LOCUS       sequence-271836-        8503 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression construct expressing sgRNA against PLK1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8503)
  AUTHORS   Swartz SZ, McKay LS, Su KC, Bury L, Padeganeh A, Maddox PS, Knouse 
            KA, Cheeseman IM
  TITLE     Quiescent Cells Actively Replenish CENP-A Nucleosomes to Maintain 
            Centromere Identity and Proliferative Potential.
  JOURNAL   Dev Cell. 2019 Oct 7;51(1):35-48.e7. doi: 
            10.1016/j.devcel.2019.07.016. Epub 2019 Aug 15.
  PUBMED    31422918
REFERENCE   2  (bases 1 to 8503)
  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"; doi: 
            "10.1016/j.devcel.2019.07.016"; journalName: "Dev Cell"; date: 
            "2019-10-7- 7"; volume: "51"; issue: "1"; pages: "35-48.e7"
COMMENT     Sequence Label: pKM328
FEATURES             Location/Qualifiers
     source          1..8503
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        47..319
                     /label=chicken beta-actin promoter
     intron          320..547
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      559..568
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             568..633
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             640..660
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             685..4785
                     /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             685..4785
                     /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             4786..4833
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(4861..4878)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4867..5074
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5083..5223
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5083..5212
                     /label=AAV2 ITR
     rep_origin      5298..5753
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5385..5404)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5595..5616
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5770..5789)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5889..5911
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5949..5967)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6035..6139
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6140..6208,6209..7000)
                     /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(6358..6377)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7171..7759
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7660..7679
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        7821..8069
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     7821..7841
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     7992..8011
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        8091..8166
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        join(8263..8503,1..45)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtcga ggtgagcccc
       61 acgttctgct tcactctccc catctccccc ccctccccac ccccaatttt gtatttattt
      121 attttttaat tattttgtgc agcgatgggg gcgggggggg gggcgcgcgc caggcggggc
      181 ggggcggggc gaggggcggg gcggggcgag gcggagaggt gcggcggcag ccaatcagag
      241 cggcgcgctc cgaaagtttc cttttatggc gaggcggcgg cggcggcggc cctataaaaa
      301 gcgaagcgcg cggcgggcgg gagtcgctgc gcgctgcctt cgccccgtgc cccgctccgc
      361 cgccgcctcg cgccgcccgc cccggctctg actgaccgcg ttactcccac aggtgagcgg
      421 gcgggacggc ccttctcctc cgggctgtaa ttagctgagc aagaggtaag ggtttaaggg
      481 atggttggtt ggtggggtat taatgtttaa ttacctggag cacctgcctg aaatcacttt
      541 ttttcaggtt ggaccggtgc caccatggac tataaggacc acgacggaga ctacaaggat
      601 catgatattg attacaaaga cgatgacgat aagatggccc caaagaagaa gcggaaggtc
      661 ggtatccacg gagtcccagc agccgacaag aagtacagca tcggcctgga catcggcacc
      721 aactctgtgg gctgggccgt gatcaccgac gagtacaagg tgcccagcaa gaaattcaag
      781 gtgctgggca acaccgaccg gcacagcatc aagaagaacc tgatcggagc cctgctgttc
      841 gacagcggcg aaacagccga ggccacccgg ctgaagagaa ccgccagaag aagatacacc
      901 agacggaaga accggatctg ctatctgcaa gagatcttca gcaacgagat ggccaaggtg
      961 gacgacagct tcttccacag actggaagag tccttcctgg tggaagagga taagaagcac
     1021 gagcggcacc ccatcttcgg caacatcgtg gacgaggtgg cctaccacga gaagtacccc
     1081 accatctacc acctgagaaa gaaactggtg gacagcaccg acaaggccga cctgcggctg
