LOCUS       sequence-272994-       10927 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  OsTIR1(WT)-P2A-mAID-EGFP-Nluc.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10927)
  AUTHORS   Yesbolatova A, Saito Y, Kitamoto N, Makino-Itou H, Ajima R, Nakano 
            R, Nakaoka H, Fukui K, Gamo K, Tominari Y, Takeuchi H, Saga Y, 
            Hayashi KI, Kanemaki MT
  TITLE     The auxin-inducible degron 2 technology provides sharp degradation 
            control in yeast, mammalian cells, and mice.
  JOURNAL   Nat Commun. 2020 Nov 11;11(1):5701. doi: 10.1038/s41467-020-19532-z.
  PUBMED    33177522
REFERENCE   2  (bases 1 to 10927)
  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: "Nat 
            Commun."; date: "2020-11-11"; volume: "11(1):5701. doi"; pages: " 
            10.1038/s41467-020-19532-z"
COMMENT     Sequence Label: pMK427 (OsTIR1(WT) mAID-EGFP-Nluc)
FEATURES             Location/Qualifiers
     source          1..10927
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    33..54
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(69..74,75..92,93..99)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    107..123
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     112..134
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     131..147
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     131..147
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     166..186
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        168..186
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    join(243..254,255..442)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     enhancer        464..843
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        845..1122
                     /label=chicken beta-actin promoter
     intron          1124..2141
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     2065..2087
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     2149..2168
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             2213..3937
                     /codon_start=1
                     /product="rice transport inhibitor response 1 protein"
                     /label=OsTIR1
                     /note="F-box protein that mediates auxin-dependent 
                     interaction of a ubiquitin ligase with an auxin-inducible 
                     degron (Tan et al., 2007)"
                     /translation="MTYFPEEVVEHIFSFLPAQRDRNTVSLVCKVWYEIERLSRRGVFV
                     GNCYAVRAGRVAARFPNVRALTVKGKPHFADFNLVPPDWGGYAGPWIEAAARGCHGLEE
                     LRMKRMVVSDESLELLARSFPRFRALVLISCEGFSTDGLAAVASHCKLLRELDLQENEV
                     EDRGPRWLSCFPDSCTSLVSLNFACIKGEVNAGSLERLVSRSPNLRSLRLNRSVSVDTL
                     AKILLRTPNLEDLGTGNLTDDFQTESYFKLTSALEKCKMLRSLSGFWDASPVCLSFIYP
                     LCAQLTGLNLSYAPTLDASDLTKMISRCVKLQRLWVLDCISDKGLQVVASSCKDLQELR
                     VFPSDFYVAGYSAVTEEGLVAVSLGCPKLNSLLYFCHQMTNAALVTVAKNCPNFTRFRL
                     CILEPGKPDVVTSQPLDEGFGAIVRECKGLQRLSISGLLTDKVFMYIGKYAKQLEMLSI
                     AFAGDSDKGMMHVMNGCKNLRKLEIRDSPFGDAALLGNFARYETMRSLWMSSCNVTLKG
                     CQVLASKMPMLNVEVINERDGSNEMEENHGDLPKVEKLYVYRTTAGARDDAPNFVKIL"
     CDS             3953..4009
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             4016..4219
                     /codon_start=1
                     /product="minimal degron from the auxin-responsive IAA17 
                     protein of Arabidopsis thaliana (Kubota et al., 2013)"
                     /label=mini-AID
                     /note="acts as an auxin-inducible degron (AID) in the 
                     presence of TIR1"
                     /translation="KEKSACPKDPAKPPAKAQVVGWPPVRSYRKNVMVSCQKSSGGPEA
                     AAFVKVSMDGAPYLRKIDLRMYK"
     CDS             join(4226..4228,4229..4231,4232..4942)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(4226..4228,4229..4231,4232..4942)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4271..4292)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4532..4551)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4879..4900
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             4979..5494
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     polyA_signal    5624..5745
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5661..5680)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5715..5734
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    5776..5809
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        5870..6369
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(5905..5928)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     6302..6321
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             6430..7449
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGYV
                     LRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPET
                     ELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQTV
