LOCUS       sequence-298835-        9253 bp    DNA     circular SYN 14-APR-2025
DEFINITION  An all-in-one bicistronic plasmid for controlling protein 
            degradation by AID technology.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9253)
  AUTHORS   Yesbolatova A, Saito Y, Kanemaki MT
  TITLE     Constructing Auxin-Inducible Degron Mutants Using an All-in-One 
            Vector.
  JOURNAL   Pharmaceuticals (Basel). 2020 May 23;13(5). pii: ph13050103. doi: 
            10.3390/ph13050103.
  PUBMED    32456235
REFERENCE   2  (bases 1 to 9253)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Pharmaceuticals (Basel)."; date: "2020-05-23"; volume: "13(5). pii:
            ph13050103. doi"; pages: " 10.3390/ph13050103"
COMMENT     Sequence Label: pAID5.1-C
FEATURES             Location/Qualifiers
     source          1..9253
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(34..462)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(144..165)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     356..375
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     589..611
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     603..620
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     604..620
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     630..649
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        630..648
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    complement(884..917)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(1474..2073)
                     /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     2054..2073
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(2113..2612)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(2161..2180)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     2554..2577
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     protein_bind    complement(2673..2706)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    2733..2854
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2744..2763)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2798..2817
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(2975..3178)
                     /codon_start=1
                     /product="minimal degron from the auxin-responsive IAA17 
                     protein of Arabidopsis thaliana (Kubota et al., 2013)"
                     /label=mini-AID
                     /note="acts as an auxin-inducible degron (AID) in the 
                     presence of TIR1"
                     /translation="KEKSACPKDPAKPPAKAQVVGWPPVRSYRKNVMVSCQKSSGGPEA
                     AAFVKVSMDGAPYLRKIDLRMYK"
     misc_feature    3191..3247
                     /label=MCS
                     /note="multiple cloning site"
     CDS             complement(3248..3304)
                     /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             complement(3320..5044)
                     /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"
     primer_bind     complement(5089..5108)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     intron          5116..6132
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     complement(5170..5192)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     promoter        complement(6133..6410)
                     /label=chicken beta-actin promoter
     enhancer        6412..6791
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     complement(7069..7089)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(7069..7087)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7108..7124)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7108..7124)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7121..7143)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7132..7148
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(7156..7162,7163..7180,7181..7186))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7201..7222
