LOCUS       sequence-254335-       10961 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A repair construct to express GFP-ARF16-PB1 and OsTIR1 under the 
            control of the CMV promoter from the human AAVS1 locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10961)
  AUTHORS   Sathyan KM, McKenna BD, Anderson WD, Duarte FM, Core L, Guertin MJ
  TITLE     An improved auxin-inducible degron system preserves native protein 
            levels and enables rapid and specific protein depletion.
  JOURNAL   Genes Dev. 2019 Oct 1;33(19-20):1441-1455. doi: 
            10.1101/gad.328237.119. Epub 2019 Aug 29.
  PUBMED    31467088
REFERENCE   2  (bases 1 to 10961)
  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: "Genes 
            Dev."; date: "2019-10-1"; volume: "33(19-20):1441-1455. doi"; pages:
            " 10.1101/gad.328237.119. Epub 2019 Aug 29"
COMMENT     Sequence Label: pMGS56 (GFP-ARF16-PB1-P2A-OsTIR1)
FEATURES             Location/Qualifiers
     source          1..10961
                     /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"
     enhancer        1155..1458
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1459..1662
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1613..1633
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1694..1703
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1700..1702,1703..1705,1706..2416)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1700..1702,1703..1705,1706..2416)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1745..1766)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2006..2025)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2353..2374
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2444..2464
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3011..3067
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             3077..4804
                     /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    4978..5099
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5015..5034)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5069..5088
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        5108..5608
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     5527..5545
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             5630..6229
                     /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(5630..5649)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6126..6146
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    6236..6460
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     misc_feature    6463..7299
                     /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(7333..7352)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7334..7352)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7359..7376)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7359..7375)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7368..7390)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             7513..7815
                     /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     7694..7713
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             8164..8958
                     /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             8164..8958
                     /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(8218..8237)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     8828..8847
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             complement(join(9208..9999,10000..10068))
                     /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     9831..9850
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10192..10780
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10681..10700
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10934..10951
                     /label=L4440
                     /note="L4440 vector, 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 ttaatagtaa tcaattacgg ggtcattagt tcatagccca
     1141 tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
     1201 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
     1261 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
     1321 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
     1381 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
     1441 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg
     1501 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
     1561 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg
     1621 ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctggtttagt gaaccgtcag
     1681 atccgctagc gctgccacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
     1741 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
     1801 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
     1861 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg
     1921 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
     1981 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
     2041 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     2101 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat
