LOCUS       sequence-272708-       10027 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 10027)
  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 10027)
  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..10027
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(34..462)
                     /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     589..611
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     603..620
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     604..620
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     630..649
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        630..648
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    join(679..690,691..853)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     protein_bind    complement(884..917)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(1474..2073)
                     /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     2054..2073
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(2113..2612)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(2161..2180)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     2554..2577
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     protein_bind    complement(2673..2706)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    2733..2854
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2744..2763)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2798..2817
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(3005..3025)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(join(3062..3772,3773..3775,3776..3778))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(3062..3772,3773..3775,3776..3778))
                     /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(3104..3125)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3453..3472
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3712..3733
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(3785..3988)
                     /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(3995..4051)
                     /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(4067..5791)
                     /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(5861..5880)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     intron          5888..6905
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     complement(5942..5964)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     promoter        complement(6907..7184)
                     /label=chicken beta-actin promoter
     enhancer        7186..7565
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     misc_feature    join(7587..7774,7775..7786)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(7843..7863)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(7843..7861)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7882..7898)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7882..7898)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7895..7917)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7906..7922
                     /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(7930..7936,7937..7954,7955..7960))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7975..7996
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8113..8130)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8284..8872)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8364..8383)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9043..9834,9835..9903))
                     /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     9666..9685
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9904..10008)
                     /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 tggcaagtgt agcggtcacg ctgcgcgtaa
      421 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg ccattcaggc
