LOCUS       sequence-323565-       11009 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses OsTIR1 under the control of the Tet promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11009)
  AUTHORS   Natsume T, Kiyomitsu T, Saga Y, Kanemaki MT
  TITLE     Rapid Protein Depletion in Human Cells by Auxin-Inducible Degron 
            Tagging with Short Homology Donors.
  JOURNAL   Cell Rep. 2016 Apr 5;15(1):210-8. doi: 10.1016/j.celrep.2016.03.001.
            Epub 2016 Mar 24.
  PUBMED    27052166
REFERENCE   2  (bases 1 to 11009)
  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"; doi: 
            "10.1016/j.celrep.2016.03.001"; journalName: "Cell Rep"; date: 
            "2016-04-5- 5"; volume: "15"; issue: "1"; pages: "210-8"
COMMENT     Sequence Label: pMK243 (Tet-OsTIR1-PURO)
FEATURES             Location/Qualifiers
     source          1..11009
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     221..240
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(262..772)
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     polyA_signal    794..875
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(1007..2734)
                     /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"
     promoter        complement(2761..3128)
                     /label=TRE3GS promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G, modified to eliminate
                     binding sites for endogenous mammalian transcription 
                     factors"
     protein_bind    2886..2904
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2922..2940
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2958..2976
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2994..3012
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3030..3048
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3066..3084
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3102..3120
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_feature    3132..3223
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     complement(3164..3183)
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     polyA_signal    3237..3285
                     /note="synthetic polyadenylation signal"
     promoter        3305..3815
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     CDS             3834..4580
                     /codon_start=1
                     /product="modified rtTA protein that binds tightly to 
                     promoters containing the tet operator in the presence of 
                     doxycycline"
                     /label=Tet-On(R) 3G
                     /translation="MSRLDKSKVINSALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHSCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLKQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     polyA_signal    4697..4778
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    4809..5612
                     /label=HA-L
                     /note="left homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     promoter        complement(5647..5665)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5686..5702)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5686..5702)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5699..5721)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5710..5726
                     /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(5734..5740,5741..5758,5759..5764))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5779..5800
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(6088..6676)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6168..6187)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(6800..6868,6869..7660)
