LOCUS       sequence-296431-       10024 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  OsTIR1(F74G)-P2A-mAID-EGFP-NLS TOL2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10024)
  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 10024)
  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: pAY15 (OsTIR1(F74G) mAID-EGFP-NLS)
FEATURES             Location/Qualifiers
     source          1..10024
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(34..461)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(144..165)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     356..375
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     588..610
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     602..619
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     603..619
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    join(678..689,690..852)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     protein_bind    complement(883..916)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(1473..2072)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APHNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     2053..2072
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(2112..2611)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(2160..2179)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     2553..2576
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     protein_bind    complement(2672..2705)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    2732..2853
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2743..2762)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2797..2816
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(3004..3024)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(join(3061..3771,3772..3774,3775..3777))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(3061..3771,3772..3774,3775..3777))
                     /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(3103..3124)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3452..3471
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3711..3732
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(3784..3987)
                     /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             complement(3994..4050)
                     /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             complement(4066..5790)
                     /codon_start=1
                     /product="rice transport inhibitor response 1 protein, 
                     mutated for use in the AID2 system of inducible degradation
                     (Yesbolatova et al., 2020)"
                     /label=OsTIR1(F74G)
                     /note="F-box protein that mediates auxin-dependent 
                     interaction of a ubiquitin ligase with an auxin-inducible 
                     degron (Tan et al., 2007)"
                     /translation="MTYFPEEVVEHIFSFLPAQRDRNTVSLVCKVWYEIERLSRRGVFV
                     GNCYAVRAGRVAARFPNVRALTVKGKPHGADFNLVPPDWGGYAGPWIEAAARGCHGLEE
                     LRMKRMVVSDESLELLARSFPRFRALVLISCEGFSTDGLAAVASHCKLLRELDLQENEV
                     EDRGPRWLSCFPDSCTSLVSLNFACIKGEVNAGSLERLVSRSPNLRSLRLNRSVSVDTL
                     AKILLRTPNLEDLGTGNLTDDFQTESYFKLTSALEKCKMLRSLSGFWDASPVCLSFIYP
                     LCAQLTGLNLSYAPTLDASDLTKMISRCVKLQRLWVLDCISDKGLQVVASSCKDLQELR
                     VFPSDFYVAGYSAVTEEGLVAVSLGCPKLNSLLYFCHQMTNAALVTVAKNCPNFTRFRL
                     CILEPGKPDVVTSQPLDEGFGAIVRECKGLQRLSISGLLTDKVFMYIGKYAKQLEMLSI
                     AFAGDSDKGMMHVMNGCKNLRKLEIRDSPFGDAALLGNFARYETMRSLWMSSCNVTLKG
                     CQVLASKMPMLNVEVINERDGSNEMEENHGDLPKVEKLYVYRTTAGARDDAPNFVKIL"
     primer_bind     complement(5860..5879)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     intron          5887..6903
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     complement(5941..5963)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     promoter        complement(6904..7181)
                     /label=chicken beta-actin promoter
     enhancer        7183..7562
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     misc_feature    join(7584..7771,7772..7783)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(7840..7860)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(7840..7858)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7879..7895)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7879..7895)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7892..7914)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7903..7919
                     /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)."
