LOCUS       sequence-271166-        8504 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses gRNA targeting CENP A 3'.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8504)
  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 8504)
  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: pKC123
FEATURES             Location/Qualifiers
     source          1..8504
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          313..540
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      552..561
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             561..626
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             633..653
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             678..4778
                     /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             678..4778
                     /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             4779..4826
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(4854..4871)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4860..5067
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5076..5216
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5076..5205
                     /label=AAV2 ITR
     rep_origin      5291..5746
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5378..5397)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5588..5609
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5763..5782)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5882..5904
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5942..5960)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6028..6132
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6133..6201,6202..6993)
                     /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(6351..6370)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7164..7752
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7653..7672
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        7814..8062
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     7814..7834
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     7985..8004
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        8084..8159
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        join(8256..8504,1..37)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 gcagtacatc tacgtattag tcatcgctat taccatggtc gaggtgagcc ccacgttctg
       61 cttcactctc cccatctccc ccccctcccc acccccaatt ttgtatttat ttatttttta
      121 attattttgt gcagcgatgg gggcgggggg gggggggggg cgcsaggcgg ggcggggcgg
      181 ggcgaggggc ggggcggggc gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg
      241 ctccgaaagt ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc
      301 gcgcggcggg cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc
      361 tcgcgccgcc cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac
      421 ggcccttctc ctccgggctg taattagctg agcaagaggt aagggtttaa gggatggttg
      481 gttggtgggg tattaatgtt taattacctg gagcacctgc ctgaaatcac tttttttcag
      541 gttggaccgg tgccaccatg gactataagg accacgacgg agactacaag gatcatgata
      601 ttgattacaa agacgatgac gataagatgg ccccaaagaa gaagcggaag gtcggtatcc
      661 acggagtccc agcagccgac aagaagtaca gcatcggcct ggacatcggc accaactctg
      721 tgggctgggc cgtgatcacc gacgagtaca aggtgcccag caagaaattc aaggtgctgg
      781 gcaacaccga ccggcacagc atcaagaaga acctgatcgg agccctgctg ttcgacagcg
      841 gcgaaacagc cgaggccacc cggctgaaga gaaccgccag aagaagatac accagacgga
      901 agaaccggat ctgctatctg caagagatct tcagcaacga gatggccaag gtggacgaca
      961 gcttcttcca cagactggaa gagtccttcc tggtggaaga ggataagaag cacgagcggc
     1021 accccatctt cggcaacatc gtggacgagg tggcctacca cgagaagtac cccaccatct
     1081 accacctgag aaagaaactg gtggacagca ccgacaaggc cgacctgcgg ctgatctatc
     1141 tggccctggc ccacatgatc aagttccggg gccacttcct gatcgagggc gacctgaacc
     1201 ccgacaacag cgacgtggac aagctgttca tccagctggt gcagacctac aaccagctgt
     1261 tcgaggaaaa ccccatcaac gccagcggcg tggacgccaa ggccatcctg tctgccagac
     1321 tgagcaagag cagacggctg gaaaatctga tcgcccagct gcccggcgag aagaagaatg
     1381 gcctgttcgg aaacctgatt gccctgagcc tgggcctgac ccccaacttc aagagcaact
