LOCUS       sequence-337692-       10902 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  deletion of a genomic locus in Prdm14 gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10902)
  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 10902)
  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-Prdm14-ex1-#1
FEATURES             Location/Qualifiers
     source          1..10902
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          148..375
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      387..396
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             396..461
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             468..488
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             513..4613
                     /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             513..4613
                     /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             4614..4661
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(4689..4706)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4695..4902
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4911..5051
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4911..5040
                     /label=AAV2 ITR
     promoter        5196..5407
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             5420..5688
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     regulatory      5717..5726
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5747..6409
                     /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"
     CDS             6479..6532
                     /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             6533..7132
                     /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(6533..6552)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7029..7049
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(7186..7206)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    7248..7369
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7285..7304)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7339..7358
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      7520..7975
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7607..7626)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7817..7838
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7992..8011)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8111..8133
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(8171..8189)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8257..8361
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8362..8430,8431..9222)
                     /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(8580..8599)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9393..9981
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9882..9901
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        10043..10291
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     10043..10063
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     10214..10233
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        10313..10388
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        10485..10770
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 gggggggggg gggggggggg gggggggggg ggggsgrggs ggrgrggkgc ggcggcagcc
       61 aatcagagcg gcgcgctccg aaagtttcct tttatggcga ggcggcggcg gcggcggccc
      121 tataaaaagc gaagcgcgcg gcgggcggga gtcgctgcgc gctgccttcg ccccgtgccc
      181 cgctccgccg ccgcctcgcg ccgcccgccc cggctctgac tgaccgcgtt actcccacag
      241 gtgagcgggc gggacggccc ttctcctccg ggctgtaatt agctgagcaa gaggtaaggg
      301 tttaagggat ggttggttgg tggggtatta atgtttaatt acctggagca cctgcctgaa
      361 atcacttttt ttcaggttgg accggtgcca ccatggacta taaggaccac gacggagact
      421 acaaggatca tgatattgat tacaaagacg atgacgataa gatggcccca aagaagaagc
      481 ggaaggtcgg tatccacgga gtcccagcag ccgacaagaa gtacagcatc ggcctggaca
      541 tcggcaccaa ctctgtgggc tgggccgtga tcaccgacga gtacaaggtg cccagcaaga
      601 aattcaaggt gctgggcaac accgaccggc acagcatcaa gaagaacctg atcggagccc