     promoter        complement(join(7927..7933,7934..7951,7952..7957))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7972..7993
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8110..8127)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8281..8869)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8361..8380)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9040..9831,9832..9900))
                     /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     9663..9682
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9901..10005)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
       61 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
      121 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
      181 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
      241 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
      301 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
      361 agaaagcgaa aggagcgggc gctagggcgc ggcgggtgtg gtggtcacgc tgcgcgtaac
      421 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcccattcgc cattcaggct
      481 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
      541 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
      601 ttgtaaaacg acggccagtg agcgcgcgta atacgactca ctatagggcg aattgggtac
      661 cggcatatgg ttcttgacag aggtgtaaaa agtactcaaa aattttactc aagtgaaagt
      721 acaagtactt agggaaaatt ttactcaatt aaaagtaaaa gtatctggct agaatcttac
      781 ttgagtaaaa gtaaaaaagt actccattaa aattgtactt gagtattaag gaagtaaaag
      841 taaaagcaag aaagatctta gtgaacctct tcgagggacc taataacttc gtatagcata
      901 cattatacga agttatatta agggttccgg atctcgacca gcttctgatg gaattagaac
      961 ttggcaaaac aatactgaga atgaagtgta tgtggaacag aggctgctga tctcgttctt
     1021 caggctatga aactgacaca tttggaaacc acagtactta gaaccacaaa gtgggaatca
     1081 agagaaaaac aatgatccca cgagagatct atagatctat agatcatgag tgggaggaat
     1141 gagctggccc ttaatttggt tttgcttgtt taaattatga tatccaacta tgaaacatta
     1201 tcataaagca atagtaaaga gccttcagta aagagcaggc atttatctaa tcccacccca
     1261 cccccacccc cgtagctcca atccttccat tcaaaatgta ggtactctgt tctcaccctt
     1321 cttaacaaag tatgacagga aaaacttcca ttttagtgga catctttatt gtttaataga
     1381 tcatcaattt ctgcatcccg gggatctgat atcatcgatg catggggtcg tgcgctcctt
     1441 tcggtcgggc gctgcgggtc gtggggcggg cgtcaggcac cgggcttgcg ggtcatgcac
     1501 caggtgcgcg gtccttcggg cacctcgacg tcggcggtga cggtgaagcc gagccgctcg
     1561 tagaagggga ggttgtgggg cgcggaggtc tccaggaagg cgggcacccc ggcgcgctcg
     1621 gccgcctcca ctccggggag cacgacggcg ctgcccagac ccttgccctg gtggtcgggc
     1681 gagacgccga cggtggccag gaaccacgcg ggctccttgg gccggtgcgg cgccaggagg
     1741 ccttccatct gttgctgcgc ggccagccgg gaaccgctca actcggccat gcgcgggccg
     1801 atctcggcga acaccgcccc cgcttcgacg ctctccggcg tggtccagac cgccaccgcg
     1861 gcgccgtcgt ccgcgaccca caccttgccg atgtcgagcc cgacgcgcgt gaggaagagt
     1921 tcttgcagct cggtgacccg ctcgatgtgg cggtccggat cgacggtgtg gcgcgtggcg
     1981 gggtagtcgg cgaacgcggc ggcgagggtg cgtacggccc tggggacgtc gtcgcgggtg
     2041 gcgaggcgca ccgtgggctt gtactcggtc atggtaagct tcagctgctc gagatctaga
     2101 tggatgcagg tcgaaaggcc cggagatgag gaagaggaga acagcgcggc agacgtgcgc
     2161 ttttgaagcg tgcagaatgc cgggcctccg gaggaccttc gggcgcccgc cccgcccctg
     2221 agcccgcccc tgagcccgcc cccggaccca ccccttccca gcctctgagc ccagaaagcg
     2281 aaggagcaaa gctgctattg gccgctgccc caaaggccta cccgcttcca ttgctcagcg
     2341 gtgctgtcca tctgcacgag actagtgaga cgtgctactt ccatttgtca cgtcctgcac
     2401 gacgcgagct gcggggcggg ggggaacttc ctgactaggg gaggagtaga aggtggcgcg
     2461 aaggggccac caaagaacgg agtcggttgg cgcctaccgg tggatgtgga atgtgtgcga
     2521 ggccagaggc cacttgtgta gcgccaagtg cccagcgggg ctgctaaagc gcatgctcca
     2581 gactgccttg ggaaaagcgc ctcccctacc cggtagaatt tcgaggtcga gatcctaagc
     2641 ttggctggac gtaaactcct cttcagacct aataacttcg tatagcatac attatacgaa
     2701 gttatattaa gggttattga atatgatcgc ttaagataca ttgatgagtt tggacaaacc
     2761 acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta
     2821 tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg
     2881 tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt
     2941 ggtatggctg attatgatca gttatctaga tccggtggat cccgggcccg cggtaccgtc
