LOCUS       sequence-484536-       10949 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  OsTIR1(F74G)-P2A-mAID-EGFP-Nluc.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10949)
  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 10949)
  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: pMK411 (OsTIR1(F74G) mAID-EGFP-Nluc)
FEATURES             Location/Qualifiers
     source          1..10949
                     /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             2236..3960
                     /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"
     CDS             3976..4032
                     /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             4039..4242
                     /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(4249..4251,4252..4254,4255..4965)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(4249..4251,4252..4254,4255..4965)
                     /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(4294..4315)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4555..4574)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4902..4923
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             5002..5517
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     polyA_signal    5647..5768
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5684..5703)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5738..5757
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    5799..5832
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        5893..6392
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(5928..5951)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     6325..6344
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             6453..7472
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGYV
                     LRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPET
                     ELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQTV
                     MDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFGD
                     SQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGNF
                     DDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE"
     CDS             6453..7472
                     /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    8035..8068
                     /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(8099..8261,8262..8273)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(8303..8322)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8304..8322)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8332..8349)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8332..8348)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8341..8363)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      8490..8944
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8576..8595)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8786..8807
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        8970..9074
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9075..9143,9144..9935)
                     /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(9293..9312)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10106..10694
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10595..10614
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10848..10865
                     /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 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 gagggctcgg 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 ttcgagctcg gtacctcgcg aatgcatcta
     2221 gatagcgctg ccaccatgac atactttcct gaagaggtcg tcgaacacat ttttagcttc
     2281 ctgcctgcac agagagatag aaacacagtg agcctggtct gcaaagtgtg gtacgagatc
     2341 gaacgcctga gccggagagg agtgttcgtc ggcaactgct atgctgtgag agcaggcagg
     2401 gtcgccgcta ggtttccaaa tgtgcgcgca ctgaccgtca aggggaaacc ccacggcgcc
     2461 gactttaacc tggtgccccc tgattgggga ggatacgccg gcccttggat cgaggcagcc
     2521 gctcgcggct gtcatggact ggaggaactg cgcatgaagc gaatggtggt ctctgacgaa
     2581 agtctggagc tgctggctcg gagcttccct aggtttcgcg cactggtgct gatttcttgc
     2641 gaaggcttca gcaccgatgg actggcagcc gtggcctccc actgtaagct gctgcgggag
     2701 ctggacctcc aggagaatga agtggaggat agaggcccca gatggctgtc ttgcttccca
     2761 gactcatgta ccagcctggt gtccctgaac tttgcctgca tcaaaggcga agtgaatgct
     2821 gggtccctgg agcggctggt ctcaagaagc cccaacctga ggtctctgcg gctgaaccgg
     2881 agcgtgagcg tggacactct ggctaagatt ctgctgagaa cccctaacct ggaggatctg
     2941 ggaaccggca atctgacaga cgatttccag acagaatcct actttaaact gacttctgcc
     3001 ctggagaagt gtaaaatgct gaggagtctg tcaggattct gggatgcttc acccgtgtgc
