LOCUS       sequence-448188-       11632 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tet-OsTIR1(F74G)-V5 donor for integration at the AAVS1 locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11632)
  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 11632)
  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: AAVS1-Tet-OsTIR1(F74G)-V5
FEATURES             Location/Qualifiers
     source          1..11632
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     149..168
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             256..855
                     /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(256..275)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     protein_bind    1412..1445
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    1476..2312
                     /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(2346..2365)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2347..2365)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2372..2389)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2372..2388)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2381..2403)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             2526..2828
                     /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     2707..2726
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             3177..3971
                     /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             3177..3971
                     /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(3231..3250)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3841..3860
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             complement(join(4221..5012,5013..5081))
                     /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     4844..4863
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5205..5793
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5694..5713
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    6081..6102
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6117..6122,6123..6140,6141..6147)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6155..6171
                     /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     6160..6182
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6179..6195
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6179..6195
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        6216..6234
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    6269..7072
                     /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    7103..7184
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(7301..8047)
                     /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(8066..8576)
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     complement(8129..8147)
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     polyA_signal    8596..8644
                     /note="synthetic polyadenylation signal"
     misc_feature    8658..8749
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     8698..8717
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        8753..9120
                     /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    8761..8779
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8797..8815
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8833..8851
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8869..8887
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8905..8923
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8941..8959
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8977..8995
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             9153..10877
                     /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             10896..10937
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     polyA_signal    11124..11205
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    11255..11288
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        join(11349..11632,1..216)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(11384..11407)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
ORIGIN
        1 aagcgggtag gcctttgggg cagcggccaa tagcagcttt gctccttcgc tttctgggct
       61 cagaggctgg gaaggggtgg gtccgggggc gggctcaggg gcgggctcag gggcggggcg
      121 ggcgcccgaa ggtcctccgg aggcccggca ttctgcacgc ttcaaaagcg cacgtctgcc
      181 gcgctgttct cctcttcctc atctccgggc ctttcgacct gcatccatct agatctcgag