                     MDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFGD
                     SQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGNF
                     DDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE"
     CDS             6430..7449
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGYV
                     LRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPET
                     ELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQTV
                     MDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFGD
                     SQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGNF
                     DDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE"
     protein_bind    8012..8045
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    join(8076..8238,8239..8250)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(8280..8299)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8281..8299)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8309..8326)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8309..8325)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8318..8340)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      8467..8922
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8554..8573)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8764..8785
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        8948..9052
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9053..9121,9122..9913)
                     /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(9271..9290)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10084..10672
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10573..10592
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10826..10843
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
       61 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
      121 caatttcaca caggaaacag ctatgaccat gattacgcca agcgcgcaat taaccctcac
      181 taaagggaac aaaagctgga gctcggatct gctgggcttg ctgaaggtag ggggtcaaga
      241 accagaggtg taaagtactt gagtaatttt acttgattac tgtacttaag tattattttt
      301 ggggattttt actttacttg agtacaatta aaaatcaata cttttacttt tacttaatta
      361 cattttttta gaaaaaaaag tactttttac tccttacaat tttatttaca gtcaaaaagt
      421 acttattttt tggagatcac ttgggccctc tagatgcatg ctcgacattg attattgact
      481 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
      541 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
      601 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
      661 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
      721 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
      781 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
      841 atggtcgagg tgagccccac gttctgcttc actctcccca tctccccccc ctccccaccc
      901 ccaattttgt atttatttat tttttaatta ttttgtgcag cgatgggggc gggggggggg
      961 ggggggcgcg cgccaggcgg ggcggggcgg ggcgaggggc ggggcggggc gaggcggaga
     1021 ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt ttccttttat ggcgaggcgg
     1081 cggcggcggc ggccctataa aaagcgaagc gcgcggcggg cggggagtcg ctgcgacgct
     1141 gccttcgccc cgtgccccgc tccgccgccg cctcgcgccg cccgccccgg ctctgactga
     1201 ccgcgttact cccacaggtg agcgggcggg acggcccttc tcctccgggc tgtaattagc
     1261 gcttggttta atgacggctt gtttcttttc tgtggctgcg tgaaagcctt gaggggctcc
     1321 gggagggccc tttgtgcggg gggagcggct cggggggtgc gtgcgtgtgt gtgtgcgtgg
     1381 ggagcgccgc gtgcggctcc gcgctgcccg gcggctgtga gcgctgcggg cgcggcgcgg
     1441 ggctttgtgc gctccgcagt gtgcgcgagg ggagcgcggc cgggggcggt gccccgcggt
     1501 gcgggggggg ctgcgagggg aacaaaggct gcgtgcgggg tgtgtgcgtg ggggggtgag
     1561 cagggggtgt gggcgcgtcg gtcgggctgc aaccccccct gcacccccct ccccgagttg
     1621 ctgagcacgg cccggcttcg ggtgcggggc tccgtacggg gcgtggcgcg gggctcgccg
     1681 tgccgggcgg ggggtggcgg caggtggggg tgccgggcgg ggcggggccg cctcgggccg
     1741 gggagggctc gggggagggg cgcggcggcc cccggagcgc cggcggctgt cgaggcgcgg
     1801 cgagccgcag ccattgcctt ttatggtaat cgtgcgagag ggcgcaggga cttcctttgt
     1861 cccaaatctg tgcggagccg aaatctggga ggcgccgccg caccccctct agcgggcgcg
     1921 gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt cgtgcgtcgc
     1981 cgcgccgccg tccccttctc cctctccagc ctcggggctg tccgcggggg gacggctgcc
     2041 ttcggggggg acggggcagg gcggggttcg gcttctggcg tgtgaccggc ggctctagag
     2101 cctctgctaa ccatgttcat gccttcttct ttttcctaca gctcctgggc aacgtgctgg
     2161 ttattgtgct gtctcatcat tttggcaaag aattcctcga agcgctgcca ccatgacata
     2221 ctttcctgaa gaggtcgtcg aacacatttt tagcttcctg cctgcacaga gagatagaaa
     2281 cacagtgagc ctggtctgca aagtgtggta cgagatcgaa cgcctgagcc ggagaggagt
     2341 gttcgtcggc aactgctatg ctgtgagagc aggcagggtc gccgctaggt ttccaaatgt
     2401 gcgcgcactg accgtcaagg ggaaacccca cttcgccgac tttaacctgg tgccccctga
     2461 ttggggagga tacgccggcc cttggatcga ggcagccgct cgcggctgtc atggactgga
     2521 ggaactgcgc atgaagcgaa tggtggtctc tgacgaaagt ctggagctgc tggctcggag
