LOCUS       sequence-337689-       10893 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 10893)
  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 10893)
  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#1
FEATURES             Location/Qualifiers
     source          1..10893
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        127..412
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     misc_RNA        complement(509..584)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(606..854)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(664..683)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(834..854)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     rep_origin      complement(916..1504)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(996..1015)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1675..2466,2467..2535))
                     /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     2298..2317
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2536..2640)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2708..2726
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2764..2786)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2886..2905
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2922..3377)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3059..3080)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3271..3290
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    3528..3649
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3539..3558)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3593..3612
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3691..3711
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3765..4364)
                     /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(3848..3868)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     4345..4364
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     CDS             complement(4365..4418)
                     /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(4488..5150)
                     /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      5171..5180
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     LTR             5209..5477
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     promoter        complement(5490..5701)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     repeat_region   5846..5986
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5857..5986
                     /label=AAV2 ITR
     polyA_signal    5995..6202
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     6191..6208
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(6236..6283)
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             complement(6284..10384)
                     /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(6284..10384)
                     /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(10409..10429)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(10436..10501)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     regulatory      10501..10510
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     intron          10522..10749
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
ORIGIN
        1 cccccccccc cccccccccg cccccatcgc tgcacaaaat aattaaaaaa taaataaata
       61 caaaattggg ggtggggagg ggggggagat ggggagagtg aagcagaacg tggggctcac
      121 ctcgaccatg gtaatagcga tgactaatac gtagatgtac tgccaagtag gaaagtccca
      181 taaggtcatg tactgggcac aatgccaggc gggccattta ccgtcattga cgtcaatagg
      241 gggcgtactt ggcatatgat acacttgatg tactgccaag tgggcagttt accgtaaata
      301 ctccacccat tgacgtcaat ggaaagtccc tattggcgtt actattgacg tcaatgggcg
      361 ggggtcgttg ggcggtcagc caggcgggcc atttaccgta agttatgtaa cgggtacctc
      421 tagagccatt tgtctgcaga attggcgcac gcgctaaaaa cggactagcc ttattttaac
      481 ttgctatttc tagctctaaa acaaaaaagc accgactcgg tgccactttt tcaagttgat
      541 aacggactag ccttatttta acttgctatt tctagctcta aaacaaggag caccgagatc
      601 gggacggtgt ttcgtccttt ccacaagata tataaagcca agaaatcgaa atactttcaa
      661 gttacggtaa gcatatgata gtccatttta aaacataatt ttaaaactgc aaactaccca
