LOCUS       sequence-483539-       15469 bp    DNA     circular SYN 15-AUG-2025
DEFINITION  Expresses TEV protease and tandemly-repeated RFP fluorescent 
            proteins to amplify the fluorescence signal produced by a single 
            transcription unit of wild-type MALAT1 ENE-mascRNA reporter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15469)
  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 15469)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 15, 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 RFP[ENE(WT)-mascRNA](pAVA2987)
FEATURES             Location/Qualifiers
     source          1..15469
                     /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             2700..4127
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGIGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     CDS             4140..4160
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             4161..5588
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGIGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     CDS             5601..5621
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             5622..7049
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGIGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     CDS             7062..7082
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             7083..8510
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGIGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     CDS             8523..8543
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             8544..9971
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGIGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     CDS             9972..9992
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             9993..10700
                     /codon_start=1
                     /product="tobacco etch virus protease (Kapust et al., 
                     2001)"
                     /label=TEV protease
                     /translation="GESLFKGPRDYNPISSTICHLTNESDGHTTSLYGIGFGPFIITNK
                     HLFRRNNGTLLVQSLHGVFKVKNTTTLQQHLIDGRDMIIIRMPKDFPPFPQKLKFREPQ
                     REERICLVTTNFQTKSMSSMVSDTSCTFPSSDGIFWKHWIQTKDGQCGSPLVSTRDGFI
                     VGIHSASNFTNTNNYFTSVPKNFMELLTNQEAQQWVSGWRLNADSVLWGGHKVFMSKPE
                     EPFQPVKEATQLMN"
     CDS             10719..10739
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     primer_bind     complement(12462..12479)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    12468..12692
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(12825..12844)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    13085..13132
                     /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             13140..14162
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             13140..14162
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     polyA_signal    14292..14413
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(14329..14348)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     14383..14402
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        14460..14564
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(14565..14633,14634..15425)
                     /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(14783..14802)
                     /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 gtcatcaaag agttcatgcg cttcaaggtg cgcatggagg
     2761 gctccatgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg
     2821 gcacccagac cgccaagctg aaggtgacca agggcggccc cctgcccttc gcctgggaca
     2881 tcctgtcccc ccagttcatg tacggctcca aggcgtacgt gaagcacccc gccgacatcc
     2941 ccgattacaa gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg
     3001 aggacggcgg tctggtgacc gtgacccagg actcctccct gcaggacggc acgctgatct
     3061 acaaggtgaa gatgcgcggc accaacttcc cccccgacgg ccccgtaatg cagaagaaga
     3121 ccatgggctg ggaggcctcc accgagcgcc tgtacccccg cgacggcgtg ctgaagggcg
     3181 agatccacca ggccctgaag ctgaaggacg gcggccacta cctggtggag ttcaagacca
     3241 tctacatggc caagaagccc gtgcaactgc ccggctacta ctacgtggac accaagctgg
     3301 acatcacctc ccacaacgag gactacacca tcgtggaaca gtacgagcgc tccgagggcc
     3361 gccaccacct gttcctgggg catggcatcg gcagcaccgg cagcggcagc tccggcaccg
     3421 cctcctccga ggacaacaac atggccgtca tcaaagagtt catgcgcttc aaggtgcgca
     3481 tggagggctc catgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct
     3541 acgagggcac ccagaccgcc aagctgaagg tgaccaaggg cggccccctg cccttcgcct
     3601 gggacatcct gtccccccag ttcatgtacg gctccaaggc gtacgtgaag caccccgccg
     3661 acatccccga ttacaagaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga
     3721 acttcgagga cggcggtctg gtgaccgtga cccaggactc ctccctgcag gacggcacgc
     3781 tgatctacaa ggtgaagatg cgcggcacca acttcccccc cgacggcccc gtaatgcaga
     3841 agaagaccat gggctgggag gcctccaccg agcgcctgta cccccgcgac ggcgtgctga
     3901 agggcgagat ccaccaggcc ctgaagctga aggacggcgg ccactacctg gtggagttca
     3961 agaccatcta catggccaag aagcccgtgc aactgcccgg ctactactac gtggacacca
     4021 agctggacat cacctcccac aacgaggact acaccatcgt ggaacagtac gagcgctccg
     4081 agggccgcca ccacctgttc ctgtacggca tggacgagct gtacaaggtc actccgtgtg
     4141 aaaacctgta cttccaaggt atggtgagca agggcgagga ggtcatcaaa gagttcatgc
     4201 gcttcaaggt gcgcatggag ggctccatga acggccacga gttcgagatc gagggcgagg
     4261 gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggcggcc
     4321 ccctgccctt cgcctgggac atcctgtccc cccagttcat gtacggctcc aaggcgtacg
     4381 tgaagcaccc cgccgacatc cccgattaca agaagctgtc cttccccgag ggcttcaagt
     4441 gggagcgcgt gatgaacttc gaggacggcg gtctggtgac cgtgacccag gactcctccc
     4501 tgcaggacgg cacgctgatc tacaaggtga agatgcgcgg caccaacttc ccccccgacg
     4561 gccccgtaat gcagaagaag accatgggct gggaggcctc caccgagcgc ctgtaccccc
     4621 gcgacggcgt gctgaagggc gagatccacc aggccctgaa gctgaaggac ggcggccact
     4681 acctggtgga gttcaagacc atctacatgg ccaagaagcc cgtgcaactg cccggctact
     4741 actacgtgga caccaagctg gacatcacct cccacaacga ggactacacc atcgtggaac
     4801 agtacgagcg ctccgagggc cgccaccacc tgttcctggg gcatggcatc ggcagcaccg
     4861 gcagcggcag ctccggcacc gcctcctccg aggacaacaa catggccgtc atcaaagagt
     4921 tcatgcgctt caaggtgcgc atggagggct ccatgaacgg ccacgagttc gagatcgagg
     4981 gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag gtgaccaagg
     5041 gcggccccct gcccttcgcc tgggacatcc tgtcccccca gttcatgtac ggctccaagg
     5101 cgtacgtgaa gcaccccgcc gacatccccg attacaagaa gctgtccttc cccgagggct
     5161 tcaagtggga gcgcgtgatg aacttcgagg acggcggtct ggtgaccgtg acccaggact
     5221 cctccctgca ggacggcacg ctgatctaca aggtgaagat gcgcggcacc aacttccccc
     5281 ccgacggccc cgtaatgcag aagaagacca tgggctggga ggcctccacc gagcgcctgt
     5341 acccccgcga cggcgtgctg aagggcgaga tccaccaggc cctgaagctg aaggacggcg
     5401 gccactacct ggtggagttc aagaccatct acatggccaa gaagcccgtg caactgcccg
     5461 gctactacta cgtggacacc aagctggaca tcacctccca caacgaggac tacaccatcg
     5521 tggaacagta cgagcgctcc gagggccgcc accacctgtt cctgtacggc atggacgagc
     5581 tgtacaagac cactgcgtgc gaaaacctgt acttccaagg tatggtgagc aagggcgagg
     5641 aggtcatcaa agagttcatg cgcttcaagg tgcgcatgga gggctccatg aacggccacg
     5701 agttcgagat cgagggcgag ggcgagggcc gcccctacga gggcacccag accgccaagc
     5761 tgaaggtgac caagggcggc cccctgccct tcgcctggga catcctgtcc ccccagttca
     5821 tgtacggctc caaggcgtac gtgaagcacc ccgccgacat ccccgattac aagaagctgt
     5881 ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc ggtctggtga
     5941 ccgtgaccca ggactcctcc ctgcaggacg gcacgctgat ctacaaggtg aagatgcgcg