     3061 ctgagcttta tctaccctct gtgtgcacag ctgacaggcc tgaacctgag ctatgcacca
     3121 accctggacg ccagtgatct gacaaagatg atctcacgct gcgtgaaact ccagcgactg
     3181 tgggtgctgg actgtatttc cgataagggg ctccaggtgg tcgccagctc ctgcaaggac
     3241 ctccaggagc tgagagtgtt cccatctgat ttttacgtgg ccggatatag tgctgtcact
     3301 gaggaaggcc tggtggcagt ctcactggga tgcccaaagc tgaacagcct gctgtatttc
     3361 tgtcatcaga tgactaatgc tgcactggtg accgtcgcca agaactgccc taatttcacc
     3421 cgatttcggc tgtgtattct ggaaccaggc aaacccgacg tggtcacatc ccagccactg
     3481 gatgaagggt ttggagctat cgtgagagag tgcaagggac tccagaggct gagcatttcc
     3541 ggcctgctga cagacaaagt gttcatgtac atcggcaagt atgctaagca gctggagatg
     3601 ctgagcattg catttgccgg agactccgat aagggcatga tgcacgtgat gaacgggtgt
     3661 aagaatctgc gaaaactgga aatccgggac agccctttcg gggatgccgc tctgctggga
     3721 aactttgcca gatacgagac aatgaggagc ctgtggatgt ctagttgcaa tgtgactctg
     3781 aagggctgtc aggtcctggc tagtaaaatg cctatgctga acgtggaagt cattaatgag
     3841 cgggacgggt ctaacgaaat ggaggaaaat catggcgacc tgccaaaggt ggagaaactg
     3901 tatgtgtatc ggaccaccgc aggggcaaga gatgatgctc ccaactttgt gaagattctg
     3961 agatcaggaa gcggagctac taacttcagc ctgctgaagc aggctggaga cgtggaggag
     4021 aaccctggac ctagatctaa ggagaagagt gcttgtccta aagatccagc caaacctccg
     4081 gccaaggcac aagttgtggg atggccaccg gtgagatcat accggaagaa cgtgatggtt
     4141 tcctgccaaa aatcaagcgg tggcccggag gcggcggcgt tcgtgaaggt atcaatggac
     4201 ggagcaccgt acttgaggaa aatcgatttg aggatgtata aagctagcat ggtgagcaag
     4261 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
     4321 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
     4381 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
     4441 ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc
     4501 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
     4561 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     4621 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     4681 aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg
     4741 aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag
     4801 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc
     4861 cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
     4921 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagggtgc aggcgcccaa
     4981 gcttcgaatt ctgcagtcga catggtcttc acactcgaag atttcgttgg ggactggcga
     5041 cagacagccg gctacaacct ggaccaagtc cttgaacagg gaggtgtgtc cagtttgttt
     5101 cagaatctcg gggtgtccgt aactccgatc caaaggattg tcctgagcgg tgaaaatggg
     5161 ctgaagatcg acatccatgt catcatcccg tatgaaggtc tgagcggcga ccaaatgggc
     5221 cagatcgaaa aaatttttaa ggtggtgtac cctgtggatg atcatcactt taaggtgatc
     5281 ctgcactatg gcacactggt aatcgacggg gttacgccga acatgatcga ctatttcgga
     5341 cggccgtatg aaggcatcgc cgtgttcgac ggcaaaaaga tcactgtaac agggaccctg
     5401 tggaacggca acaaaattat cgacgagcgc ctgatcaacc ccgacggctc cctgctgttc
     5461 cgagtaacca tcaacggagt gaccggctgg cggctgtgcg aacgcattct ggcgtaagga
     5521 tccaccggat ctagataact gatcataatc agccatacca catttgtaga ggttttactt
     5581 gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt
     5641 gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     5701 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     5761 gtatcttaac gcgtcgatca tattcaataa cccttaatat aacttcgtat aatgtatgct
     5821 atacgaagtt attaggtctg aagaggagtt tacgtccagc caagcttagg atctcgacct
     5881 cgaaattcta ccgggtaggg gaggcgcttt tcccaaggca gtctggagca tgcgctttag
     5941 cagccccgct gggcacttgg cgctacacaa gtggcctctg gcctcgcaca cattccacat
     6001 ccaccggtag gcgccaaccg actccgttct ttggtggccc cttcgcgcca ccttctactc
     6061 ctcccctagt caggaagttc ccccccgccc cgcagctcgc gtcgtgcagg acgtgacaaa
     6121 tggaagtagc acgtctcact agtctcgtgc agatggacag caccgctgag caatggaagc
     6181 gggtaggcct ttggggcagc ggccaatagc agctttgctc cttcgctttc tgggctcaga
     6241 ggctgggaag gggtgggtcc gggggcgggc tcaggggcgg gctcaggggc ggggcgggcg
     6301 cccgaaggtc ctccggaggc ccggcattct gcacgcttca aaagcgcacg tctgccgcgc
     6361 tgttctcctc ttcctcatct ccgggccttt cgacctgcat ccatctagat ctcgagcagc
     6421 tgaagcttac cgctagcatg gatagatccg gaaagcctga actcaccgcg acgtctgtcg
     6481 agaagtttct gatcgaaaag ttcgacagcg tctccgacct gatgcagctc tcggagggcg
     6541 aagaatctcg tgctttcagc ttcgatgtag gagggcgtgg atatgtcctg cgggtaaata
     6601 gctgcgccga tggtttctac aaagatcgtt atgtttatcg gcactttgca tcggccgcgc
     6661 tcccgattcc ggaagtgctt gacattgggg aattcagcga gagcctgacc tattgcatct
     6721 cccgccgtgc acagggtgtc acgttgcaag acctgcctga aaccgaactg cccgctgttc
     6781 tgcagccggt cgcggaggcc atggatgcga tcgctgcggc cgatcttagc cagacgagcg
