LOCUS       sequence-399553-       10592 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Frame +0 selector for NHEJ knock-in.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10592)
  AUTHORS   Shi Y, Kopparapu N, Ohler L, Dickinson DJ
  TITLE     Efficient and rapid fluorescent protein knock-in with universal 
            donors in mouse embryonic stem cells.
  JOURNAL   Development. 2023 May 15;150(10):dev201367. doi: 10.1242/dev.201367.
            Epub 2023 May 25.
  PUBMED    37129004
REFERENCE   2  (bases 1 to 10592)
  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: 
            "Development."; date: "2023-05-15"; volume: "150(10):dev201367. 
            doi"; pages: " 10.1242/dev.201367. Epub 2023 May 25"
COMMENT     Sequence Label: pDD425
FEATURES             Location/Qualifiers
     source          1..10592
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             8..477
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     CDS             946..1362
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1292..1314
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1330..1346
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     promoter        1373..1621
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1373..1393
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     1544..1563
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        1642..1717
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        1770..2952
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2001..2943
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     2900..2920
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      2955..2964
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2964..2987
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2988..3692
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(3130..3148)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3398..3417
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             3717..3770
                     /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             3771..7874
                     /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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             3771..7874
                     /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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             7887..7907
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     LTR             8086..8555
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(8685..8702)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8856..9444)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8936..8955)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9604..10395,10396..10464))
                     /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     10227..10246
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10465..10569)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctgcagcctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca
       61 gggccaagaa cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt
      121 tcctgccccg gctcagggcc aagaacagat ggtccccaga tgcggtccag ccctcagcag
      181 tttctagaga accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc
      241 ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga
      301 gctcaataaa agagcccaca acccctcact cggggcgcca gtcctccgat tgactgagtc
      361 gcccgggtac ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc
      421 tgttccttgg gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcattt
      481 gggggctcgt ccgggatcgg gagacccctg cccagggacc accgacccac caccgggagg
      541 caagctggcc agcaacttat ctgtgtctgt ccgattgtct agtgtctatg actgatttta
      601 tgcgcctgcg tcggtactag ctagttagct aactagctct gtatctggcg gacccgtggt
      661 ggaactgacg agttctgaac acccggccgc aaccctggga gacgtcccag ggactttggg
      721 ggccgttttt gtggcccgac ctgaggaagg gagtcgatgt ggaatccgac cccgtcagga