      481 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga
      541 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac
      601 gttgtaaaac gacggccagt gagcgcgcgt aatacgactc actatagggc gaattgggta
      661 ccggcatatg gttcttgaca gaggtgtaaa aagtactcaa aaattttact caagtgaaag
      721 tacaagtact tagggaaaat tttactcaat taaaagtaaa agtatctggc tagaatctta
      781 cttgagtaaa agtaaaaaag tactccatta aaattgtact tgagtattaa ggaagtaaaa
      841 gtaaaagcaa gaaagatctt agtgaacctc ttcgagggac ctaataactt cgtatagcat
      901 acattatacg aagttatatt aagggttccg gatctcgacc agcttctgat ggaattagaa
      961 cttggcaaaa caatactgag aatgaagtgt atgtggaaca gaggctgctg atctcgttct
     1021 tcaggctatg aaactgacac atttggaaac cacagtactt agaaccacaa agtgggaatc
     1081 aagagaaaaa caatgatccc acgagagatc tatagatcta tagatcatga gtgggaggaa
     1141 tgagctggcc cttaatttgg ttttgcttgt ttaaattatg atatccaact atgaaacatt
     1201 atcataaagc aatagtaaag agccttcagt aaagagcagg catttatcta atcccacccc
     1261 acccccaccc ccgtagctcc aatccttcca ttcaaaatgt aggtactctg ttctcaccct
     1321 tcttaacaaa gtatgacagg aaaaacttcc attttagtgg acatctttat tgtttaatag
     1381 atcatcaatt tctgcatccc ggggatctga tatcatcgat gcatggggtc gtgcgctcct
     1441 ttcggtcggg cgctgcgggt cgtggggcgg gcgtcaggca ccgggcttgc gggtcatgca
     1501 ccaggtgcgc ggtccttcgg gcacctcgac gtcggcggtg acggtgaagc cgagccgctc
     1561 gtagaagggg aggttgtggg gcgcggaggt ctccaggaag gcgggcaccc cggcgcgctc
     1621 ggccgcctcc actccgggga gcacgacggc gctgcccaga cccttgccct ggtggtcggg
     1681 cgagacgccg acggtggcca ggaaccacgc gggctccttg ggccggtgcg gcgccaggag
     1741 gccttccatc tgttgctgcg cggccagccg ggaaccgctc aactcggcca tgcgcgggcc
     1801 gatctcggcg aacaccgccc ccgcttcgac gctctccggc gtggtccaga ccgccaccgc
     1861 ggcgccgtcg tccgcgaccc acaccttgcc gatgtcgagc ccgacgcgcg tgaggaagag
     1921 ttcttgcagc tcggtgaccc gctcgatgtg gcggtccgga tcgacggtgt ggcgcgtggc
     1981 ggggtagtcg gcgaacgcgg cggcgagggt gcgtacggcc ctggggacgt cgtcgcgggt
     2041 ggcgaggcgc accgtgggct tgtactcggt catggtaagc ttcagctgct cgagatctag
     2101 atggatgcag gtcgaaaggc ccggagatga ggaagaggag aacagcgcgg cagacgtgcg
     2161 cttttgaagc gtgcagaatg ccgggcctcc ggaggacctt cgggcgcccg ccccgcccct
     2221 gagcccgccc ctgagcccgc ccccggaccc accccttccc agcctctgag cccagaaagc
     2281 gaaggagcaa agctgctatt ggccgctgcc ccaaaggcct acccgcttcc attgctcagc
     2341 ggtgctgtcc atctgcacga gactagtgag acgtgctact tccatttgtc acgtcctgca
     2401 cgacgcgagc tgcggggcgg gggggaactt cctgactagg ggaggagtag aaggtggcgc
     2461 gaaggggcca ccaaagaacg gagtcggttg gcgcctaccg gtggatgtgg aatgtgtgcg
     2521 aggccagagg ccacttgtgt agcgccaagt gcccagcggg gctgctaaag cgcatgctcc
     2581 agactgcctt gggaaaagcg cctcccctac ccggtagaat ttcgaggtcg agatcctaag
     2641 cttggctgga cgtaaactcc tcttcagacc taataacttc gtatagcata cattatacga
     2701 agttatatta agggttattg aatatgatcg cttaagatac attgatgagt ttggacaaac
     2761 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     2821 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     2881 gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg
     2941 tggtatggct gattatgatc agttatctag atccggtgga tcccgggccc gcggtaccgt
     3001 cgatgacctt tcttttcttc ttggggtcga ctgcagaatt cgaagcttgg gcgcctgcac
     3061 ccttgtacag ctcgtccatg ccgagagtga tcccggcggc ggtcacgaac tccagcagga
     3121 ccatgtgatc gcgcttctcg ttggggtctt tgctcagggc ggactgggtg ctcaggtagt
     3181 ggttgtcggg cagcagcacg gggccgtcgc cgatgggggt gttctgctgg tagtggtcgg
     3241 cgagctgcac gctgccgtcc tcgatgttgt ggcggatctt gaagttcacc ttgatgccgt
     3301 tcttctgctt gtcggccatg atatagacgt tgtggctgtt gtagttgtac tccagcttgt
     3361 gccccaggat gttgccgtcc tccttgaagt cgatgccctt cagctcgatg cggttcacca
     3421 gggtgtcgcc ctcgaacttc acctcggcgc gggtcttgta gttgccgtcg tccttgaaga
     3481 agatggtgcg ctcctggacg tagccttcgg gcatggcgga cttgaagaag tcgtgctgct
