LOCUS       sequence-233793-       11572 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  OsTIR1 introduction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11572)
  AUTHORS   Yesbolatova A, Natsume T, Hayashi KI, Kanemaki MT
  TITLE     Generation of conditional auxin-inducible degron (AID) cells and 
            tight control of degron-fused proteins using the degradation 
            inhibitor auxinole.
  JOURNAL   Methods. 2019 Apr 23. pii: S1046-2023(18)30331-1. doi: 
            10.1016/j.ymeth.2019.04.010.
  PUBMED    31026591
REFERENCE   2  (bases 1 to 11572)
  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: "Methods. 
            2019 Apr 23. pii: S1046-2023(18)30331-1. doi: 
            10.1016/j.ymeth.2019.04.010."
COMMENT     Sequence Label: pMK365 (Tet-OsTIR1-loxP-PURO-loxP)
FEATURES             Location/Qualifiers
     source          1..11572
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             152..1879
                     /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"
     polyA_signal    2063..2144
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    2194..2227
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2288..2787
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(2323..2346)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     2720..2739
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             2827..3426
                     /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     complement(2827..2846)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     protein_bind    3983..4016
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    4047..4883
                     /label=HA-R
                     /note="right homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     primer_bind     complement(4917..4936)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4918..4936)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4943..4960)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4943..4959)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4952..4974)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             5097..5399
                     /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     5278..5297
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             5748..6542
                     /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             5748..6542
                     /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(5802..5821)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6412..6431
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             complement(join(6792..7583,7584..7652))
                     /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     7415..7434
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7776..8364
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8265..8284
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    8652..8673
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8688..8693,8694..8711,8712..8718)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8726..8742
                     /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     8731..8753
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8750..8766
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8750..8766
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        8787..8805
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    8840..9643
                     /label=HA-L
                     /note="left homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     polyA_signal    9674..9755
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(9872..10618)
                     /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"
     promoter        complement(10637..11147)
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     complement(10700..10718)
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     polyA_signal    11167..11215
                     /note="synthetic polyadenylation signal"
     misc_feature    11229..11320
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     11269..11288
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        join(11324..11572,1..119)
                     /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    11332..11350
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    11368..11386
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    11404..11422
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    11440..11458
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    11476..11494
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    11512..11530
