LOCUS       sequence-496528-       13070 bp    DNA     circular SYN 07-JAN-2026
DEFINITION  Overexpression vector for eGFP tagged GR or NR3C1. Monitor 
            post-translational degradation of GR via EGFP:mCherry ratio. 
            Contains neomycin-resistance selection marker..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13070)
  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 13070)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 7, 2026 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: pSquash2_SFFV-eGFP-NR3C1-IRES-mCherry_NeomycinR
FEATURES             Location/Qualifiers
     source          1..13070
                     /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(2517..2519,2520..2522,2523..3233)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2517..2519,2520..2522,2523..3233)
                     /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(2562..2583)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2823..2842)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3170..3191
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     RBS             5455..5463
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     misc_feature    join(5624..6198,6199..6201,6202..6210)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5791..5808)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     6018..6037
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     regulatory      6220..6229
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6226..6936
                     /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(6371..6389)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     6639..6658
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    6992..7109
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        7243..8421
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          7474..8412
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     8369..8389
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             8434..9225
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(8488..8507)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     9098..9117
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     misc_feature    9253..9841
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9306..9326)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9724..9735)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9913..10146
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    10224..10345
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10261..10280)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10315..10334
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      10385..10520
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(10540..10559)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(10541..10559)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(10569..10586)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(10569..10585)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10578..10600)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      10727..11182
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10814..10833)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11024..11045
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        11208..11312
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(11313..11381,11382..12173)
                     /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(11531..11550)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      12344..12932
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     12833..12852
                     /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 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
     2581 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
     2641 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
     2701 tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc
     2761 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
     2821 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
     2881 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
     2941 tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca
     3001 tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc
     3061 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     3121 tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     3181 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagggcggcg
     3241 gcggctccgg cggcggcggc tccatggact ctaaagagtc tctgacgccg gggagagagg
     3301 aaaatccttc atcagtgctc gcgcaggagc gcggagatgt catggatttt tataagacgc
     3361 tcaggggcgg ggcgacggtg aaagtgtccg catcctctcc cagcttggcg gtcgcgtccc
     3421 aatctgactc taagcagcgc agattgctcg tggacttccc taaagggtct gtaagcaatg
     3481 cccagcaacc ggatctgtca aaagctgtca gtctctcaat gggcctgtac atgggcgaaa
     3541 ccgaaaccaa agtgatgggc aacgacctcg gttttcccca gcagggccaa atttcactgt
     3601 ctagcggtga gacagatctt aagctccttg aagaatcaat tgcgaacctc aatcgctcta
     3661 cctcagtccc ggagaatccc aaaagtagtg cctctacggc ggtgagcgcc gcaccaacag
     3721 agaaagaatt cccgaaaaca cattccgacg tatccagtga gcagcaacat cttaagggac
     3781 agaccggaac aaatggtggt aatgtcaaac tttataccac agaccagtca acgtttgaca
     3841 ttctccagga tctggaattt agtagtggat cccccgggaa ggaaacaaac gagtcaccgt
     3901 ggcgctctga cctcctgatc gatgagaatt gccttctgtc accgctggcg ggggaggatg
