LOCUS       sequence-167283-       10782 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This plasmid should be used for Cre/loxP-based 
            Supernova-CRISPR/Cas9..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10782)
  AUTHORS   Luo W, Mizuno H, Iwata R, Nakazawa S, Yasuda K, Itohara S, Iwasato T
  TITLE     Supernova: A Versatile Vector System for Single-Cell Labeling and 
            Gene Function Studies in vivo.
  JOURNAL   Sci Rep. 2016 Oct 24;6:35747. doi: 10.1038/srep35747.
  PUBMED    27775045
REFERENCE   2  (bases 1 to 10782)
  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"; journalName: "Sci Rep."; 
            date: "2016-10-24"; volume: "6:35747. doi"; pages: " 
            10.1038/srep35747"
COMMENT     Sequence Label: pK237.U6-Chimeric-CAG-loxP-stop-loxP-hspCas9
            (...
FEATURES             Location/Qualifiers
     source          1..10782
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..249
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1..21
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     172..191
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        268..343
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        440..725
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        728..1003
                     /label=chicken beta-actin promoter
     intron          1004..2012
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1936..1958
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     protein_bind    complement(2069..2102)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     intron          2636..2701
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             2831..2851
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    3276..3410
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3313..3332)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3367..3386
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(3452..3485)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      3521..3530
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3530..3595
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3602..3622
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3647..7747
                     /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             3647..7747
                     /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             7748..7795
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(7823..7840)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7829..8036
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   8045..8185
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   8045..8174
                     /label=AAV2 ITR
     rep_origin      8260..8715
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8347..8366)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8557..8578
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8732..8751)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8851..8873
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(8911..8929)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8997..9101
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9102..9170,9171..9962)
                     /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(9320..9339)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10133..10721
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10622..10641
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag
       61 ataattggaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga
      121 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat
      181 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga
      241 cgaaacaccg ggtcttcgag aagacctgtt ttagagctag aaatagcaag ttaaaataag
      301 gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgcttttttg ttttagagct
      361 agaaatagca agttaaaata aggctagtcc gtttttagcg cgtgcgccaa ttctgcagac
      421 aaatggctct agaggtaccc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      481 ccaacgaccc ccgcccattg acgtcaatag taacgccaat agggactttc cattgacgtc