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7339..7356)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7510..8098)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7590..7609)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8269..9060,9061..9129))
                     /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     8892..8911
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9130..9234)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
       61 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
      121 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
      181 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
      241 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
      301 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
      361 agaaagcgaa aggagcgggc gctagggcgc nggcgggtgt ggtggtcacg ctgcgcgtaa
      421 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg ccattcaggc
      481 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga
      541 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac
      601 gttgtaaaac gacggccagt gagcgcgcgt aatacgactc actatagggc gaattgggta
      661 ccggcatatg gttcttgaca gaggtgtaaa aagtactcaa aaattttact caagtgaaag
      721 tacaagtact tagggaaaat tttactcaat taaaagtaaa agtatctggc tagaatctta
      781 cttgagtaaa agtaaaaaag tactccatta aaattgtact tgagtattaa ggaagtaaaa
      841 gtaaaagcaa gaaagatctt agtgaacctc ttcgagggac ctaataactt cgtatagcat
      901 acattatacg aagttatatt aagggttccg gatctcgacc agcttctgat ggaattagaa
      961 cttggcaaaa caatactgag aatgaagtgt atgtggaaca gaggctgctg atctcgttct
     1021 tcaggctatg aaactgacac atttggaaac cacagtactt agaaccacaa agtgggaatc
     1081 aagagaaaaa caatgatccc acgagagatc tatagatcta tagatcatga gtgggaggaa
     1141 tgagctggcc cttaatttgg ttttgcttgt ttaaattatg atatccaact atgaaacatt
     1201 atcataaagc aatagtaaag agccttcagt aaagagcagg catttatcta atcccacccc
     1261 acccccaccc ccgtagctcc aatccttcca ttcaaaatgt aggtactctg ttctcaccct
     1321 tcttaacaaa gtatgacagg aaaaacttcc attttagtgg acatctttat tgtttaatag
     1381 atcatcaatt tctgcatccc ggggatctga tatcatcgat gcatggggtc gtgcgctcct
     1441 ttcggtcggg cgctgcgggt cgtggggcgg gcgtcaggca ccgggcttgc gggtcatgca
     1501 ccaggtgcgc ggtccttcgg gcacctcgac gtcggcggtg acggtgaagc cgagccgctc
     1561 gtagaagggg aggttgtggg gcgcggaggt ctccaggaag gcgggcaccc cggcgcgctc
     1621 ggccgcctcc actccgggga gcacgacggc gctgcccaga cccttgccct ggtggtcggg
     1681 cgagacgccg acggtggcca ggaaccacgc gggctccttg ggccggtgcg gcgccaggag
     1741 gccttccatc tgttgctgcg cggccagccg ggaaccgctc aactcggcca tgcgcgggcc
     1801 gatctcggcg aacaccgccc ccgcttcgac gctctccggc gtggtccaga ccgccaccgc
     1861 ggcgccgtcg tccgcgaccc acaccttgcc gatgtcgagc ccgacgcgcg tgaggaagag
     1921 ttcttgcagc tcggtgaccc gctcgatgtg gcggtccgga tcgacggtgt ggcgcgtggc
     1981 ggggtagtcg gcgaacgcgg cggcgagggt gcgtacggcc ctggggacgt cgtcgcgggt
     2041 ggcgaggcgc accgtgggct tgtactcggt catggtaagc ttcagctgct cgagatctag
     2101 atggatgcag gtcgaaaggc ccggagatga ggaagaggag aacagcgcgg cagacgtgcg
     2161 cttttgaagc gtgcagaatg ccgggcctcc ggaggacctt cgggcgcccg ccccgcccct
     2221 gagcccgccc ctgagcccgc ccccggaccc accccttccc agcctctgag cccagaaagc
     2281 gaaggagcaa agctgctatt ggccgctgcc ccaaaggcct acccgcttcc attgctcagc
     2341 ggtgctgtcc atctgcacga gactagtgag acgtgctact tccatttgtc acgtcctgca
     2401 cgacgcgagc tgcggggcgg gggggaactt cctgactagg ggaggagtag aaggtggcgc
     2461 gaaggggcca ccaaagaacg gagtcggttg gcgcctaccg gtggatgtgg aatgtgtgcg
     2521 aggccagagg ccacttgtgt agcgccaagt gcccagcggg gctgctaaag cgcatgctcc
     2581 agactgcctt gggaaaagcg cctcccctac ccggtagaat ttcgaggtcg agatcctaag
     2641 cttggctgga cgtaaactcc tcttcagacc taataacttc gtatagcata cattatacga
     2701 agttatatta agggttattg aatatgatcg cttaagatac attgatgagt ttggacaaac
