LOCUS       sequence-296198-       10924 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 10924)
  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 10924)
  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..10924
                     /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          1123..2139
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     2063..2085
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     2147..2166
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             2211..3935
                     /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             3951..4007
                     /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             4014..4217
                     /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(4224..4226,4227..4229,4230..4940)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(4224..4226,4227..4229,4230..4940)
                     /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(4269..4290)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4530..4549)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4877..4898
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             4977..5492
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     polyA_signal    5622..5743
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5659..5678)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5713..5732
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    5774..5807
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        5868..6367
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(5903..5926)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     6300..6319
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             6428..7447
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGYV
                     LRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPET
                     ELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQTV
                     MDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFGD
                     SQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGNF
                     DDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE"
     CDS             6428..7447
                     /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    8010..8043
                     /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(8074..8236,8237..8248)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(8278..8297)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8279..8297)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8307..8324)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8307..8323)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8316..8338)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      8465..8919
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8551..8570)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8761..8782
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        8945..9049
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9050..9118,9119..9910)
                     /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(9268..9287)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10081..10669
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10570..10589
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10823..10840
                     /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 cgcsrggsgg gggggggsgg ggsgrggggs ggggsggggs gaggcggaga
     1021 ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt ttccttttat ggcgaggcgg
     1081 cggcggcggc ggccctataa aaagcgaagc gcgcggcggg cgggagtcgc tgcgcgctgc
     1141 cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc cgccccggct ctgactgacc
     1201 gcgttactcc cacaggtgag cgggcgggac ggcccttctc ctccgggctg taattagcgc
     1261 ttggtttaat gacggcttgt ttcttttctg tggctgcgtg aaagccttga ggggctccgg
     1321 gagggccctt tgtgcggggg gagcggctcg gggggtgcgt gcgtgtgtgt gtgcgtgggg
     1381 agcgccgcgt gcggctccgc gctgcccggc ggctgtgagc gctgcgggcg cggcgcgggg
     1441 ctttgtgcgc tccgcagtgt gcgcgagggg agcgcggccg ggggcggtgc cccgcggtgc
     1501 ggggggggct gcgaggggaa caaaggctgc gtgcggggtg tgtgcgtggg ggggtgagca
     1561 gggggtgtgg gcgcgtcggt cgggctgcaa ccccccctgc acccccctcc ccgagttgct
     1621 gagcacggcc cggcttcggg tgcggggctc cgtacggggc gtggcgcggg gctcgccgtg
     1681 ccgggcgggg ggtggcggca ggtgggggtg ccgggcgggg cggggccgcc tcgggccggg
     1741 grgggctcgg gggaggggcg cggcggcccc cggagcgccg gcggctgtcg aggcgcggcg
     1801 agccgcagcc attgcctttt atggtaatcg tgcgagaggg cgcagggact tcctttgtcc
     1861 caaatctgtg cggagccgaa atctgggagg cgccgccgca ccccctctag cgggcgcggg
     1921 gcgaagcggt gcggcgccgg caggaaggaa atgggcgggg agggccttcg tgcgtcgccg
     1981 cgccgccgtc cccttctccc tctccagcct cggggctgtc cgcgggggga cggctgcctt
     2041 cgggggggac ggggcagggc ggggttcggc ttctggcgtg tgaccggcgg ctctagagcc
     2101 tctgctaacc atgttcatgc cttcttcttt ttcctacagc tcctgggcaa cgtgctggtt
     2161 attgtgctgt ctcatcattt tggcaaagaa ttcctcgaag cgctgccacc atgacatact
     2221 ttcctgaaga ggtcgtcgaa cacattttta gcttcctgcc tgcacagaga gatagaaaca
