LOCUS       sequence-484638-       11623 bp    DNA     circular SYN 26-AUG-2025
DEFINITION  Expresses TEV protease and tandemly-repeated GFP fluorescent 
            proteins to amplify the fluorescence signal produced by a single 
            transcription unit of MALAT1 ENE(WT)-mascRNA(mut 8356-8370) 
            reporter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11623)
  AUTHORS   Che R, Panah M, Mirani B, Knowles K, Ostapovich A, Majumdar D, Chen 
            X, DeSimone J, White W, Noonan M, Luo H, Alexandrov A
  TITLE     Identification of human pathways acting on nuclear non-coding RNAs 
            using the Mirror forward genetic approach.
  JOURNAL   Nat Commun. 2025 May 22;16(1):4741. doi: 10.1038/s41467-025-59998-3.
  PUBMED    40399278
REFERENCE   2  (bases 1 to 11623)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 26, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2025-05-22"; volume: "16(1):4741. doi"; pages: " 
            10.1038/s41467-025-59998-3"
COMMENT     Sequence Label: Mirror GFP[ENE(WT)-mascRNA(mut 8356-8370)](pAV...
FEATURES             Location/Qualifiers
     source          1..11623
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      127..715
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     616..635
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     869..886
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1003..1024
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        1462..2644
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          1693..2635
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     2592..2612
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      2694..2703
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2700..2702,2703..2705,2706..3416)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2700..2702,2703..2705,2706..3416)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             3429..3449
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             join(3450..3452,3453..3455,3456..4166)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3450..3452,3453..3455,3456..4166)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             4179..4199
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             join(4200..4202,4203..4205,4206..4916)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(4200..4202,4203..4205,4206..4916)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             4929..4949
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             join(4950..4952,4953..4955,4956..5666)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(4950..4952,4953..4955,4956..5666)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             5679..5699
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             join(5700..5702,5703..5705,5706..6416)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(5700..5702,5703..5705,5706..6416)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             6417..6437
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             6438..7145
                     /codon_start=1
                     /product="tobacco etch virus protease (Kapust et al., 
                     2001)"
                     /label=TEV protease
                     /translation="GESLFKGPRDYNPISSTICHLTNESDGHTTSLYGIGFGPFIITNK
                     HLFRRNNGTLLVQSLHGVFKVKNTTTLQQHLIDGRDMIIIRMPKDFPPFPQKLKFREPQ
                     REERICLVTTNFQTKSMSSMVSDTSCTFPSSDGIFWKHWIQTKDGQCGSPLVSTRDGFI
                     VGIHSASNFTNTNNYFTSVPKNFMELLTNQEAQQWVSGWRLNADSVLWGGHKVFMSKPE
                     EPFQPVKEATQLMN"
     CDS             7164..7184
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     polyA_signal    8913..9137
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(9270..9289)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    9530..9577
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             9732..9752
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             9753..10349
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="TEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERV
                     TELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAA
                     QQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSA
                     PRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     10246..10266
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    10375..10582
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        10614..10718
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10719..10787,10788..11579)
                     /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(10937..10956)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
       61 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
      121 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
      181 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
      241 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
      301 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
      361 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
      421 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
      481 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
      541 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
      601 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
      661 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
      721 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
      781 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
      841 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
      901 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
      961 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac
     1021 tcattaggca ccccaggcac gcgaagcgtt gttggtttgt agtgcccggt ttcgaaccgg
     1081 ggacctttcg cgtgttaggc gaacgtgata accactacac tacggaaacc aacggtgcta
     1141 gacgcgtatt gtctattctg actcggatcc gtacgaattc tcatgtttga cagcttatca
     1201 tcgattagct ttggagctaa gccagcaatg gtagagggaa gattctgcac gtcccttcca
