LOCUS       sequence-337993-       10837 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  deletion of a genomic locus in T(Brachyury) gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10837)
  AUTHORS   Aramaki S, Kagiwada S, Wu G, Obridge D, Adachi K, Kutejova E, 
            Lickert H, Hubner K, Scholer HR
  TITLE     Residual pluripotency is required for inductive germ cell 
            segregation.
  JOURNAL   EMBO Rep. 2021 Aug 4;22(8):e52553. doi: 10.15252/embr.202152553. 
            Epub 2021 Jun 22.
  PUBMED    34156139
REFERENCE   2  (bases 1 to 10837)
  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: "EMBO Rep.";
            date: "2021-08-4"; volume: "22(8):e52553. doi"; pages: " 
            10.15252/embr.202152553. Epub 2021 Jun 22"
COMMENT     Sequence Label: pX330A-T#2
FEATURES             Location/Qualifiers
     source          1..10837
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        109..394
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     misc_RNA        complement(491..566)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(588..836)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(646..665)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(816..836)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     rep_origin      complement(898..1486)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(978..997)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1657..2448,2449..2517))
                     /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     2280..2299
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2518..2622)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2690..2708
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2746..2768)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2868..2887
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2904..3359)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3041..3062)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3253..3272
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    3510..3631
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3521..3540)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3575..3594
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3673..3693
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3747..4346)
                     /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(3830..3850)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     4327..4346
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     CDS             complement(4347..4400)
                     /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             complement(4470..5132)
                     /codon_start=1
                     /product="green fluorescent protein 2 from Pontellina 
                     plumata, also known as ppluGFP2 (Shagin et al., 2004)"
                     /label=CopGFP
                     /translation="PAMEIECRITGTLNGVEFELVGGGEGTPKQGRMTNKMKSTKGALT
                     FSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYRY
                     EAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDGG
                     YYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA"
     regulatory      5153..5162
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     LTR             5191..5459
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     promoter        complement(5472..5683)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     repeat_region   5828..5968