     3001 gatgaccttt cttttcttct tggggtcgac tgcagaattc gaagcttggg cgcctgcacc
     3061 cttgtacagc tcgtccatgc cgagagtgat cccggcggcg gtcacgaact ccagcaggac
     3121 catgtgatcg cgcttctcgt tggggtcttt gctcagggcg gactgggtgc tcaggtagtg
     3181 gttgtcgggc agcagcacgg ggccgtcgcc gatgggggtg ttctgctggt agtggtcggc
     3241 gagctgcacg ctgccgtcct cgatgttgtg gcggatcttg aagttcacct tgatgccgtt
     3301 cttctgcttg tcggccatga tatagacgtt gtggctgttg tagttgtact ccagcttgtg
     3361 ccccaggatg ttgccgtcct ccttgaagtc gatgcccttc agctcgatgc ggttcaccag
     3421 ggtgtcgccc tcgaacttca cctcggcgcg ggtcttgtag ttgccgtcgt ccttgaagaa
     3481 gatggtgcgc tcctggacgt agccttcggg catggcggac ttgaagaagt cgtgctgctt
     3541 catgtggtcg gggtagcggc tgaagcactg cacgccgtag gtcagggtgg tcacgagggt
     3601 gggccagggc acgggcagct tgccggtggt gcagatgaac ttcagggtca gcttgccgta
     3661 ggtggcatcg ccctcgccct cgccggacac gctgaacttg tggccgttta cgtcgccgtc
     3721 cagctcgacc aggatgggca ccaccccggt gaacagctcc tcgcccttgc tcaccatgct
     3781 agctttatac atcctcaaat cgattttcct caagtacggt gctccgtcca ttgatacctt
     3841 cacgaacgcc gccgcctccg ggccaccgct tgatttttgg caggaaacca tcacgttctt
     3901 ccggtatgat ctcaccggtg gccatcccac aacttgtgcc ttggccggag gtttggctgg
     3961 atctttagga caagcactct tctccttaga tctaggtcca gggttctcct ccacgtctcc
     4021 agcctgcttc agcaggctga agttagtagc tccgcttcct gatctcagaa tcttcacaaa
     4081 gttgggagca tcatctcttg cccctgcggt ggtccgatac acatacagtt tctccacctt
     4141 tggcaggtcg ccatgatttt cctccatttc gttagacccg tcccgctcat taatgacttc
     4201 cacgttcagc ataggcattt tactagccag gacctgacag cccttcagag tcacattgca
     4261 actagacatc cacaggctcc tcattgtctc gtatctggca aagtttccca gcagagcggc
     4321 atccccgaaa gggctgtccc ggatttccag ttttcgcaga ttcttacacc cgttcatcac
     4381 gtgcatcatg cccttatcgg agtctccggc aaatgcaatg ctcagcatct ccagctgctt
     4441 agcatacttg ccgatgtaca tgaacacttt gtctgtcagc aggccggaaa tgctcagcct
     4501 ctggagtccc ttgcactctc tcacgatagc tccaaaccct tcatccagtg gctgggatgt
     4561 gaccacgtcg ggtttgcctg gttccagaat acacagccga aatcgggtga aattagggca
     4621 gttcttggcg acggtcacca gtgcagcatt agtcatctga tgacagaaat acagcaggct
     4681 gttcagcttt gggcatccca gtgagactgc caccaggcct tcctcagtga cagcactata
     4741 tccggccacg taaaaatcag atgggaacac tctcagctcc tggaggtcct tgcaggagct
     4801 ggcgaccacc tggagcccct tatcggaaat acagtccagc acccacagtc gctggagttt
     4861 cacgcagcgt gagatcatct ttgtcagatc actggcgtcc agggttggtg catagctcag
     4921 gttcaggcct gtcagctgtg cacacagagg gtagataaag ctcaggcaca cgggtgaagc
     4981 atcccagaat cctgacagac tcctcagcat tttacacttc tccagggcag aagtcagttt
     5041 aaagtaggat tctgtctgga aatcgtctgt cagattgccg gttcccagat cctccaggtt
     5101 aggggttctc agcagaatct tagccagagt gtccacgctc acgctccggt tcagccgcag
     5161 agacctcagg ttggggcttc ttgagaccag ccgctccagg gacccagcat tcacttcgcc
     5221 tttgatgcag gcaaagttca gggacaccag gctggtacat gagtctggga agcaagacag
     5281 ccatctgggg cctctatcct ccacttcatt ctcctggagg tccagctccc gcagcagctt
     5341 acagtgggag gccacggctg ccagtccatc ggtgctgaag ccttcgcaag aaatcagcac
     5401 cagtgcgcga aacctaggga agctccgagc cagcagctcc agactttcgt cagagaccac
     5461 cattcgcttc atgcgcagtt cctccagtcc atgacagccg cgagcggctg cctcgatcca
     5521 agggccggcg tatcctcccc aatcaggggg caccaggtta aagtcggcgc cgtggggttt
     5581 ccccttgacg gtcagtgcgc gcacatttgg aaacctagcg gcgaccctgc ctgctctcac
     5641 agcatagcag ttgccgacga acactcctct ccggctcagg cgttcgatct cgtaccacac
     5701 tttgcagacc aggctcactg tgtttctatc tctctgtgca ggcaggaagc taaaaatgtg
     5761 ttcgacgacc tcttcaggaa agtatgtcat ggtggcagcg ctatctagat gcattcgcga
     5821 ggtaccgagc tcgaattctt tgccaaaatg atgagacagc acaataacca gcacgttgcc
     5881 caggagctgt aggaaaaaga agaaggcatg aacatggtta gcagaggctc tagagccgcc
     5941 ggtcacacgc cagaagccga accccgccct gccccgtccc ccccgaaggc agccgtcccc
     6001 ccgcggacag ccccgaggct ggagagggag aaggggacgg cggcgcggcg acgcacgaag
     6061 gccctccccg cccatttcct tcctgccggc gccgcaccgc ttcgccccgc gcccgctaga
     6121 gggggtgcgg cggcgcctcc cagatttcgg ctccgcacag atttgggaca aaggaagtcc
     6181 ctgcgccctc tcgcacgatt accataaaag gcaatggctg cggctcgccg cgcctcgaca
     6241 gccgccggcg ctccgggggc cgccgcgccc ctcccccgag cccyccccgg cccgaggcgg
     6301 ccccgccccg cccggcaccc ccacctgccg ccaccccccg cccggcacgg cgagccccgc