      241 cagctgaagc ttaccatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac
      301 gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac
      361 accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg
      421 cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc
      481 tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg
      541 gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg
      601 caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag
      661 ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg
      721 gtgcccgcct tcctggagac ctccgcgccc cacaacctcc ccttctacga gcggctcggc
      781 ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc
      841 aagcccggtg cctgacgccc gccccacgac ccgcagcgcc cgaccgaaag gagcgcacga
      901 ccccatgcat cgatgatatc agatccccgg gatgcagaaa ttgatgatct attaaacaat
      961 aaagatgtcc actaaaatgg aagtttttcc tgtcatactt tgttaagaag ggtgagaaca
     1021 gagtacctac attttgaatg gaaggattgg agctacgggg gtgggggtgg ggtgggatta
     1081 gataaatgcc tgctctttac tgaaggctct ttactattgc tttatgataa tgtttcatag
     1141 ttggatatca taatttaaac aagcaaaacc aaattaaggg ccagctcatt cctcccactc
     1201 atgatctata gatctataga tctctcgtgg gatcattgtt tttctcttga ttcccacttt
     1261 gtggttctaa gtactgtggt ttccaaatgt gtcagtttca tagcctgaag aacgagatca
     1321 gcagcctctg ttccacatac acttcattct cagtattgtt ttgccaagtt ctaattccat
     1381 cagaagctgg tcgagatccg gaacccttaa tataacttcg tataatgtat gctatacgaa
     1441 gttattaggt ccctcgaaga ggttcactaa gctttactag ggacaggatt ggtgacagaa
     1501 aagccccatc cttaggcctc ctccttccta gtctcctgat attgggtcta acccccacct
     1561 cctgttaggc agattcctta tctggtgaca cacccccatt tcctggagcc atctctctcc
     1621 ttgccagaac ctctaaggtt tgcttacgat ggagccagag aggatcctgg gagggagagc
     1681 ttggcagggg gtgggaggga agggggggat gcgtgacctg cccggttctc agtggccacc
     1741 ctgcgctacc ctctcccaga acctgagctg ctctgacgcg gctgtctggt gcgtttcact
     1801 gatcctggtg ctgcagcttc cttacacttc ccaagaggag aagcagtttg gaaaaacaaa
     1861 atcagaataa gttggtcctg agttctaact ttggctcttc acctttctag tccccaattt
     1921 atattgttcc tccgtgcgtc agttttacct gtgagataag gccagtagcc agccccgtcc
     1981 tggcagggct gtggtgagga ggggggtgtc cgtgtggaaa actccctttg tgagaatggt
     2041 gcgtcctagg tgttcaccag gtcgtggccg cctctactcc ctttctcttt ctccatcctt
     2101 ctttccttaa agagtcccca gtgctatctg ggacatattc ctccgcccag agcagggtcc
     2161 cgcttcccta aggccctgct ctgggcttct gggtttgagt ccttggcaag cccaggagag
     2221 gcgctcaggc ttccctgtcc cccttcctcg tccaccatct catgcccctg gctctcctgc
     2281 cccttcccta caggggttcc tggctctgct ctaagggcga attcgcggcc gctaaattca
     2341 attcgcccta tagtgagtcg tattacaatt cactggccgt cgttttacaa cgtcgtgact
     2401 gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct
     2461 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctatacg
     2521 tacggcagtt taaggtttac acctataaaa gagagagccg ttatcgtctg tttgtggatg
     2581 tacagagtga tattattgac acgccggggc gacggatggt gatccccctg gccagtgcac
     2641 gtctgctgtc agataaagtc tcccgtgaac tttacccggt ggtgcatatc ggggatgaaa
     2701 gctggcgcat gatgaccacc gatatggcca gtgtgccggt ctccgttatc ggggaagaag
     2761 tggctgatct cagccaccgc gaaaatgaca tcaaaaacgc cattaacctg atgttctggg
     2821 gaatataaat gtcaggcatg agattatcaa aaaggatctt cacctagatc cttttcacgt
     2881 agaaagccag tccgcagaaa cggtgctgac cccggatgaa tgtcagctac tgggctatct
     2941 ggacaaggga aaacgcaagc gcaaagagaa agcaggtagc ttgcagtggg cttacatggc
     3001 gatagctaga ctgggcggtt ttatggacag caagcgaacc ggaattgcca gctggggcgc
     3061 cctctggtaa ggttgggaag ccctgcaaag taaactggat ggctttcttg ccgccaagga
     3121 tctgatggcg caggggatca agctctgatc aagagacagg atgaggatcg tttcgcatga
     3181 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     3241 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     3301 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag
     3361 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     3421 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     3481 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     3541 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     3601 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     3661 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg
     3721 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     3781 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     3841 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     3901 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     3961 agttcttctg aattattaac gcttacaatt tcctgatgcg gtattttctc cttacgcatc
     4021 tgtgcggtat ttcacaccgc atcaggtggc acttttcggg gaaatgtgcg cggaacccct