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5839..5968
                     /label=AAV2 ITR
     polyA_signal    5977..6184
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     6173..6190
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(6218..6265)
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             complement(6266..10366)
                     /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             complement(6266..10366)
                     /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             complement(10391..10411)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(10418..10483)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     regulatory      10483..10492
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     intron          10504..10731
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
ORIGIN
        1 cgcccccatc gctgcacaaa ataattaaaa aataaataaa tacaaaattg ggggtgggga
       61 ggggggggag atggggagag tgaagcagaa cgtggggctc acctcgacca tggtaatagc
      121 gatgactaat acgtagatgt actgccaagt aggaaagtcc cataaggtca tgtactgggc
      181 acaatgccag gcgggccatt taccgtcatt gacgtcaata gggggcgtac ttggcatatg
      241 atacacttga tgtactgcca agtgggcagt ttaccgtaaa tactccaccc attgacgtca
      301 atggaaagtc cctattggcg ttactattga cgtcaatggg cgggggtcgt tgggcggtca
      361 gccaggcggg ccatttaccg taagttatgt aacgggtacc tctagagcca tttgtctgca
      421 gaattggcgc acgcgctaaa aacggactag ccttatttta acttgctatt tctagctcta
      481 aaacaaaaaa gcaccgactc ggtgccactt tttcaagttg ataacggact agccttattt
      541 taacttgcta tttctagctc taaaaccgag gcgaggggca ctcgcccggt gtttcgtcct
      601 ttccacaaga tatataaagc caagaaatcg aaatactttc aagttacggt aagcatatga
      661 tagtccattt taaaacataa ttttaaaact gcaaactacc caagaaatta ttactttcta
      721 cgtcacgtat tttgtactaa tatctttgtg tttacagtca aattaattcc aattatctct
      781 ctaacagcct tgtatcgtat atgcaaatat gaaggaatca tgggaaatag gccctcacat
      841 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
      901 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
      961 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     1021 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     1081 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     1141 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     1201 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     1261 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     1321 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     1381 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     1441 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     1501 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     1561 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     1621 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     1681 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     1741 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcggct
     1801 tccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     1861 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     1921 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     1981 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     2041 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     2101 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     2161 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     2221 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     2281 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     2341 