LOCUS       sequence-426332-       11572 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tet-OsTIR1(F74G) (AAVS1 insertion donor).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11572)
  AUTHORS   Hatoyama Y, Islam M, Bond AG, Hayashi KI, Ciulli A, Kanemaki MT
  TITLE     Combination of AID2 and BromoTag expands the utility of degron-based
            protein knockdowns.
  JOURNAL   EMBO Rep. 2024 Sep;25(9):4062-4077. doi: 10.1038/s44319-024-00224-4.
            Epub 2024 Aug 23.
  PUBMED    39179892
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"; doi: 
            "10.1038/s44319-024-00224-4"; journalName: "EMBO Rep"; date: 
            "2024-09"; volume: "25"; issue: "9"; pages: "4062-4077"
COMMENT     Sequence Label: pMK382
FEATURES             Location/Qualifiers
     source          1..11572
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        242..260
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    295..1098
                     /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    1129..1210
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(1327..2073)
                     /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(2092..2602)
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     complement(2155..2173)
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     polyA_signal    2622..2670
                     /note="synthetic polyadenylation signal"
     misc_feature    2684..2775
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     2724..2743
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        2779..3146
                     /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    2787..2805
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2823..2841
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2859..2877
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2895..2913
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2931..2949
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2967..2985
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3003..3021
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             3179..4906
                     /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"
     polyA_signal    5090..5171
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    5221..5254
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        5315..5814
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(5350..5373)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     5747..5766
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             5854..6453
                     /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(5854..5873)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     protein_bind    7010..7043
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    7074..7910
                     /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(7944..7963)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7945..7963)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7970..7987)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7970..7986)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7979..8001)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             8124..8426
                     /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     8305..8324
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             8775..9569
                     /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             8775..9569
                     /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(8829..8848)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     9439..9458
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             complement(join(9819..10610,10611..10679))
                     /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     10442..10461
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10803..11391
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11292..11311
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctca
      241 gaattaaccc tcactaaagg gactagtcct gcaggtttaa acgaattcgc cctttgcttt
      301 ctctgaccag cattctctcc cctgggcctg tgccgctttc tgtctgcagc ttgtggcctg
      361 ggtcacctct acggctggcc cagatccttc cctgccgcct ccttcaggtt ccgtcttcct
      421 ccactccctc ttccccttgc tctctgctgt gttgctgccc aaggatgctc tttccggagc
      481 acttccttct cggcgctgca ccacgtgatg tcctctgagc ggatcctccc cgtgtctggg
