagtga acaccgagat caccaaggcc
     2041 cccctgagcg cctctatgat caagagatac gacgagcacc accaggacct gaccctgctg
     2101 aaagctctcg tgcggcagca gctgcctgag aagtacaaag agattttctt cgaccagagc
     2161 aagaacggct acgccggcta cattgacggc ggagccagcc aggaagagtt ctacaagttc
     2221 atcaagccca tcctggaaaa gatggacggc accgaggaac tgctcgtgaa gctgaacaga
     2281 gaggacctgc tgcggaagca gcggaccttc gacaacggca gcatccccca ccagatccac
     2341 ctgggagagc tgcacgccat tctgcggcgg caggaagatt tttacccatt cctgaaggac
     2401 aaccgggaaa agatcgagaa gatcctgacc ttccgcatcc cctactacgt gggccctctg
     2461 gccaggggaa acagcagatt cgcctggatg accagaaaga gcgaggaaac catcaccccc
     2521 tggaacttcg aggaagtggt ggacaagggc gcttccgccc agagcttcat cgagcggatg
     2581 accaacttcg ataagaacct gcccaacgag aaggtgctgc ccaagcacag cctgctgtac
     2641 gagtacttca ccgtgtataa cgagctgacc aaagtgaaat acgtgaccga gggaatgaga
     2701 aagcccgcct tcctgagcgg cgagcagaaa aaggccatcg tggacctgct gttcaagacc
     2761 aaccggaaag tgaccgtgaa gcagctgaaa gaggactact tcaagaaaat cgagtgcttc
     2821 gactccgtgg aaatctccgg cgtggaagat cggttcaacg cctccctggg cacataccac
     2881 gatctgctga aaattatcaa ggacaaggac ttcctggaca atgaggaaaa cgaggacatt
     2941 ctggaagata tcgtgctgac cctgacactg tttgaggaca gagagatgat cgaggaacgg
     3001 ctgaaaacct atgcccacct gttcgacgac aaagtgatga agcagctgaa gcggcggaga
     3061 tacaccggct ggggcaggct gagccggaag ctgatcaacg gcatccggga caagcagtcc
     3121 ggcaagacaa tcctggattt cctgaagtcc gacggcttcg ccaacagaaa cttcatgcag
     3181 ctgatccacg acgacagcct gacctttaaa gaggacatcc agaaagccca ggtgtccggc
     3241 cagggcgata gcctgcacga gcacattgcc aatctggccg gcagccccgc cattaagaag
     3301 ggcatcctgc agacagtgaa ggtggtggac gagctcgtga aagtgatggg ccggcacaag
     3361 cccgagaaca tcgtgatcga aatggccaga gagaaccaga ccacccagaa gggacagaag
     3421 aacagccgcg agagaatgaa gcggatcgaa gagggcatca aagagctggg cagccagatc
     3481 ctgaaagaac accccgtgga aaacacccag ctgcagaacg agaagctgta cctgtactac
     3541 ctgcagaatg ggcgggatat gtacgtggac caggaactgg acatcaaccg gctgtccgac
     3601 tacgatgtgg accatatcgt gcctcagagc tttctgaagg acgactccat cgacaacaag
     3661 gtgctgacca gaagcgacaa gaaccggggc aagagcgaca acgtgccctc cgaagaggtc
     3721 gtgaagaaga tgaagaacta ctggcggcag ctgctgaacg ccaagctgat tacccagaga
     3781 aagttcgaca atctgaccaa ggccgagaga ggcggcctga gcgaactgga taaggccggc
     3841 ttcatcaaga gacagctggt ggaaacccgg cagatcacaa agcacgtggc acagatcctg
     3901 gactcccgga tgaacactaa gtacgacgag aatgacaagc tgatccggga agtgaaagtg
     3961 atcaccctga agtccaagct ggtgtccgat ttccggaagg atttccagtt ttacaaagtg
     4021 cgcgagatca acaactacca ccacgcccac gacgcctacc tgaacgccgt cgtgggaacc
     4081 gccctgatca aaaagtaccc taagctggaa agcgagttcg tgtacggcga ctacaaggtg
     4141 tacgacgtgc ggaagatgat cgccaagagc gagcaggaaa tcggcaaggc taccgccaag
     4201 tacttcttct acagcaacat catgaacttt ttcaagaccg agattaccct ggccaacggc
     4261 gagatccgga agcggcctct gatcgagaca aacggcgaaa ccggggagat cgtgtgggat
     4321 aagggccggg attttgccac cgtgcggaaa gtgctgagca tgccccaagt gaatatcgtg
     4381 aaaaagaccg aggtgcagac aggcggcttc agcaaagagt ctatcctgcc caagaggaac
     4441 agcgataagc tgatcgccag aaagaaggac tgggacccta agaagtacgg cggcttcgac
     4501 agccccaccg tggcctattc tgtgctggtg gtggccaaag tggaaaaggg caagtccaag
     4561 aaactgaaga gtgtgaaaga gctgctgggg atcaccatca tggaaagaag cagcttcgag
     4621 aagaatccca tcgactttct ggaagccaag ggctacaaag aagtgaaaaa ggacctgatc
     4681 atcaagctgc ctaagtactc cctgttcgag ctggaaaacg gccggaagag aatgctggcc
     4741 tctgccggcg aactgcagaa gggaaacgaa ctggccctgc cctccaaata tgtgaacttc
     4801 ctgtacctgg ccagccacta tgagaagctg aagggctccc ccgaggataa tgagcagaaa
     4861 cagctgtttg tggaacagca caagcactac ctggacgaga tcatcgagca gatcagcgag
     4921 ttctccaaga gagtgatcct ggccgacgct aatctggaca aagtgctgtc cgcctacaac
     4981 aagcaccggg ataagcccat cagagagcag gccgagaata tcatccacct gtttaccctg
     5041 accaatctgg gagcccctgc cgccttcaag tactttgaca ccaccatcga ccggaagagg
     5101 tacaccagca ccaaagaggt gctggacgcc accctgatcc accagagcat caccggcctg
     5161 tacgagacac ggatcgacct gtctcagctg ggaggcgaca aaaggccggc ggccacgaaa
     5221 aaggccggcc aggcaaaaaa gaaaaaggaa ttcggcagtg gagagggcag aggaagtctg
     5281 ctaacatgcg gtgacgtcga ggagaatcct ggcccaatga ccgagtacaa gcccacggtg