     6361 gccacgcccc gtacggagcc ccgcacccga agccgggccg tgctcagcaa ctcggggagg
     6421 ggggtgcagg gggggttgca gcccgaccga cgcgcccaca ccccctgctc acccccccac
     6481 gcacacaccc cgcacgcagc ctttgttccc ctcgcagccc cccccgcacc gcggggcacc
     6541 gcccccggcc gcgctcccct cgcgcacact gcggagcgca caaagccccg cgccgcgccc
     6601 gcagcgctca cagccgccgg gcagcgcgga gccgcacgcg gcgctcccca cgcacacaca
     6661 cacgcacgca ccccccgagc cgctcccccc gcacaaaggg ccctcccgga gcccctcaag
     6721 gctttcacgc agccacagaa aagaaacaag ccgtcattaa accaagcgct aattacagcc
     6781 cggaggagaa gggccgtccc gcccgctcac ctgtgggagt aacgcggtca gtcagagccg
     6841 gggcgggcgg cgcgaggcgg cggcggagcg gggcacgggg cgaaggcagc gcgcagcgac
     6901 tcccgcccgc cgcgcgcttc gctttttata gggccgccgc cgccgccgcc tcgccataaa
     6961 aggaaacttt cggagcgcgc cgctctgatt ggctgccgcc gcacctctcc gcctcgcccc
     7021 sccccscccc ycsccccscc ccsccccscc yggcgcgcgc cccccccccc cccccgcccc
     7081 catcgctgca caaaataatt aaaaaataaa taaatacaaa attgggggtg gggagggggg
     7141 ggagatgggg agagtgaagc agaacgtggg gctcacctcg accatggtaa tagcgatgac
     7201 taatacgtag atgtactgcc aagtaggaaa gtcccataag gtcatgtact gggcataatg
     7261 ccaggcgggc catttaccgt cattgacgtc aatagggggc gtacttggca tatgatacac
     7321 ttgatgtact gccaagtggg cagtttaccg taaatactcc acccattgac gtcaatggaa
     7381 agtccctatt ggcgttacta tgggaacata cgtcattatt gacgtcaatg ggcgggggtc
     7441 gttgggcggt cagccaggcg ggccatttac cgtaagttat gtaacgcgga actccatata
     7501 tgggctatga actaatgacc ccgtaattga ttactattaa taactagtca ataatcaatg
     7561 tcgagcatgc atctagaggg cccaagtgat ctccaaaaaa taagtacttt ttgactgtaa
     7621 ataaaattgt aaggagtaaa aagtactttt ttttctaaaa aaatgtaatt aagtaaaagt
     7681 aaaagtattg atttttaatt gtactcaagt aaagtaaaaa tccccaaaaa taatacttaa
     7741 gtacagtaat caagtaaaat tactcaagta ctttacacct ctggttcttg accccctacc
     7801 ttcagcaagc ccagcagatc cgagctccag cttttgttcc ctttagtgag ggttaattgc
     7861 gcgcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     7921 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     7981 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     8041 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     8101 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     8161 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     8221 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     8281 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     8341 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     8401 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     8461 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     8521 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     8581 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     8641 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     8701 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     8761 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     8821 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     8881 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     8941 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     9001 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     9061 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     9121 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     9181 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     9241 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     9301 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     9361 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     9421 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     9481 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     9541 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     9601 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     9661 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     9721 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     9781 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     9841 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     9901 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     9961 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
    10021 agtg
//