      781 tatgtggttc tggtaggaga cgagaaccta aaacagttcc cgcctccgtc tgaatttttg
      841 ctttcggttt ggaaccgaag ccgcgcgtct tgtctgctgc agcgctgcag catcgttctg
      901 tgttgtctct gtctgactgt gtttctgtat ttgtctgaaa attagggcca gactgttacc
      961 actcccttaa gtttgacctt aggtcactgg aaagatgtcg agcggatcgc tcacaaccag
     1021 tcggtagatg tcaagaagag acgttgggtt accttctgct ctgcagaatg gccaaccttt
     1081 aacgtcggat ggccgcgaga cggcaccttt aaccgagacc tcatcaccca ggttaagatc
     1141 aaggtctttt cacctggccc gcatggacac ccagaccagg tcccctacat cgtgacctgg
     1201 gaagccttgg cttttgaccc ccctccctgg gtcaagccct ttgtacaccc taagcctccg
     1261 cctcctcttc ctccatccgc cccgtctctc ccccttgaac ctcctcgttc gaccccgcct
     1321 cgatcctccc tttatccagc cctcactcct tctctaggcg ccggccggat ccgagggcct
     1381 atttcccatg attccttcat atttgcatat acgatacaag gctgttagag agataattgg
     1441 aattaatttg actgtaaaca caaagatatt agtacaaaat acgtgacgta gaaagtaata
     1501 atttcttggg tagtttgcag ttttaaaatt atgttttaaa atggactatc atatgcttac
     1561 cgtaacttga aagtatttcg atttcttggc tttatatatc ttgtggaaag gacgaaacac
     1621 cgccagtacc caaaaagcgg ggttttagag ctagaaatag caagttaaaa taaggctagt
     1681 ccgttatcaa cttgaaaaag tggcaccgag tcggtgcttt ttttcgaatt ccgatgtacg
     1741 ggccagatat acgcgttatc atcgcgtgag gctccggtgc ccgtcagtgg gcagagcgca
     1801 catcgcccac agtccccgag aagttggggg gaggggtcgg caattgaacc ggtgcctaga
     1861 gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg
     1921 agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg
     1981 ggtttgccgc cagaacacag gtaagtgccg tgtgtggttc ccgcgggcct ggcctcttta
     2041 cgggttatgg cccttgcgtg ccttgaatta cttccacgcc cctggctgca gtacgtgatt
     2101 cttgatcccg agcttcgggt tggaagtggg tgggagagtt cgaggccttg cgcttaagga
     2161 gccccttcgc ctcgtgcttg agttgaggcc tggcctgggc gctggggccg ccgcgtgcga
     2221 atctggtggc accttcgcgc ctgtctcgct gctttcgata agtctctagc catttaaaat
     2281 ttttgatgac ctgctgcgac gctttttttc tggcaagata gtcttgtaaa tgcgggccaa
     2341 gatctgcaca ctggtatttc ggtttttggg gccgcgggcg gcgacggggc ccgtgcgtcc
     2401 cagcgcacat gttcggcgag gcggggcctg cgagcgcggc caccgagaat cggacggggg
     2461 tagtctcaag ctggccggcc tgctctggtg cctggcctcg cgccgccgtg tatcgccccg
     2521 ccctgggcgg caaggctggc ccggtcggca ccagttgcgt gagcggaaag atggccgctt
     2581 cccggccctg ctgcagggag ctcaaaatgg aggacgcggc gctcgggaga gcgggcgggt
     2641 gagtcaccca cacaaaggaa aagggccttt ccgtcctcag ccgtcgcttc atgtgactcc
     2701 acggagtacc gggcgccgtc caggcacctc gattagttct cgagcttttg gagtacgtcg
     2761 tctttaggtt ggggggaggg gttttatgcg atggagtttc cccacactga gtgggtggag
     2821 actgaagtta ggccagcttg gcacttgatg taattctcct tggaatttgc cctttttgag
     2881 tttggatctt ggttcattct caagcctcag acagtggttc aaagtttttt tcttccattt
     2941 caggtgtcgt gagggccacc atggattata aggatgacga cgataaggtg agcaagggcg
     3001 aggaggataa catggccatc atcaaggagt tcatgcgctt caaggtgcac atggagggct
     3061 ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca
     3121 cccagaccgc caagctgaag gtgaccaagg gtggccccct gcccttcgcc tgggacatcc
     3181 tgtcccctca gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc gacatccccg
     3241 actacttgaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg
     3301 acggcggcgt ggtgaccgtg acccaggact cctccctgca ggacggcgag ttcatctaca
     3361 aggtgaagct gcgcggcacc aacttcccct ccgacggccc cgtaatgcag aagaagacca
     3421 tgggctggga ggcctcctcc gagcggatgt accccgagga cggcgccctg aagggcgaga
     3481 tcaagcagag gctgaagctg aaggacggcg gccactacga cgctgaggtc aagaccacct
     3541 acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc aagttggaca
     3601 tcacctccca caacgaggac tacaccatcg tggaacagta cgaacgcgcc gagggccgcc
     3661 actccaccgg cggcatggac gagctgtaca aggtcgctac catcgaggga tccggagagg
     3721 gcagaggaag tctgctaaca tgcggtgacg tcgaggagaa tcctggccca atggacaaga
     3781 agtacagcat cggcctggac atcggcacca acagcgtggg ctgggccgtg atcaccgacg
     3841 agtacaaggt gcccagcaag aagttcaagg tgctgggcaa caccgaccgc cacagcatca
     3901 agaagaacct gatcggcgcc ctgctgttcg acagcggcga gaccgccgag gccacccgcc