     6841 ggttcggccc attcggaccg caaggaatcg gtcaatacac tacatggcgt gatttcatat
     6901 gcgcgattgc tgatccccat gtgtatcact ggcaaactgt gatggacgac accgtcagtg
     6961 cgtccgtcgc gcaggctctc gatgagctga tgctttgggc cgaggactgc cccgaagtcc
     7021 ggcacctcgt gcacgcggat ttcggctcca acaatgtcct gacggacaat ggccgcataa
     7081 cagcggtcat tgactggagc gaggcgatgt tcggggattc ccaatacgag gtcgccaaca
     7141 tcttcttctg gaggccgtgg ttggcttgta tggagcagca gacgcgctac ttcgagcgga
     7201 ggcatccgga gcttgcagga tcgccgcggc tccgggcgta tatgctccgc attggtcttg
     7261 accaactcta tcagagcttg gttgacggca atttcgatga tgcagcttgg gcgcagggtc
     7321 gatgcgacgc aatcgtccga tccggagccg ggactgtcgg gcgtacacaa atcgcccgca
     7381 gaagcgcggc cgtctggacc gatggctgtg tagaagtact cgccgatagt ggaaaccgac
     7441 gccccagcac tcgtccgagg gcaaaggaat aggctagccg cccgccccac gacccgcagc
     7501 gcccgaccga aaggagcgca cgaccccatg catcgatgat atcagatccc cgggatgcag
     7561 aaattgatga tctattaaac aataaagatg tccactaaaa tggaagtttt tcctgtcata
     7621 ctttgttaag aagggtgaga acagagtacc tacattttga atggaaggat tggagctacg
     7681 ggggtggggg tggggtggga ttagataaat gcctgctctt tactgaaggc tctttactat
     7741 tgctttatga taatgtttca tagttggata tcataattta aacaagcaaa accaaattaa
     7801 gggccagctc attcctccca ctcatgatct atagatctat agatctctcg tgggatcatt
     7861 gtttttctct tgattcccac tttgtggttc taagtactgt ggtttccaaa tgtgtcagtt
     7921 tcatagcctg aagaacgaga tcagcagcct ctgttccaca tacacttcat tctcagtatt
     7981 gttttgccaa gttctaattc catcagaagc tggtcgagat ccggaaccct taatataact
     8041 tcgtataatg tatgctatac gaagttatta ggtccctcga agaggttcac taagatcttt
     8101 cttgctttta cttttacttc cttaatactc aagtacaatt ttaatggagt acttttttac
     8161 ttttactcaa gtaagattct agccagatac ttttactttt aattgagtaa aattttccct
     8221 aagtacttgt actttcactt gagtaaaatt tttgagtact ttttacacct ctgtcaagaa
     8281 ccatatgccg gtacccaatt cgccctatag tgagtcgtat tacgcgcgct cactggccgt
     8341 cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc
     8401 acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca
     8461 acagttgcgc agcctgaatg gcgaatggga cgcgccctgt agcggcgcat taagcgcggc
     8521 gggtgtggtg gttacgcgca gcgtgaccac cacacccgcc gcgccctagc gcccgctcct
     8581 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     8641 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     8701 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     8761 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     8821 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     8881 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca
     8941 atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     9001 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     9061 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     9121 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     9181 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     9241 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     9301 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     9361 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     9421 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     9481 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     9541 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     9601 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     9661 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     9721 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     9781 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     9841 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     9901 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     9961 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
    10021 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
    10081 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
    10141 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
    10201 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
    10261 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
    10321 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
    10381 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
    10441 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
    10501 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
    10561 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
    10621 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
    10681 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
    10741 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
    10801 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
    10861 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
    10921 attaatgcag ctggcacgac aggtttccc
//