      661 tgctgttcga cagcggcgaa acagccgagg ccacccggct gaagagaacc gccagaagaa
      721 gatacaccag acggaagaac cggatctgct atctgcaaga gatcttcagc aacgagatgg
      781 ccaaggtgga cgacagcttc ttccacagac tggaagagtc cttcctggtg gaagaggata
      841 agaagcacga gcggcacccc atcttcggca acatcgtgga cgaggtggcc taccacgaga
      901 agtaccccac catctaccac ctgagaaaga aactggtgga cagcaccgac aaggccgacc
      961 tgcggctgat ctatctggcc ctggcccaca tgatcaagtt ccggggccac ttcctgatcg
     1021 agggcgacct gaaccccgac aacagcgacg tggacaagct gttcatccag ctggtgcaga
     1081 cctacaacca gctgttcgag gaaaacccca tcaacgccag cggcgtggac gccaaggcca
     1141 tcctgtctgc cagactgagc aagagcagac ggctggaaaa tctgatcgcc cagctgcccg
     1201 gcgagaagaa gaatggcctg ttcggaaacc tgattgccct gagcctgggc ctgaccccca
     1261 acttcaagag caacttcgac ctggccgagg atgccaaact gcagctgagc aaggacacct
     1321 acgacgacga cctggacaac ctgctggccc agatcggcga ccagtacgcc gacctgtttc
     1381 tggccgccaa gaacctgtcc gacgccatcc tgctgagcga catcctgaga gtgaacaccg
     1441 agatcaccaa ggcccccctg agcgcctcta tgatcaagag atacgacgag caccaccagg
     1501 acctgaccct gctgaaagct ctcgtgcggc agcagctgcc tgagaagtac aaagagattt
     1561 tcttcgacca gagcaagaac ggctacgccg gctacattga cggcggagcc agccaggaag
     1621 agttctacaa gttcatcaag cccatcctgg aaaagatgga cggcaccgag gaactgctcg
     1681 tgaagctgaa cagagaggac ctgctgcgga agcagcggac cttcgacaac ggcagcatcc
     1741 cccaccagat ccacctggga gagctgcacg ccattctgcg gcggcaggaa gatttttacc
     1801 cattcctgaa ggacaaccgg gaaaagatcg agaagatcct gaccttccgc atcccctact
     1861 acgtgggccc tctggccagg ggaaacagca gattcgcctg gatgaccaga aagagcgagg
     1921 aaaccatcac cccctggaac ttcgaggaag tggtggacaa gggcgcttcc gcccagagct
     1981 tcatcgagcg gatgaccaac ttcgataaga acctgcccaa cgagaaggtg ctgcccaagc
     2041 acagcctgct gtacgagtac ttcaccgtgt ataacgagct gaccaaagtg aaatacgtga
     2101 ccgagggaat gagaaagccc gccttcctga gcggcgagca gaaaaaggcc atcgtggacc
     2161 tgctgttcaa gaccaaccgg aaagtgaccg tgaagcagct gaaagaggac tacttcaaga
     2221 aaatcgagtg cttcgactcc gtggaaatct ccggcgtgga agatcggttc aacgcctccc
     2281 tgggcacata ccacgatctg ctgaaaatta tcaaggacaa ggacttcctg gacaatgagg
     2341 aaaacgagga cattctggaa gatatcgtgc tgaccctgac actgtttgag gacagagaga
     2401 tgatcgagga acggctgaaa acctatgccc acctgttcga cgacaaagtg atgaagcagc
     2461 tgaagcggcg gagatacacc ggctggggca ggctgagccg gaagctgatc aacggcatcc
     2521 gggacaagca gtccggcaag acaatcctgg atttcctgaa gtccgacggc ttcgccaaca
     2581 gaaacttcat gcagctgatc cacgacgaca gcctgacctt taaagaggac atccagaaag
     2641 cccaggtgtc cggccagggc gatagcctgc acgagcacat tgccaatctg gccggcagcc
     2701 ccgccattaa gaagggcatc ctgcagacag tgaaggtggt ggacgagctc gtgaaagtga
     2761 tgggccggca caagcccgag aacatcgtga tcgaaatggc cagagagaac cagaccaccc
     2821 agaagggaca gaagaacagc cgcgagagaa tgaagcggat cgaagagggc atcaaagagc
     2881 tgggcagcca gatcctgaaa gaacaccccg tggaaaacac ccagctgcag aacgagaagc
     2941 tgtacctgta ctacctgcag aatgggcggg atatgtacgt ggaccaggaa ctggacatca
     3001 accggctgtc cgactacgat gtggaccata tcgtgcctca gagctttctg aaggacgact
     3061 ccatcgacaa caaggtgctg accagaagcg acaagaaccg gggcaagagc gacaacgtgc
     3121 cctccgaaga ggtcgtgaag aagatgaaga actactggcg gcagctgctg aacgccaagc
     3181 tgattaccca gagaaagttc gacaatctga ccaaggccga gagaggcggc ctgagcgaac
     3241 tggataaggc cggcttcatc aagagacagc tggtggaaac ccggcagatc acaaagcacg
     3301 tggcacagat cctggactcc cggatgaaca ctaagtacga cgagaatgac aagctgatcc
     3361 gggaagtgaa agtgatcacc ctgaagtcca agctggtgtc cgatttccgg aaggatttcc
     3421 agttttacaa agtgcgcgag atcaacaact accaccacgc ccacgacgcc tacctgaacg
     3481 ccgtcgtggg aaccgccctg atcaaaaagt accctaagct ggaaagcgag ttcgtgtacg
     3541 gcgactacaa ggtgtacgac gtgcggaaga tgatcgccaa gagcgagcag gaaatcggca
     3601 aggctaccgc caagtacttc ttctacagca acatcatgaa ctttttcaag accgagatta
     3661 ccctggccaa cggcgagatc cggaagcggc ctctgatcga gacaaacggc gaaaccgggg
     3721 agatcgtgtg ggataagggc cgggattttg ccaccgtgcg gaaagtgctg agcatgcccc
     3781 aagtgaatat cgtgaaaaag accgaggtgc agacaggcgg cttcagcaaa gagtctatcc
     3841 tgcccaagag gaacagcgat aagctgatcg ccagaaagaa ggactgggac cctaagaagt