     5341 cgcctcgcca cccgcgacga cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc
     5401 gactaccccg ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag
     5461 ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac
     5521 gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc
     5581 gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag
     5641 atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc
     5701 ggagtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg
     5761 gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc
     5821 cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg
     5881 cgcacctggt gcatgacccg caagcccggt gcctgagaat tctaactaga gctcgctgat
     5941 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     6001 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     6061 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     6121 gggaggattg ggaagagaat agcaggcatg ctggggagcg gccgcaggaa cccctagtga
     6181 tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg
     6241 tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc
     6301 tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     6361 tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     6421 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     6481 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     6541 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg
     6601 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     6661 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     6721 gggctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     6781 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg
     6841 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
     6901 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     6961 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     7021 gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     7081 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     7141 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     7201 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     7261 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     7321 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     7381 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     7441 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     7501 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     7561 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     7621 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     7681 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     7741 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     7801 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     7861 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     7921 tggagccggt gagcgtggaa gccgcggtat cattgcagca ctggggccag atggtaagcc
     7981 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     8041 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     8101 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     8161 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     8221 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     8281 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     8341 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     8401 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     8461 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     8521 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     8581 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     8641 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     8701 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     8761 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     8821 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     8881 ttgctggcct tttgctcaca tgtgagggcc tatttcccat gattccttca tatttgcata
     8941 tacgatacaa ggctgttaga gagataattg gaattaattt gactgtaaac acaaagatat
     9001 tagtacaaaa tacgtgacgt agaaagtaat aatttcttgg gtagtttgca gttttaaaat
     9061 tatgttttaa aatggactat catatgctta ccgtaacttg aaagtatttc gatttcttgg
     9121 ctttatatat cttgtggaaa ggacgaaaca cctgatgcct tcctcctccc cggttt
//