                     /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(7018..7037)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             complement(7910..8704)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(7910..8704)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(8021..8040)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     8631..8650
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     CDS             complement(9053..9355)
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
                     SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     complement(9155..9174)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     primer_bind     9478..9500
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     9492..9509
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     9493..9509
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9516..9535
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        9516..9534
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    9569..10405
                     /label=HA-R
                     /note="right homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     polyA_signal    10419..10643
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     CDS             complement(join(10650..11009,1..240))
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(10733..10753)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
ORIGIN
        1 ctccggcgtg gtccagaccg ccaccgcggc gccgtcgtcc gcgacccaca ccttgccgat
       61 gtcgagcccg acgcgcgtga ggaagagttc ttgcagctcg gtgacccgct cgatgtggcg
      121 gtccggatcg acggtgtggc gcgtggcggg gtagtcggcg aacgcggcgg cgagggtgcg
      181 tacggccctg gggacgtcgt cgcgggtggc gaggcgcacc gtgggcttgt actcggtcat
      241 ggtaagctcc ggtggatccc cctggggaga gaggtcggtg attcggtcaa cgagggagcc
      301 gactgccgac gtgcgctccg gaggcttgca gaatgcggaa caccgcgcgg gcaggaacag
      361 ggcccacact accgccccac accccgcctc ccgcaccgcc ccttcccggc cgctgctctc
      421 ggcgcgccct gctgagcagc cgctattggc cacagcccat cgcggtcggc gcgctgccat
      481 tgctccctgg cgctgtccgt ctgcgagggt actagtgaga cgtgcggctt ccgtttgtca
      541 cgtccggcac gccgcgaacc gcaaggaacc ttcccgactt aggggcggag caggaagcgt
      601 cgccgggggg cccacaaggg tagcggcgaa gatccgggtg acgctgcgaa cggacgtgaa
      661 gaatgtgcga gacccagggt cggcgccgct gcgtttcccg gaaccacgcc cagagcagcc
      721 gcgtccctgc gcaaacccag ggctgccttg gaaaaggcgc aaccccaacc ccgaaaagtg
      781 ccacctgacg tcggcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt
      841 tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta
      901 tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat
      961 gtggtatggc tgattatgat cctctagaca tatgctgcag agatcttcac agaatcttca
     1021 caaagttggg agcatcatct cttgcccctg cggtggtccg atacacatac agtttctcca
     1081 cctttggcag gtcgccatga ttttcctcca tttcgttaga cccgtcccgc tcattaatga
     1141 cttccacgtt cagcataggc attttactag ccaggacctg acagcccttc agagtcacat
     1201 tgcaactaga catccacagg ctcctcattg tctcgtatct ggcaaagttt cccagcagag
     1261 cggcatcccc gaaagggctg tcccggattt ccagttttcg cagattctta cacccgttca
     1321 tcacgtgcat catgccctta tcggagtctc cggcaaatgc aatgctcagc atctccagct
     1381 gcttagcata cttgccgatg tacatgaaca ctttgtctgt cagcaggccg gaaatgctca
     1441 gcctctggag tcccttgcac tctctcacga tagctccaaa cccttcatcc agtggctggg
     1501 atgtgaccac gtcgggtttg cctggttcca gaatacacag ccgaaatcgg gtgaaattag
     1561 ggcagttctt ggcgacggtc accagtgcag cattagtcat ctgatgacag aaatacagca
     1621 ggctgttcag ctttgggcat cccagtgaga ctgccaccag gccttcctca gtgacagcac
     1681 tatatccggc cacgtaaaaa tcagatggga acactctcag ctcctggagg tccttgcagg
     1741 agctggcgac cacctggagc cccttatcgg aaatacagtc cagcacccac agtcgctgga