     3961 acagctttct gttggaaggt aattccaatg aagactgcaa acctcttata ctgcctgata
     4021 ctaagccgaa aatcaaggac aatggagatc tcgtactcag ttcaccgagt aacgtaactc
     4081 tccctcaggt gaagactgaa aaggaggatt tcatagagct ctgcacgcct ggtgtaatca
     4141 agcaggagaa gctcggtacc gtgtactgcc aggcttcctt tccaggagcg aatataattg
     4201 gaaataaaat gagtgcaatt agtgtacatg gggtatcaac aagcggaggt caaatgtacc
     4261 attatgatat gaatactgca tctttgtcac agcaacagga ccaaaaacct atatttaatg
     4321 tcatcccccc tatacccgta gggtcagaga actggaatcg gtgtcaaggc tcaggtgacg
     4381 acaacctgac ctcattgggg acattgaatt ttcccggaag aacggttttt tctaacgggt
     4441 atagtagtcc ttccatgaga ccagacgtgt ctagtccccc ttcttccagt agtactgcca
     4501 ctacaggccc gcctcctaaa ttgtgtctgg tgtgctctga cgaggcttcc ggttgtcatt
     4561 atggtgtatt gacctgtggt agctgtaaag ttttttttaa aagagctgta gaaggtcaac
     4621 acaattacct ctgtgccgga cgaaatgact gcatcatcga taaaattcgg cggaaaaact
     4681 gcccggcgtg ccggtatcgg aaatgcctgc aagctggtat gaatcttgaa gcacgcaaga
     4741 ctaagaagaa gattaaaggg atccagcagg cgacaaccgg agtctcccaa gagacttccg
     4801 agaacccagg taataaaacc atcgtaccag cgacgcttcc acaactcaca ccaactcttg
     4861 tgtctttgct tgaggtgatc gagccagaag tcttgtatgc tgggtacgac tcttcagttc
     4921 ctgatagcac ttggagaatt atgactaccc ttaacatgct tggaggccga caagttattg
     4981 cagcggtcaa gtgggccaaa gctatcccag gctttcgaaa tctccacctc gacgaccaga
     5041 tgacgttgtt gcaatactcc tggatgtttc tcatggcgtt cgccttgggc tggagatcct
     5101 acagacagag cagtgcgaac ttgctctgtt ttgcgccaga cttgataatc aatgaacagc
     5161 gaatgacatt gccctgtatg tatgatcaat gcaaacatat gctgtatgtt tcttctgaat
     5221 tgcatcggct gcaagttagt tacgaggagt atctgtgcat gaagaccctt ttgcttctct
     5281 catcagtacc aaaagacggt ttgaagagcc aggagctctt tgatgagatc agaatgactt
     5341 atattaagga gctcggcaag gccattgtaa aacgagaagg caacagtagt caaaattggc
     5401 aaagattcta ccagctgact aagctcctcg attcaatgca cgaagtggtt gagaacctcc
     5461 ttaattactg cttccagacg ttcctcgata agacaatgag tatagaattc ccggagatgc
     5521 tcgcggaaat catcaccaat cagattccca agtactcaaa cggtaatata aaaaaacttc
     5581 tgttccacca aaaatgataa atgcatgagt aactgaggat ccgcccctct ccctcccccc
     5641 cccctaacgt tactggccga agccgcttgg aataaggccg gtgtgcgttt gtctatatgt
     5701 tattttccac catattgccg tcttttggca atgtgagggc ccggaaacct ggccctgtct
     5761 tcttgacgag cattcctagg ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga
     5821 atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag acaaacaacg tctgtagcga
     5881 ccctttgcag gcagcggaac cccccacctg gcgacaggtg cctctgcggc caaaagccac
     5941 gtgtataaga tacacctgca aaggcggcac aaccccagtg ccacgttgtg agttggatag
     6001 ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg aaggatgccc
     6061 agaaggtacc ccattgtatg ggatctgatc tggggcctcg gtacacatgc tttacatgtg
     6121 tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg gttttccttt
     6181 gaaaaacacg atgataatat ggccacaacc ctggaattcg ccaccatggt gagcaagggc
     6241 gaggaggata acatggccat catcaaggag ttcatgcgct tcaaggtgca catggagggc
     6301 tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc
     6361 acccagaccg ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc
     6421 ctgtcccctc agttcatgta cggctccaag gcctacgtga agcaccccgc cgacatcccc
     6481 gactacttga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag
     6541 gacggcggcg tggtgaccgt gacccaggac tcctccctgc aggacggcga gttcatctac
     6601 aaggtgaagc tgcgcggcac caacttcccc tccgacggcc ccgtaatgca gaagaagacc
     6661 atgggctggg aggcctcctc cgagcggatg taccccgagg acggcgccct gaagggcgag
     6721 atcaagcaga ggctgaagct gaaggacggc ggccactacg acgctgaggt caagaccacc
     6781 tacaaggcca agaagcccgt gcagctgccc ggcgcctaca acgtcaacat caagttggac
     6841 atcacctccc acaacgagga ctacaccatc gtggaacagt acgaacgcgc cgagggccgc
     6901 cactccaccg gcggcatgga cgagctgtac aagtaaacta gtaagcttgg cgtaactaga
     6961 tcttgagaca aatggcagta ttcatccaca attttaaaag aaaagggggg attggggggt
     7021 acagtgcagg ggaaagaata gtagacataa tagcaacaga catacaaact aaagaattac
     7081 aaaaacaaat tacaaaaatt caaaattttc gggtttatta cagggacagc agagatccac
     7141 tttgggctcg agggggcccg ggtgcaaaga tggataaagt tttaaacaga gaggaatctt
     7201 tgcagctaat ggaccttcta ggtcttgaaa ggagtgggaa ttggctccgg tgcccgtcag
     7261 tgggcagagc gcacatcgcc cacagtcccc gagaagttgg ggggaggggt cggcaattga
     7321 tccggtgcct agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt gtactggctc
     7381 cgcctttttc ccgagggtgg gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt
     7441 ctttttcgca acgggtttgc cgccagaaca caggtaagtg ccgtgtgtgg ttcccgcggg
     7501 cctggcctct ttacgggtta tggcccttgc gtgccttgaa ttacttccac ctggctgcag
     7561 tacgtgattc ttgatcccga gcttcgggtt ggaagtgggt gggagagttc gaggccttgc
     7621 gcttaaggag ccccttcgcc tcgtgcttga gttgaggcct ggcctgggcg ctggggccgc
     7681 cgcgtgcgaa tctggtggca ccttcgcgcc tgtctcgctg ctttcgataa gtctctagcc
     7741 atttaaaatt tttgatgacc tgctgcgacg ctttttttct ggcaagatag tcttgtaaat
     7801 gcgggccaag atctgcacac tggtatttcg gtttttgggg ccgcgggcgg cgacggggcc
     7861 cgtgcgtccc agcgcacatg ttcggcgagg cggggcctgc gagcgcggcc accgagaatc
     7921 ggacgggggt agtctcaagc tggccggcct gctctggtgc ctggcctcgc gccgccgtgt
     7981 atcgccccgc cctgggcggc aaggctggcc cggtcggcac cagttgcgtg agcggaaaga
     8041 tggccgcttc ccggccctgc tgcagggagc tcaaaatgga ggacgcggcg ctcgggagag
     8101 cgggcgggtg agtcacccac acaaaggaaa agggcctttc cgtcctcagc cgtcgcttca
     8161 tgtgactcca cggagtaccg ggcgccgtcc aggcacctcg attagttctc gagcttttgg