     3901 acggcggctt cgacagcccc accgtggcct attctgtgct ggtggtggcc aaagtggaaa
     3961 agggcaagtc caagaaactg aagagtgtga aagagctgct ggggatcacc atcatggaaa
     4021 gaagcagctt cgagaagaat cccatcgact ttctggaagc caagggctac aaagaagtga
     4081 aaaaggacct gatcatcaag ctgcctaagt actccctgtt cgagctggaa aacggccgga
     4141 agagaatgct ggcctctgcc ggcgaactgc agaagggaaa cgaactggcc ctgccctcca
     4201 aatatgtgaa cttcctgtac ctggccagcc actatgagaa gctgaagggc tcccccgagg
     4261 ataatgagca gaaacagctg tttgtggaac agcacaagca ctacctggac gagatcatcg
     4321 agcagatcag cgagttctcc aagagagtga tcctggccga cgctaatctg gacaaagtgc
     4381 tgtccgccta caacaagcac cgggataagc ccatcagaga gcaggccgag aatatcatcc
     4441 acctgtttac cctgaccaat ctgggagccc ctgccgcctt caagtacttt gacaccacca
     4501 tcgaccggaa gaggtacacc agcaccaaag aggtgctgga cgccaccctg atccaccaga
     4561 gcatcaccgg cctgtacgag acacggatcg acctgtctca gctgggaggc gacaaaaggc
     4621 cggcggccac gaaaaaggcc ggccaggcaa aaaagaaaaa gtaagaattc ctagagctcg
     4681 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     4741 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     4801 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     4861 caagggggag gattgggaag agaatagcag gcatgctggg gagcggccgc aggaacccct
     4921 agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc
     4981 aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag
     5041 ctgcctgcag gaaagagccc acaacccctc acttggtggg ccagtcctct gatagactgt
     5101 gtcccctgga tacccgtatt ctagagctag cgaattcgaa tttaaatcgg atccgcggcc
     5161 gcgaaggatc tgcgatcgct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt
     5221 ccccgagaag ttggggggag gggtcggcaa ttgaacgggt gcctagagaa ggtggcgcgg
     5281 ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga
     5341 accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag
     5401 aacacagctg aagcttcgag gggctcgcat ctctccttca cgcgcccgcc gccctacctg
     5461 aggccgccat ccacgccggt tgagtcgcgt tctgccgcct cccgcctgtg gtgcctcctg
     5521 aactgcgtcc gccgtctagg taagtttaaa gctcaggtcg agaccgggcc tttgtccggc
     5581 gctcccttgg agcctaccta gactcagccg gctctccacg ctttgcctga ccctgcttgc
     5641 tcaactctac gtctttgttt cgttttctgt tctgcgccgt tacagatcca agctgtgacc
     5701 ggcgcctacg ctagacgcca ccatggagag cgacgagagc ggcctgcccg ccatggagat
     5761 cgagtgccgc atcaccggca ccctgaacgg cgtggagttc gagctggtgg gcggcggaga
     5821 gggcaccccc aagcagggcc gcatgaccaa caagatgaag agcaccaaag gcgccctgac
     5881 cttcagcccc tacctgctga gccacgtgat gggctacggc ttctaccact tcggcaccta
     5941 ccccagcggc tacgagaacc ccttcctgca cgccatcaac aacggcggct acaccaacac
     6001 ccgcatcgag aagtacgagg acggcggcgt gctgcacgtg agcttcagct accgctacga
     6061 ggccggccgc gtgatcggcg acttcaaggt ggtgggcacc ggcttccccg aggacagcgt
     6121 gatcttcacc gacaagatca tccgcagcaa cgccaccgtg gagcacctgc accccatggg
     6181 cgataacgtg ctggtgggca gcttcgcccg caccttcagc ctgcgcgacg gcggctacta
     6241 cagcttcgtg gtggacagcc acatgcactt caagagcgcc atccacccca gcatcctgca
     6301 gaacgggggc cccatgttcg ccttccgccg cgtggaggag ctgcacagca acaccgagct
     6361 gggcatcgtg gagtaccagc acgccttcaa gacccccatc gccttcgcca gatcccgcgc
     6421 tcagtcgtcc aattctgccg tggacggcac cgccggaccc ggctccaccg gatctcgcga
     6481 gggcagagga agtcttctaa catgcggtga cgtggaggag aatcccggcc ctatgaccga
     6541 gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct
     6601 cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat
     6661 cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa
     6721 ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga
     6781 agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct
     6841 ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg
     6901 gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt
     6961 cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc
     7021 cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga
     7081 ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gaaatcaacc
     7141 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     7201 gctatgtgga tacgctgctt taatgccttt gtatcatgcg ttaactaaac ttgtttattg