      541 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
      601 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tgtgcccagt
      661 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
      721 ccatgggtcg aggtgagccc cacgttctgc ttcactctcc ccatctcccc cccctcccca
      781 cccccaattt tgtatttatt tattttttaa ttattttgtg cagcgatggg ggcggggggg
      841 gggggggcgc gcgccaggcg gggcggggcg gggcgagggg cggggcgggg cgaggcggag
      901 aggtgcggcg gcagccaatc agagcggcgc gctccgaaag tttcctttta tggcgaggcg
      961 gcggcggcgg cggccctata aaaagcgaag cgcgcggcgg gcgggagtcg ctgcgttgcc
     1021 ttcgccccgt gccccgctcc gcgccgcctc gcgccgcccg ccccggctct gactgaccgc
     1081 gttactccca caggtgagcg ggcgggacgg cccttctcct ccgggctgta attagcgctt
     1141 ggtttaatga cggctcgttt cttttctgtg gctgcgtgaa agccttaaag ggctccggga
     1201 gggccctttg tgcggggggg agcggctcgg ggggtgcgtg cgtgtgtgtg tgcgtgggga
     1261 gcgccgcgtg cggcccgcgc tgcccggcgg ctgtgagcgc tgcgggcgcg gcgcggggct
     1321 ttgtgcgctc cgcgtgtgcg cgaggggagc gcggccgggg gcggtgcccc gcggtgcggg
     1381 ggggctgcga ggggaacaaa ggctgcgtgc ggggtgtgtg cgtggggggg tgagcagggg
     1441 gtgtgggcgc ggcggtcggg ctgtaacccc cccctgcacc cccctccccg agttgctgag
     1501 cacggcccgg cttcgggtgc ggggctccgt gcggggcgtg gcgcggggct cgccgtgccg
     1561 ggcggggggt ggcggcaggt gggggtgccg ggcggggcgg ggccgcctcg ggccggggag
     1621 ggctcggggg aggggcgcgg cggccccgga gcgccggcgg ctgtcgaggc gcggcgagcc
     1681 gcagccattg ccttttatgg taatcgtgcg agagggcgca gggacttcct ttgtcccaaa
     1741 tctggcggag ccgaaatctg ggaggcgccg ccgcaccccc tctagcgggc gcgggcgaag
     1801 cggtgcggcg ccggcaggaa ggaaatgggc ggggagggcc ttcgtgcgtc gccgcgccgc
     1861 cgtccccttc tccatctcca gcctcggggc tgccgcaggg ggacggctgc cttcgggggg
     1921 gacggggcag ggcggggttc ggcttctggc gtgtgaccgg cggctctaga gcctctgcta
     1981 accatgttca tgccttcttc tttttcctac agctcctggg caacgtgctg gttgttgtgc
     2041 tgtctcatca ttttggcaaa gaattctaat aacttcgtat agcatacatt atacgaagtt
     2101 atattaaggg ttccggatcc tcggggacac caaatatggc gatctcggcc ttttcgtttc
     2161 ttggagctgg gacatgtttg ccatcgatcc atctaccacc agaacggccg ttagatctgc
     2221 tgccaccgtt gtttccaccg aagaaaccac cgttgccgta accaccacga cggttgttgc
     2281 taaagaagct gccaccgcca cggccaccgt tgtagccgcc gttgttgtta ttgtagttgc
     2341 tactgttatt tctggcactt cttggttttc ctcttaagtg aggaggaaca taaccattct
     2401 cgttgttgtc gttgatgctt aaattttgca cttgttcgct cagttcagcc ataatatgaa
     2461 atgcttttct tgttgttctt acggaatacc acttgccacc tatcaccaca actaactttt
     2521 tcccgttcct ccatctcttt tatatttttt ttctcgaggg atctttgtga aggaacctta
     2581 cttctgtggt gtgacataat tggacaaact acctacagag atttaaagct ctaaggtaaa
     2641 tataaaattt ttaagtgtat aatgtgttaa actactgatt ctaattgttt gtgtatttta
     2701 gattccaacc tatggaactg atgaatggga gcagtggtgg aatgccttta atgaggaaaa
     2761 cctgttttgc tcagaagaaa tgccatctag tgatgatgag gctactgctg actctcaaca
     2821 ttctactcct ccaaaaaaga agagaaaggt agaagacccc aaggactttc cttcagaatt
     2881 gctaagtttt ttgagtcatg ctgtgtttag taatagaact cttgcttgct ttgctattta
     2941 caccacaaag gaaaaagctg cactgctata caagaaaatt atggaaaaat attctgtaac
     3001 ctttataagt aggcataaca gttataatca taacatactg ttttttctta ctccacacag
     3061 gcatagagtg tctgctatta ataactatgc tcaaaaattg tgtaccttta gctttttaat
     3121 ttgtaaaggg gttaataagg aatatttgat gtatagtgcc ttgactagag atcataatca
     3181 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga
     3241 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg
     3301 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     3361 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc tgacatggta
     3421 agtaagcttg ggctgcaggt cgagggacct aataacttcg tatagcatac attatacgaa
     3481 gttatatgtc gacggtaccg cgggcccggg atccaccggt gccaccatgg actataagga
     3541 ccacgacgga gactacaagg atcatgatat tgattacaaa gacgatgacg ataagatggc
     3601 cccaaagaag aagcggaagg tcggtatcca cggagtccca gcagccgaca agaagtacag
     3661 catcggcctg gacatcggca ccaactctgt gggctgggcc gtgatcaccg acgagtacaa
     3721 ggtgcccagc aagaaattca aggtgctggg caacaccgac cggcacagca tcaagaagaa
     3781 cctgatcgga gccctgctgt tcgacagcgg cgaaacagcc gaggccaccc ggctgaagag