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     2401 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     2461 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     2521 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     2581 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     2641 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     2701 cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     2761 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     2821 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
     2881 gattgtactg agagtgcacc ataaaattgt aaacgttaat attttgttaa aattcgcgtt
     2941 aaatttttgt taaatcagct cattttttaa ccaatagacc gaaatcggca aaatccctta
     3001 taaatcaaaa gaatagcccg agatagagtt gagtgttgtt ccagtttgga acaagagtcc
     3061 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     3121 cccactacgt gaaccatcac ccaaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     3181 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     3241 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct aaggcgctgg caagtgtagc
     3301 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgta
     3361 ctatggttgc tttgacgtat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
     3421 cgcatcaggc gcccctgcag gcccggaagg gagaccagat gagcttctga tagactaatt
     3481 gacatcattt gagtcaattc cagacatgat aagatacatt gatgagtttg gacaaaccac
     3541 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     3601 tgtaaccatt ataagctgca ataaacaagt ttagttaacg catgatacaa aggcattaaa
     3661 gcagcgtatc cacatagcgt aaaaggagca acatagttaa gaataccagt caatctttca
     3721 caaattttgt aatccagagg ttgatttcag gcaccgggct tgcgggtcat gcaccaggtg
     3781 cgcggtcctt cgggcacctc gacgtcggcg gtgacggtga agccgagccg ctcgtagaag
     3841 gggaggttgc ggggcgcgga ggtctccagg aaggcgggca ccccggcgcg ctcggccgcc
     3901 tccactccgg ggagcacgac ggcgctgccc agacccttgc cctggtggtc gggcgagacg
     3961 ccgacggtgg ccaggaacca cgcgggctcc ttgggccggt gcggcgccag gaggccttcc
     4021 atctgttgct gcgcggccag ccgggaaccg ctcaactcgg ccatgcgcgg gccgatctcg
     4081 gcgaacaccg cccccgcttc gacgctctcc ggcgtggtcc agaccgccac cgcggcgccg
     4141 tcgtccgcga cccacacctt gccgatgtcg agcccgacgc gcgtgaggaa gagttcttgc
     4201 agctcggtga cccgctcgat gtggcggtcc ggatcgacgg tgtggcgcgt ggcggggtag
     4261 tcggcgaacg cggcggcgag ggtgcgtacg gccctgggga cgtcgtcgcg ggtggcgagg
     4321 cgcaccgtgg gcttgtactc ggtcataggg ccgggattct cctccacgtc accgcatgtt
     4381 agaagacttc ctctgccctc gcgagatccg gtggagccgg gtccggcggt gccgtccacg
     4441 gcagaattgg acgactgagc gcgggatctg gcgaaggcga tgggggtctt gaaggcgtgc
     4501 tggtactcca cgatgcccag ctcggtgttg ctgtgcagct cctccacgcg gcggaaggcg
     4561 aacatggggc ccccgttctg caggatgctg gggtggatgg cgctcttgaa gtgcatgtgg
     4621 ctgtccacca cgaagctgta gtagccgccg tcgcgcaggc tgaaggtgcg ggcgaagctg
     4681 cccaccagca cgttatcgcc catggggtgc aggtgctcca cggtggcgtt gctgcggatg
     4741 atcttgtcgg tgaagatcac gctgtcctcg gggaagccgg tgcccaccac cttgaagtcg
     4801 ccgatcacgc ggccggcctc gtagcggtag ctgaagctca cgtgcagcac gccgccgtcc
     4861 tcgtacttct cgatgcgggt gttggtgtag ccgccgttgt tgatggcgtg caggaagggg
     4921 ttctcgtagc cgctggggta ggtgccgaag tggtagaagc cgtagcccat cacgtggctc
     4981 agcaggtagg ggctgaaggt cagggcgcct ttggtgctct tcatcttgtt ggtcatgcgg
     5041 ccctgcttgg gggtgccctc tccgccgccc accagctcga actccacgcc gttcagggtg
     5101 ccggtgatgc ggcactcgat ctccatggcg ggcaggccgc tctcgtcgct ctccatggtg
     5161 gcgtctagcg taggcgccgg tcacagcttg gatctgtaac ggcgcagaac agaaaacgaa