     6001 gcaccaactt cccccccgac ggccccgtaa tgcagaagaa gaccatgggc tgggaggcct
     6061 ccaccgagcg cctgtacccc cgcgacggcg tgctgaaggg cgagatccac caggccctga
     6121 agctgaagga cggcggccac tacctggtgg agttcaagac catctacatg gccaagaagc
     6181 ccgtgcaact gcccggctac tactacgtgg acaccaagct ggacatcacc tcccacaacg
     6241 aggactacac catcgtggaa cagtacgagc gctccgaggg ccgccaccac ctgttcctgg
     6301 ggcatggcat cggcagcacc ggcagcggca gctccggcac cgcctcctcc gaggacaaca
     6361 acatggccgt catcaaagag ttcatgcgct tcaaggtgcg catggagggc tccatgaacg
     6421 gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg
     6481 ccaagctgaa ggtgaccaag ggcggccccc tgcccttcgc ctgggacatc ctgtcccccc
     6541 agttcatgta cggctccaag gcgtacgtga agcaccccgc cgacatcccc gattacaaga
     6601 agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggtc
     6661 tggtgaccgt gacccaggac tcctccctgc aggacggcac gctgatctac aaggtgaaga
     6721 tgcgcggcac caacttcccc cccgacggcc ccgtaatgca gaagaagacc atgggctggg
     6781 aggcctccac cgagcgcctg tacccccgcg acggcgtgct gaagggcgag atccaccagg
     6841 ccctgaagct gaaggacggc ggccactacc tggtggagtt caagaccatc tacatggcca
     6901 agaagcccgt gcaactgccc ggctactact acgtggacac caagctggac atcacctccc
     6961 acaacgagga ctacaccatc gtggaacagt acgagcgctc cgagggccgc caccacctgt
     7021 tcctgtacgg catggacgag ctgtacaagg tcactacgtg tgaaaacctg tacttccaag
     7081 gtatggtgag caagggcgag gaggtcatca aagagttcat gcgcttcaag gtgcgcatgg
     7141 agggctccat gaacggccac gagttcgaga tcgagggcga gggcgagggc cgcccctacg
     7201 agggcaccca gaccgccaag ctgaaggtga ccaagggcgg ccccctgccc ttcgcctggg
     7261 acatcctgtc cccccagttc atgtacggct ccaaggcgta cgtgaagcac cccgccgaca
     7321 tccccgatta caagaagctg tccttccccg agggcttcaa gtgggagcgc gtgatgaact
     7381 tcgaggacgg cggtctggtg accgtgaccc aggactcctc cctgcaggac ggcacgctga
     7441 tctacaaggt gaagatgcgc ggcaccaact tcccccccga cggccccgta atgcagaaga
     7501 agaccatggg ctgggaggcc tccaccgagc gcctgtaccc ccgcgacggc gtgctgaagg
     7561 gcgagatcca ccaggccctg aagctgaagg acggcggcca ctacctggtg gagttcaaga
     7621 ccatctacat ggccaagaag cccgtgcaac tgcccggcta ctactacgtg gacaccaagc
     7681 tggacatcac ctcccacaac gaggactaca ccatcgtgga acagtacgag cgctccgagg
     7741 gccgccacca cctgttcctg gggcatggca tcggcagcac cggcagcggc agctccggca
     7801 ccgcctcctc cgaggacaac aacatggccg tcatcaaaga gttcatgcgc ttcaaggtgc
     7861 gcatggaggg ctccatgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc
     7921 cctacgaggg cacccagacc gccaagctga aggtgaccaa gggcggcccc ctgcccttcg
     7981 cctgggacat cctgtccccc cagttcatgt acggctccaa ggcgtacgtg aagcaccccg
     8041 ccgacatccc cgattacaag aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga
     8101 tgaacttcga ggacggcggt ctggtgaccg tgacccagga ctcctccctg caggacggca
     8161 cgctgatcta caaggtgaag atgcgcggca ccaacttccc ccccgacggc cccgtaatgc
     8221 agaagaagac catgggctgg gaggcctcca ccgagcgcct gtacccccgc gacggcgtgc
     8281 tgaagggcga gatccaccag gccctgaagc tgaaggacgg cggccactac ctggtggagt
     8341 tcaagaccat ctacatggcc aagaagcccg tgcaactgcc cggctactac tacgtggaca
     8401 ccaagctgga catcacctcc cacaacgagg actacaccat cgtggaacag tacgagcgct
     8461 ccgagggccg ccaccacctg ttcctgtacg gcatggacga gctgtacaag accacttcgt
     8521 gcgaaaacct gtacttccaa ggtatggtga gcaagggcga ggaggtcatc aaagagttca
     8581 tgcgcttcaa ggtgcgcatg gagggctcca tgaacggcca cgagttcgag atcgagggcg
     8641 agggcgaggg ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggcg
     8701 gccccctgcc cttcgcctgg gacatcctgt ccccccagtt catgtacggc tccaaggcgt
     8761 acgtgaagca ccccgccgac atccccgatt acaagaagct gtccttcccc gagggcttca
     8821 agtgggagcg cgtgatgaac ttcgaggacg gcggtctggt gaccgtgacc caggactcct
     8881 ccctgcagga cggcacgctg atctacaagg tgaagatgcg cggcaccaac ttcccccccg
     8941 acggccccgt aatgcagaag aagaccatgg gctgggaggc ctccaccgag cgcctgtacc
