LOCUS       sequence-495610-       11915 bp    DNA     circular SYN 17-DEC-2025
DEFINITION  Overexpression vector for eGFP tagged RARA or NR1B1. Monitor 
            post-translational degradation of RARA via EGFP:mCherry ratio. 
            Contains puromycin-resistance selection marker..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11915)
  AUTHORS   Tsai JM, Aguirre JD, Li YD, Brown J, Focht V, Kater L, Kempf G, 
            Sandoval B, Schmitt S, Rutter JC, Galli P, Sandate CR, Cutler JA, 
            Zou C, Donovan KA, Lumpkin RJ, Cavadini S, Park PMC, Sievers Q, 
            Hatton C, Ener E, Regalado BD, Sperling MT, Slabicki M, Kim J, Zon 
            R, Zhang Z, Miller PG, Belizaire R, Sperling AS, Fischer ES, 
            Irizarry R, Armstrong SA, Thoma NH, Ebert BL
  TITLE     UBR5 forms ligand-dependent complexes on chromatin to regulate 
            nuclear hormone receptor stability.
  JOURNAL   Mol Cell. 2023 Aug 3;83(15):2753-2767.e10. doi: 
            10.1016/j.molcel.2023.06.028. Epub 2023 Jul 20.
  PUBMED    37478846
REFERENCE   2  (bases 1 to 11915)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 17, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.molcel.2023.06.028"; journalName: "Mol Cell"; date: 
            "2023-08-3- 3"; volume: "83"; issue: "15"; pages: "2753-2767.e10"
COMMENT     Sequence Label: pZucchini2_SFFV-RARA-eGFP-IRES-mCherry_PuroR
FEATURES             Location/Qualifiers
     source          1..11915
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     16..33
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    150..171
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(186..191,192..209,210..216)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    224..240
                     /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     229..251
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     248..264
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     248..264
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     283..303
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        285..303
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        331..557
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             558..738
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    785..910
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1403..1636
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1821..1865
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     promoter        2067..2473
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     regulatory      2511..2520
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3933..3935,3936..4649)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(3933..3935,3936..4646)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(3975..3996)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4236..4255)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4583..4604
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    join(4673..5247,5248..5250,5251..5259)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4840..4857)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5067..5086
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     regulatory      5269..5278
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5275..5985
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(5420..5438)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     5688..5707
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    6041..6158
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        6292..7470
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          6523..7461
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     7418..7438
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             7483..8082
                     /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(7483..7502)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7979..7999
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    8098..8686
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8151..8171)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8569..8580)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8758..8991
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    9069..9190
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9106..9125)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9160..9179
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      9230..9365
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(9385..9404)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(9386..9404)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(9414..9431)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(9414..9430)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9423..9445)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      9572..10027