     5221 acaaagacgt agagttgagc aagcagggtc aggcaaagcg tggagagccg gctgagtcta
     5281 ggtaggctcc aagggagcgc cggacaaagg cccggtctcg acctgagctt taaacttacc
     5341 tagacggcgg acgcagttca ggaggcacca caggcgggag gcggcagaac gcgactcaac
     5401 cggcgtggat ggcggcctca ggtagggcgg cgggcgcgtg aaggagagat gcgagcccct
     5461 cgaagcttca gctgtgttct ggcggcaaac ccgttgcgaa aaagaacgtt cacggcgact
     5521 actgcactta tatacggttc tcccccaccc tcgggaaaaa ggcggagcca gtacacgaca
     5581 tcactttccc agtttacccc gcgccacctt ctctaggcac ccgttcaatt gccgacccct
     5641 ccccccaact tctcggggac tgtgggcgat gtgcgctctg cccactgacg ggcaccggag
     5701 cgatcgcaga tccttcgcgg ccgcggatcc gatttaaatt cgaattcgct agctctagaa
     5761 tacgggtatc caggggacac agtctatcag aggactggcc caccaagtga ggggttgtgg
     5821 gctctttcct gcaggcagct gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc
     5881 gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga
     5941 gtggccaact ccatcactag gggttcctgc ggccgctccc cagcatgcct gctattctct
     6001 tcccaatcct cccccttgct gtcctgcccc accccacccc ccagaataga atgacaccta
     6061 ctcagacaat gcgatgcaat ttcctcattt tattaggaaa ggacagtggg agtggcacct
     6121 tccagggtca aggaaggcac gggggagggg caaacaacag atggctggca actagaaggc
     6181 acagtcgagg ctgatcagcg agctctagga attcttactt tttctttttt gcctggccgg
     6241 cctttttcgt ggccgccggc cttttgtcgc ctcccagctg agacaggtcg atccgtgtct
     6301 cgtacaggcc ggtgatgctc tggtggatca gggtggcgtc cagcacctct ttggtgctgg
     6361 tgtacctctt ccggtcgatg gtggtgtcaa agtacttgaa ggcggcaggg gctcccagat
     6421 tggtcagggt aaacaggtgg atgatattct cggcctgctc tctgatgggc ttatcccggt
     6481 gcttgttgta ggcggacagc actttgtcca gattagcgtc ggccaggatc actctcttgg
     6541 agaactcgct gatctgctcg atgatctcgt ccaggtagtg cttgtgctgt tccacaaaca
     6601 gctgtttctg ctcattatcc tcgggggagc ccttcagctt ctcatagtgg ctggccaggt
     6661 acaggaagtt cacatatttg gagggcaggg ccagttcgtt tcccttctgc agttcgccgg
     6721 cagaggccag cattctcttc cggccgtttt ccagctcgaa cagggagtac ttaggcagct
     6781 tgatgatcag gtcctttttc acttctttgt agcccttggc ttccagaaag tcgatgggat
     6841 tcttctcgaa gctgcttctt tccatgatgg tgatccccag cagctctttc acactcttca
     6901 gtttcttgga cttgcccttt tccactttgg ccaccaccag cacagaatag gccacggtgg
     6961 ggctgtcgaa gccgccgtac ttcttagggt cccagtcctt ctttctggcg atcagcttat
     7021 cgctgttcct cttgggcagg atagactctt tgctgaagcc gcctgtctgc acctcggtct
     7081 ttttcacgat attcacttgg ggcatgctca gcactttccg cacggtggca aaatcccggc
     7141 ccttatccca cacgatctcc ccggtttcgc cgtttgtctc gatcagaggc cgcttccgga
     7201 tctcgccgtt ggccagggta atctcggtct tgaaaaagtt catgatgttg ctgtagaaga
     7261 agtacttggc ggtagccttg ccgatttcct gctcgctctt ggcgatcatc ttccgcacgt
     7321 cgtacacctt gtagtcgccg tacacgaact cgctttccag cttagggtac tttttgatca
     7381 gggcggttcc cacgacggcg ttcaggtagg cgtcgtgggc gtggtggtag ttgttgatct
     7441 cgcgcacttt gtaaaactgg aaatccttcc ggaaatcgga caccagcttg gacttcaggg
     7501 tgatcacttt cacttcccgg atcagcttgt cattctcgtc gtacttagtg ttcatccggg
     7561 agtccaggat ctgtgccacg tgctttgtga tctgccgggt ttccaccagc tgtctcttga
     7621 tgaagccggc cttatccagt tcgctcaggc cgcctctctc ggccttggtc agattgtcga
     7681 actttctctg ggtaatcagc ttggcgttca gcagctgccg ccagtagttc ttcatcttct
     7741 tcacgacctc ttcggagggc acgttgtcgc tcttgccccg gttcttgtcg cttctggtca
     7801 gcaccttgtt gtcgatggag tcgtccttca gaaagctctg aggcacgata tggtccacat
     7861 cgtagtcgga cagccggttg atgtccagtt cctggtccac gtacatatcc cgcccattct
     7921 gcaggtagta caggtacagc ttctcgttct gcagctgggt gttttccacg gggtgttctt
     7981 tcaggatctg gctgcccagc tctttgatgc cctcttcgat ccgcttcatt ctctcgcggc