     1141 atctatctgg ccctggccca catgatcaag ttccggggcc acttcctgat cgagggcgac
     1201 ctgaaccccg acaacagcga cgtggacaag ctgttcatcc agctggtgca gacctacaac
     1261 cagctgttcg aggaaaaccc catcaacgcc agcggcgtgg acgccaaggc catcctgtct
     1321 gccagactga gcaagagcag acggctggaa aatctgatcg cccagctgcc cggcgagaag
     1381 aagaatggcc tgttcggaaa cctgattgcc ctgagcctgg gcctgacccc caacttcaag
     1441 agcaacttcg acctggccga ggatgccaaa ctgcagctga gcaaggacac ctacgacgac
     1501 gacctggaca acctgctggc ccagatcggc gaccagtacg ccgacctgtt tctggccgcc
     1561 aagaacctgt ccgacgccat cctgctgagc gacatcctga gagtgaacac cgagatcacc
     1621 aaggcccccc tgagcgcctc tatgatcaag agatacgacg agcaccacca ggacctgacc
     1681 ctgctgaaag ctctcgtgcg gcagcagctg cctgagaagt acaaagagat tttcttcgac
     1741 cagagcaaga acggctacgc cggctacatt gacggcggag ccagccagga agagttctac
     1801 aagttcatca agcccatcct ggaaaagatg gacggcaccg aggaactgct cgtgaagctg
     1861 aacagagagg acctgctgcg gaagcagcgg accttcgaca acggcagcat cccccaccag
     1921 atccacctgg gagagctgca cgccattctg cggcggcagg aagattttta cccattcctg
     1981 aaggacaacc gggaaaagat cgagaagatc ctgaccttcc gcatccccta ctacgtgggc
     2041 cctctggcca ggggaaacag cagattcgcc tggatgacca gaaagagcga ggaaaccatc
     2101 accccctgga acttcgagga agtggtggac aagggcgctt ccgcccagag cttcatcgag
     2161 cggatgacca acttcgataa gaacctgccc aacgagaagg tgctgcccaa gcacagcctg
     2221 ctgtacgagt acttcaccgt gtataacgag ctgaccaaag tgaaatacgt gaccgaggga
     2281 atgagaaagc ccgccttcct gagcggcgag cagaaaaagg ccatcgtgga cctgctgttc
     2341 aagaccaacc ggaaagtgac cgtgaagcag ctgaaagagg actacttcaa gaaaatcgag
     2401 tgcttcgact ccgtggaaat ctccggcgtg gaagatcggt tcaacgcctc cctgggcaca
     2461 taccacgatc tgctgaaaat tatcaaggac aaggacttcc tggacaatga ggaaaacgag
     2521 gacattctgg aagatatcgt gctgaccctg acactgtttg aggacagaga gatgatcgag
     2581 gaacggctga aaacctatgc ccacctgttc gacgacaaag tgatgaagca gctgaagcgg
     2641 cggagataca ccggctgggg caggctgagc cggaagctga tcaacggcat ccgggacaag
     2701 cagtccggca agacaatcct ggatttcctg aagtccgacg gcttcgccaa cagaaacttc
     2761 atgcagctga tccacgacga cagcctgacc tttaaagagg acatccagaa agcccaggtg
     2821 tccggccagg gcgatagcct gcacgagcac attgccaatc tggccggcag ccccgccatt
     2881 aagaagggca tcctgcagac agtgaaggtg gtggacgagc tcgtgaaagt gatgggccgg
     2941 cacaagcccg agaacatcgt gatcgaaatg gccagagaga accagaccac ccagaaggga
     3001 cagaagaaca gccgcgagag aatgaagcgg atcgaagagg gcatcaaaga gctgggcagc
     3061 cagatcctga aagaacaccc cgtggaaaac acccagctgc agaacgagaa gctgtacctg
     3121 tactacctgc agaatgggcg ggatatgtac gtggaccagg aactggacat caaccggctg
     3181 tccgactacg atgtggacca tatcgtgcct cagagctttc tgaaggacga ctccatcgac
     3241 aacaaggtgc tgaccagaag cgacaagaac cggggcaaga gcgacaacgt gccctccgaa
     3301 gaggtcgtga agaagatgaa gaactactgg cggcagctgc tgaacgccaa gctgattacc
     3361 cagagaaagt tcgacaatct gaccaaggcc gagagaggcg gcctgagcga actggataag
     3421 gccggcttca tcaagagaca gctggtggaa acccggcaga tcacaaagca cgtggcacag
     3481 atcctggact cccggatgaa cactaagtac gacgagaatg acaagctgat ccgggaagtg
     3541 aaagtgatca ccctgaagtc caagctggtg tccgatttcc ggaaggattt ccagttttac
     3601 aaagtgcgcg agatcaacaa ctaccaccac gcccacgacg cctacctgaa cgccgtcgtg
     3661 ggaaccgccc tgatcaaaaa gtaccctaag ctggaaagcg agttcgtgta cggcgactac
     3721 aaggtgtacg acgtgcggaa gatgatcgcc aagagcgagc aggaaatcgg caaggctacc
     3781 gccaagtact tcttctacag caacatcatg aactttttca agaccgagat taccctggcc
     3841 aacggcgaga tccggaagcg gcctctgatc gagacaaacg gcgaaaccgg ggagatcgtg
     3901 tgggataagg gccgggattt tgccaccgtg cggaaagtgc tgagcatgcc ccaagtgaat
     3961 atcgtgaaaa agaccgaggt gcagacaggc ggcttcagca aagagtctat cctgcccaag
     4021 aggaacagcg ataagctgat cgccagaaag aaggactggg accctaagaa gtacggcggc
     4081 ttcgacagcc ccaccgtggc ctattctgtg ctggtggtgg ccaaagtgga aaagggcaag
     4141 tccaagaaac tgaagagtgt gaaagagctg ctggggatca ccatcatgga aagaagcagc
     4201 ttcgagaaga atcccatcga ctttctggaa gccaagggct acaaagaagt gaaaaaggac
     4261 ctgatcatca agctgcctaa gtactccctg ttcgagctgg aaaacggccg gaagagaatg
     4321 ctggcctctg ccggcgaact gcagaaggga aacgaactgg ccctgccctc caaatatgtg
     4381 aacttcctgt acctggccag ccactatgag aagctgaagg gctcccccga ggataatgag
     4441 cagaaacagc tgtttgtgga acagcacaag cactacctgg acgagatcat cgagcagatc
     4501 agcgagttct ccaagagagt gatcctggcc gacgctaatc tggacaaagt gctgtccgcc
     4561 tacaacaagc accgggataa gcccatcaga gagcaggccg agaatatcat ccacctgttt