     3961 tgaagcgcac cgcccgccgc cgctacaccc gccgcaagaa ccgcatctgc tacctgcagg
     4021 agatcttcag caacgagatg gccaaggtgg acgacagctt cttccaccgc ctggaggaga
     4081 gcttcctggt ggaggaggac aagaagcacg agcgccaccc catcttcggc aacatcgtgg
     4141 acgaggtggc ctaccacgag aagtacccca ccatctacca cctgcgcaag aagctggtgg
     4201 acagcaccga caaggccgac ctgcgcctga tctacctggc cctggcccac atgatcaagt
     4261 tccgcggcca cttcctgatc gagggcgacc tgaaccccga caacagcgac gtggacaagc
     4321 tgttcatcca gctggtgcag acctacaacc agctgttcga ggagaacccc atcaacgcca
     4381 gcggcgtgga cgccaaggcc atcctgagcg cccgcctgag caagagccgc cgcctggaga
     4441 acctgatcgc ccagctgccc ggcgagaaga agaacggcct gttcggcaac ctgatcgccc
     4501 tgagcctggg cctgaccccc aacttcaaga gcaacttcga cctggccgag gacgccaagc
     4561 tgcagctgag caaggacacc tacgacgacg acctggacaa cctgctggcc cagatcggcg
     4621 accagtacgc cgacctgttc ctggccgcca agaacctgag cgacgccatc ctgctgagcg
     4681 acatcctgcg cgtgaacacc gagatcacca aggcccccct gagcgccagc atgatcaagc
     4741 gctacgacga gcaccaccag gacctgaccc tgctgaaggc cctggtgcgc cagcagctgc
     4801 ccgagaagta caaggagatc ttcttcgacc agagcaagaa cggctacgcc ggctacatcg
     4861 acggcggcgc cagccaggag gagttctaca agttcatcaa gcccatcctg gagaagatgg
     4921 acggcaccga ggagctgctg gtgaagctga accgcgagga cctgctgcgc aagcagcgca
     4981 ccttcgacaa cggcagcatc ccccaccaga tccacctggg cgagctgcac gccatcctgc
     5041 gccgccagga ggacttctac cccttcctga aggacaaccg cgagaagatc gagaagatcc
     5101 tgaccttccg catcccctac tacgtcggcc ccctggcccg cggcaacagc cgcttcgcct
     5161 ggatgacccg caagagcgag gagaccatca ccccctggaa cttcgaggag gtggtggaca
     5221 agggcgccag cgcccagagc ttcatcgagc gcatgaccaa cttcgacaag aacctgccca
     5281 acgagaaggt gctgcccaag cacagcctgc tgtacgagta cttcaccgtg tacaacgagc
     5341 tgaccaaggt gaagtacgtg accgagggca tgcgcaagcc cgccttcctg agcggcgagc
     5401 agaagaaggc catcgtggac ctgctgttca agaccaaccg caaggtgacc gtgaagcagc
     5461 tgaaggagga ctacttcaag aagatcgagt gcttcgacag cgtggagatc agcggcgtgg
     5521 aggaccgctt caacgccagc ctgggcacct accacgacct gctgaagatc atcaaggaca
     5581 aggacttcct ggacaacgag gagaacgagg acatcctgga ggacatcgtg ctgaccctga
     5641 ccctgttcga ggaccgcgag atgatcgagg agcgcctgaa gacctacgcc cacctgttcg
     5701 acgacaaggt gatgaagcag ctgaagcgcc gccgctacac cggctggggc cgcctgagcc
     5761 gcaagctgat caacggcatc cgcgacaagc agagcggcaa gaccatcctg gacttcctga
     5821 agagcgacgg cttcgccaac cgcaacttca tgcagctgat ccacgacgac agcctgacct
     5881 tcaaggagga catccagaag gcccaggtga gcggccaggg cgacagcctg cacgagcaca
     5941 tcgccaacct ggccggcagc cccgccatca agaagggcat cctgcagacc gtgaaggtgg
     6001 tggacgagct ggtgaaggtg atgggccgcc acaagcccga gaacatcgtg atcgagatgg
     6061 cccgcgagaa ccagaccacc cagaagggcc agaagaacag ccgcgagcgc atgaagcgca
     6121 tcgaggaggg catcaaggag ctgggcagcc agatcctgaa ggagcacccc gtggagaaca
     6181 cccagctgca gaacgagaag ctgtacctgt actacctgca gaacggccgc gacatgtacg
     6241 tggaccagga gctggacatc aaccgcctga gcgactacga cgtggaccac atcgtgcccc
     6301 agagcttcct gaaggacgac agcatcgaca acaaggtgct gacccgcagc gacaagaacc
     6361 gcggcaagag cgacaacgtg cccagcgagg aggtggtgaa gaagatgaag aactactggc
     6421 gccagctgct gaacgccaag ctgatcaccc agcgcaagtt cgacaacctg accaaggccg
     6481 agcgcggcgg cctgagcgag ctggacaagg ccggcttcat caagcgccag ctggtggaga
     6541 cccgccagat caccaagcac gtggcccaga tcctggacag ccgcatgaac accaagtacg
     6601 acgagaacga caagctgatc cgcgaggtga aggtgatcac cctgaagagc aagctggtga
     6661 gcgacttccg caaggacttc cagttctaca aggtgcgcga gatcaacaac taccaccacg
     6721 cccacgacgc ctacctgaac gccgtggtgg gcaccgccct gatcaagaag taccccaagc
     6781 tggagagcga gttcgtgtac ggcgactaca aggtgtacga cgtgcgcaag atgatcgcca
     6841 agagcgagca ggagatcggc aaggccaccg ccaagtactt cttctacagc aacatcatga
     6901 acttcttcaa gaccgagatc accctggcca acggcgagat ccgcaagcgc cccctgatcg
     6961 agaccaacgg cgagaccggc gagatcgtgt gggacaaggg ccgcgacttc gccaccgtgc
     7021 gcaaggtgct gagcatgccc caggtgaaca tcgtgaagaa gaccgaggtg cagaccggcg
     7081 gcttcagcaa ggagagcatc ctgcccaagc gcaacagcga caagctgatc gcccgcaaga