     8041 tgttcttctg tcccttctgg gtggtctggt tctctctggc catttcgatc acgatgttct
     8101 cgggcttgtg ccggcccatc actttcacga gctcgtccac caccttcact gtctgcagga
     8161 tgcccttctt aatggcgggg ctgccggcca gattggcaat gtgctcgtgc aggctatcgc
     8221 cctggccgga cacctgggct ttctggatgt cctctttaaa ggtcaggctg tcgtcgtgga
     8281 tcagctgcat gaagtttctg ttggcgaagc cgtcggactt caggaaatcc aggattgtct
     8341 tgccggactg cttgtcccgg atgccgttga tcagcttccg gctcagcctg ccccagccgg
     8401 tgtatctccg ccgcttcagc tgcttcatca ctttgtcgtc gaacaggtgg gcataggttt
     8461 tcagccgttc ctcgatcatc tctctgtcct caaacagtgt cagggtcagc acgatatctt
     8521 ccagaatgtc ctcgttttcc tcattgtcca ggaagtcctt gtccttgata attttcagca
     8581 gatcgtggta tgtgcccagg gaggcgttga accgatcttc cacgccggag atttccacgg
     8641 agtcgaagca ctcgattttc ttgaagtagt cctctttcag ctgcttcacg gtcactttcc
     8701 ggttggtctt gaacagcagg tccacgatgg cctttttctg ctcgccgctc aggaaggcgg
     8761 gctttctcat tccctcggtc acgtatttca ctttggtcag ctcgttatac acggtgaagt
     8821 actcgtacag caggctgtgc ttgggcagca ccttctcgtt gggcaggttc ttatcgaagt
     8881 tggtcatccg ctcgatgaag ctctgggcgg aagcgccctt gtccaccact tcctcgaagt
     8941 tccagggggt gatggtttcc tcgctctttc tggtcatcca ggcgaatctg ctgtttcccc
     9001 tggccagagg gcccacgtag taggggatgc ggaaggtcag gatcttctcg atcttttccc
     9061 ggttgtcctt caggaatggg taaaaatctt cctgccgccg cagaatggcg tgcagctctc
     9121 ccaggtggat ctggtggggg atgctgccgt tgtcgaaggt ccgctgcttc cgcagcaggt
     9181 cctctctgtt cagcttcacg agcagttcct cggtgccgtc catcttttcc aggatgggct
     9241 tgatgaactt gtagaactct tcctggctgg ctccgccgtc aatgtagccg gcgtagccgt
     9301 tcttgctctg gtcgaagaaa atctctttgt acttctcagg cagctgctgc cgcacgagag
     9361 ctttcagcag ggtcaggtcc tggtggtgct cgtcgtatct cttgatcata gaggcgctca
     9421 ggggggcctt ggtgatctcg gtgttcactc tcaggatgtc gctcagcagg atggcgtcgg
     9481 acaggttctt ggcggccaga aacaggtcgg cgtactggtc gccgatctgg gccagcaggt
     9541 tgtccaggtc gtcgtcgtag gtgtccttgc tcagctgcag tttggcatcc tcggccaggt
     9601 cgaagttgct cttgaagttg ggggtcaggc ccaggctcag ggcaatcagg tttccgaaca
     9661 ggccattctt cttctcgccg ggcagctggg cgatcagatt ttccagccgt ctgctcttgc
     9721 tcagtctggc agacaggatg gccttggcgt ccacgccgct ggcgttgatg gggttttcct
     9781 cgaacagctg gttgtaggtc tgcaccagct ggatgaacag cttgtccacg tcgctgttgt
     9841 cggggttcag gtcgccctcg atcaggaagt ggccccggaa cttgatcatg tgggccaggg
     9901 ccagatagat cagccgcagg tcggccttgt cggtgctgtc caccagtttc tttctcaggt
     9961 ggtagatggt ggggtacttc tcgtggtagg ccacctcgtc cacgatgttg ccgaagatgg
    10021 ggtgccgctc gtgcttctta tcctcttcca ccaggaagga ctcttccagt ctgtggaaga
    10081 agctgtcgtc caccttggcc atctcgttgc tgaagatctc ttgcagatag cagatccggt
    10141 tcttccgtct ggtgtatctt cttctggcgg ttctcttcag ccgggtggcc tcggctgttt
    10201 cgccgctgtc gaacagcagg gctccgatca ggttcttctt gatgctgtgc cggtcggtgt
    10261 tgcccagcac cttgaatttc ttgctgggca ccttgtactc gtcggtgatc acggcccagc
    10321 ccacagagtt ggtgccgatg tccaggccga tgctgtactt cttgtcggct gctgggactc
    10381 cgtggatacc gaccttccgc ttcttctttg gggccatctt atcgtcatcg tctttgtaat
    10441 caatatcatg atccttgtag tctccgtcgt ggtccttata gtccatggtg gcaccggtcc
    10501 aacctgaaaa aaagtgattt caggcaggtg ctccaggtaa ttaaacatta ataccccacc
    10561 aaccaaccat cccttaaacc cttacctctt gctcagctaa ttacagcccg gaggagaagg
    10621 gccgtcccgc ccgctcacct gtgggagtaa cgcggtcagt cagagccggg gcgggcggcg
    10681 cgaggcggcg gcggagcggg gcacggggcg aaggcagcgc gcagcgactc ccgcccgccg
    10741 cgcgcttcgc tttttatagg gccgccgccg ccgccgcctc gccataaaag gaaactttcg
    10801 gagcgcgccg ctctgattgg ctgccgccgc acctctc
//