     4081 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagatt atcaaaaagg
     4141 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     4201 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     4261 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     4321 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     4381 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     4441 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     4501 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     4561 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     4621 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     4681 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     4741 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     4801 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     4861 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     4921 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     4981 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     5041 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     5101 tattgaagca tttatcaggg ttattgtctc atgaccaaaa tcccttaacg tgagttttcg
     5161 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     5221 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     5281 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     5341 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     5401 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     5461 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     5521 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     5581 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     5641 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     5701 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     5761 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     5821 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     5881 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     5941 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
     6001 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
     6061 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta
     6121 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
     6181 ggaaacagct atgaccatga ttacgccaag ctcagaatta accctcacta aagggactag
     6241 tcctgcaggt ttaaacgaat tcgccctttg ctttctctga ccagcattct ctcccctggg
     6301 cctgtgccgc tttctgtctg cagcttgtgg cctgggtcac ctctacggct ggcccagatc
     6361 cttccctgcc gcctccttca ggttccgtct tcctccactc cctcttcccc ttgctctctg
     6421 ctgtgttgct gcccaaggat gctctttccg gagcacttcc ttctcggcgc tgcaccacgt
     6481 gatgtcctct gagcggatcc tccccgtgtc tgggtcctct ccgggcatct ctcctccctc
     6541 acccaacccc atgccgtctt cactcgctgg gttccctttt ccttctcctt ctggggcctg
     6601 tgccatctct cgtttcttag gatggccttc tccgacggat gtctcccttg cgtcccgcct
     6661 ccccttcttg taggcctgca tcatcaccgt ttttctggac aaccccaaag taccccgtct
     6721 ccctggcttt agccacctct ccatcctctt gctttctttg cctggacacc ccgttctcct
     6781 gtggattcgg gtcacctctc actcctttca tttgggcagc tcccctaccc cccttacctc
     6841 tctagtctgt gctagctctt ccagccccct gtcatggcat cttccagggg tccgagagct
     6901 cagctagtct tcttcctcca acccgggccc ctatgtccac ttcaggacag catgtttgct
     6961 gcctccaggg atcctgtgtc cccgagctgg gaccacctta tattcccagg gccggttaat
     7021 gtggctctgg ttctgggtac ttttatctgt cccctccacc ccacagtggg gcaagctgag
     7081 tcagtgagcg aggaagctcg gggcagtgaa aaaaatgctt tatttgtgaa atttgtgatg
     7141 ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca
     7201 ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc
     7261 tctacaaatg tggtatggct gattatgatc ctctggagat ttacccgggg agcatgtcaa
     7321 ggtcaaaatc gtcaagagcg tcagcaggca gcatatcaag gtcaaagtcg tcaagggcat
     7381 cggctgggag catgtctaag tcaaaatcgt caagggcgtc ggtcggcccg ccgctttcgc
     7441 actttagctg tttctccagg ccacatatga ttagttccag gccgaaaagg aaggcaggtt
     7501 cggctccctg ccggtcgaac agctcaattg cttgtttcag aagtgggggc atagaatcgg
     7561 tggtaggtgt ctctctttcc tcttttgcta cttgatgctc ctgttcctcc aatacgcagc
     7621 ccagtgtaaa gtggcccacg gcggacagag cgtacagtgc gttctccagg gagaagcctt
     7681 gctgacacag gaacgcgagc tgattttcca gggtttcgta ctgtttctct gttgggcggg
     7741 tgccgagatg cactttagcc ccgtcgcgat gtgagaggag agcacagcgg tatgacttgg
     7801 cgttgttccg cagaaagtct tgccatgact cgccttccag ggggcaggag tgggtatgat
     7861 gcctgtccag catctcgatt ggcagggcat cgagcagggc ccgcttgttc ttcacgtgcc
     7921 agtacagggt aggctgctca actcccagct tttgagcgag tttccttgtc gtcaggcctt
     7981 cgataccgac tccattgagt aattccagag cagagtttat gactttgctc ttgtccagtc