     7141 aggactggga ccccaagaag tacggcggct tcgacagccc caccgtggcc tacagcgtgc
     7201 tggtggtggc caaggtggag aagggcaaga gcaagaagct gaagagcgtg aaggagctgc
     7261 tgggcatcac catcatggag cgcagcagct tcgagaagaa ccccatcgac ttcctggagg
     7321 ccaagggcta caaggaggtg aagaaggacc tgatcatcaa gctgcccaag tacagcctgt
     7381 tcgagctgga gaacggccgc aagcgcatgc tggccagcgc cggcgagctg cagaagggca
     7441 acgagctggc cctgcccagc aagtacgtga acttcctgta cctggccagc cactacgaga
     7501 agctgaaggg cagccccgag gacaacgagc agaagcagct gttcgtggag cagcacaagc
     7561 actacctgga cgagatcatc gagcagatca gcgagttcag caagcgcgtg atcctggccg
     7621 acgccaacct ggacaaggtg ctgagcgcct acaacaagca ccgcgacaag cccatccgcg
     7681 agcaggccga gaacatcatc cacctgttca ccctgaccaa cctgggcgcc cccgccgcct
     7741 tcaagtactt cgacaccacc atcgaccgca agcgctacac cagcaccaag gaggtgctgg
     7801 acgccaccct gatccaccag agcatcaccg gcctgtacga gacccgcatc gacctgagcc
     7861 agctgggcgg cgacagccgc gccgacccca agaagaagcg caaggtgtga gcggccgcat
     7921 cgataaaata aaagatttta tttagtctcc agaaaaaggg gggaatgaaa gaccccacct
     7981 gtaggtttgg caagctagct taagtaacgc cattttgcaa ggcatggaaa aatacataac
     8041 tgagaataga gaagttcaga tcaaggtcag gaacagatgg aacagctgaa tatgggccaa
     8101 acaggatatc tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gatggaacag
     8161 ctgaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
     8221 gaacagatgg tccccagatg cggtccagcc ctcagcagtt tctagagaac catcagatgt
     8281 ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt
     8341 tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac
     8401 ccctcactcg gggcgccagt cctccgattg actgagtcgc ccgggtaccc gtgtatccaa
     8461 taaaccctct tgcagttgca tccgacttgt ggtctcgctg ttccttggga gggtctcctc
     8521 tgagtgattg actacccgtc agcgggggtc tttcacatgc agcatgtatc aaaattaatt
     8581 tggttttttt tcttaagtat ttacattaaa tggccatagt tgcattaatg aatcggccaa
     8641 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     8701 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     8761 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     8821 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     8881 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     8941 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     9001 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     9061 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     9121 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     9181 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     9241 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     9301 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     9361 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     9421 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     9481 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     9541 acctagatcc ttttgcggcc gcaaatcaat ctaaagtata tatgagtaaa cttggtctga
     9601 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     9661 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     9721 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     9781 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     9841 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     9901 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     9961 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
    10021 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
    10081 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
    10141 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
    10201 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
    10261 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
    10321 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
    10381 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
    10441 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
    10501 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
    10561 ggttccgcgc acatttcccc gaaaagtgcc ac
//