     2161 ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga
     2221 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     2281 gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga
     2341 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     2401 ggacgagctg tacaagtccg gccggactca gatctcgagc atgcctaaga agaagcggaa
     2461 ggtgggcagc acactgactt ccgggcagcg ggatcaccgg gacgtcgaga acgagctgag
     2521 ctccgccgcc ttcagctccc agagcttcgg ggtgcccgac atgagcttta agcccggctg
     2581 cagcagcgac gtggccgtga ccgacgccgg gatgcccagc caggggctgt ggaacaacca
     2641 gacccagcgg atgaggacat tcacaaaggt gcagaagcgg gggagcgtgg gccggagcat
     2701 cgatatcacc aggtacaggg attacgatga gctgaggcat gatctcgcct gtatgttcgg
     2761 catccagggc cagctggagg acccctaccg gatggactgg aagctggtgt acgtggacca
     2821 cgagaacgat atcctgctgg tgggcgatga cccctgggag gagttcgtgg gctgcgtgaa
     2881 gagcattaag attctgtctg ctgctgaggt gcagcagatg agcctggatg gagatctggg
     2941 aggagtgcca cctcagactc aggcctgtag tgctagtgat gatgctaacg cctggagagg
     3001 gggaagcgga gctactaact tcagcctgct gaagcaggct ggagacgtgg aggagaaccc
     3061 tggacctgct gccaccatga catactttcc tgaagaggtc gtcgaacaca tttttagctt
     3121 cctgcctgca cagagagata gaaacacagt gagcctggtc tgcaaagtgt ggtacgagat
     3181 cgaacgcctg agccggagag gagtgttcgt cggcaactgc tatgctgtga gagcaggcag
     3241 ggtcgccgct aggtttccaa atgtgcgcgc actgaccgtc aaggggaaac cccacttcgc
     3301 cgactttaac ctggtgcccc ctgattgggg aggatacgcc ggcccttgga tcgaggcagc
     3361 cgctcgcggc tgtcatggac tggaggaact gcgcatgaag cgaatggtgg tctctgacga
     3421 aagtctggag ctgctggctc ggagcttccc taggtttcgc gcactggtgc tgatttcttg
     3481 cgaaggcttc agcaccgatg gactggcagc cgtggcctcc cactgtaagc tgctgcggga
     3541 gctggacctc caggagaatg aagtggagga tagaggcccc agatggctgt cttgcttccc
     3601 agactcatgt accagcctgg tgtccctgaa ctttgcctgc atcaaaggcg aagtgaatgc
     3661 tgggtccctg gagcggctgg tctcaagaag ccccaacctg aggtctctgc ggctgaaccg
     3721 gagcgtgagc gtggacactc tggctaagat tctgctgaga acccctaacc tggaggatct
     3781 gggaaccggc aatctgacag acgatttcca gacagaatcc tactttaaac tgacttctgc
     3841 cctggagaag tgtaaaatgc tgaggagtct gtcaggattc tgggatgctt cacccgtgtg
     3901 cctgagcttt atctaccctc tgtgtgcaca gctgacaggc ctgaacctga gctatgcacc
     3961 aaccctggac gccagtgatc tgacaaagat gatctcacgc tgcgtgaaac tccagcgact
     4021 gtgggtgctg gactgtattt ccgataaggg gctccaggtg gtcgccagct cctgcaagga
     4081 cctccaggag ctgagagtgt tcccatctga tttttacgtg gccggatata gtgctgtcac
     4141 tgaggaaggc ctggtggcag tctcactggg atgcccaaag ctgaacagcc tgctgtattt
     4201 ctgtcatcag atgactaatg ctgcactggt gaccgtcgcc aagaactgcc ctaatttcac
     4261 ccgatttcgg ctgtgtattc tggaaccagg caaacccgac gtggtcacat cccagccact
     4321 ggatgaaggg tttggagcta tcgtgagaga gtgcaaggga ctccagaggc tgagcatttc
     4381 cggcctgctg acagacaaag tgttcatgta catcggcaag tatgctaagc agctggagat
     4441 gctgagcatt gcatttgccg gagactccga taagggcatg atgcacgtga tgaacgggtg
     4501 taagaatctg cgaaaactgg aaatccggga cagccctttc ggggatgccg ctctgctggg
     4561 aaactttgcc agatacgaga caatgaggag cctgtggatg tctagttgca atgtgactct
     4621 gaagggctgt caggtcctgg ctagtaaaat gcctatgctg aacgtggaag tcattaatga
     4681 gcgggacggg tctaacgaaa tggaggaaaa tcatggcgac ctgccaaagg tggagaaact
     4741 gtatgtgtat cggaccaccg caggggcaag agatgatgct cccaactttg tgaagattct
     4801 gtgaggatcc atcggatccc gggcccgtcg acggtaccgc gggcccggga tccaccgggg
     4861 ccgcgactct agatcataat cagccatacc acatttgtag aggttttact tgctttaaaa
     4921 aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac
     4981 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     5041 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa
     5101 gttggggttg cgccttttcc aaggcagccc tgggtttgcg cagggacgcg gctgctctgg
     5161 gcgtggttcc gggaaacgca gcggcgccga ccctgggtct cgcacattct tcacgtccgt
     5221 tcgcagcgtc acccggatct tcgccgctac ccttgtgggc cccccggcga cgcttcctgc
     5281 tccgccccta agtcgggaag gttccttgcg gttcgcggcg tgccggacgt gacaaacgga
     5341 agccgcacgt ctcactagta ccctcgcaga cggacagcgc cagggagcaa tggcagcgcg
     5401 ccgaccgcga tgggctgtgg ccaatagcgg ctgctcagca gggcgcgccg agagcagcgg
     5461 ccgggaaggg gcggtgcggg aggcggggtg tggggcggta gtgtgggccc tgttcctgcc
     5521 cgcgcggtgt tccgcattct gcaagcctcc ggagcgcacg tcggcagtcg gctccctcgt
     5581 tgaccgaatc accgacctct ctccccaggg ggatccaccg gagcttacca tgaccgagta
     5641 caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc
     5701 cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga
     5761 gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt
     5821 gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc
     5881 gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc
     5941 cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt
     6001 cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt
     6061 gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc
     6121 gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt
     6181 gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgag gtaccctgtg
     6241 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     6301 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     6361 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     6421 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg ttactaggga caggattggt