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9659..9678)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9869..9890
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        10053..10157
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10158..10226,10227..11018)
                     /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     complement(10376..10395)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      11189..11777
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11678..11697
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
       61 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
      121 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc
      181 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa
      241 tttcacacag gaaacagcta tgaccatgat tacgccaagc gcgcaattaa ccctcactaa
      301 agggaacaaa agctggagct gcaagcttaa tgtagtctta tgcaatactc ttgtagtctt
      361 gcaacatggt aacgatgagt tagcaacatg ccttacaagg agagaaaaag caccgtgcat
      421 gccgattggt ggaagtaagg tggtacgatc gtgccttatt aggaaggcaa cagacgggtc
      481 tgacatggat tggacgaacc actgaattgc cgcattgcag agatattgta tttaagtgcc
      541 tagctcgata cataaacggg tctctctggt tagaccagat ctgagcctgg gagctctctg
      601 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
      661 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
      721 tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga
      781 ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg
      841 gcgactggtg agtacgccaa aaattttgac tagcggaggc tagaaggaga gagatgggtg
      901 cgagagcgtc agtattaagc gggggagaat tagatcgcga tgggaaaaaa ttcggttaag
      961 gccaggggga aagaaaaaat ataaattaaa acatatagta tgggcaagca gggagctaga
     1021 acgattcgca gttaatcctg gcctgttaga aacatcagaa ggctgtagac aaatactggg
     1081 acagctacaa ccatcccttc agacaggatc agaagaactt agatcattat ataatacagt
     1141 agcaaccctc tattgtgtgc atcaaaggat agagataaaa gacaccaagg aagctttaga
     1201 caagatagag gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg ccgctgatct
     1261 tcagacctgg aggaggagat atgagggaca attggagaag tgaattatat aaatataaag
     1321 tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga
     1381 gagaaaaaag agcagtggga ataggagctt tgttccttgg gttcttggga gcagcaggaa
     1441 gcactatggg cgcagcgtca atgacgctga cggtacaggc cagacaatta ttgtctggta
     1501 tagtgcagca gcagaacaat ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac
     1561 tcacagtctg gggcatcaag cagctccagg caagaatcct ggctgtggaa agatacctaa
     1621 aggatcaaca gctcctgggg atttggggtt gctctggaaa actcatttgc accactgctg
     1681 tgccttggaa tgctagttgg agtaataaat ctctggaaca gatttggaat cacacgacct
     1741 ggatggagtg ggacagagaa attaacaatt acacaagctt aatacactcc ttaattgaag
     1801 aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt ggaattagat aaatgggcaa
     1861 gtttgtggaa ttggtttaac ataacaaatt ggctgtggta tataaaatta ttcataatga
     1921 tagtaggagg cttggtaggt ttaagaatag tttttgctgt actttctata gtgaatagag
     1981 ttaggcaggg atattcacca ttatcgtttc agacccacct cccaaccccg aggggaccct
     2041 attccagcac atatgagcta gctgcagtaa cgccattttg caaggcatgg aaaaatacca
     2101 aaccaagaat agagaagttc agatcaaggg cgggtacatg aaaatagcta acgttgggcc
     2161 aaacaggata tctgcggtga gcagtttcgg ccccggcccg gggccaagaa cagatggtca
     2221 ccgcagtttc ggccccggcc cgaggccaag aacagatggt ccccagatat ggcccaaccc
     2281 tcagcagttt cttaagaccc atcagatgtt tccaggctcc cccaaggacc tgaaatgacc
     2341 ctgcgcctta tttgaattaa ccaatcagcc tgcttctcgc ttctgttcgc gcgcttctgc
     2401 ttcccgagct ctataaaaga gctcacaacc cctcactcgg cgcgccagtc ctccgacaga
     2461 ctgagtcggc cggtcgaatc aagcttatcg ataccgtcga ctccggaata gccaccatgg
     2521 cctctaattc ttcttcttgc ccaactccag gcggtggcca tttgaatggg tatcccgtgc
     2581 caccctatgc gttcttcttt cctccaatgc tgggtggcct gtcacctccg ggtgctctca
     2641 caactctcca acatcaactc cccgtttcag gctatagcac gccgagcccg gcgacaatcg
     2701 aaacgcaatc ttcaagctca gaggagatcg ttccctctcc gccaagtccg ccccccttgc
     2761 cgcgcatata caaaccttgt ttcgtgtgtc aggacaaatc ctcaggatat cattacgggg
     2821 tttccgcctg cgagggatgc aagggcttct tccgacgatc aatccaaaag aatatggtct
     2881 acacttgtca ccgagataag aactgcatca tcaacaaagt cacgcggaat aggtgccaat
     2941 actgtcgact gcagaaatgc tttgaagtag gaatgagcaa agagagcgtt cgcaacgaca
     3001 gaaataagaa gaaaaaggaa gttccgaaac ccgaatgtag tgagtcatat actctgaccc
     3061 ctgaggtcgg cgaactcata gagaaagttc gaaaggcgca tcaagagaca tttcccgccc
     3121 tttgtcaact ggggaaatat accacaaata atagttctga gcaacgagtt tccctggata
     3181 tcgatttgtg ggataagttt tccgagctgt ctactaagtg cattataaag acagtcgagt
     3241 tcgcaaaaca gctgcccggg tttactacgt tgaccatagc agatcagata acgctcttga
     3301 aagcggcgtg cctcgatatt ctcatactgc ggatctgtac gagatatacg ccagagcaag
     3361 atacaatgac tttttccgat gggctcaccc ttaacagaac gcaaatgcat aatgcgggct
     3421 ttggcccatt gaccgacctc gtatttgcct tcgctaatca acttctccct ctcgagatgg
     3481 acgatgccga aaccggcctg ctgtccgcta tatgcttgat ctgcggagat cgccaggacc