      541 tcctctccgg gcatctctcc tccctcaccc aaccccatgc cgtcttcact cgctgggttc
      601 ccttttcctt ctccttctgg ggcctgtgcc atctctcgtt tcttaggatg gccttctccg
      661 acggatgtct cccttgcgtc ccgcctcccc ttcttgtagg cctgcatcat caccgttttt
      721 ctggacaacc ccaaagtacc ccgtctccct ggctttagcc acctctccat cctcttgctt
      781 tctttgcctg gacaccccgt tctcctgtgg attcgggtca cctctcactc ctttcatttg
      841 ggcagctccc ctacccccct tacctctcta gtctgtgcta gctcttccag ccccctgtca
      901 tggcatcttc caggggtccg agagctcagc tagtcttctt cctccaaccc gggcccctat
      961 gtccacttca ggacagcatg tttgctgcct ccagggatcc tgtgtccccg agctgggacc
     1021 accttatatt cccagggccg gttaatgtgg ctctggttct gggtactttt atctgtcccc
     1081 tccaccccac agtggggcaa gctgagtcag tgagcgagga agctcggggc agtgaaaaaa
     1141 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     1201 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg
     1261 ggaggttttt taaagcaagt aaaacctcta caaatgtggt atggctgatt atgatcctct
     1321 ggagatttac ccggggagca tgtcaaggtc aaaatcgtca agagcgtcag caggcagcat
     1381 atcaaggtca aagtcgtcaa gggcatcggc tgggagcatg tctaagtcaa aatcgtcaag
     1441 ggcgtcggtc ggcccgccgc tttcgcactt tagctgtttc tccaggccac atatgattag
     1501 ttccaggccg aaaaggaagg caggttcggc tccctgccgg tcgaacagct caattgcttg
     1561 tttcagaagt gggggcatag aatcggtggt aggtgtctct ctttcctctt ttgctacttg
     1621 atgctcctgt tcctccaata cgcagcccag tgtaaagtgg cccacggcgg acagagcgta
     1681 cagtgcgttc tccagggaga agccttgctg acacaggaac gcgagctgat tttccagggt
     1741 ttcgtactgt ttctctgttg ggcgggtgcc gagatgcact ttagccccgt cgcgatgtga
     1801 gaggagagca cagcggtatg acttggcgtt gttccgcaga aagtcttgcc atgactcgcc
     1861 ttccaggggg caggagtggg tatgatgcct gtccagcatc tcgattggca gggcatcgag
     1921 cagggcccgc ttgttcttca cgtgccagta cagggtaggc tgctcaactc ccagcttttg
     1981 agcgagtttc cttgtcgtca ggccttcgat accgactcca ttgagtaatt ccagagcaga
     2041 gtttatgact ttgctcttgt ccagtctaga catggtaatt cgatgatccc cctggggaga
     2101 gaggtcggtg attcggtcaa cgagggagcc gactgccgac gtgcgctccg gaggcttgca
     2161 gaatgcggaa caccgcgcgg gcaggaacag ggcccacact accgccccac accccgcctc
     2221 ccgcaccgcc ccttcccggc cgctgctctc ggcgcgccct gctgagcagc cgctattggc
     2281 cacagcccat cgcggtcggc gcgctgccat tgctccctgg cgctgtccgt ctgcgagggt
     2341 actagtgaga cgtgcggctt ccgtttgtca cgtccggcac gccgcgaacc gcaaggaacc
     2401 ttcccgactt aggggcggag caggaagcgt cgccgggggg cccacaaggg tagcggcgaa
     2461 gatccgggtg acgctgcgaa cggacgtgaa gaatgtgcga gacccagggt cggcgccgct
     2521 gcgtttcccg gaaccacgcc cagagcagcc gcgtccctgc gcaaacccag ggctgccttg
     2581 gaaaaggcgc aaccccaacc ccgtggaatt atcacctcga gaataaaata tctttatttt
     2641 cattacatct gtgtgttggt tttttgtgtg aatcgatagt actaacatac gctctccatc
     2701 aaaacaaaac gaaacaaaac aaactagcaa aataggctgt ccccagtgca agtgcaggtg
     2761 ccagaacatt tctctctcga gtttactccc tatcagtgat agagaacgta tgaagagttt
     2821 actccctatc agtgatagag aacgtatgca gactttactc cctatcagtg atagagaacg
     2881 tataaggagt ttactcccta tcagtgatag agaacgtatg accagtttac tccctatcag
     2941 tgatagagaa cgtatctaca gtttactccc tatcagtgat agagaacgta tatccagttt
     3001 actccctatc agtgatagag aacgtataag ctttgcttat gtaaaccagg gcgcctataa
     3061 aagagtgctg attttttgag taaacttcaa ttccacaaca cttttgtctt ataccaactt
     3121 tccgtaccac ttcctaccct cgtaaagaat tcgcggccgc acgcgaagcg ctgccaccat
     3181 gacatacttt cctgaagagg tcgtcgaaca catttttagc ttcctgcctg cacagagaga
     3241 tagaaacaca gtgagcctgg tctgcaaagt gtggtacgag atcgaacgcc tgagccggag
     3301 aggagtgttc gtcggcaact gctatgctgt gagagcaggc agggtcgccg ctaggtttcc
     3361 aaatgtgcgc gcactgaccg tcaaggggaa accccacggc gccgacttta acctggtgcc
     3421 ccctgattgg ggaggatacg ccggcccttg gatcgaggca gccgctcgcg gctgtcatgg
     3481 actggaggaa ctgcgcatga agcgaatggt ggtctctgac gaaagtctgg agctgctggc
     3541 tcggagcttc cctaggtttc gcgcactggt gctgatttct tgcgaaggct tcagcaccga
     3601 tggactggca gccgtggcct cccactgtaa gctgctgcgg gagctggacc tccaggagaa
     3661 tgaagtggag gatagaggcc ccagatggct gtcttgcttc ccagactcat gtaccagcct
     3721 ggtgtccctg aactttgcct gcatcaaagg cgaagtgaat gctgggtccc tggagcggct
     3781 ggtctcaaga agccccaacc tgaggtctct gcggctgaac cggagcgtga gcgtggacac
     3841 tctggctaag attctgctga gaacccctaa cctggaggat ctgggaaccg gcaatctgac
     3901 agacgatttc cagacagaat cctactttaa actgacttct gccctggaga agtgtaaaat
     3961 gctgaggagt ctgtcaggat tctgggatgc ttcacccgtg tgcctgagct ttatctaccc
     4021 tctgtgtgca cagctgacag gcctgaacct gagctatgca ccaaccctgg acgccagtga