     4621 accctgacca atctgggagc ccctgccgcc ttcaagtact ttgacaccac catcgaccgg
     4681 aagaggtaca ccagcaccaa agaggtgctg gacgccaccc tgatccacca gagcatcacc
     4741 ggcctgtacg agacacggat cgacctgtct cagctgggag gcgacaaaag gccggcggcc
     4801 acgaaaaagg ccggccaggc aaaaaagaaa aagtaagaat tcctagagct cgctgatcag
     4861 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
     4921 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc
     4981 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
     5041 aggattggga agagaatagc aggcatgctg gggagcggcc gcaggaaccc ctagtgatgg
     5101 agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg
     5161 cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc
     5221 aggggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac
     5281 gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     5341 tacgcgcagc gtgaccgcta cacttgccag cgccttagcg cccgctcctt tcgctttctt
     5401 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     5461 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga
     5521 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     5581 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaact ctatctcggg
     5641 ctattctttt gatttataag ggattttgcc gatttcggtc tattggttaa aaaatgagct
     5701 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa ttttatggtg
     5761 cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac
     5821 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     5881 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     5941 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     6001 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     6061 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     6121 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     6181 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     6241 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     6301 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     6361 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     6421 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     6481 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     6541 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     6601 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     6661 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     6721 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     6781 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     6841 agccggtgag cgtggaagcc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     6901 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     6961 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     7021 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     7081 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     7141 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     7201 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     7261 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     7321 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     7381 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     7441 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     7501 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     7561 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     7621 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     7681 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     7741 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     7801 ctggcctttt gctcacatgt gagggcctat ttcccatgat tccttcatat ttgcatatac
     7861 gatacaaggc tgttagagag ataattggaa ttaatttgac tgtaaacaca aagatattag
     7921 tacaaaatac gtgacgtaga aagtaataat ttcttgggta gtttgcagtt ttaaaattat
     7981 gttttaaaat ggactatcat atgcttaccg taacttgaaa gtatttcgat ttcttggctt
     8041 tatatatctt gtggaaagga cgaaacaacg tttttgtaca tgttcgggtg gttttagagc
     8101 tagaaatagc aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt
     8161 cggtgctttt ttgttttaga gctagaaata gcaagttaaa ataaggctag tccgttttta
     8221 gcgcgtgcgc caattctgca gacaaatggc tctagaggta cccgttacat aacttacggt
     8281 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa tagtaacgcc
     8341 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
     8401 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
     8461 gcccgcctgg cattgtgccc agtacatgac cttatgggac ttt
//