     3541 tcatgtggtc ggggtagcgg ctgaagcact gcacgccgta ggtcagggtg gtcacgaggg
     3601 tgggccaggg cacgggcagc ttgccggtgg tgcagatgaa cttcagggtc agcttgccgt
     3661 aggtggcatc gccctcgccc tcgccggaca cgctgaactt gtggccgttt acgtcgccgt
     3721 ccagctcgac caggatgggc accaccccgg tgaacagctc ctcgcccttg ctcaccatgc
     3781 tagctttata catcctcaaa tcgattttcc tcaagtacgg tgctccgtcc attgatacct
     3841 tcacgaacgc cgccgcctcc gggccaccgc ttgatttttg gcaggaaacc atcacgttct
     3901 tccggtatga tctcaccggt ggccatccca caacttgtgc cttggccgga ggtttggctg
     3961 gatctttagg acaagcactc ttctccttag atctaggtcc agggttctcc tccacgtctc
     4021 cagcctgctt cagcaggctg aagttagtag ctccgcttcc tgatctcaga atcttcacaa
     4081 agttgggagc atcatctctt gcccctgcgg tggtccgata cacatacagt ttctccacct
     4141 ttggcaggtc gccatgattt tcctccattt cgttagaccc gtcccgctca ttaatgactt
     4201 ccacgttcag cataggcatt ttactagcca ggacctgaca gcccttcaga gtcacattgc
     4261 aactagacat ccacaggctc ctcattgtct cgtatctggc aaagtttccc agcagagcgg
     4321 catccccgaa agggctgtcc cggatttcca gttttcgcag attcttacac ccgttcatca
     4381 cgtgcatcat gcccttatcg gagtctccgg caaatgcaat gctcagcatc tccagctgct
     4441 tagcatactt gccgatgtac atgaacactt tgtctgtcag caggccggaa atgctcagcc
     4501 tctggagtcc cttgcactct ctcacgatag ctccaaaccc ttcatccagt ggctgggatg
     4561 tgaccacgtc gggtttgcct ggttccagaa tacacagccg aaatcgggtg aaattagggc
     4621 agttcttggc gacggtcacc agtgcagcat tagtcatctg atgacagaaa tacagcaggc
     4681 tgttcagctt tgggcatccc agtgagactg ccaccaggcc ttcctcagtg acagcactat
     4741 atccggccac gtaaaaatca gatgggaaca ctctcagctc ctggaggtcc ttgcaggagc
     4801 tggcgaccac ctggagcccc ttatcggaaa tacagtccag cacccacagt cgctggagtt
     4861 tcacgcagcg tgagatcatc tttgtcagat cactggcgtc cagggttggt gcatagctca
     4921 ggttcaggcc tgtcagctgt gcacacagag ggtagataaa gctcaggcac acgggtgaag
     4981 catcccagaa tcctgacaga ctcctcagca ttttacactt ctccagggca gaagtcagtt
     5041 taaagtagga ttctgtctgg aaatcgtctg tcagattgcc ggttcccaga tcctccaggt
     5101 taggggttct cagcagaatc ttagccagag tgtccacgct cacgctccgg ttcagccgca
     5161 gagacctcag gttggggctt cttgagacca gccgctccag ggacccagca ttcacttcgc
     5221 ctttgatgca ggcaaagttc agggacacca ggctggtaca tgagtctggg aagcaagaca
     5281 gccatctggg gcctctatcc tccacttcat tctcctggag gtccagctcc cgcagcagct
     5341 tacagtggga ggccacggct gccagtccat cggtgctgaa gccttcgcaa gaaatcagca
     5401 ccagtgcgcg aaacctaggg aagctccgag ccagcagctc cagactttcg tcagagacca
     5461 ccattcgctt catgcgcagt tcctccagtc catgacagcc gcgagcggct gcctcgatcc
     5521 aagggccggc gtatcctccc caatcagggg gcaccaggtt aaagtcggcg ccgtggggtt
     5581 tccccttgac ggtcagtgcg cgcacatttg gaaacctagc ggcgaccctg cctgctctca
     5641 cagcatagca gttgccgacg aacactcctc tccggctcag gcgttcgatc tcgtaccaca
     5701 ctttgcagac caggctcact gtgtttctat ctctctgtgc aggcaggaag ctaaaaatgt
     5761 gttcgacgac ctcttcagga aagtatgtca tggtggcagc gctatctaga tgcattcgcg
     5821 aggtaccgag ctcgaattct ttgccaaaat gatgagacag cacaataacc agcacgttgc
     5881 ccaggagctg taggaaaaag aagaaggcat gaacatggtt agcagaggct ctagagccgc
     5941 cggtcacacg ccagaagccg aaccccgccc tgccccgtcc cccccgaagg cagccgtccc
     6001 cccgcggaca gccccgaggc tggagaggga gaaggggacg gcggcgcggc gacgcacgaa
     6061 ggccctcccc gcccatttcc ttcctgccgg cgccgcaccg cttcgccccg cgcccgctag
     6121 agggggtgcg gcggcgcctc ccagatttcg gctccgcaca gatttgggac aaaggaagtc
     6181 cctgcgccct ctcgcacgat taccataaaa ggcaatggct gcggctcgcc gcgcctcgac
     6241 agccgccggc gctccggggg ccgccgcgcc cctcccccga gccctccccg gcccgaggcg
     6301 gccccgcccc gcccggcacc cccacctgcc gccacccccc gcccggcacg gcgagccccg
     6361 cgccacgccc cgtacggagc cccgcacccg aagccgggcc gtgctcagca actcggggag
     6421 gggggtgcag ggggggttgc agcccgaccg acgcgcccac accccctgct caccccccca
     6481 cgcacacacc ccgcacgcag cctttgttcc cctcgcagcc ccccccgcac cgcggggcac
     6541 cgcccccggc cgcgctcccc tcgcgcacac tgcggagcgc acaaagcccc gcgccgcgcc