     2581 cttccctagg tttcgcgcac tggtgctgat ttcttgcgaa ggcttcagca ccgatggact
     2641 ggcagccgtg gcctcccact gtaagctgct gcgggagctg gacctccagg agaatgaagt
     2701 ggaggataga ggccccagat ggctgtcttg cttcccagac tcatgtacca gcctggtgtc
     2761 cctgaacttt gcctgcatca aaggcgaagt gaatgctggg tccctggagc ggctggtctc
     2821 aagaagcccc aacctgaggt ctctgcggct gaaccggagc gtgagcgtgg acactctggc
     2881 taagattctg ctgagaaccc ctaacctgga ggatctggga accggcaatc tgacagacga
     2941 tttccagaca gaatcctact ttaaactgac ttctgccctg gagaagtgta aaatgctgag
     3001 gagtctgtca ggattctggg atgcttcacc cgtgtgcctg agctttatct accctctgtg
     3061 tgcacagctg acaggcctga acctgagcta tgcaccaacc ctggacgcca gtgatctgac
     3121 aaagatgatc tcacgctgcg tgaaactcca gcgactgtgg gtgctggact gtatttccga
     3181 taaggggctc caggtggtcg ccagctcctg caaggacctc caggagctga gagtgttccc
     3241 atctgatttt tacgtggccg gatatagtgc tgtcactgag gaaggcctgg tggcagtctc
     3301 actgggatgc ccaaagctga acagcctgct gtatttctgt catcagatga ctaatgctgc
     3361 actggtgacc gtcgccaaga actgccctaa tttcacccga tttcggctgt gtattctgga
     3421 accaggcaaa cccgacgtgg tcacatccca gccactggat gaagggtttg gagctatcgt
     3481 gagagagtgc aagggactcc agaggctgag catttccggc ctgctgacag acaaagtgtt
     3541 catgtacatc ggcaagtatg ctaagcagct ggagatgctg agcattgcat ttgccggaga
     3601 ctccgataag ggcatgatgc acgtgatgaa cgggtgtaag aatctgcgaa aactggaaat
     3661 ccgggacagc cctttcgggg atgccgctct gctgggaaac tttgccagat acgagacaat
     3721 gaggagcctg tggatgtcta gttgcaatgt gactctgaag ggctgtcagg tcctggctag
     3781 taaaatgcct atgctgaacg tggaagtcat taatgagcgg gacgggtcta acgaaatgga
     3841 ggaaaatcat ggcgacctgc caaaggtgga gaaactgtat gtgtatcgga ccaccgcagg
     3901 ggcaagagat gatgctccca actttgtgaa gattctgaga tcaggaagcg gagctactaa
     3961 cttcagcctg ctgaagcagg ctggagacgt ggaggagaac cctggaccta gatctaagga
     4021 gaagagtgct tgtcctaaag atccagccaa acctccggcc aaggcacaag ttgtgggatg
     4081 gccaccggtg agatcatacc ggaagaacgt gatggtttcc tgccaaaaat caagcggtgg
     4141 cccggaggcg gcggcgttcg tgaaggtatc aatggacgga gcaccgtact tgaggaaaat
     4201 cgatttgagg atgtataaag ctagcatggt gagcaagggc gaggagctgt tcaccggggt
     4261 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg
     4321 cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg
     4381 caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt
     4441 cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg
     4501 ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga
     4561 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa
     4621 ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta
     4681 tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat
     4741 cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg
     4801 ccccgtgctg ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc
     4861 caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct
     4921 cggcatggac gagctgtaca agggtgcagg cgcccaagct tcgaattctg cagtcgacat
     4981 ggtcttcaca ctcgaagatt tcgttgggga ctggcgacag acagccggct acaacctgga
     5041 ccaagtcctt gaacagggag gtgtgtccag tttgtttcag aatctcgggg tgtccgtaac
     5101 tccgatccaa aggattgtcc tgagcggtga aaatgggctg aagatcgaca tccatgtcat
     5161 catcccgtat gaaggtctga gcggcgacca aatgggccag atcgaaaaaa tttttaaggt
     5221 ggtgtaccct gtggatgatc atcactttaa ggtgatcctg cactatggca cactggtaat
     5281 cgacggggtt acgccgaaca tgatcgacta tttcggacgg ccgtatgaag gcatcgccgt
     5341 gttcgacggc aaaaagatca ctgtaacagg gaccctgtgg aacggcaaca aaattatcga
     5401 cgagcgcctg atcaaccccg acggctccct gctgttccga gtaaccatca acggagtgac
     5461 cggctggcgg ctgtgcgaac gcattctggc gtaaggatcc accggatcta gataactgat
     5521 cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct
     5581 ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc
     5641 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     5701 actgcattct agttgtggtt tgtccaaact catcaatgta tcttaacgcg tcgatcatat
     5761 tcaataaccc ttaatataac ttcgtataat gtatgctata cgaagttatt aggtctgaag
     5821 aggagtttac gtccagccaa gcttaggatc tcgacctcga aattctaccg ggtaggggag
     5881 gcgcttttcc caaggcagtc tggagcatgc gctttagcag ccccgctggg cacttggcgc
     5941 tacacaagtg gcctctggcc tcgcacacat tccacatcca ccggtaggcg ccaaccgact
     6001 ccgttctttg gtggcccctt cgcgccacct tctactcctc ccctagtcag gaagttcccc
     6061 cccgccccgc agctcgcgtc gtgcaggacg tgacaaatgg aagtagcacg tctcactagt
     6121 ctcgtgcaga tggacagcac cgctgagcaa tggaagcggg taggcctttg gggcagcggc
     6181 caatagcagc tttgctcctt cgctttctgg gctcagaggc tgggaagggg tgggtccggg
     6241 ggcgggctca ggggcgggct caggggcggg gcgggcgccc gaaggtcctc cggaggcccg
     6301 gcattctgca cgcttcaaaa gcgcacgtct gccgcgctgt tctcctcttc ctcatctccg
     6361 ggcctttcga cctgcatcca tctagatctc gagcagctga agcttaccgc tagcatggat
     6421 agatccggaa agcctgaact caccgcgacg tctgtcgaga agtttctgat cgaaaagttc
     6481 gacagcgtct ccgacctgat gcagctctcg gagggcgaag aatctcgtgc tttcagcttc
     6541 gatgtaggag ggcgtggata tgtcctgcgg gtaaatagct gcgccgatgg tttctacaaa