     4081 tctgacaaag atgatctcac gctgcgtgaa actccagcga ctgtgggtgc tggactgtat
     4141 ttccgataag gggctccagg tggtcgccag ctcctgcaag gacctccagg agctgagagt
     4201 gttcccatct gatttttacg tggccggata tagtgctgtc actgaggaag gcctggtggc
     4261 agtctcactg ggatgcccaa agctgaacag cctgctgtat ttctgtcatc agatgactaa
     4321 tgctgcactg gtgaccgtcg ccaagaactg ccctaatttc acccgatttc ggctgtgtat
     4381 tctggaacca ggcaaacccg acgtggtcac atcccagcca ctggatgaag ggtttggagc
     4441 tatcgtgaga gagtgcaagg gactccagag gctgagcatt tccggcctgc tgacagacaa
     4501 agtgttcatg tacatcggca agtatgctaa gcagctggag atgctgagca ttgcatttgc
     4561 cggagactcc gataagggca tgatgcacgt gatgaacggg tgtaagaatc tgcgaaaact
     4621 ggaaatccgg gacagccctt tcggggatgc cgctctgctg ggaaactttg ccagatacga
     4681 gacaatgagg agcctgtgga tgtctagttg caatgtgact ctgaagggct gtcaggtcct
     4741 ggctagtaaa atgcctatgc tgaacgtgga agtcattaat gagcgggacg ggtctaacga
     4801 aatggaggaa aatcatggcg acctgccaaa ggtggagaaa ctgtatgtgt atcggaccac
     4861 cgcaggggca agagatgatg ctcccaactt tgtgaagatt ctgtgaggat ccatcggatc
     4921 ccgggcccgt cgacacgcgt atcgataccg gtggatccag atctctgcag catatgtcta
     4981 gaggatcata atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc
     5041 acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat
     5101 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     5161 tttttcactg ccgacgtcag gtggcacttt tcggggcgat catattcaat aacccttaat
     5221 ataacttcgt ataatgtatg ctatacgaag ttattaggtc tgaagaggag tttacgtcca
     5281 gccaagctta ggatctcgac ctcgaaattc taccgggtag gggaggcgct tttcccaagg
     5341 cagtctggag catgcgcttt agcagccccg ctgggcactt ggcgctacac aagtggcctc
     5401 tggcctcgca cacattccac atccaccggt aggcgccaac cgactccgtt ctttggtggc
     5461 cccttcgcgc caccttctac tcctccccta gtcaggaagt tcccccccgc cccgcagctc
     5521 gcgtcgtgca ggacgtgaca aatggaagta gcacgtctca ctagtctcgt gcagatggac
     5581 agcaccgctg agcaatggaa gcgggtaggc ctttggggca gcggccaata gcagctttgc
     5641 tccttcgctt tctgggctca gaggctggga aggggtgggt ccgggggcgg gctcaggggc
     5701 gggctcaggg gcggggcggg cgcccgaagg tcctccggag gcccggcatt ctgcacgctt
     5761 caaaagcgca cgtctgccgc gctgttctcc tcttcctcat ctccgggcct ttcgacctgc
     5821 atccatctag atctcgagca gctgaagctt accatgaccg agtacaagcc cacggtgcgc
     5881 ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac
     5941 taccccgcca cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt caccgagctg
     6001 caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt cgcggacgac
     6061 ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc
     6121 gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca gcaacagatg
     6181 gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc caccgtcggc
     6241 gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc cggagtggag
     6301 gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgcccca caacctcccc
     6361 ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc
     6421 acctggtgca tgacccgcaa gcccggtgcc tgacgcccgc cccacgaccc gcagcgcccg
     6481 accgaaagga gcgcacgacc ccatgcatcg atgatatcag atccccggga tgcagaaatt
     6541 gatgatctat taaacaataa agatgtccac taaaatggaa gtttttcctg tcatactttg
     6601 ttaagaaggg tgagaacaga gtacctacat tttgaatgga aggattggag ctacgggggt
     6661 gggggtgggg tgggattaga taaatgcctg ctctttactg aaggctcttt actattgctt
     6721 tatgataatg tttcatagtt ggatatcata atttaaacaa gcaaaaccaa attaagggcc
     6781 agctcattcc tcccactcat gatctataga tctatagatc tctcgtggga tcattgtttt
     6841 tctcttgatt cccactttgt ggttctaagt actgtggttt ccaaatgtgt cagtttcata
     6901 gcctgaagaa cgagatcagc agcctctgtt ccacatacac ttcattctca gtattgtttt
     6961 gccaagttct aattccatca gaagctggtc gagatccgga acccttaata taacttcgta
     7021 taatgtatgc tatacgaagt tattaggtcc ctcgaagagg ttcactaagc tttactaggg
     7081 acaggattgg tgacagaaaa gccccatcct taggcctcct ccttcctagt ctcctgatat
     7141 tgggtctaac ccccacctcc tgttaggcag attccttatc tggtgacaca cccccatttc
     7201 ctggagccat ctctctcctt gccagaacct ctaaggtttg cttacgatgg agccagagag
     7261 gatcctggga gggagagctt ggcagggggt gggagggaag ggggggatgc gtgacctgcc
     7321 cggttctcag tggccaccct gcgctaccct ctcccagaac ctgagctgct ctgacgcggc
     7381 tgtctggtgc gtttcactga tcctggtgct gcagcttcct tacacttccc aagaggagaa
     7441 gcagtttgga aaaacaaaat cagaataagt tggtcctgag ttctaacttt ggctcttcac
     7501 ctttctagtc cccaatttat attgttcctc cgtgcgtcag ttttacctgt gagataaggc
     7561 cagtagccag ccccgtcctg gcagggctgt ggtgaggagg ggggtgtccg tgtggaaaac