     6481 gacagaaaag ccccatcctt aggcctcctc cttcctagtc tcctgatatt gggtctaacc
     6541 cccacctcct gttaggcaga ttccttatct ggtgacacac ccccatttcc tggagccatc
     6601 tctctccttg ccagaacctc taaggtttgc ttacgatgga gccagagagg atcctgggag
     6661 ggagagcttg gcagggggtg ggagggaagg gggggatgcg tgacctgccc ggttctcagt
     6721 ggccaccctg cgctaccctc tcccagaacc tgagctgctc tgacgcggct gtctggtgcg
     6781 tttcactgat cctggtgctg cagcttcctt acacttccca agaggagaag cagtttggaa
     6841 aaacaaaatc agaataagtt ggtcctgagt tctaactttg gctcttcacc tttctagtcc
     6901 ccaatttata ttgttcctcc gtgcgtcagt tttacctgtg agataaggcc agtagccagc
     6961 cccgtcctgg cagggctgtg gtgaggaggg gggtgtccgt gtggaaaact ccctttgtga
     7021 gaatggtgcg tcctaggtgt tcaccaggtc gtggccgcct ctactccctt tctctttctc
     7081 catccttctt tccttaaaga gtccccagtg ctatctggga catattcctc cgcccagagc
     7141 agggtcccgc ttccctaagg ccctgctctg ggcttctggg tttgagtcct tggcaagccc
     7201 aggagaggcg ctcaggcttc cctgtccccc ttcctcgtcc accatctcat gcccctggct
     7261 ctcctgcccc ttccctacag gggttcctgg ctctgctcta agggcgaatt cgcggccgct
     7321 aaattcaatt cgccctatag tgagtcgtat tacaattcac tggccgtcgt tttacaacgt
     7381 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc
     7441 gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc
     7501 ctatacgtac ggcagtttaa ggtttacacc tataaaagag agagccgtta tcgtctgttt
     7561 gtggatgtac agagtgatat tattgacacg ccggggcgac ggatggtgat ccccctggcc
     7621 agtgcacgtc tgctgtcaga taaagtctcc cgtgaacttt acccggtggt gcatatcggg
     7681 gatgaaagct ggcgcatgat gaccaccgat atggccagtg tgccggtctc cgttatcggg
     7741 gaagaagtgg ctgatctcag ccaccgcgaa aatgacatca aaaacgccat taacctgatg
     7801 ttctggggaa tataaatgtc aggcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     7861 ttcacgtaga aagccagtcc gcagaaacgg tgctgacccc ggatgaatgt cagctactgg
     7921 gctatctgga caagggaaaa cgcaagcgca aagagaaagc aggtagcttg cagtgggctt
     7981 acatggcgat agctagactg ggcggtttta tggacagcaa gcgaaccgga attgccagct
     8041 ggggcgccct ctggtaaggt tgggaagccc tgcaaagtaa actggatggc tttcttgccg
     8101 ccaaggatct gatggcgcag gggatcaagc tctgatcaag agacaggatg aggatcgttt
     8161 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     8221 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     8281 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     8341 ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     8401 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     8461 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     8521 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     8581 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     8641 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc
     8701 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     8761 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     8821 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     8881 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     8941 cttgacgagt tcttctgaat tattaacgct tacaatttcc tgatgcggta ttttctcctt
     9001 acgcatctgt gcggtatttc acaccgcatc aggtggcact tttcggggaa atgtgcgcgg
     9061 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagattatc
     9121 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     9181 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     9241 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     9301 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     9361 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     9421 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     9481 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     9541 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     9601 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     9661 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     9721 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     9781 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     9841 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     9901 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     9961 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
    10021 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
    10081 tcaatattat tgaagcattt atcagggtta ttgtctcatg accaaaatcc cttaacgtga
    10141 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
    10201 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
    10261 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
    10321 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
    10381 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
    10441 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
    10501 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
    10561 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
    10621 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
    10681 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
    10741 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
    10801 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
    10861 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
    10921 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa g
//