     2281 cagtgagcct ggtctgcaaa gtgtggtacg agatcgaacg cctgagccgg agaggagtgt
     2341 tcgtcggcaa ctgctatgct gtgagagcag gcagggtcgc cgctaggttt ccaaatgtgc
     2401 gcgcactgac cgtcaagggg aaaccccact tcgccgactt taacctggtg ccccctgatt
     2461 ggggaggata cgccggccct tggatcgagg cagccgctcg cggctgtcat ggactggagg
     2521 aactgcgcat gaagcgaatg gtggtctctg acgaaagtct ggagctgctg gctcggagct
     2581 tccctaggtt tcgcgcactg gtgctgattt cttgcgaagg cttcagcacc gatggactgg
     2641 cagccgtggc ctcccactgt aagctgctgc gggagctgga cctccaggag aatgaagtgg
     2701 aggatagagg ccccagatgg ctgtcttgct tcccagactc atgtaccagc ctggtgtccc
     2761 tgaactttgc ctgcatcaaa ggcgaagtga atgctgggtc cctggagcgg ctggtctcaa
     2821 gaagccccaa cctgaggtct ctgcggctga accggagcgt gagcgtggac actctggcta
     2881 agattctgct gagaacccct aacctggagg atctgggaac cggcaatctg acagacgatt
     2941 tccagacaga atcctacttt aaactgactt ctgccctgga gaagtgtaaa atgctgagga
     3001 gtctgtcagg attctgggat gcttcacccg tgtgcctgag ctttatctac cctctgtgtg
     3061 cacagctgac aggcctgaac ctgagctatg caccaaccct ggacgccagt gatctgacaa
     3121 agatgatctc acgctgcgtg aaactccagc gactgtgggt gctggactgt atttccgata
     3181 aggggctcca ggtggtcgcc agctcctgca aggacctcca ggagctgaga gtgttcccat
     3241 ctgattttta cgtggccgga tatagtgctg tcactgagga aggcctggtg gcagtctcac
     3301 tgggatgccc aaagctgaac agcctgctgt atttctgtca tcagatgact aatgctgcac
     3361 tggtgaccgt cgccaagaac tgccctaatt tcacccgatt tcggctgtgt attctggaac
     3421 caggcaaacc cgacgtggtc acatcccagc cactggatga agggtttgga gctatcgtga
     3481 gagagtgcaa gggactccag aggctgagca tttccggcct gctgacagac aaagtgttca
     3541 tgtacatcgg caagtatgct aagcagctgg agatgctgag cattgcattt gccggagact
     3601 ccgataaggg catgatgcac gtgatgaacg ggtgtaagaa tctgcgaaaa ctggaaatcc
     3661 gggacagccc tttcggggat gccgctctgc tgggaaactt tgccagatac gagacaatga
     3721 ggagcctgtg gatgtctagt tgcaatgtga ctctgaaggg ctgtcaggtc ctggctagta
     3781 aaatgcctat gctgaacgtg gaagtcatta atgagcggga cgggtctaac gaaatggagg
     3841 aaaatcatgg cgacctgcca aaggtggaga aactgtatgt gtatcggacc accgcagggg
     3901 caagagatga tgctcccaac tttgtgaaga ttctgagatc aggaagcgga gctactaact
     3961 tcagcctgct gaagcaggct ggagacgtgg aggagaaccc tggacctaga tctaaggaga
     4021 agagtgcttg tcctaaagat ccagccaaac ctccggccaa ggcacaagtt gtgggatggc
     4081 caccggtgag atcataccgg aagaacgtga tggtttcctg ccaaaaatca agcggtggcc
     4141 cggaggcggc ggcgttcgtg aaggtatcaa tggacggagc accgtacttg aggaaaatcg
     4201 atttgaggat gtataaagct agcatggtga gcaagggcga ggagctgttc accggggtgg
     4261 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg
     4321 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca
     4381 agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca
     4441 gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
     4501 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
     4561 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
     4621 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata
     4681 tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg
     4741 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     4801 ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca
     4861 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
     4921 gcatggacga gctgtacaag ggtgcaggcg cccaagcttc gaattctgca gtcgacatgg
     4981 tcttcacact cgaagatttc gttggggact ggcgacagac agccggctac aacctggacc
     5041 aagtccttga acagggaggt gtgtccagtt tgtttcagaa tctcggggtg tccgtaactc
     5101 cgatccaaag gattgtcctg agcggtgaaa atgggctgaa gatcgacatc catgtcatca
     5161 tcccgtatga aggtctgagc ggcgaccaaa tgggccagat cgaaaaaatt tttaaggtgg
     5221 tgtaccctgt ggatgatcat cactttaagg tgatcctgca ctatggcaca ctggtaatcg
     5281 acggggttac gccgaacatg atcgactatt tcggacggcc gtatgaaggc atcgccgtgt
     5341 tcgacggcaa aaagatcact gtaacaggga ccctgtggaa cggcaacaaa attatcgacg
     5401 agcgcctgat caaccccgac ggctccctgc tgttccgagt aaccatcaac ggagtgaccg
     5461 gctggcggct gtgcgaacgc attctggcgt aaggatccac cggatctaga taactgatca
     5521 taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc
     5581 ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt
     5641 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     5701 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaacgcgtc gatcatattc
     5761 aataaccctt aatataactt cgtataatgt atgctatacg aagttattag gtctgaagag
     5821 gagtttacgt ccagccaagc ttaggatctc gacctcgaaa ttctaccggg taggggaggc
     5881 gcttttccca aggcagtctg gagcatgcgc tttagcagcc ccgctgggca cttggcgcta
     5941 cacaagtggc ctctggcctc gcacacattc cacatccacc ggtaggcgcc aaccgactcc
     6001 gttctttggt ggccccttcg cgccaccttc tactcctccc ctagtcagga agttcccccc
     6061 cgccccgcag ctcgcgtcgt gcaggacgtg acaaatggaa gtagcacgtc tcactagtct
     6121 cgtgcagatg gacagcaccg ctgagcaatg gaagcgggta ggcctttggg gcagcggcca
     6181 atagcagctt tgctccttcg ctttctgggc tcagaggctg ggaaggggtg ggtccggggg
     6241 cgggctcagg ggcgggctca ggggcggggc gggcgcccga aggtcctccg gaggcccggc
     6301 attctgcacg cttcaaaagc gcacgtctgc cgcgctgttc tcctcttcct catctccggg
     6361 cctttcgacc tgcatccatc tagatctcga gcagctgaag cttaccgcta gcatggatag
     6421 atccggaaag cctgaactca ccgcgacgtc tgtcgagaag tttctgatcg aaaagttcga