     1801 gtttcacgca gcgtgagatc atctttgtca gatcactggc gtccagggtt ggtgcatagc
     1861 tcaggttcag gcctgtcagc tgtgcacaca gagggtagat aaagctcagg cacacgggtg
     1921 aagcatccca gaatcctgac agactcctca gcattttaca cttctccagg gcagaagtca
     1981 gtttaaagta ggattctgtc tggaaatcgt ctgtcagatt gccggttccc agatcctcca
     2041 ggttaggggt tctcagcaga atcttagcca gagtgtccac gctcacgctc cggttcagcc
     2101 gcagagacct caggttgggg cttcttgaga ccagccgctc cagggaccca gcattcactt
     2161 cgcctttgat gcaggcaaag ttcagggaca ccaggctggt acatgagtct gggaagcaag
     2221 acagccatct ggggcctcta tcctccactt cattctcctg gaggtccagc tcccgcagca
     2281 gcttacagtg ggaggccacg gctgccagtc catcggtgct gaagccttcg caagaaatca
     2341 gcaccagtgc gcgaaaccta gggaagctcc gagccagcag ctccagactt tcgtcagaga
     2401 ccaccattcg cttcatgcgc agttcctcca gtccatgaca gccgcgagcg gctgcctcga
     2461 tccaagggcc ggcgtatcct ccccaatcag ggggcaccag gttaaagtcg gcgaagtggg
     2521 gtttcccctt gacggtcagt gcgcgcacat ttggaaacct agcggcgacc ctgcctgctc
     2581 tcacagcata gcagttgccg acgaacactc ctctccggct caggcgttcg atctcgtacc
     2641 acactttgca gaccaggctc actgtgtttc tatctctctg tgcaggcagg aagctaaaaa
     2701 tgtgttcgac gacctcttca ggaaagtatg tcatggtggc acgcgtgcgg ccgcgaattc
     2761 tttacgaggg taggaagtgg tacggaaagt tggtataaga caaaagtgtt gtggaattga
     2821 agtttactca aaaaatcagc actcttttat aggcgccctg gtttacataa gcaaagctta
     2881 tacgttctct atcactgata gggagtaaac tggatatacg ttctctatca ctgataggga
     2941 gtaaactgta gatacgttct ctatcactga tagggagtaa actggtcata cgttctctat
     3001 cactgatagg gagtaaactc cttatacgtt ctctatcact gatagggagt aaagtctgca
     3061 tacgttctct atcactgata gggagtaaac tcttcatacg ttctctatca ctgataggga
     3121 gtaaactcga gagagaaatg ttctggcacc tgcacttgca ctggggacag cctattttgc
     3181 tagtttgttt tgtttcgttt tgttttgatg gagagcgtat gttagtacta tcgattcaca
     3241 caaaaaacca acacacagat gtaatgaaaa taaagatatt ttattctcga ggtgataatt
     3301 ccacggggtt ggggttgcgc cttttccaag gcagccctgg gtttgcgcag ggacgcggct
     3361 gctctgggcg tggttccggg aaacgcagcg gcgccgaccc tgggtctcgc acattcttca
     3421 cgtccgttcg cagcgtcacc cggatcttcg ccgctaccct tgtgggcccc ccggcgacgc
     3481 ttcctgctcc gcccctaagt cgggaaggtt ccttgcggtt cgcggcgtgc cggacgtgac
     3541 aaacggaagc cgcacgtctc actagtaccc tcgcagacgg acagcgccag ggagcaatgg
     3601 cagcgcgccg accgcgatgg gctgtggcca atagcggctg ctcagcaggg cgcgccgaga
     3661 gcagcggccg ggaaggggcg gtgcgggagg cggggtgtgg ggcggtagtg tgggccctgt
     3721 tcctgcccgc gcggtgttcc gcattctgca agcctccgga gcgcacgtcg gcagtcggct
     3781 ccctcgttga ccgaatcacc gacctctctc cccaggggga tcatcgaatt accatgtcta
     3841 gactggacaa gagcaaagtc ataaactctg ctctggaatt actcaatgga gtcggtatcg
     3901 aaggcctgac gacaaggaaa ctcgctcaaa agctgggagt tgagcagcct accctgtact
     3961 ggcacgtgaa gaacaagcgg gccctgctcg atgccctgcc aatcgagatg ctggacaggc
     4021 atcataccca ctcctgcccc ctggaaggcg agtcatggca agactttctg cggaacaacg
     4081 ccaagtcata ccgctgtgct ctcctctcac atcgcgacgg ggctaaagtg catctcggca
     4141 cccgcccaac agagaaacag tacgaaaccc tggaaaatca gctcgcgttc ctgtgtcagc
     4201 aaggcttctc cctggagaac gcactgtacg ctctgtccgc cgtgggccac tttacactgg
     4261 gctgcgtatt ggaggaacag gagcatcaag tagcaaaaga ggaaagagag acacctacca
     4321 ccgattctat gcccccactt ctgaaacaag caattgagct gttcgaccgg cagggagccg
     4381 aacctgcctt ccttttcggc ctggaactaa tcatatgtgg cctggagaaa cagctaaagt
     4441 gcgaaagcgg cgggccgacc gacgcccttg acgattttga cttagacatg ctcccagccg
     4501 atgcccttga cgactttgac cttgatatgc tgcctgctga cgctcttgac gattttgacc
     4561 ttgacatgct ccccgggtaa atctccagag gatcataatc agccatacca catttgtaga
     4621 ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa
     4681 tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     4741 catcacaaat ttcacaaata aagcattttt ttcactgccc cgagcttcct cgctcactga