     1441 tcgacctggc cgaggatgcc aaactgcagc tgagcaagga cacctacgac gacgacctgg
     1501 acaacctgct ggcccagatc ggcgaccagt acgccgacct gtttctggcc gccaagaacc
     1561 tgtccgacgc catcctgctg agcgacatcc tgagagtgaa caccgagatc accaaggccc
     1621 ccctgagcgc ctctatgatc aagagatacg acgagcacca ccaggacctg accctgctga
     1681 aagctctcgt gcggcagcag ctgcctgaga agtacaaaga gattttcttc gaccagagca
     1741 agaacggcta cgccggctac attgacggcg gagccagcca ggaagagttc tacaagttca
     1801 tcaagcccat cctggaaaag atggacggca ccgaggaact gctcgtgaag ctgaacagag
     1861 aggacctgct gcggaagcag cggaccttcg acaacggcag catcccccac cagatccacc
     1921 tgggagagct gcacgccatt ctgcggcggc aggaagattt ttacccattc ctgaaggaca
     1981 accgggaaaa gatcgagaag atcctgacct tccgcatccc ctactacgtg ggccctctgg
     2041 ccaggggaaa cagcagattc gcctggatga ccagaaagag cgaggaaacc atcaccccct
     2101 ggaacttcga ggaagtggtg gacaagggcg cttccgccca gagcttcatc gagcggatga
     2161 ccaacttcga taagaacctg cccaacgaga aggtgctgcc caagcacagc ctgctgtacg
     2221 agtacttcac cgtgtataac gagctgacca aagtgaaata cgtgaccgag ggaatgagaa
     2281 agcccgcctt cctgagcggc gagcagaaaa aggccatcgt ggacctgctg ttcaagacca
     2341 accggaaagt gaccgtgaag cagctgaaag aggactactt caagaaaatc gagtgcttcg
     2401 actccgtgga aatctccggc gtggaagatc ggttcaacgc ctccctgggc acataccacg
     2461 atctgctgaa aattatcaag gacaaggact tcctggacaa tgaggaaaac gaggacattc
     2521 tggaagatat cgtgctgacc ctgacactgt ttgaggacag agagatgatc gaggaacggc
     2581 tgaaaaccta tgcccacctg ttcgacgaca aagtgatgaa gcagctgaag cggcggagat
     2641 acaccggctg gggcaggctg agccggaagc tgatcaacgg catccgggac aagcagtccg
     2701 gcaagacaat cctggatttc ctgaagtccg acggcttcgc caacagaaac ttcatgcagc
     2761 tgatccacga cgacagcctg acctttaaag aggacatcca gaaagcccag gtgtccggcc
     2821 agggcgatag cctgcacgag cacattgcca atctggccgg cagccccgcc attaagaagg
     2881 gcatcctgca gacagtgaag gtggtggacg agctcgtgaa agtgatgggc cggcacaagc
     2941 ccgagaacat cgtgatcgaa atggccagag agaaccagac cacccagaag ggacagaaga
     3001 acagccgcga gagaatgaag cggatcgaag agggcatcaa agagctgggc agccagatcc
     3061 tgaaagaaca ccccgtggaa aacacccagc tgcagaacga gaagctgtac ctgtactacc
     3121 tgcagaatgg gcgggatatg tacgtggacc aggaactgga catcaaccgg ctgtccgact
     3181 acgatgtgga ccatatcgtg cctcagagct ttctgaagga cgactccatc gacaacaagg
     3241 tgctgaccag aagcgacaag aaccggggca agagcgacaa cgtgccctcc gaagaggtcg
     3301 tgaagaagat gaagaactac tggcggcagc tgctgaacgc caagctgatt acccagagaa
     3361 agttcgacaa tctgaccaag gccgagagag gcggcctgag cgaactggat aaggccggct
     3421 tcatcaagag acagctggtg gaaacccggc agatcacaaa gcacgtggca cagatcctgg
     3481 actcccggat gaacactaag tacgacgaga atgacaagct gatccgggaa gtgaaagtga
     3541 tcaccctgaa gtccaagctg gtgtccgatt tccggaagga tttccagttt tacaaagtgc
     3601 gcgagatcaa caactaccac cacgcccacg acgcctacct gaacgccgtc gtgggaaccg
     3661 ccctgatcaa aaagtaccct aagctggaaa gcgagttcgt gtacggcgac tacaaggtgt
     3721 acgacgtgcg gaagatgatc gccaagagcg agcaggaaat cggcaaggct accgccaagt
     3781 acttcttcta cagcaacatc atgaactttt tcaagaccga gattaccctg gccaacggcg
     3841 agatccggaa gcggcctctg atcgagacaa acggcgaaac cggggagatc gtgtgggata
     3901 agggccggga ttttgccacc gtgcggaaag tgctgagcat gccccaagtg aatatcgtga
     3961 aaaagaccga ggtgcagaca ggcggcttca gcaaagagtc tatcctgccc aagaggaaca
     4021 gcgataagct gatcgccaga aagaaggact gggaccctaa gaagtacggc ggcttcgaca
     4081 gccccaccgt ggcctattct gtgctggtgg tggccaaagt ggaaaagggc aagtccaaga
     4141 aactgaagag tgtgaaagag ctgctgggga tcaccatcat ggaaagaagc agcttcgaga
     4201 agaatcccat cgactttctg gaagccaagg gctacaaaga agtgaaaaag gacctgatca
     4261 tcaagctgcc taagtactcc ctgttcgagc tggaaaacgg ccggaagaga atgctggcct
     4321 ctgccggcga actgcagaag ggaaacgaac tggccctgcc ctccaaatat gtgaacttcc
     4381 tgtacctggc cagccactat gagaagctga agggctcccc cgaggataat gagcagaaac
     4441 agctgtttgt ggaacagcac aagcactacc tggacgagat catcgagcag atcagcgagt
     4501 tctccaagag agtgatcctg gccgacgcta atctggacaa agtgctgtcc gcctacaaca
     4561 agcaccggga taagcccatc agagagcagg ccgagaatat catccacctg tttaccctga
     4621 ccaatctggg agcccctgcc gccttcaagt actttgacac caccatcgac cggaagaggt
     4681 acaccagcac caaagaggtg ctggacgcca ccctgatcca ccagagcatc accggcctgt