     7261 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     7321 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga
     7381 attgactcaa atgatgtcaa ttagtctatc agaagctcat ctggtctccc ttccgggcct
     7441 gcaggggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     7501 acgtcaaagc aaccatagta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     7561 gttacgcgca gcgtgaccgc tacacttgcc agcgccttag cgcccgctcc tttcgctttc
     7621 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     7681 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt
     7741 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     7801 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ctctatctcg
     7861 ggctattctt ttgatttata agggattttg ccgatttcgg tctattggtt aaaaaatgag
     7921 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aattttatgg
     7981 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca
     8041 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     8101 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     8161 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     8221 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     8281 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     8341 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     8401 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     8461 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     8521 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     8581 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     8641 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     8701 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     8761 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     8821 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     8881 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     8941 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     9001 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     9061 ggagccggtg agcgtggaag ccgcggtatc attgcagcac tggggccaga tggtaagccc
     9121 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     9181 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     9241 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     9301 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     9361 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     9421 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     9481 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     9541 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     9601 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     9661 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     9721 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     9781 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     9841 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     9901 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     9961 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
    10021 tgctggcctt ttgctcacat gtgagggcct atttcccatg attccttcat atttgcatat
    10081 acgatacaag gctgttagag agataattgg aattaatttg actgtaaaca caaagatatt
    10141 agtacaaaat acgtgacgta gaaagtaata atttcttggg tagtttgcag ttttaaaatt
    10201 atgttttaaa atggactatc atatgcttac cgtaacttga aagtatttcg atttcttggc
    10261 tttatatatc ttgtggaaag gacgaaacac cggagaagtg tgcgctctcg gtgttttaga
    10321 gctagaaata gcaagttaaa ataaggctag tccgttatca acttgaaaaa gtggcaccga
    10381 gtcggtgctt ttttgtttta gagctagaaa tagcaagtta aaataaggct agtccgtttt
    10441 tagcgcgtgc gccaattctg cagacaaatg gctctagagg tacccgttac ataacttacg
    10501 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aatagtaacg
    10561 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
    10621 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
    10681 tggcccgcct ggcattgtgc ccagtacatg accttatggg actttcctac ttggcagtac
    10741 atctacgtat tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact
    10801 ctccccatct cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt
    10861 tgtgcagcga tgggggcggg gggggggggg gggcgcgcgc cg
//