     3841 aaccgccaga agaagataca ccagacggaa gaaccggatc tgctatctgc aagagatctt
     3901 cagcaacgag atggccaagg tggacgacag cttcttccac agactggaag agtccttcct
     3961 ggtggaagag gataagaagc acgagcggca ccccatcttc ggcaacatcg tggacgaggt
     4021 ggcctaccac gagaagtacc ccaccatcta ccacctgaga aagaaactgg tggacagcac
     4081 cgacaaggcc gacctgcggc tgatctatct ggccctggcc cacatgatca agttccgggg
     4141 ccacttcctg atcgagggcg acctgaaccc cgacaacagc gacgtggaca agctgttcat
     4201 ccagctggtg cagacctaca accagctgtt cgaggaaaac cccatcaacg ccagcggcgt
     4261 ggacgccaag gccatcctgt ctgccagact gagcaagagc agacggctgg aaaatctgat
     4321 cgcccagctg cccggcgaga agaagaatgg cctgttcgga aacctgattg ccctgagcct
     4381 gggcctgacc cccaacttca agagcaactt cgacctggcc gaggatgcca aactgcagct
     4441 gagcaaggac acctacgacg acgacctgga caacctgctg gcccagatcg gcgaccagta
     4501 cgccgacctg tttctggccg ccaagaacct gtccgacgcc atcctgctga gcgacatcct
     4561 gagagtgaac accgagatca ccaaggcccc cctgagcgcc tctatgatca agagatacga
     4621 cgagcaccac caggacctga ccctgctgaa agctctcgtg cggcagcagc tgcctgagaa
     4681 gtacaaagag attttcttcg accagagcaa gaacggctac gccggctaca ttgacggcgg
     4741 agccagccag gaagagttct acaagttcat caagcccatc ctggaaaaga tggacggcac
     4801 cgaggaactg ctcgtgaagc tgaacagaga ggacctgctg cggaagcagc ggaccttcga
     4861 caacggcagc atcccccacc agatccacct gggagagctg cacgccattc tgcggcggca
     4921 ggaagatttt tacccattcc tgaaggacaa ccgggaaaag atcgagaaga tcctgacctt
     4981 ccgcatcccc tactacgtgg gccctctggc caggggaaac agcagattcg cctggatgac
     5041 cagaaagagc gaggaaacca tcaccccctg gaacttcgag gaagtggtgg acaagggcgc
     5101 ttccgcccag agcttcatcg agcggatgac caacttcgat aagaacctgc ccaacgagaa
     5161 ggtgctgccc aagcacagcc tgctgtacga gtacttcacc gtgtataacg agctgaccaa
     5221 agtgaaatac gtgaccgagg gaatgagaaa gcccgccttc ctgagcggcg agcagaaaaa
     5281 ggccatcgtg gacctgctgt tcaagaccaa ccggaaagtg accgtgaagc agctgaaaga
     5341 ggactacttc aagaaaatcg agtgcttcga ctccgtggaa atctccggcg tggaagatcg
     5401 gttcaacgcc tccctgggca cataccacga tctgctgaaa attatcaagg acaaggactt
     5461 cctggacaat gaggaaaacg aggacattct ggaagatatc gtgctgaccc tgacactgtt
     5521 tgaggacaga gagatgatcg aggaacggct gaaaacctat gcccacctgt tcgacgacaa
     5581 agtgatgaag cagctgaagc ggcggagata caccggctgg ggcaggctga gccggaagct
     5641 gatcaacggc atccgggaca agcagtccgg caagacaatc ctggatttcc tgaagtccga
     5701 cggcttcgcc aacagaaact tcatgcagct gatccacgac gacagcctga cctttaaaga
     5761 ggacatccag aaagcccagg tgtccggcca gggcgatagc ctgcacgagc acattgccaa
     5821 tctggccggc agccccgcca ttaagaaggg catcctgcag acagtgaagg tggtggacga
     5881 gctcgtgaaa gtgatgggcc ggcacaagcc cgagaacatc gtgatcgaaa tggccagaga
     5941 gaaccagacc acccagaagg gacagaagaa cagccgcgag agaatgaagc ggatcgaaga
     6001 gggcatcaaa gagctgggca gccagatcct gaaagaacac cccgtggaaa acacccagct
     6061 gcagaacgag aagctgtacc tgtactacct gcagaatggg cgggatatgt acgtggacca
     6121 ggaactggac atcaaccggc tgtccgacta cgatgtggac catatcgtgc ctcagagctt
     6181 tctgaaggac gactccatcg acaacaaggt gctgaccaga agcgacaaga accggggcaa
     6241 gagcgacaac gtgccctccg aagaggtcgt gaagaagatg aagaactact ggcggcagct
     6301 gctgaacgcc aagctgatta cccagagaaa gttcgacaat ctgaccaagg ccgagagagg
     6361 cggcctgagc gaactggata aggccggctt catcaagaga cagctggtgg aaacccggca
     6421 gatcacaaag cacgtggcac agatcctgga ctcccggatg aacactaagt acgacgagaa
     6481 tgacaagctg atccgggaag tgaaagtgat caccctgaag tccaagctgg tgtccgattt
     6541 ccggaaggat ttccagtttt acaaagtgcg cgagatcaac aactaccacc acgcccacga
     6601 cgcctacctg aacgccgtcg tgggaaccgc cctgatcaaa aagtacccta agctggaaag
     6661 cgagttcgtg tacggcgact acaaggtgta cgacgtgcgg aagatgatcg ccaagagcga
     6721 gcaggaaatc ggcaaggcta ccgccaagta cttcttctac agcaacatca tgaacttttt
     6781 caagaccgag attaccctgg ccaacggcga gatccggaag cggcctctga tcgagacaaa
     6841 cggcgaaacc ggggagatcg tgtgggataa gggccgggat tttgccaccg tgcggaaagt
     6901 gctgagcatg ccccaagtga atatcgtgaa aaagaccgag gtgcagacag gcggcttcag
     6961 caaagagtct atcctgccca agaggaacag cgataagctg atcgccagaa agaaggactg
     7021 ggaccctaag aagtacggcg gcttcgacag ccccaccgtg gcctattctg tgctggtggt
     7081 ggccaaagtg gaaaagggca agtccaagaa actgaagagt gtgaaagagc tgctggggat