     8041 tagacatggt aattcgatga tccccctggg gagagaggtc ggtgattcgg tcaacgaggg
     8101 agccgactgc cgacgtgcgc tccggaggct tgcagaatgc ggaacaccgc gcgggcagga
     8161 acagggccca cactaccgcc ccacaccccg cctcccgcac cgccccttcc cggccgctgc
     8221 tctcggcgcg ccctgctgag cagccgctat tggccacagc ccatcgcggt cggcgcgctg
     8281 ccattgctcc ctggcgctgt ccgtctgcga gggtactagt gagacgtgcg gcttccgttt
     8341 gtcacgtccg gcacgccgcg aaccgcaagg aaccttcccg acttaggggc ggagcaggaa
     8401 gcgtcgccgg ggggcccaca agggtagcgg cgaagatccg ggtgacgctg cgaacggacg
     8461 tgaagaatgt gcgagaccca gggtcggcgc cgctgcgttt cccggaacca cgcccagagc
     8521 agccgcgtcc ctgcgcaaac ccagggctgc cttggaaaag gcgcaacccc aaccccgtgg
     8581 aattatcacc tcgagaataa aatatcttta ttttcattac atctgtgtgt tggttttttg
     8641 tgtgaatcga tagtactaac atacgctctc catcaaaaca aaacgaaaca aaacaaacta
     8701 gcaaaatagg ctgtccccag tgcaagtgca ggtgccagaa catttctctc tcgagtttac
     8761 tccctatcag tgatagagaa cgtatgaaga gtttactccc tatcagtgat agagaacgta
     8821 tgcagacttt actccctatc agtgatagag aacgtataag gagtttactc cctatcagtg
     8881 atagagaacg tatgaccagt ttactcccta tcagtgatag agaacgtatc tacagtttac
     8941 tccctatcag tgatagagaa cgtatatcca gtttactccc tatcagtgat agagaacgta
     9001 taagctttgc ttatgtaaac cagggcgcct ataaaagagt gctgattttt tgagtaaact
     9061 tcaattccac aacacttttg tcttatacca actttccgta ccacttccta ccctcgtaaa
     9121 gaattcgcgg ccgcacgcga agcgctgcca ccatgacata ctttcctgaa gaggtcgtcg
     9181 aacacatttt tagcttcctg cctgcacaga gagatagaaa cacagtgagc ctggtctgca
     9241 aagtgtggta cgagatcgaa cgcctgagcc ggagaggagt gttcgtcggc aactgctatg
     9301 ctgtgagagc aggcagggtc gccgctaggt ttccaaatgt gcgcgcactg accgtcaagg
     9361 ggaaacccca cggcgccgac tttaacctgg tgccccctga ttggggagga tacgccggcc
     9421 cttggatcga ggcagccgct cgcggctgtc atggactgga ggaactgcgc atgaagcgaa
     9481 tggtggtctc tgacgaaagt ctggagctgc tggctcggag cttccctagg tttcgcgcac
     9541 tggtgctgat ttcttgcgaa ggcttcagca ccgatggact ggcagccgtg gcctcccact
     9601 gtaagctgct gcgggagctg gacctccagg agaatgaagt ggaggataga ggccccagat
     9661 ggctgtcttg cttcccagac tcatgtacca gcctggtgtc cctgaacttt gcctgcatca
     9721 aaggcgaagt gaatgctggg tccctggagc ggctggtctc aagaagcccc aacctgaggt
     9781 ctctgcggct gaaccggagc gtgagcgtgg acactctggc taagattctg ctgagaaccc
     9841 ctaacctgga ggatctggga accggcaatc tgacagacga tttccagaca gaatcctact
     9901 ttaaactgac ttctgccctg gagaagtgta aaatgctgag gagtctgtca ggattctggg
     9961 atgcttcacc cgtgtgcctg agctttatct accctctgtg tgcacagctg acaggcctga
    10021 acctgagcta tgcaccaacc ctggacgcca gtgatctgac aaagatgatc tcacgctgcg
    10081 tgaaactcca gcgactgtgg gtgctggact gtatttccga taaggggctc caggtggtcg
    10141 ccagctcctg caaggacctc caggagctga gagtgttccc atctgatttt tacgtggccg
    10201 gatatagtgc tgtcactgag gaaggcctgg tggcagtctc actgggatgc ccaaagctga
    10261 acagcctgct gtatttctgt catcagatga ctaatgctgc actggtgacc gtcgccaaga
    10321 actgccctaa tttcacccga tttcggctgt gtattctgga accaggcaaa cccgacgtgg
    10381 tcacatccca gccactggat gaagggtttg gagctatcgt gagagagtgc aagggactcc
    10441 agaggctgag catttccggc ctgctgacag acaaagtgtt catgtacatc ggcaagtatg
    10501 ctaagcagct ggagatgctg agcattgcat ttgccggaga ctccgataag ggcatgatgc
    10561 acgtgatgaa cgggtgtaag aatctgcgaa aactggaaat ccgggacagc cctttcgggg
    10621 atgccgctct gctgggaaac tttgccagat acgagacaat gaggagcctg tggatgtcta
    10681 gttgcaatgt gactctgaag ggctgtcagg tcctggctag taaaatgcct atgctgaacg
    10741 tggaagtcat taatgagcgg gacgggtcta acgaaatgga ggaaaatcat ggcgacctgc
    10801 caaaggtgga gaaactgtat gtgtatcgga ccaccgcagg ggcaagagat gatgctccca
    10861 actttgtgaa gattctggga tccggtgcag gcgccggtaa gcctatccct aaccctctcc
    10921 tcggtctcga ttctacgtga ggatccatcg gatcccgggc ccgtcgacac gcgtatcgat
    10981 accggtggat ccagatctct gcagcatatg tctagaggat cataatcagc cataccacat
    11041 ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata
    11101 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa
    11161 gcaatagcat cacaaatttc acaaataaag catttttttc actgccgacg tcaggtggca
    11221 cttttcgggg cgatcatatt caataaccct taatataact tcgtataatg tatgctatac
    11281 gaagttatta ggtctgaaga ggagtttacg tccagccaag cttaggatct cgacctcgaa
    11341 attctaccgg gtaggggagg cgcttttccc aaggcagtct ggagcatgcg ctttagcagc
    11401 cccgctgggc acttggcgct acacaagtgg cctctggcct cgcacacatt ccacatccac
    11461 cggtaggcgc caaccgactc cgttctttgg tggccccttc gcgccacctt ctactcctcc
    11521 cctagtcagg aagttccccc ccgccccgca gctcgcgtcg tgcaggacgt gacaaatgga
    11581 agtagcacgt ctcactagtc tcgtgcagat ggacagcacc gctgagcaat gg
//