      721 agaaattatt actttctacg tcacgtattt tgtactaata tctttgtgtt tacagtcaaa
      781 ttaattccaa ttatctctct aacagccttg tatcgtatat gcaaatatga aggaatcatg
      841 ggaaataggc cctcacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
      901 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
      961 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     1021 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     1081 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     1141 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     1201 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     1261 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     1321 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     1381 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     1441 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     1501 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     1561 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     1621 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     1681 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     1741 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     1801 tgcaatgata ccgcggcttc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     1861 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     1921 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     1981 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     2041 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     2101 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     2161 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     2221 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     2281 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     2341 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     2401 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     2461 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     2521 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     2581 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     2641 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     2701 ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga
     2761 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
     2821 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa
     2881 ctatgcggca tcagagcaga ttgtactgag agtgcaccat aaaattgtaa acgttaatat
     2941 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aatagaccga
     3001 aatcggcaaa atcccttata aatcaaaaga atagcccgag atagagttga gtgttgttcc
     3061 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     3121 cgtctatcag ggcgatggcc cactacgtga accatcaccc aaatcaagtt ttttggggtc
     3181 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     3241 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctaa
     3301 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     3361 gccgctacag ggcgcgtact atggttgctt tgacgtatgc ggtgtgaaat accgcacaga
     3421 tgcgtaagga gaaaataccg catcaggcgc ccctgcaggc ccggaaggga gaccagatga
     3481 gcttctgata gactaattga catcatttga gtcaattcca gacatgataa gatacattga
     3541 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
     3601 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagttt agttaacgca
     3661 tgatacaaag gcattaaagc agcgtatcca catagcgtaa aaggagcaac atagttaaga
     3721 ataccagtca atctttcaca aattttgtaa tccagaggtt gatttcaggc accgggcttg
     3781 cgggtcatgc accaggtgcg cggtccttcg ggcacctcga cgtcggcggt gacggtgaag
     3841 ccgagccgct cgtagaaggg gaggttgcgg ggcgcggagg tctccaggaa ggcgggcacc
     3901 ccggcgcgct cggccgcctc cactccgggg agcacgacgg cgctgcccag acccttgccc
     3961 tggtggtcgg gcgagacgcc gacggtggcc aggaaccacg cgggctcctt gggccggtgc
     4021 ggcgccagga ggccttccat ctgttgctgc gcggccagcc gggaaccgct caactcggcc
     4081 atgcgcgggc cgatctcggc gaacaccgcc cccgcttcga cgctctccgg cgtggtccag
     4141 accgccaccg cggcgccgtc gtccgcgacc cacaccttgc cgatgtcgag cccgacgcgc
     4201 gtgaggaaga gttcttgcag ctcggtgacc cgctcgatgt ggcggtccgg atcgacggtg
     4261 tggcgcgtgg cggggtagtc ggcgaacgcg gcggcgaggg tgcgtacggc cctggggacg