     1261 ggcggcctcc ccgtcaccac cccccccaac ccgccccgac cggagctgag agtaattcat
     1321 acaaaaggac tcgcccctgc cttggggaat cccagggacc gtcgttaaac tcccactaac
     1381 gtagaaccca gagatcgctg cgttcccgcc ccctcacccg cccgctctcg tcatcactga
     1441 ggtggagaag agcatgcgtg aggctccggt gcccgtcagt gggcagagcg cacatcgccc
     1501 acagtccccg agaagttggg gggaggggtc ggcaattgaa ccggtgccta gagaaagtgg
     1561 cgcggggtaa actgggaaag tgatgtcgtg tactggctcc gcctttttcc cgagggtggg
     1621 ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc
     1681 gccagaacac aggtaagtgc cgtgtgtggt tcccgcgggc ctggcctctt tacgggttat
     1741 ggcccttgcg tgccttgaat tacttccacg cccctggctg cagtacgtga ttcttgatcc
     1801 cgagcttcgg gttggaagtg ggtgggagag ttcgaggcct tgcgcttaag gagccccttc
     1861 gcctcgtgct tgagttgagg cctggcctgg gcgctggggc cgccgcgtgc gaatctggtg
     1921 gcaccttcgc gcctgtctcg ctgctttcga taagtctcta gccatttaaa atttttgatg
     1981 acctgctgcg acgctttttt tctggcaaga tagtcttgta aatgcgggcc aagatctgca
     2041 cactggtatt tcggtttttg gggccgcggg cggcgacggg gcccgtgcgt cccagcgcac
     2101 atgttcggcg aggcggggcc tgcgagcgcg gccaccgaga atcggacggg ggtagtctca
     2161 agctggccgg cctgctctgg tgcctggcct cgcgccgccg tgtatcgccc cgccctgggc
     2221 ggcaaggctg gcccggtcgg caccagttgc gtgagcggaa agatggccgc ttcccggccc
     2281 tgctgcaggg agctcaaaat ggaggacgcg gcgctcggga gagcgggcgg gtgagtcacc
     2341 cacacaaagg aaaagggcct ttccgtcctc agccgtcgct tcatgtgact ccacggagta
     2401 ccgggcgccg tccaggcacc tcgattagtt ctcgagcttt tggagtacgt cgtctttagg
     2461 ttggggggag gggttttatg cgatggagtt tccccacact gagtgggtgg agactgaagt
     2521 taggccagct tggcacttga tgtaattctc cttggaattt gccctttttg agtttggatc
     2581 ttggttcatt ctcaagcctc agacagtggt tcaaagtttt tttcttccat ttcaggtgtc
     2641 gtgaaaactc tagcgtttcc gctctagaac tagtccagct agcgctaccg gtcgccacca
     2701 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
     2761 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg
     2821 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc
     2881 tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc
     2941 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
     3001 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
     3061 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
     3121 agctggagta caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg
     3181 gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg
     3241 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
     3301 acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc
     3361 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaaggtca
     3421 ctccgtgtga aaacctgtac ttccaaggta tggtgagcaa gggcgaggag ctgttcaccg
     3481 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt
     3541 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca
     3601 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt
     3661 gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
     3721 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg
     3781 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
     3841 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg
     3901 tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca
     3961 acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
     4021 acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag
     4081 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca
     4141 ctctcggcat ggacgagctg tacaagacca ctgcgtgcga aaacctgtac ttccaaggta
     4201 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
     4261 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg
     4321 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc
     4381 tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc
     4441 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
     4501 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
     4561 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
     4621 agctggagta caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg
     4681 gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg
     4741 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
     4801 acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc
     4861 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaaggtca
     4921 ctacgtgtga aaacctgtac ttccaaggta tggtgagcaa gggcgaggag ctgttcaccg
     4981 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt
     5041 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca
     5101 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt
     5161 gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
     5221 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg
     5281 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
     5341 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg
     5401 tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca
     5461 acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
     5521 acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag
     5581 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca
     5641 ctctcggcat ggacgagctg tacaagacca cttcgtgcga aaacctgtac ttccaaggta
     5701 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
     5761 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg
     5821 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc
     5881 tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc
     5941 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
     6001 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
     6061 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
     6121 agctggagta caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg
     6181 gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg
     6241 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
     6301 acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc
     6361 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaaggaaa
     6421 acctgtactt ccaaagtgga gaaagcttgt ttaagggacc acgtgattac aacccgatat