     4801 ctcagcttgc cccactgtgg ggtggagggg acagataaaa gtacccagaa ccagagccac
     4861 attaaccggc cctgggaata taaggtggtc ccagctcggg gacacaggat ccctggaggc
     4921 agcaaacatg ctgtcctgaa gtggacatag gggcccgggt tggaggaaga agactagctg
     4981 agctctcgga cccctggaag atgccatgac agggggctgg aagagctagc acagactaga
     5041 gaggtaaggg gggtagggga gctgcccaaa tgaaaggagt gagaggtgac ccgaatccac
     5101 aggagaacgg ggtgtccagg caaagaaagc aagaggatgg agaggtggct aaagccaggg
     5161 agacggggta ctttggggtt gtccagaaaa acggtgatga tgcaggccta caagaagggg
     5221 aggcgggacg caagggagac atccgtcgga gaaggccatc ctaagaaacg agagatggca
     5281 caggccccag aaggagaagg aaaagggaac ccagcgagtg aagacggcat ggggttgggt
     5341 gagggaggag agatgcccgg agaggaccca gacacgggga ggatccgctc agaggacatc
     5401 acgtggtgca gcgccgagaa ggaagtgctc cggaaagagc atccttgggc agcaacacag
     5461 cagagagcaa ggggaagagg gagtggagga agacggaacc tgaaggaggc ggcagggaag
     5521 gatctgggcc agccgtagag gtgacccagg ccacaagctg cagacagaaa gcggcacagg
     5581 cccaggggag agaatgctgg tcagagaaag caaagggcga attcgtttaa acctgcagga
     5641 ctagtccctt tagtgagggt taattctgag cttggcgtaa tcatggtcat agctgtttcc
     5701 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     5761 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     5821 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     5881 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     5941 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     6001 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     6061 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     6121 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     6181 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     6241 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     6301 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     6361 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     6421 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     6481 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     6541 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     6601 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     6661 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     6721 cgaaaactca cgttaaggga ttttggtcat gagacaataa ccctgataaa tgcttcaata
     6781 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     6841 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     6901 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     6961 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     7021 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     7081 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     7141 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     7201 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     7261 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     7321 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     7381 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     7441 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     7501 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     7561 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     7621 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     7681 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     7741 cctttttgat aatctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     7801 aggggttccg cgcacatttc cccgaaaagt gccacctgat gcggtgtgaa ataccgcaca
     7861 gatgcgtaag gagaaaatac cgcatcagga aattgtaagc gttaataatt cagaagaact
     7921 cgtcaagaag gcgatagaag gcgatgcgct gcgaatcggg agcggcgata ccgtaaagca
     7981 cgaggaagcg gtcagcccat tcgccgccaa gctcttcagc aatatcacgg gtagccaacg