     2761 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     2821 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     2881 gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg
     2941 tggtatggct gattatgatc agttatctag attatttata catcctcaaa tcgattttcc
     3001 tcaagtacgg tgctccgtcc attgatacct tcacgaacgc cgccgcctcc gggccaccgc
     3061 ttgatttttg gcaggaaacc atcacgttct tccggtatga tctcaccggt ggccatccca
     3121 caacttgtgc cttggccgga ggtttggctg gatctttagg acaagcactc ttctccttgg
     3181 cgcctgcacc ggatcccggg cccgcggtac cgtcgactgc agaattcgaa gcttgagctc
     3241 gagatctagg tccagggttc tcctccacgt ctccagcctg cttcagcagg ctgaagttag
     3301 tagctccgct tcctgatctc agaatcttca caaagttggg agcatcatct cttgcccctg
     3361 cggtggtccg atacacatac agtttctcca cctttggcag gtcgccatga ttttcctcca
     3421 tttcgttaga cccgtcccgc tcattaatga cttccacgtt cagcataggc attttactag
     3481 ccaggacctg acagcccttc agagtcacat tgcaactaga catccacagg ctcctcattg
     3541 tctcgtatct ggcaaagttt cccagcagag cggcatcccc gaaagggctg tcccggattt
     3601 ccagttttcg cagattctta cacccgttca tcacgtgcat catgccctta tcggagtctc
     3661 cggcaaatgc aatgctcagc atctccagct gcttagcata cttgccgatg tacatgaaca
     3721 ctttgtctgt cagcaggccg gaaatgctca gcctctggag tcccttgcac tctctcacga
     3781 tagctccaaa cccttcatcc agtggctggg atgtgaccac gtcgggtttg cctggttcca
     3841 gaatacacag ccgaaatcgg gtgaaattag ggcagttctt ggcgacggtc accagtgcag
     3901 cattagtcat ctgatgacag aaatacagca ggctgttcag ctttgggcat cccagtgaga
     3961 ctgccaccag gccttcctca gtgacagcac tatatccggc cacgtaaaaa tcagatggga
     4021 acactctcag ctcctggagg tccttgcagg agctggcgac cacctggagc cccttatcgg
     4081 aaatacagtc cagcacccac agtcgctgga gtttcacgca gcgtgagatc atctttgtca
     4141 gatcactggc gtccagggtt ggtgcatagc tcaggttcag gcctgtcagc tgtgcacaca
     4201 gagggtagat aaagctcagg cacacgggtg aagcatccca gaatcctgac agactcctca
     4261 gcattttaca cttctccagg gcagaagtca gtttaaagta ggattctgtc tggaaatcgt
     4321 ctgtcagatt gccggttccc agatcctcca ggttaggggt tctcagcaga atcttagcca
     4381 gagtgtccac gctcacgctc cggttcagcc gcagagacct caggttgggg cttcttgaga
     4441 ccagccgctc cagggaccca gcattcactt cgcctttgat gcaggcaaag ttcagggaca
     4501 ccaggctggt acatgagtct gggaagcaag acagccatct ggggcctcta tcctccactt
     4561 cattctcctg gaggtccagc tcccgcagca gcttacagtg ggaggccacg gctgccagtc
     4621 catcggtgct gaagccttcg caagaaatca gcaccagtgc gcgaaaccta gggaagctcc
     4681 gagccagcag ctccagactt tcgtcagaga ccaccattcg cttcatgcgc agttcctcca
     4741 gtccatgaca gccgcgagcg gctgcctcga tccaagggcc ggcgtatcct ccccaatcag
     4801 ggggcaccag gttaaagtcg gcgaagtggg gtttcccctt gacggtcagt gcgcgcacat
     4861 ttggaaacct agcggcgacc ctgcctgctc tcacagcata gcagttgccg acgaacactc
     4921 ctctccggct caggcgttcg atctcgtacc acactttgca gaccaggctc actgtgtttc
     4981 tatctctctg tgcaggcagg aagctaaaaa tgtgttcgac gacctcttca ggaaagtatg
     5041 tcatggtggc agcgcttcga ggaattcttt gccaaaatga tgagacagca caataaccag
     5101 cacgttgccc aggagctgta ggaaaaagaa gaaggcatga acatggttag cagaggctct
     5161 agagccgccg gtcacacgcc agaagccgaa ccccgccctg ccccgtcccc cccgaaggca
     5221 gccgtccccc cgcggacagc cccgaggctg gagagggaga aggggacggc ggcgcggcga
     5281 cgcacgaagg ccctccccgc ccatttcctt cctgccggcg ccgcaccgct tcgccccgcg
     5341 cccgctagag ggggtgcggc ggcgcctccc agatttcggc tccgcacaga tttgggacaa
     5401 aggaagtccc tgcgccctct cgcacgatta ccataaaagg caatggctgc ggctcgccgc
     5461 gcctcgacag ccgccggcgc tccgggggcc gccgcgcccc tcccccgagc ccyccccggc
     5521 ccgaggcggc cccgccccgc ccggcacccc cacctgccgc caccccccgc ccggcacggc
     5581 gagccccgcg ccacgccccg tacggagccc cgcacccgaa gccgggccgt gctcagcaac
     5641 tcggggaggg gggtgcaggg ggggttgcag cccgaccgac gcgcccacac cccctgctca
     5701 cccccccacg cacacacccc gcacgcagcc tttgttcccc tcgcagcccc ccccgcaccg
     5761 cggggcaccg cccccggccg cgctcccctc gcgcacactg cggagcgcac aaagccccgc