     7141 caccatcatg gaaagaagca gcttcgagaa gaatcccatc gactttctgg aagccaaggg
     7201 ctacaaagaa gtgaaaaagg acctgatcat caagctgcct aagtactccc tgttcgagct
     7261 ggaaaacggc cggaagagaa tgctggcctc tgccggcgaa ctgcagaagg gaaacgaact
     7321 ggccctgccc tccaaatatg tgaacttcct gtacctggcc agccactatg agaagctgaa
     7381 gggctccccc gaggataatg agcagaaaca gctgtttgtg gaacagcaca agcactacct
     7441 ggacgagatc atcgagcaga tcagcgagtt ctccaagaga gtgatcctgg ccgacgctaa
     7501 tctggacaaa gtgctgtccg cctacaacaa gcaccgggat aagcccatca gagagcaggc
     7561 cgagaatatc atccacctgt ttaccctgac caatctggga gcccctgccg ccttcaagta
     7621 ctttgacacc accatcgacc ggaagaggta caccagcacc aaagaggtgc tggacgccac
     7681 cctgatccac cagagcatca ccggcctgta cgagacacgg atcgacctgt ctcagctggg
     7741 aggcgacaaa aggccggcgg ccacgaaaaa ggccggccag gcaaaaaaga aaaagtaaga
     7801 attcctagag ctcgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     7861 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     7921 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     7981 gtggggcagg acagcaaggg ggaggattgg gaagagaata gcaggcatgc tggggagcgg
     8041 ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact
     8101 gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc
     8161 gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt attttctcct tacgcatctg
     8221 tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt agcggcgcat
     8281 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     8341 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     8401 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     8461 ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     8521 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     8581 caacactcaa ccctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg
     8641 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     8701 taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     8761 gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     8821 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     8881 cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta
     8941 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     9001 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     9061 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     9121 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     9181 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     9241 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     9301 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     9361 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     9421 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     9481 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     9541 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     9601 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     9661 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     9721 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     9781 ggtttattgc tgataaatct ggagccggtg agcgtggaag ccgcggtatc attgcagcac
     9841 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     9901 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     9961 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
    10021 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
    10081 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
    10141 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
    10201 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
    10261 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
    10321 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
    10381 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
    10441 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
    10501 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
    10561 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
    10621 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
    10681 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
    10741 ttttacggtt cctggccttt tgctggcctt ttgctcacat gt
//