     6601 cgcagcgctc acagccgccg ggcagcgcgg agccgcacgc ggcgctcccc acgcacacac
     6661 acacgcacgc accccccgag ccgctccccc cgcacaaagg gccctcccgg agcccctcaa
     6721 ggctttcacg cagccacaga aaagaaacaa gccgtcatta aaccaagcgc taattacagc
     6781 ccggaggaga agggccgtcc cgcccgctca cctgtgggag taacgcggtc agtcagagcc
     6841 ggggcgggcg gcgcgaggcg gcggcggagc ggggcacggg gcgaaggcag cgtcgcagcg
     6901 actccccgcc cgccgcgcgc ttcgcttttt atagggccgc cgccgccgcc gcctcgccat
     6961 aaaaggaaac tttcggagcg cgccgctctg attggctgcc gccgcacctc tccgcctcgc
     7021 cccgccccgc ccctcgcccc gccccgcccc gcctggcgcg cgcccccccc ccccccccgc
     7081 ccccatcgct gcacaaaata attaaaaaat aaataaatac aaaattgggg gtggggaggg
     7141 gggggagatg gggagagtga agcagaacgt ggggctcacc tcgaccatgg taatagcgat
     7201 gactaatacg tagatgtact gccaagtagg aaagtcccat aaggtcatgt actgggcata
     7261 atgccaggcg ggccatttac cgtcattgac gtcaataggg ggcgtacttg gcatatgata
     7321 cacttgatgt actgccaagt gggcagttta ccgtaaatac tccacccatt gacgtcaatg
     7381 gaaagtccct attggcgtta ctatgggaac atacgtcatt attgacgtca atgggcgggg
     7441 gtcgttgggc ggtcagccag gcgggccatt taccgtaagt tatgtaacgc ggaactccat
     7501 atatgggcta tgaactaatg accccgtaat tgattactat taataactag tcaataatca
     7561 atgtcgagca tgcatctaga gggcccaagt gatctccaaa aaataagtac tttttgactg
     7621 taaataaaat tgtaaggagt aaaaagtact tttttttcta aaaaaatgta attaagtaaa
     7681 agtaaaagta ttgattttta attgtactca agtaaagtaa aaatccccaa aaataatact
     7741 taagtacagt aatcaagtaa aattactcaa gtactttaca cctctggttc ttgaccccct
     7801 accttcagca agcccagcag atccgagctc cagcttttgt tccctttagt gagggttaat
     7861 tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     7921 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     7981 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     8041 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     8101 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     8161 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     8221 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     8281 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     8341 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     8401 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     8461 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     8521 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     8581 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     8641 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     8701 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     8761 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     8821 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     8881 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     8941 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     9001 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     9061 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     9121 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     9181 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     9241 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     9301 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     9361 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     9421 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     9481 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     9541 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     9601 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     9661 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     9721 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     9781 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     9841 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     9901 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     9961 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
    10021 aaaagtg
//