     5821 gccgcgcccg cagcgctcac agccgccggg cagcgcggag ccgcacgcgg cgctccccac
     5881 gcacacacac acgcacgcac cccccgagcc gctccccccg cacaaagggc cctcccggag
     5941 cccctcaagg ctttcacgca gccacagaaa agaaacaagc cgtcattaaa ccaagcgcta
     6001 attacagccc ggaggagaag ggccgtcccg cccgctcacc tgtgggagta acgcggtcag
     6061 tcagagccgg ggcgggcggc gcgaggcggc ggcggagcgg ggcacggggc gaaggcagcg
     6121 cgcagcgact cccgcccgcc gcgcgcttcg ctttttatag ggccgccgcc gccgccgcct
     6181 cgccataaaa ggaaactttc ggagcgcgcc gctctgattg gctgccgccg cacctctccg
     6241 cctcgccccs ccccsccccy csccccsccc csccccsccy ggcgcgcgcc cccccccccc
     6301 ccccgccccc atcgctgcac aaaataatta aaaaataaat aaatacaaaa ttgggggtgg
     6361 ggaggggggg gagatgggga gagtgaagca gaacgtgggg ctcacctcga ccatggtaat
     6421 agcgatgact aatacgtaga tgtactgcca agtaggaaag tcccataagg tcatgtactg
     6481 ggcataatgc caggcgggcc atttaccgtc attgacgtca atagggggcg tacttggcat
     6541 atgatacact tgatgtactg ccaagtgggc agtttaccgt aaatactcca cccattgacg
     6601 tcaatggaaa gtccctattg gcgttactat gggaacatac gtcattattg acgtcaatgg
     6661 gcgggggtcg ttgggcggtc agccaggcgg gccatttacc gtaagttatg taacgcggaa
     6721 ctccatatat gggctatgaa ctaatgaccc cgtaattgat tactattaat aactagtcaa
     6781 taatcaatgt cgagcatgca tctagagggc ccaagtgatc tccaaaaaat aagtactttt
     6841 tgactgtaaa taaaattgta aggagtaaaa agtacttttt tttctaaaaa aatgtaatta
     6901 agtaaaagta aaagtattga tttttaattg tactcaagta aagtaaaaat ccccaaaaat
     6961 aatacttaag tacagtaatc aagtaaaatt actcaagtac tttacacctc tggttcttga
     7021 ccccctacct tcagcaagcc cagcagatcc gagctccagc ttttgttccc tttagtgagg
     7081 gttaattgcg cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     7141 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     7201 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     7261 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     7321 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     7381 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     7441 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     7501 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     7561 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     7621 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     7681 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     7741 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     7801 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     7861 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     7921 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     7981 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     8041 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     8101 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     8161 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     8221 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     8281 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     8341 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     8401 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     8461 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     8521 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     8581 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     8641 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     8701 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     8761 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     8821 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     8881 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     8941 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     9001 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     9061 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     9121 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     9181 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     9241 tccccgaaaa gtg
//