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    11548..11566
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
ORIGIN
        1 aagctttgct tatgtaaacc agggcgccta taaaagagtg ctgatttttt gagtaaactt
       61 caattccaca acacttttgt cttataccaa ctttccgtac cacttcctac cctcgtaaag
      121 aattcgcggc cgcacgcgaa gcgctgccac catgacatac tttcctgaag aggtcgtcga
      181 acacattttt agcttcctgc ctgcacagag agatagaaac acagtgagcc tggtctgcaa
      241 agtgtggtac gagatcgaac gcctgagccg gagaggagtg ttcgtcggca actgctatgc
      301 tgtgagagca ggcagggtcg ccgctaggtt tccaaatgtg cgcgcactga ccgtcaaggg
      361 gaaaccccac ttcgccgact ttaacctggt gccccctgat tggggaggat acgccggccc
      421 ttggatcgag gcagccgctc gcggctgtca tggactggag gaactgcgca tgaagcgaat
      481 ggtggtctct gacgaaagtc tggagctgct ggctcggagc ttccctaggt ttcgcgcact
      541 ggtgctgatt tcttgcgaag gcttcagcac cgatggactg gcagccgtgg cctcccactg
      601 taagctgctg cgggagctgg acctccagga gaatgaagtg gaggatagag gccccagatg
      661 gctgtcttgc ttcccagact catgtaccag cctggtgtcc ctgaactttg cctgcatcaa
      721 aggcgaagtg aatgctgggt ccctggagcg gctggtctca agaagcccca acctgaggtc
      781 tctgcggctg aaccggagcg tgagcgtgga cactctggct aagattctgc tgagaacccc
      841 taacctggag gatctgggaa ccggcaatct gacagacgat ttccagacag aatcctactt
      901 taaactgact tctgccctgg agaagtgtaa aatgctgagg agtctgtcag gattctggga
      961 tgcttcaccc gtgtgcctga gctttatcta ccctctgtgt gcacagctga caggcctgaa
     1021 cctgagctat gcaccaaccc tggacgccag tgatctgaca aagatgatct cacgctgcgt
     1081 gaaactccag cgactgtggg tgctggactg tatttccgat aaggggctcc aggtggtcgc
     1141 cagctcctgc aaggacctcc aggagctgag agtgttccca tctgattttt acgtggccgg
     1201 atatagtgct gtcactgagg aaggcctggt ggcagtctca ctgggatgcc caaagctgaa
     1261 cagcctgctg tatttctgtc atcagatgac taatgctgca ctggtgaccg tcgccaagaa
     1321 ctgccctaat ttcacccgat ttcggctgtg tattctggaa ccaggcaaac ccgacgtggt
     1381 cacatcccag ccactggatg aagggtttgg agctatcgtg agagagtgca agggactcca
     1441 gaggctgagc atttccggcc tgctgacaga caaagtgttc atgtacatcg gcaagtatgc
     1501 taagcagctg gagatgctga gcattgcatt tgccggagac tccgataagg gcatgatgca
     1561 cgtgatgaac gggtgtaaga atctgcgaaa actggaaatc cgggacagcc ctttcgggga
     1621 tgccgctctg ctgggaaact ttgccagata cgagacaatg aggagcctgt ggatgtctag
     1681 ttgcaatgtg actctgaagg gctgtcaggt cctggctagt aaaatgccta tgctgaacgt
     1741 ggaagtcatt aatgagcggg acgggtctaa cgaaatggag gaaaatcatg gcgacctgcc
     1801 aaaggtggag aaactgtatg tgtatcggac caccgcaggg gcaagagatg atgctcccaa
     1861 ctttgtgaag attctgtgag gatccatcgg atcccgggcc cgtcgacacg cgtatcgata
     1921 ccggtggatc cagatctctg cagcatatgt ctagaggatc ataatcagcc ataccacatt
     1981 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
     2041 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag
     2101 caatagcatc acaaatttca caaataaagc atttttttca ctgccgacgt caggtggcac
     2161 ttttcggggc gatcatattc aataaccctt aatataactt cgtataatgt atgctatacg
     2221 aagttattag gtctgaagag gagtttacgt ccagccaagc ttaggatctc gacctcgaaa
     2281 ttctaccggg taggggaggc gcttttccca aggcagtctg gagcatgcgc tttagcagcc
     2341 ccgctgggca cttggcgcta cacaagtggc ctctggcctc gcacacattc cacatccacc
     2401 ggtaggcgcc aaccgactcc gttctttggt ggccccttcg cgccaccttc tactcctccc
     2461 ctagtcagga agttcccccc cgccccgcag ctcgcgtcgt gcaggacgtg acaaatggaa
     2521 gtagcacgtc tcactagtct cgtgcagatg gacagcaccg ctgagcaatg gaagcgggta
     2581 ggcctttggg gcagcggcca atagcagctt tgctccttcg ctttctgggc tcagaggctg
     2641 ggaaggggtg ggtccggggg cgggctcagg ggcgggctca ggggcggggc gggcgcccga
     2701 aggtcctccg gaggcccggc attctgcacg cttcaaaagc gcacgtctgc cgcgctgttc
     2761 tcctcttcct catctccggg cctttcgacc tgcatccatc tagatctcga gcagctgaag
     2821 cttaccatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtccccagg
     2881 gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat
     2941 ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg
     3001 ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg
     3061 ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg
     3121 agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc
     3181 aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt
     3241 ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc
     3301 ttcctggaga cctccgcgcc ccacaacctc cccttctacg agcggctcgg cttcaccgtc
     3361 accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt
     3421 gcctgacgcc cgccccacga cccgcagcgc ccgaccgaaa ggagcgcacg accccatgca
     3481 tcgatgatat cagatccccg ggatgcagaa attgatgatc tattaaacaa taaagatgtc
     3541 cactaaaatg gaagtttttc ctgtcatact ttgttaagaa gggtgagaac agagtaccta
     3601 cattttgaat ggaaggattg gagctacggg ggtgggggtg gggtgggatt agataaatgc