     9001 cccgcgacgg cgtgctgaag ggcgagatcc accaggccct gaagctgaag gacggcggcc
     9061 actacctggt ggagttcaag accatctaca tggccaagaa gcccgtgcaa ctgcccggct
     9121 actactacgt ggacaccaag ctggacatca cctcccacaa cgaggactac accatcgtgg
     9181 aacagtacga gcgctccgag ggccgccacc acctgttcct ggggcatggc atcggcagca
     9241 ccggcagcgg cagctccggc accgcctcct ccgaggacaa caacatggcc gtcatcaaag
     9301 agttcatgcg cttcaaggtg cgcatggagg gctccatgaa cggccacgag ttcgagatcg
     9361 agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg aaggtgacca
     9421 agggcggccc cctgcccttc gcctgggaca tcctgtcccc ccagttcatg tacggctcca
     9481 aggcgtacgt gaagcacccc gccgacatcc ccgattacaa gaagctgtcc ttccccgagg
     9541 gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg tctggtgacc gtgacccagg
     9601 actcctccct gcaggacggc acgctgatct acaaggtgaa gatgcgcggc accaacttcc
     9661 cccccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc accgagcgcc
     9721 tgtacccccg cgacggcgtg ctgaagggcg agatccacca ggccctgaag ctgaaggacg
     9781 gcggccacta cctggtggag ttcaagacca tctacatggc caagaagccc gtgcaactgc
     9841 ccggctacta ctacgtggac accaagctgg acatcacctc ccacaacgag gactacacca
     9901 tcgtggaaca gtacgagcgc tccgagggcc gccaccacct gttcctgtac ggcatggacg
     9961 agctgtacaa ggaaaacctg tacttccaaa gtggagaaag cttgtttaag ggaccacgtg
    10021 attacaaccc gatatcgagc accatttgtc atttgacgaa tgaatctgat gggcacacaa
    10081 catcgttgta tggtattgga tttggtccct tcatcattac aaacaagcac ttgtttagaa
    10141 gaaataatgg aacactgttg gtccaatcac tacatggtgt attcaaggtc aagaacacca
    10201 cgactttgca acaacacctc attgatggga gggacatgat aattattcgc atgcctaagg
    10261 atttcccacc atttcctcaa aagctgaaat ttagagagcc acaaagggaa gagcgcatat
    10321 gtcttgtgac aaccaacttc caaactaaga gcatgtctag catggtgtca gacactagtt
    10381 gcacattccc ttcatctgat ggcatattct ggaagcattg gattcaaacc aaggatgggc
    10441 agtgtggcag tccattagta tcaactagag atgggttcat tgttggtata cactcagcat
    10501 cgaatttcac caacacaaac aattatttca caagcgtgcc gaaaaacttc atggaattgt
    10561 tgacaaatca ggaggcgcag cagtgggtta gtggttggcg attaaatgct gactcagtat
    10621 tgtggggggg ccataaagtt ttcatgagca aacctgaaga gccttttcag ccagttaagg
    10681 aagcgactca actcatgaat gaattggtgt actcgcaaga aaacctgtac ttccaaagta
    10741 tgcatagatc acgagatatc agccatggct tcccgccggc ggtggcggcg caggatgatg
    10801 gcacgctgcc catgtcttgt gcccaggaga gcgggatgga ccgtcaccct gcagcctgtg
    10861 cttctgctag gatcaatgtg gagctcgtgc acctgactcc tgaggagaag tctgccgtta
    10921 ctgccctgtg gggcaaggtg aacgtggatg aagttggtgg tgaggccctg ggcaggctgc
    10981 tggtggtcta cccttggacc cagaggttct ttgagtcctt tggggatctg tccactcctg
    11041 atgctgttat gggcaaccct aaggtgaagg ctcatggcaa gaaagtgctc ggtgccttta
    11101 gtgatggcct ggctcacctg gacaacctca agggcacctt tgccacactg agtgagctgc
    11161 actgtgacaa gctgcacgtg gatcctgaga acttcagggt gagtctatgg gacccttgat
    11221 gttttctttc cccttctttt ctatggttaa gttcatgtca taggaagggg ataagtaaca
    11281 gggtacagtt tagaatggga aacagacgaa tgattgcatc agtgtggaag tctcaggatc
    11341 gttttagttt cttttatttg ctgttcataa caattgtttt cttttgttta attcttgctt
    11401 tctttttttt tcttctccgc aatttttact attatactta atgccttaac attgtgtata
    11461 acaaaaggaa atatctctga gatacattaa gtaacttaaa aaaaaaactt tacacagtct
    11521 gcctagtaca ttactatttg gaatatatgt gtgcttattt gcatattcat aatctcccta
    11581 ctttattttc ttttattttt aattgataca taatcattat acatatttat gggttaaagt
    11641 gtaatgtttt aatatgtgta cacatattga ccaaatcagg gtaattttgc atttgtaatt
    11701 ttaaaaaatg ctttcttctt ttaatatact tttttgttta tcttatttct aatactttcc
    11761 ctaatctctt tctttcaggg caataatgat acaatgtatc atgcctcttt gcaccattct
    11821 aaagaataac agtgataatt tctgggttaa ggcaatagca atatttctgc atataaatat
    11881 ttctgcatat aaattgtaac tgatgtaaga ggtttcatat tgctaatagc agctacaatc