     8221 agtacgtcgt ctttaggttg gggggagggg ttttatgcga tggagtttcc ccacactgag
     8281 tgggtggaga ctgaagttag gccagcttgg cacttgatgt aattctcctt ggaatttgcc
     8341 ctttttgagt ttggatcttg gttcattctc aagcctcaga cagtggttca aagttttttt
     8401 cttccatttc aggtgtcgtg acgtacggcc accatgattg aacaagatgg attgcacgca
     8461 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     8521 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     8581 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg
     8641 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     8701 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     8761 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     8821 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     8881 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     8941 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc
     9001 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     9061 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     9121 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     9181 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttcctgca gggataaacg
     9241 cgttaagtcg acaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     9301 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     9361 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
     9421 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
     9481 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
     9541 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     9601 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt
     9661 ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
     9721 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
     9781 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
     9841 cgtcgacttt aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag
     9901 aaaagggggg actggaaggg ctaattcact cccaacgaag acaagatctg ctttttgctt
     9961 gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc taactaggga
    10021 acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc
    10081 tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct
    10141 ctagcagtac gtatagtagt tcatgtcatc ttattattca gtatttataa cttgcaaaga
    10201 aatgaatatc agagagtgag aggaacttgt ttattgcagc ttataatggt tacaaataaa
    10261 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
    10321 tgtccaaact catcaatgta tcttatcatg tctggctcta gctatcccgc ccctaactcc
    10381 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
    10441 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
    10501 aggaggcttt tttggaggcc tagggacgta cccaattcgc cctatagtga gtcgtattac
    10561 gcgcgctcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
    10621 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
    10681 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc gccctgtagc
    10741 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
    10801 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
    10861 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
    10921 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
    10981 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
    11041 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
    11101 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
    11161 aaaatattaa cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta
    11221 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
    11281 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
    11341 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
    11401 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
    11461 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
    11521 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
    11581 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
    11641 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
    11701 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
    11761 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
    11821 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
    11881 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
    11941 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
    12001 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
    12061 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
    12121 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
    12181 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
    12241 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
    12301 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
    12361 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
    12421 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
    12481 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
    12541 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
    12601 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
    12661 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
    12721 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
    12781 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
    12841 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
    12901 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
    12961 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
    13021 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
//