     3661 ctgctcttta ctgaaggctc tttactattg ctttatgata atgtttcata gttggatatc
     3721 ataatttaaa caagcaaaac caaattaagg gccagctcat tcctcccact catgatctat
     3781 agatctatag atctctcgtg ggatcattgt ttttctcttg attcccactt tgtggttcta
     3841 agtactgtgg tttccaaatg tgtcagtttc atagcctgaa gaacgagatc agcagcctct
     3901 gttccacata cacttcattc tcagtattgt tttgccaagt tctaattcca tcagaagctg
     3961 gtcgagatcc ggaaccctta atataacttc gtataatgta tgctatacga agttattagg
     4021 tccctcgaag aggttcacta agctttacta gggacaggat tggtgacaga aaagccccat
     4081 ccttaggcct cctccttcct agtctcctga tattgggtct aacccccacc tcctgttagg
     4141 cagattcctt atctggtgac acacccccat ttcctggagc catctctctc cttgccagaa
     4201 cctctaaggt ttgcttacga tggagccaga gaggatcctg ggagggagag cttggcaggg
     4261 ggtgggaggg aaggggggga tgcgtgacct gcccggttct cagtggccac cctgcgctac
     4321 cctctcccag aacctgagct gctctgacgc ggctgtctgg tgcgtttcac tgatcctggt
     4381 gctgcagctt ccttacactt cccaagagga gaagcagttt ggaaaaacaa aatcagaata
     4441 agttggtcct gagttctaac tttggctctt cacctttcta gtccccaatt tatattgttc
     4501 ctccgtgcgt cagttttacc tgtgagataa ggccagtagc cagccccgtc ctggcagggc
     4561 tgtggtgagg aggggggtgt ccgtgtggaa aactcccttt gtgagaatgg tgcgtcctag
     4621 gtgttcacca ggtcgtggcc gcctctactc cctttctctt tctccatcct tctttcctta
     4681 aagagtcccc agtgctatct gggacatatt cctccgccca gagcagggtc ccgcttccct
     4741 aaggccctgc tctgggcttc tgggtttgag tccttggcaa gcccaggaga ggcgctcagg
     4801 cttccctgtc ccccttcctc gtccaccatc tcatgcccct ggctctcctg ccccttccct
     4861 acaggggttc ctggctctgc tctaagggcg aattcgcggc cgctaaattc aattcgccct
     4921 atagtgagtc gtattacaat tcactggccg tcgttttaca acgtcgtgac tgggaaaacc
     4981 ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata
     5041 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctatac gtacggcagt
     5101 ttaaggttta cacctataaa agagagagcc gttatcgtct gtttgtggat gtacagagtg
     5161 atattattga cacgccgggg cgacggatgg tgatccccct ggccagtgca cgtctgctgt
     5221 cagataaagt ctcccgtgaa ctttacccgg tggtgcatat cggggatgaa agctggcgca
     5281 tgatgaccac cgatatggcc agtgtgccgg tctccgttat cggggaagaa gtggctgatc
     5341 tcagccaccg cgaaaatgac atcaaaaacg ccattaacct gatgttctgg ggaatataaa
     5401 tgtcaggcat gagattatca aaaaggatct tcacctagat ccttttcacg tagaaagcca
     5461 gtccgcagaa acggtgctga ccccggatga atgtcagcta ctgggctatc tggacaaggg
     5521 aaaacgcaag cgcaaagaga aagcaggtag cttgcagtgg gcttacatgg cgatagctag
     5581 actgggcggt tttatggaca gcaagcgaac cggaattgcc agctggggcg ccctctggta
     5641 aggttgggaa gccctgcaaa gtaaactgga tggctttctt gccgccaagg atctgatggc
     5701 gcaggggatc aagctctgat caagagacag gatgaggatc gtttcgcatg attgaacaag
     5761 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     5821 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     5881 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag
     5941 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     6001 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     6061 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     6121 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     6181 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     6241 tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg
     6301 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     6361 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     6421 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     6481 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     6541 gaattattaa cgcttacaat ttcctgatgc ggtattttct ccttacgcat ctgtgcggta
     6601 tttcacaccg catcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     6661 tttttctaaa tacattcaaa tatgtatccg ctcatgagat tatcaaaaag gatcttcacc
     6721 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     6781 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     6841 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     6901 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     6961 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     7021 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     7081 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     7141 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     7201 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     7261 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     7321 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     7381 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     7441 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     7501 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     7561 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     7621 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     7681 atttatcagg gttattgtct catgaccaaa atcccttaac gtgagttttc gttccactga