     3541 tggaacagcc cgatcgcgta gacatgctcc aggaacccct tcttgaagcc ttgaaagtgt
     3601 acgtgcggaa gcggaggcca tcacggccac acatgtttcc caaaatgttg atgaagatta
     3661 ccgatctgag aagcatcagt gcgaagggcg ctgagagggt gatcacgctc aaaatggaga
     3721 tccccggttc tatgccgcca cttatacaag agatgttgga gaattctgag gggctggaca
     3781 cattgtctgg acagcctgga ggtggcggcc gagatggagg gggactggca ccaccccccg
     3841 gaagttgttc cccaagcttg agcccctctt caaatcgaag cagccctgca actcactctc
     3901 caggcggcgg cggctccggc ggcggcggct ccgtgagcaa gggcgaggag ctgttcaccg
     3961 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt
     4021 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca
     4081 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt
     4141 gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
     4201 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg
     4261 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
     4321 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg
     4381 tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca
     4441 acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
     4501 acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag
     4561 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca
     4621 ctctcggcat ggacgagctg tacaagtaaa tgcatgagta actgaggatc cgcccctctc
     4681 cctccccccc ccctaacgtt actggccgaa gccgcttgga ataaggccgg tgtgcgtttg
     4741 tctatatgtt attttccacc atattgccgt cttttggcaa tgtgagggcc cggaaacctg
     4801 gccctgtctt cttgacgagc attcctaggg gtctttcccc tctcgccaaa ggaatgcaag
     4861 gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga caaacaacgt
     4921 ctgtagcgac cctttgcagg cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc
     4981 aaaagccacg tgtataagat acacctgcaa aggcggcaca accccagtgc cacgttgtga
     5041 gttggatagt tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac aaggggctga
     5101 aggatgccca gaaggtaccc cattgtatgg gatctgatct ggggcctcgg tacacatgct
     5161 ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg
     5221 ttttcctttg aaaaacacga tgataatatg gccacaaccc tggaattcgc caccatggtg
     5281 agcaagggcg aggaggataa catggccatc atcaaggagt tcatgcgctt caaggtgcac
     5341 atggagggct ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc
     5401 tacgagggca cccagaccgc caagctgaag gtgaccaagg gtggccccct gcccttcgcc
     5461 tgggacatcc tgtcccctca gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc
     5521 gacatccccg actacttgaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg
     5581 aacttcgagg acggcggcgt ggtgaccgtg acccaggact cctccctgca ggacggcgag
     5641 ttcatctaca aggtgaagct gcgcggcacc aacttcccct ccgacggccc cgtaatgcag
     5701 aagaagacca tgggctggga ggcctcctcc gagcggatgt accccgagga cggcgccctg
     5761 aagggcgaga tcaagcagag gctgaagctg aaggacggcg gccactacga cgctgaggtc
     5821 aagaccacct acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc
     5881 aagttggaca tcacctccca caacgaggac tacaccatcg tggaacagta cgaacgcgcc
     5941 gagggccgcc actccaccgg cggcatggac gagctgtaca agtaaactag taagcttggc
     6001 gtaactagat cttgagacaa atggcagtat tcatccacaa ttttaaaaga aaagggggga
     6061 ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac atacaaacta
     6121 aagaattaca aaaacaaatt acaaaaattc aaaattttcg ggtttattac agggacagca
     6181 gagatccact ttgggctcga gggggcccgg gtgcaaagat ggataaagtt ttaaacagag
     6241 aggaatcttt gcagctaatg gaccttctag gtcttgaaag gagtgggaat tggctccggt
     6301 gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc
     6361 ggcaattgat ccggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg
     6421 tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc
     6481 gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc cgtgtgtggt
     6541 tcccgcgggc ctggcctctt tacgggttat ggcccttgcg tgccttgaat tacttccacc
     6601 tggctgcagt acgtgattct tgatcccgag cttcgggttg gaagtgggtg ggagagttcg
     6661 aggccttgcg cttaaggagc cccttcgcct cgtgcttgag ttgaggcctg gcctgggcgc
     6721 tggggccgcc gcgtgcgaat ctggtggcac cttcgcgcct gtctcgctgc tttcgataag
     6781 tctctagcca tttaaaattt ttgatgacct gctgcgacgc tttttttctg gcaagatagt
     6841 cttgtaaatg cgggccaaga tctgcacact ggtatttcgg tttttggggc cgcgggcggc
     6901 gacggggccc gtgcgtccca gcgcacatgt tcggcgaggc ggggcctgcg agcgcggcca
     6961 ccgagaatcg gacgggggta gtctcaagct ggccggcctg ctctggtgcc tggcctcgcg
     7021 ccgccgtgta tcgccccgcc ctgggcggca aggctggccc ggtcggcacc agttgcgtga
     7081 gcggaaagat ggccgcttcc cggccctgct gcagggagct caaaatggag gacgcggcgc
     7141 tcgggagagc gggcgggtga gtcacccaca caaaggaaaa gggcctttcc gtcctcagcc
     7201 gtcgcttcat gtgactccac ggagtaccgg gcgccgtcca ggcacctcga ttagttctcg
     7261 agcttttgga gtacgtcgtc tttaggttgg ggggaggggt tttatgcgat ggagtttccc
     7321 cacactgagt gggtggagac tgaagttagg ccagcttggc acttgatgta attctccttg
     7381 gaatttgccc tttttgagtt tggatcttgg ttcattctca agcctcagac agtggttcaa
     7441 agtttttttc ttccatttca ggtgtcgtga cgtacggcca ccatgaccga gtacaagccc