     6481 cagcgtctcc gacctgatgc agctctcgga gggcgaagaa tctcgtgctt tcagcttcga
     6541 tgtaggaggg cgtggatatg tcctgcgggt aaatagctgc gccgatggtt tctacaaaga
     6601 tcgttatgtt tatcggcact ttgcatcggc cgcgctcccg attccggaag tgcttgacat
     6661 tggggaattc agcgagagcc tgacctattg catctcccgc cgtgcacagg gtgtcacgtt
     6721 gcaagacctg cctgaaaccg aactgcccgc tgttctgcag ccggtcgcgg aggccatgga
     6781 tgcgatcgct gcggccgatc ttagccagac gagcgggttc ggcccattcg gaccgcaagg
     6841 aatcggtcaa tacactacat ggcgtgattt catatgcgcg attgctgatc cccatgtgta
     6901 tcactggcaa actgtgatgg acgacaccgt cagtgcgtcc gtcgcgcagg ctctcgatga
     6961 gctgatgctt tgggccgagg actgccccga agtccggcac ctcgtgcacg cggatttcgg
     7021 ctccaacaat gtcctgacgg acaatggccg cataacagcg gtcattgact ggagcgaggc
     7081 gatgttcggg gattcccaat acgaggtcgc caacatcttc ttctggaggc cgtggttggc
     7141 ttgtatggag cagcagacgc gctacttcga gcggaggcat ccggagcttg caggatcgcc
     7201 gcggctccgg gcgtatatgc tccgcattgg tcttgaccaa ctctatcaga gcttggttga
     7261 cggcaatttc gatgatgcag cttgggcgca gggtcgatgc gacgcaatcg tccgatccgg
     7321 agccgggact gtcgggcgta cacaaatcgc ccgcagaagc gcggccgtct ggaccgatgg
     7381 ctgtgtagaa gtactcgccg atagtggaaa ccgacgcccc agcactcgtc cgagggcaaa
     7441 ggaataggct agccgcccgc cccacgaccc gcagcgcccg accgaaagga gcgcacgacc
     7501 ccatgcatcg atgatatcag atccccggga tgcagaaatt gatgatctat taaacaataa
     7561 agatgtccac taaaatggaa gtttttcctg tcatactttg ttaagaaggg tgagaacaga
     7621 gtacctacat tttgaatgga aggattggag ctacgggggt gggggtgggg tgggattaga
     7681 taaatgcctg ctctttactg aaggctcttt actattgctt tatgataatg tttcatagtt
     7741 ggatatcata atttaaacaa gcaaaaccaa attaagggcc agctcattcc tcccactcat
     7801 gatctataga tctatagatc tctcgtggga tcattgtttt tctcttgatt cccactttgt
     7861 ggttctaagt actgtggttt ccaaatgtgt cagtttcata gcctgaagaa cgagatcagc
     7921 agcctctgtt ccacatacac ttcattctca gtattgtttt gccaagttct aattccatca
     7981 gaagctggtc gagatccgga acccttaata taacttcgta taatgtatgc tatacgaagt
     8041 tattaggtcc ctcgaagagg ttcactaaga tctttcttgc ttttactttt acttccttaa
     8101 tactcaagta caattttaat ggagtacttt tttactttta ctcaagtaag attctagcca
     8161 gatactttta cttttaattg agtaaaattt tccctaagta cttgtacttt cacttgagta
     8221 aaatttttga gtacttttta cacctctgtc aagaaccata tgccggtacc caattcgccc
     8281 tatagtgagt cgtattacgc gcgctcactg gccgtcgttt tacaacgtcg tgactgggaa
     8341 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
     8401 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     8461 tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     8521 accaccacac ccgccgcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     8581 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     8641 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     8701 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     8761 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     8821 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     8881 ttaacgcgaa ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa
     8941 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     9001 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     9061 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     9121 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     9181 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     9241 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     9301 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     9361 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     9421 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     9481 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     9541 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     9601 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     9661 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     9721 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     9781 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     9841 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     9901 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     9961 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
    10021 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
    10081 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
    10141 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
    10201 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
    10261 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
    10321 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
    10381 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
    10441 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
    10501 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
    10561 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
    10621 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
    10681 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
    10741 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
    10801 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
    10861 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
    10921 tccc
//