     7741 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     7801 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     7861 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     7921 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     7981 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     8041 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     8101 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     8161 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     8221 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     8281 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     8341 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     8401 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     8461 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     8521 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt
     8581 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag
     8641 cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg
     8701 cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc
     8761 tatgaccatg attacgccaa gctcagaatt aaccctcact aaagggacta gtcctgcagg
     8821 tttaaacgaa ttcgcccttt gctttctctg accagcattc tctcccctgg gcctgtgccg
     8881 ctttctgtct gcagcttgtg gcctgggtca cctctacggc tggcccagat ccttccctgc
     8941 cgcctccttc aggttccgtc ttcctccact ccctcttccc cttgctctct gctgtgttgc
     9001 tgcccaagga tgctctttcc ggagcacttc cttctcggcg ctgcaccacg tgatgtcctc
     9061 tgagcggatc ctccccgtgt ctgggtcctc tccgggcatc tctcctccct cacccaaccc
     9121 catgccgtct tcactcgctg ggttcccttt tccttctcct tctggggcct gtgccatctc
     9181 tcgtttctta ggatggcctt ctccgacgga tgtctccctt gcgtcccgcc tccccttctt
     9241 gtaggcctgc atcatcaccg tttttctgga caaccccaaa gtaccccgtc tccctggctt
     9301 tagccacctc tccatcctct tgctttcttt gcctggacac cccgttctcc tgtggattcg
     9361 ggtcacctct cactcctttc atttgggcag ctcccctacc ccccttacct ctctagtctg
     9421 tgctagctct tccagccccc tgtcatggca tcttccaggg gtccgagagc tcagctagtc
     9481 ttcttcctcc aacccgggcc cctatgtcca cttcaggaca gcatgtttgc tgcctccagg
     9541 gatcctgtgt ccccgagctg ggaccacctt atattcccag ggccggttaa tgtggctctg
     9601 gttctgggta cttttatctg tcccctccac cccacagtgg ggcaagctga gtcagtgagc
     9661 gaggaagctc ggggcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt
     9721 tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta
     9781 tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat
     9841 gtggtatggc tgattatgat cctctggaga tttacccggg gagcatgtca aggtcaaaat
     9901 cgtcaagagc gtcagcaggc agcatatcaa ggtcaaagtc gtcaagggca tcggctggga
     9961 gcatgtctaa gtcaaaatcg tcaagggcgt cggtcggccc gccgctttcg cactttagct
    10021 gtttctccag gccacatatg attagttcca ggccgaaaag gaaggcaggt tcggctccct
    10081 gccggtcgaa cagctcaatt gcttgtttca gaagtggggg catagaatcg gtggtaggtg
    10141 tctctctttc ctcttttgct acttgatgct cctgttcctc caatacgcag cccagtgtaa
    10201 agtggcccac ggcggacaga gcgtacagtg cgttctccag ggagaagcct tgctgacaca
    10261 ggaacgcgag ctgattttcc agggtttcgt actgtttctc tgttgggcgg gtgccgagat
    10321 gcactttagc cccgtcgcga tgtgagagga gagcacagcg gtatgacttg gcgttgttcc
    10381 gcagaaagtc ttgccatgac tcgccttcca gggggcagga gtgggtatga tgcctgtcca
    10441 gcatctcgat tggcagggca tcgagcaggg cccgcttgtt cttcacgtgc cagtacaggg
    10501 taggctgctc aactcccagc ttttgagcga gtttccttgt cgtcaggcct tcgataccga
    10561 ctccattgag taattccaga gcagagttta tgactttgct cttgtccagt ctagacatgg
    10621 taattcgatg atccccctgg ggagagaggt cggtgattcg gtcaacgagg gagccgactg
    10681 ccgacgtgcg ctccggaggc ttgcagaatg cggaacaccg cgcgggcagg aacagggccc
    10741 acactaccgc cccacacccc gcctcccgca ccgccccttc ccggccgctg ctctcggcgc
    10801 gccctgctga gcagccgcta ttggccacag cccatcgcgg tcggcgcgct gccattgctc
    10861 cctggcgctg tccgtctgcg agggtactag tgagacgtgc ggcttccgtt tgtcacgtcc
    10921 ggcacgccgc gaaccgcaag gaaccttccc gacttagggg cggagcagga agcgtcgccg
    10981 gggggcccac aagggtagcg gcgaagatcc gggtgacgct gcgaacggac gtgaagaatg
    11041 tgcgagaccc agggtcggcg ccgctgcgtt tcccggaacc acgcccagag cagccgcgtc
    11101 cctgcgcaaa cccagggctg ccttggaaaa ggcgcaaccc caaccccgtg gaattatcac
    11161 ctcgagaata aaatatcttt attttcatta catctgtgtg ttggtttttt gtgtgaatcg
    11221 atagtactaa catacgctct ccatcaaaac aaaacgaaac aaaacaaact agcaaaatag
    11281 gctgtcccca gtgcaagtgc aggtgccaga acatttctct ctcgagttta ctccctatca
    11341 gtgatagaga acgtatgaag agtttactcc ctatcagtga tagagaacgt atgcagactt
    11401 tactccctat cagtgataga gaacgtataa ggagtttact ccctatcagt gatagagaac
    11461 gtatgaccag tttactccct atcagtgata gagaacgtat ctacagttta ctccctatca
    11521 gtgatagaga acgtatatcc agtttactcc ctatcagtga tagagaacgt at
//