     7621 tccctttgtg agaatggtgc gtcctaggtg ttcaccaggt cgtggccgcc tctactccct
     7681 ttctctttct ccatccttct ttccttaaag agtccccagt gctatctggg acatattcct
     7741 ccgcccagag cagggtcccg cttccctaag gccctgctct gggcttctgg gtttgagtcc
     7801 ttggcaagcc caggagaggc gctcaggctt ccctgtcccc cttcctcgtc caccatctca
     7861 tgcccctggc tctcctgccc cttccctaca ggggttcctg gctctgctct aagggcgaat
     7921 tcgcggccgc taaattcaat tcgccctata gtgagtcgta ttacaattca ctggccgtcg
     7981 ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac
     8041 atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
     8101 agttgcgcag cctatacgta cggcagttta aggtttacac ctataaaaga gagagccgtt
     8161 atcgtctgtt tgtggatgta cagagtgata ttattgacac gccggggcga cggatggtga
     8221 tccccctggc cagtgcacgt ctgctgtcag ataaagtctc ccgtgaactt tacccggtgg
     8281 tgcatatcgg ggatgaaagc tggcgcatga tgaccaccga tatggccagt gtgccggtct
     8341 ccgttatcgg ggaagaagtg gctgatctca gccaccgcga aaatgacatc aaaaacgcca
     8401 ttaacctgat gttctgggga atataaatgt caggcatgag attatcaaaa aggatcttca
     8461 cctagatcct tttcacgtag aaagccagtc cgcagaaacg gtgctgaccc cggatgaatg
     8521 tcagctactg ggctatctgg acaagggaaa acgcaagcgc aaagagaaag caggtagctt
     8581 gcagtgggct tacatggcga tagctagact gggcggtttt atggacagca agcgaaccgg
     8641 aattgccagc tggggcgccc tctggtaagg ttgggaagcc ctgcaaagta aactggatgg
     8701 ctttcttgcc gccaaggatc tgatggcgca ggggatcaag ctctgatcaa gagacaggat
     8761 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     8821 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     8881 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     8941 ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     9001 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     9061 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     9121 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     9181 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     9241 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     9301 cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     9361 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     9421 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     9481 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     9541 tctatcgcct tcttgacgag ttcttctgaa ttattaacgc ttacaatttc ctgatgcggt
     9601 attttctcct tacgcatctg tgcggtattt cacaccgcat caggtggcac ttttcgggga
     9661 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     9721 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     9781 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     9841 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     9901 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     9961 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
    10021 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
    10081 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
    10141 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
    10201 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
    10261 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
    10321 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
    10381 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
    10441 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
    10501 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
    10561 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
    10621 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
    10681 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gaccaaaatc
    10741 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
    10801 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
    10861 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
    10921 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
    10981 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
    11041 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
    11101 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
    11161 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
    11221 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
    11281 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
    11341 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
    11401 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
    11461 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
    11521 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga ag
//