     4741 acgagacacg gatcgacctg tctcagctgg gaggcgacaa aaggccggcg gccacgaaaa
     4801 aggccggcca ggcaaaaaag aaaaagtaag aattcctaga gctcgctgat cagcctcgac
     4861 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     4921 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     4981 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     5041 ggaagagaat agcaggcatg ctggggagcg gccgcaggaa cccctagtga tggagttggc
     5101 cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg
     5161 cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg
     5221 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag
     5281 caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     5341 agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc
     5401 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     5461 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca
     5521 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     5581 tttaatagtg gactcttgtt ccaaactgga acaacactca actctatctc gggctattct
     5641 tttgatttat aagggatttt gccgatttcg gtctattggt taaaaaatga gctgatttaa
     5701 caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc
     5761 agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct
     5821 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     5881 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag
     5941 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     6001 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     6061 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     6121 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     6181 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     6241 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     6301 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     6361 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     6421 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     6481 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     6541 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     6601 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     6661 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     6721 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     6781 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     6841 gagcgtggaa gccgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     6901 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     6961 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     7021 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     7081 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     7141 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     7201 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     7261 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     7321 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     7381 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     7441 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     7501 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     7561 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     7621 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     7681 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     7741 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     7801 tttgctcaca tgtgagggcc tatttcccat gattccttca tatttgcata tacgatacaa
     7861 ggctgttaga gagataattg gaattaattt gactgtaaac acaaagatat tagtacaaaa
     7921 tacgtgacgt agaaagtaat aatttcttgg gtagtttgca gttttaaaat tatgttttaa
     7981 aatggactat catatgctta ccgtaacttg aaagtatttc gatttcttgg ctttatatat
     8041 cttgtggaaa ggacgaaaca ccgtgtcatc agtgttgcca tcggttttag agctagaaat
     8101 agcaagttaa aataaggcta gtccgttatc aacttgaaaa agtggcaccg agtcggtgct
     8161 tttttgtttt agagctagaa atagcaagtt aaaataaggc tagtccgttt ttagcgcgtg
     8221 cgccaattct gcagacaaat ggctctagag gtacccgtta cataacttac ggtaaatggc
     8281 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caatagtaac gccaataggg
     8341 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
     8401 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
     8461 tggcattgtg cccagtacat gaccttatgg gactttccta cttg
//