     6601 gatcgttatg tttatcggca ctttgcatcg gccgcgctcc cgattccgga agtgcttgac
     6661 attggggaat tcagcgagag cctgacctat tgcatctccc gccgtgcaca gggtgtcacg
     6721 ttgcaagacc tgcctgaaac cgaactgccc gctgttctgc agccggtcgc ggaggccatg
     6781 gatgcgatcg ctgcggccga tcttagccag acgagcgggt tcggcccatt cggaccgcaa
     6841 ggaatcggtc aatacactac atggcgtgat ttcatatgcg cgattgctga tccccatgtg
     6901 tatcactggc aaactgtgat ggacgacacc gtcagtgcgt ccgtcgcgca ggctctcgat
     6961 gagctgatgc tttgggccga ggactgcccc gaagtccggc acctcgtgca cgcggatttc
     7021 ggctccaaca atgtcctgac ggacaatggc cgcataacag cggtcattga ctggagcgag
     7081 gcgatgttcg gggattccca atacgaggtc gccaacatct tcttctggag gccgtggttg
     7141 gcttgtatgg agcagcagac gcgctacttc gagcggaggc atccggagct tgcaggatcg
     7201 ccgcggctcc gggcgtatat gctccgcatt ggtcttgacc aactctatca gagcttggtt
     7261 gacggcaatt tcgatgatgc agcttgggcg cagggtcgat gcgacgcaat cgtccgatcc
     7321 ggagccggga ctgtcgggcg tacacaaatc gcccgcagaa gcgcggccgt ctggaccgat
     7381 ggctgtgtag aagtactcgc cgatagtgga aaccgacgcc ccagcactcg tccgagggca
     7441 aaggaatagg ctagccgccc gccccacgac ccgcagcgcc cgaccgaaag gagcgcacga
     7501 ccccatgcat cgatgatatc agatccccgg gatgcagaaa ttgatgatct attaaacaat
     7561 aaagatgtcc actaaaatgg aagtttttcc tgtcatactt tgttaagaag ggtgagaaca
     7621 gagtacctac attttgaatg gaaggattgg agctacgggg gtgggggtgg ggtgggatta
     7681 gataaatgcc tgctctttac tgaaggctct ttactattgc tttatgataa tgtttcatag
     7741 ttggatatca taatttaaac aagcaaaacc aaattaaggg ccagctcatt cctcccactc
     7801 atgatctata gatctataga tctctcgtgg gatcattgtt tttctcttga ttcccacttt
     7861 gtggttctaa gtactgtggt ttccaaatgt gtcagtttca tagcctgaag aacgagatca
     7921 gcagcctctg ttccacatac acttcattct cagtattgtt ttgccaagtt ctaattccat
     7981 cagaagctgg tcgagatccg gaacccttaa tataacttcg tataatgtat gctatacgaa
     8041 gttattaggt ccctcgaaga ggttcactaa gatctttctt gcttttactt ttacttcctt
     8101 aatactcaag tacaatttta atggagtact tttttacttt tactcaagta agattctagc
     8161 cagatacttt tacttttaat tgagtaaaat tttccctaag tacttgtact ttcacttgag
     8221 taaaattttt gagtactttt tacacctctg tcaagaacca tatgccggta cccaattcgc
     8281 cctatagtga gtcgtattac gcgcgctcac tggccgtcgt tttacaacgt cgtgactggg
     8341 aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc
     8401 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
     8461 aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     8521 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     8581 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     8641 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     8701 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     8761 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     8821 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     8881 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttaggtggca cttttcgggg
     8941 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     9001 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     9061 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     9121 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     9181 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     9241 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     9301 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     9361 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     9421 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     9481 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     9541 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     9601 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     9661 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     9721 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     9781 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     9841 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     9901 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     9961 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
    10021 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
    10081 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
    10141 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
    10201 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
    10261 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
    10321 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
    10381 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
    10441 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
    10501 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
    10561 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
    10621 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
    10681 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
    10741 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
    10801 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
    10861 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
    10921 gtttccc
//