     8041 ctatgtcctg atagcggtcc gccacaccca gccggccaca gtcgatgaat ccagaaaagc
     8101 ggccattttc caccatgata ttcggcaagc aggcatcgcc atgggtcacg acgagatcct
     8161 cgccgtcggg catgctcgcc ttgagcctgg cgaacagttc ggctggcgcg agcccctgat
     8221 gctcttcgtc cagatcatcc tgatcgacaa gaccggcttc catccgagta cgtgctcgct
     8281 cgatgcgatg tttcgcttgg tggtcgaatg ggcaggtagc cggatcaagc gtatgcagcc
     8341 gccgcattgc atcagccatg atggatactt tctcggcagg agcaaggtga gatgacagga
     8401 gatcctgccc cggcacttcg cccaatagca gccagtccct tcccgcttca gtgacaacgt
     8461 cgagcacagc tgcgcaagga acgcccgtcg tggccagcca cgatagccgc gctgcctcgt
     8521 cttgcagttc attcagggca ccggacaggt cggtcttgac aaaaagaacc gggcgcccct
     8581 gcgctgacag ccggaacacg gcggcatcag agcagccgat tgtctgttgt gcccagtcat
     8641 agccgaatag cctctccacc caagcggccg gagaacctgc gtgcaatcca tcttgttcaa
     8701 tcatgcgaaa cgatcctcat cctgtctctt gatcagagct tgatcccctg cgccatcaga
     8761 tccttggcgg caagaaagcc atccagttta ctttgcaggg cttcccaacc ttaccagagg
     8821 gcgccccagc tggcaattcc ggttcgcttg ctgtccataa aaccgcccag tctagctatc
     8881 gccatgtaag cccactgcaa gctacctgct ttctctttgc gcttgcgttt tcccttgtcc
     8941 agatagccca gtagctgaca ttcatccggg gtcagcaccg tttctgcgga ctggctttct
     9001 acgtgaaaag gatctaggtg aagatccttt ttgataatct catgcctgac atttatattc
     9061 cccagaacat caggttaatg gcgtttttga tgtcattttc gcggtggctg agatcagcca
     9121 cttcttcccc gataacggag accggcacac tggccatatc ggtggtcatc atgcgccagc
     9181 tttcatcccc gatatgcacc accgggtaaa gttcacggga gactttatct gacagcagac
     9241 gtgcactggc cagggggatc accatccgtc gccccggcgt gtcaataata tcactctgta
     9301 catccacaaa cagacgataa cggctctctc ttttataggt gtaaacctta aactgccgta
     9361 cgtataggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
     9421 agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc
     9481 agtcacgacg ttgtaaaacg acggccagtg aattgtaata cgactcacta tagggcgaat
     9541 tgaatttagc ggccgcgaat tcgcccttag agcagagcca ggaacccctg tagggaaggg
     9601 gcaggagagc caggggcatg agatggtgga cgaggaaggg ggacagggaa gcctgagcgc
     9661 ctctcctggg cttgccaagg actcaaaccc agaagcccag agcagggcct tagggaagcg
     9721 ggaccctgct ctgggcggag gaatatgtcc cagatagcac tggggactct ttaaggaaag
     9781 aaggatggag aaagagaaag ggagtagagg cggccacgac ctggtgaaca cctaggacgc
     9841 accattctca caaagggagt tttccacacg gacacccccc tcctcaccac agccctgcca
     9901 ggacggggct ggctactggc cttatctcac aggtaaaact gacgcacgga ggaacaatat
     9961 aaattgggga ctagaaaggt gaagagccaa agttagaact caggaccaac ttattctgat
    10021 tttgtttttc caaactgctt ctcctcttgg gaagtgtaag gaagctgcag caccaggatc
    10081 agtgaaacgc accagacagc cgcgtcagag cagctcaggt tctgggagag ggtagcgcag
    10141 ggtggccact gagaaccggg caggtcacgc atccccccct tccctcccac cccctgccaa
    10201 gctctccctc ccaggatcct ctctggctcc atcgtaagca aaccttagag gttctggcaa
    10261 ggagagagat ggctccagga aatgggggtg tgtcaccaga taaggaatct gcctaacagg
    10321 aggtgggggt tagacccaat atcaggagac taggaaggag gaggcctaag gatggggctt
    10381 ttctgtcacc aatcctgtcc ctagtaaagc ttggtacccc atagagccca ccgcatcccc
    10441 agcatgcctg ctattgtctt cccaatcctc ccccttgctg tcctgcccca ccccaccccc
    10501 cagaatagaa tgacacctac tcagacaatg cgatgcaatt tcctcatttt attaggaaag
    10561 gacagtggga gtggcacctt ccagggtcaa ggaaggcacg ggggaggggc aaacaacaga
    10621 tggctggcaa ctagaaggca cagggtacct caggcaccgg gcttgcgggt catgcaccag
    10681 gtgcgcggtc cttcgggcac ctcgacgtcg gcggtgacgg tgaagccgag ccgctcgtag
    10741 aaggggaggt tgcggggcgc ggaggtctcc aggaaggcgg gcaccccggc gcgctcggcc
    10801 gcctccactc cggggagcac gacggcgctg cccagaccct tgccctggtg gtcgggcgag
    10861 acgccgacgg tggccaggaa ccacgcgggc tccttgggcc ggtgcggcgc caggaggcct
    10921 tccatctgtt gctgcgcggc cagccgggaa ccgctcaact cggccatgcg cgggccgatc
    10981 tcggcgaaca ccgcccccgc ttcgacgct
//