     7501 acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg
     7561 ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc
     7621 accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc
     7681 gcggacgacg gcgccgccgt ggcggtctgg accacgccgg agagcgtcga agcgggggcg
     7741 gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag
     7801 caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc
     7861 accgtcggag tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc
     7921 ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc
     7981 aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa
     8041 ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gaacgcgtta agtcgacaat
     8101 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
     8161 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
     8221 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg
     8281 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt
     8341 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt
     8401 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     8461 ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc
     8521 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat
     8581 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc
     8641 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcgtcg actttaagac
     8701 caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg
     8761 aagggctaat tcactcccaa cgaagacaag atctgctttt tgcttgtact gggtctctct
     8821 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
     8881 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
     8941 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agtacgtata
     9001 gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga
     9061 gtgagaggaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     9121 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     9181 atgtatctta tcatgtctgg ctctagctat cccgccccta actccgccca tcccgcccct
     9241 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     9301 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     9361 aggcctaggg acgtacccaa ttcgccctat agtgagtcgt attacgcgcg ctcactggcc
     9421 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca
     9481 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
     9541 caacagttgc gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg
     9601 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     9661 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     9721 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     9781 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     9841 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     9901 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     9961 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt
    10021 acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
    10081 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
    10141 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
    10201 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
    10261 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
    10321 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
    10381 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
    10441 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
    10501 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
    10561 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
    10621 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
    10681 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
    10741 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
    10801 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
    10861 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
    10921 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
    10981 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
    11041 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
    11101 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
    11161 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
    11221 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
    11281 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
    11341 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
    11401 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
    11461 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
    11521 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
    11581 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
    11641 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
    11701 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
    11761 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
    11821 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
    11881 ccgcctttga gtgagctgat accgctcgcc gcagc
//