     4321 tcgtcgcggg tggcgaggcg caccgtgggc ttgtactcgg tcatagggcc gggattctcc
     4381 tccacgtcac cgcatgttag aagacttcct ctgccctcgc gagatccggt ggagccgggt
     4441 ccggcggtgc cgtccacggc agaattggac gactgagcgc gggatctggc gaaggcgatg
     4501 ggggtcttga aggcgtgctg gtactccacg atgcccagct cggtgttgct gtgcagctcc
     4561 tccacgcggc ggaaggcgaa catggggccc ccgttctgca ggatgctggg gtggatggcg
     4621 ctcttgaagt gcatgtggct gtccaccacg aagctgtagt agccgccgtc gcgcaggctg
     4681 aaggtgcggg cgaagctgcc caccagcacg ttatcgccca tggggtgcag gtgctccacg
     4741 gtggcgttgc tgcggatgat cttgtcggtg aagatcacgc tgtcctcggg gaagccggtg
     4801 cccaccacct tgaagtcgcc gatcacgcgg ccggcctcgt agcggtagct gaagctcacg
     4861 tgcagcacgc cgccgtcctc gtacttctcg atgcgggtgt tggtgtagcc gccgttgttg
     4921 atggcgtgca ggaaggggtt ctcgtagccg ctggggtagg tgccgaagtg gtagaagccg
     4981 tagcccatca cgtggctcag caggtagggg ctgaaggtca gggcgccttt ggtgctcttc
     5041 atcttgttgg tcatgcggcc ctgcttgggg gtgccctctc cgccgcccac cagctcgaac
     5101 tccacgccgt tcagggtgcc ggtgatgcgg cactcgatct ccatggcggg caggccgctc
     5161 tcgtcgctct ccatggtggc gtctagcgta ggcgccggtc acagcttgga tctgtaacgg
     5221 cgcagaacag aaaacgaaac aaagacgtag agttgagcaa gcagggtcag gcaaagcgtg
     5281 gagagccggc tgagtctagg taggctccaa gggagcgccg gacaaaggcc cggtctcgac
     5341 ctgagcttta aacttaccta gacggcggac gcagttcagg aggcaccaca ggcgggaggc
     5401 ggcagaacgc gactcaaccg gcgtggatgg cggcctcagg tagggcggcg ggcgcgtgaa
     5461 ggagagatgc gagcccctcg aagcttcagc tgtgttctgg cggcaaaccc gttgcgaaaa
     5521 agaacgttca cggcgactac tgcacttata tacggttctc ccccaccctc gggaaaaagg
     5581 cggagccagt acacgacatc actttcccag tttaccccgc gccaccttct ctaggcaccc
     5641 gttcaattgc cgacccctcc ccccaacttc tcggggactg tgggcgatgt gcgctctgcc
     5701 cactgacggg caccggagcg atcgcagatc cttcgcggcc gcggatccga tttaaattcg
     5761 aattcgctag ctctagaata cgggtatcca ggggacacag tctatcagag gactggccca
     5821 ccaagtgagg ggttgtgggc tctttcctgc aggcagctgc gcgctcgctc gctcactgag
     5881 gccgcccggg caaagcccgg gcgtcgggcg acctttggtc gcccggcctc agtgagcgag
     5941 cgagcgcgca gagagggagt ggccaactcc atcactaggg gttcctgcgg ccgctcccca
     6001 gcatgcctgc tattctcttc ccaatcctcc cccttgctgt cctgccccac cccacccccc
     6061 agaatagaat gacacctact cagacaatgc gatgcaattt cctcatttta ttaggaaagg
     6121 acagtgggag tggcaccttc cagggtcaag gaaggcacgg gggaggggca aacaacagat
     6181 ggctggcaac tagaaggcac agtcgaggct gatcagcgag ctctaggaat tcttactttt
     6241 tcttttttgc ctggccggcc tttttcgtgg ccgccggcct tttgtcgcct cccagctgag
     6301 acaggtcgat ccgtgtctcg tacaggccgg tgatgctctg gtggatcagg gtggcgtcca
     6361 gcacctcttt ggtgctggtg tacctcttcc ggtcgatggt ggtgtcaaag tacttgaagg
     6421 cggcaggggc tcccagattg gtcagggtaa acaggtggat gatattctcg gcctgctctc
     6481 tgatgggctt atcccggtgc ttgttgtagg cggacagcac tttgtccaga ttagcgtcgg
     6541 ccaggatcac tctcttggag aactcgctga tctgctcgat gatctcgtcc aggtagtgct
     6601 tgtgctgttc cacaaacagc tgtttctgct cattatcctc gggggagccc ttcagcttct
     6661 catagtggct ggccaggtac aggaagttca catatttgga gggcagggcc agttcgtttc
     6721 ccttctgcag ttcgccggca gaggccagca ttctcttccg gccgttttcc agctcgaaca
     6781 gggagtactt aggcagcttg atgatcaggt cctttttcac ttctttgtag cccttggctt
     6841 ccagaaagtc gatgggattc ttctcgaagc tgcttctttc catgatggtg atccccagca
     6901 gctctttcac actcttcagt ttcttggact tgcccttttc cactttggcc accaccagca
     6961 cagaataggc cacggtgggg ctgtcgaagc cgccgtactt cttagggtcc cagtccttct
     7021 ttctggcgat cagcttatcg ctgttcctct tgggcaggat agactctttg ctgaagccgc
     7081 ctgtctgcac ctcggtcttt ttcacgatat tcacttgggg catgctcagc actttccgca
     7141 cggtggcaaa atcccggccc ttatcccaca cgatctcccc ggtttcgccg tttgtctcga
     7201 tcagaggccg cttccggatc tcgccgttgg ccagggtaat ctcggtcttg aaaaagttca
     7261 tgatgttgct gtagaagaag tacttggcgg tagccttgcc gatttcctgc tcgctcttgg
     7321 cgatcatctt ccgcacgtcg tacaccttgt agtcgccgta cacgaactcg ctttccagct
     7381 tagggtactt tttgatcagg gcggttccca cgacggcgtt caggtaggcg tcgtgggcgt
     7441 ggtggtagtt gttgatctcg cgcactttgt aaaactggaa atccttccgg aaatcggaca
     7501 ccagcttgga cttcagggtg atcactttca cttcccggat cagcttgtca ttctcgtcgt
     7561 acttagtgtt catccgggag tccaggatct gtgccacgtg ctttgtgatc tgccgggttt