     6481 cgagcaccat ttgtcatttg acgaatgaat ctgatgggca cacaacatcg ttgtatggta
     6541 ttggatttgg tcccttcatc attacaaaca agcacttgtt tagaagaaat aatggaacac
     6601 tgttggtcca atcactacat ggtgtattca aggtcaagaa caccacgact ttgcaacaac
     6661 acctcattga tgggagggac atgataatta ttcgcatgcc taaggatttc ccaccatttc
     6721 ctcaaaagct gaaatttaga gagccacaaa gggaagagcg catatgtctt gtgacaacca
     6781 acttccaaac taagagcatg tctagcatgg tgtcagacac tagttgcaca ttcccttcat
     6841 ctgatggcat attctggaag cattggattc aaaccaagga tgggcagtgt ggcagtccat
     6901 tagtatcaac tagagatggg ttcattgttg gtatacactc agcatcgaat ttcaccaaca
     6961 caaacaatta tttcacaagc gtgccgaaaa acttcatgga attgttgaca aatcaggagg
     7021 cgcagcagtg ggttagtggt tggcgattaa atgctgactc agtattgtgg gggggccata
     7081 aagttttcat gagcaaacct gaagagcctt ttcagccagt taaggaagcg actcaactca
     7141 tgaatgaatt ggtgtactcg caagaaaacc tgtacttcca aagtatgcat agatcacgag
     7201 atatcagcca tggcttcccg ccggcggtgg cggcgcagga tgatggcacg ctgcccatgt
     7261 cttgtgccca ggagagcggg atggaccgtc accctgcagc ctgtgcttct gctaggatca
     7321 atgtggagct cgtgcacctg actcctgagg agaagtctgc cgttactgcc ctgtggggca
     7381 aggtgaacgt ggatgaagtt ggtggtgagg ccctgggcag gctgctggtg gtctaccctt
     7441 ggacccagag gttctttgag tcctttgggg atctgtccac tcctgatgct gttatgggca
     7501 accctaaggt gaaggctcat ggcaagaaag tgctcggtgc ctttagtgat ggcctggctc
     7561 acctggacaa cctcaagggc acctttgcca cactgagtga gctgcactgt gacaagctgc
     7621 acgtggatcc tgagaacttc agggtgagtc tatgggaccc ttgatgtttt ctttcccctt
     7681 cttttctatg gttaagttca tgtcatagga aggggataag taacagggta cagtttagaa
     7741 tgggaaacag acgaatgatt gcatcagtgt ggaagtctca ggatcgtttt agtttctttt
     7801 atttgctgtt cataacaatt gttttctttt gtttaattct tgctttcttt ttttttcttc
     7861 tccgcaattt ttactattat acttaatgcc ttaacattgt gtataacaaa aggaaatatc
     7921 tctgagatac attaagtaac ttaaaaaaaa aactttacac agtctgccta gtacattact
     7981 atttggaata tatgtgtgct tatttgcata ttcataatct ccctacttta ttttctttta
     8041 tttttaattg atacataatc attatacata tttatgggtt aaagtgtaat gttttaatat
     8101 gtgtacacat attgaccaaa tcagggtaat tttgcatttg taattttaaa aaatgctttc
     8161 ttcttttaat atactttttt gtttatctta tttctaatac tttccctaat ctctttcttt
     8221 cagggcaata atgatacaat gtatcatgcc tctttgcacc attctaaaga ataacagtga
     8281 taatttctgg gttaaggcaa tagcaatatt tctgcatata aatatttctg catataaatt
     8341 gtaactgatg taagaggttt catattgcta atagcagcta caatccagct accattctgc
     8401 ttttatttta tggttgggat aaggctggat tattctgagt ccaagctagg cccttttgct
     8461 aatcatgttc atacctctta tcttcctccc acagctcctg ggcaacgtgc tggtctgtgt
     8521 gctggcccat cactttggca aagaattcac cccaccagtg caggctgcct atcagaaagt
     8581 ggtggctggt gtggctaatg ccctggccca caagtatcac taagcggccg ctcgctttct
     8641 tgctgtccaa tttctattaa aggttccttt gttccctaag tccaactact aaactggggg
     8701 atattatgaa gggcctcgag tagggtcatg aaggtttttc ttttcctgag aaaacaacac
     8761 gtattgtttt ctcaggtttt gctttttggc ctttttctag cttaaaaaaa aaaaaagcaa
     8821 aactacgacc accaacccac tcctggtttc caggacgggg ttcaaatccc tgcggcgtct
     8881 ttgctttgac tctcgaggct gatcagcctc gactgtgcct tctagttgcc agccatctgt
     8941 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     9001 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     9061 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
     9121 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc
     9181 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     9241 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     9301 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     9361 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtaggta
     9421 cgctagcacc gttggtttcc gtagtgtagt ggttatcacg ttcgcctaac acgcgaaagg
     9481 tccccggttc gaaaccgggc actacaaacc aacaacgctg gtacccctag aagttcctat
     9541 tccgaagttc ctattctcta gaaagtatag gaacttcctt ggccaaaaag cctgaaagat
     9601 cacgagatat cagccatggc ttcccgccgg cggtggcggc gcaggatgat ggcacgctgc
     9661 ccacgtcttg tgcccaggag agcgggacgg accgtcaccc tgcagcctgt gcttctgcta
     9721 ggatcaatgt ggaaaacctg tacttccaag gtaccgagta caagcccacg gtgcgcctcg
     9781 ccacccgcga cgacgtcccc agggccgtcc gcaccctcgc cgccgcgttc gccgactacc
     9841 ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag
     9901 aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg
     9961 ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga
    10021 tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag
    10081 gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct
    10141 cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg
    10201 ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct
    10261 acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct
    10321 ggtgcatgac ccgcaagccc ggtgcctgag gtaccgctcg ctgatcagcc tcgactgtgc
    10381 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
    10441 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
    10501 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
    10561 acaatagcag gcatgctggg gagtcgacag gtggcacttt tcggggaaat gtgcgcggaa
    10621 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
    10681 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
    10741 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
    10801 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
    10861 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
    10921 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
    10981 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
    11041 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
    11101 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
    11161 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
    11221 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
    11281 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
    11341 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
    11401 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
    11461 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
    11521 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
    11581 tgtcagacca agtttactca tatatacttt agattgattt aaa
//