     7621 ccaccagctg tctcttgatg aagccggcct tatccagttc gctcaggccg cctctctcgg
     7681 ccttggtcag attgtcgaac tttctctggg taatcagctt ggcgttcagc agctgccgcc
     7741 agtagttctt catcttcttc acgacctctt cggagggcac gttgtcgctc ttgccccggt
     7801 tcttgtcgct tctggtcagc accttgttgt cgatggagtc gtccttcaga aagctctgag
     7861 gcacgatatg gtccacatcg tagtcggaca gccggttgat gtccagttcc tggtccacgt
     7921 acatatcccg cccattctgc aggtagtaca ggtacagctt ctcgttctgc agctgggtgt
     7981 tttccacggg gtgttctttc aggatctggc tgcccagctc tttgatgccc tcttcgatcc
     8041 gcttcattct ctcgcggctg ttcttctgtc ccttctgggt ggtctggttc tctctggcca
     8101 tttcgatcac gatgttctcg ggcttgtgcc ggcccatcac tttcacgagc tcgtccacca
     8161 ccttcactgt ctgcaggatg cccttcttaa tggcggggct gccggccaga ttggcaatgt
     8221 gctcgtgcag gctatcgccc tggccggaca cctgggcttt ctggatgtcc tctttaaagg
     8281 tcaggctgtc gtcgtggatc agctgcatga agtttctgtt ggcgaagccg tcggacttca
     8341 ggaaatccag gattgtcttg ccggactgct tgtcccggat gccgttgatc agcttccggc
     8401 tcagcctgcc ccagccggtg tatctccgcc gcttcagctg cttcatcact ttgtcgtcga
     8461 acaggtgggc ataggttttc agccgttcct cgatcatctc tctgtcctca aacagtgtca
     8521 gggtcagcac gatatcttcc agaatgtcct cgttttcctc attgtccagg aagtccttgt
     8581 ccttgataat tttcagcaga tcgtggtatg tgcccaggga ggcgttgaac cgatcttcca
     8641 cgccggagat ttccacggag tcgaagcact cgattttctt gaagtagtcc tctttcagct
     8701 gcttcacggt cactttccgg ttggtcttga acagcaggtc cacgatggcc tttttctgct
     8761 cgccgctcag gaaggcgggc tttctcattc cctcggtcac gtatttcact ttggtcagct
     8821 cgttatacac ggtgaagtac tcgtacagca ggctgtgctt gggcagcacc ttctcgttgg
     8881 gcaggttctt atcgaagttg gtcatccgct cgatgaagct ctgggcggaa gcgcccttgt
     8941 ccaccacttc ctcgaagttc cagggggtga tggtttcctc gctctttctg gtcatccagg
     9001 cgaatctgct gtttcccctg gccagagggc ccacgtagta ggggatgcgg aaggtcagga
     9061 tcttctcgat cttttcccgg ttgtccttca ggaatgggta aaaatcttcc tgccgccgca
     9121 gaatggcgtg cagctctccc aggtggatct ggtgggggat gctgccgttg tcgaaggtcc
     9181 gctgcttccg cagcaggtcc tctctgttca gcttcacgag cagttcctcg gtgccgtcca
     9241 tcttttccag gatgggcttg atgaacttgt agaactcttc ctggctggct ccgccgtcaa
     9301 tgtagccggc gtagccgttc ttgctctggt cgaagaaaat ctctttgtac ttctcaggca
     9361 gctgctgccg cacgagagct ttcagcaggg tcaggtcctg gtggtgctcg tcgtatctct
     9421 tgatcataga ggcgctcagg ggggccttgg tgatctcggt gttcactctc aggatgtcgc
     9481 tcagcaggat ggcgtcggac aggttcttgg cggccagaaa caggtcggcg tactggtcgc
     9541 cgatctgggc cagcaggttg tccaggtcgt cgtcgtaggt gtccttgctc agctgcagtt
     9601 tggcatcctc ggccaggtcg aagttgctct tgaagttggg ggtcaggccc aggctcaggg
     9661 caatcaggtt tccgaacagg ccattcttct tctcgccggg cagctgggcg atcagatttt
     9721 ccagccgtct gctcttgctc agtctggcag acaggatggc cttggcgtcc acgccgctgg
     9781 cgttgatggg gttttcctcg aacagctggt tgtaggtctg caccagctgg atgaacagct
     9841 tgtccacgtc gctgttgtcg gggttcaggt cgccctcgat caggaagtgg ccccggaact
     9901 tgatcatgtg ggccagggcc agatagatca gccgcaggtc ggccttgtcg gtgctgtcca
     9961 ccagtttctt tctcaggtgg tagatggtgg ggtacttctc gtggtaggcc acctcgtcca
    10021 cgatgttgcc gaagatgggg tgccgctcgt gcttcttatc ctcttccacc aggaaggact
    10081 cttccagtct gtggaagaag ctgtcgtcca ccttggccat ctcgttgctg aagatctctt
    10141 gcagatagca gatccggttc ttccgtctgg tgtatcttct tctggcggtt ctcttcagcc
    10201 gggtggcctc ggctgtttcg ccgctgtcga acagcagggc tccgatcagg ttcttcttga
    10261 tgctgtgccg gtcggtgttg cccagcacct tgaatttctt gctgggcacc ttgtactcgt
    10321 cggtgatcac ggcccagccc acagagttgg tgccgatgtc caggccgatg ctgtacttct
    10381 tgtcggctgc tgggactccg tggataccga ccttccgctt cttctttggg gccatcttat
    10441 cgtcatcgtc tttgtaatca atatcatgat ccttgtagtc tccgtcgtgg tccttatagt
    10501 ccatggtggc accggtccaa cctgaaaaaa agtgatttca ggcaggtgct ccaggtaatt
    10561 aaacattaat accccaccaa ccaaccatcc cttaaaccct tacctcttgc tcagctaatt
    10621 acagcccgga ggagaagggc cgtcccgccc gctcacctgt gggagtaacg cggtcagtca
    10681 gagccggggc gggcggcgcg aggcggcggc ggagcggggc acggggcgaa ggcagcgcgc
    10741 agcgactccc gcccgccgcg cgcttcgctt tttatagggc cgccgccgcc gccgcctcgc
    10801 cataaaagga aactttcgga gcgcgccgct ctgattggct gccgccgcac ctctccgcct
    10861 cgccccgccc cgcccctcgc cccgccccgc ccc
//