    11941 cagctaccat tctgctttta ttttatggtt gggataaggc tggattattc tgagtccaag
    12001 ctaggccctt ttgctaatca tgttcatacc tcttatcttc ctcccacagc tcctgggcaa
    12061 cgtgctggtc tgtgtgctgg cccatcactt tggcaaagaa ttcaccccac cagtgcaggc
    12121 tgcctatcag aaagtggtgg ctggtgtggc taatgccctg gcccacaagt atcactaagc
    12181 ggccgctcgc tttcttgctg tccaatttct attaaaggtt cctttgttcc ctaagtccaa
    12241 ctactaaact gggggatatt atgaagggcc tcgagtaggg tcatgaaggt ttttcttttc
    12301 ctgagaaaac aacacgtatt gttttctcag gttttgcttt ttggcctttt tctagcttaa
    12361 aaaaaaaaaa agcaaaagat gctggtggtt ggcactcctg gtttccagga cggggttcaa
    12421 atccctgcgg cgtctttgct ttgactctcg aggctgatca gcctcgactg tgccttctag
    12481 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
    12541 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
    12601 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
    12661 caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg
    12721 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
    12781 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
    12841 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
    12901 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
    12961 tggttcacgt aggtacgcta gcaccgttgg tttccgtagt gtagtggtta tcacgttcgc
    13021 ctaacacgcg aaaggtcccc ggttcgaaac cgggcactac aaaccaacaa cgctggtacc
    13081 cctagaagtt cctattccga agttcctatt ctctagaaag tataggaact tccttggcca
    13141 aaaagcctga actcaccgcg acgtctgtcg agaagtttct gatcgaaaag ttcgacagcg
    13201 tctccgacct gatgcagctc tcggagggcg aagaatctcg tgctttcagc ttcgatgtag
    13261 gagggcgtgg atatgtcctg cgggtaaata gctgcgccga tggtttctac aaagatcgtt
    13321 atgtttatcg gcactttgca tcggccgcgc tcccgattcc ggaagtgctt gacattgggg
    13381 aattcagcga gagcctgacc tattgcatct cccgccgtgc acagggtgtc acgttgcaag
    13441 acctgcctga aaccgaactg cccgctgttc tgcagccggt cgcggaggcc atggatgcga
    13501 tcgctgcggc cgatcttagc cagacgagcg ggttcggccc attcggaccg caaggaatcg
    13561 gtcaatacac tacatggcgt gatttcatat gcgcgattgc tgatccccat gtgtatcact
    13621 ggcaaactgt gatggacgac accgtcagtg cgtccgtcgc gcaggctctc gatgagctga
    13681 tgctttgggc cgaggactgc cccgaagtcc ggcacctcgt gcacgcggat ttcggctcca
    13741 acaatgtcct gacggacaat ggccgcataa cagcggtcat tgactggagc gaggcgatgt
    13801 tcggggattc ccaatacgag gtcgccaaca tcttcttctg gaggccgtgg ttggcttgta
    13861 tggagcagca gacgcgctac ttcgagcgga ggcatccgga gcttgcagga tcgccgcggc
    13921 tccgggcgta tatgctccgc attggtcttg accaactcta tcagagcttg gttgacggca
    13981 atttcgatga tgcagcttgg gcgcagggtc gatgcgacgc aatcgtccga tccggagccg
    14041 ggactgtcgg gcgtacacaa atcgcccgca gaagcgcggc cgtctggacc gatggctgtg
    14101 tagaagtact cgccgatagt ggaaaccgac gccccagcac tcgtccgagg gcaaaggaat
    14161 agcacgtact acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
    14221 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
    14281 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
    14341 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
    14401 tcaatgtatc ttatcatgtc tgtataccgt cgacaggtgg cacttttcgg ggaaatgtgc
    14461 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
    14521 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
    14581 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
    14641 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
    14701 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
    14761 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
    14821 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
    14881 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
    14941 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
    15001 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
    15061 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
    15121 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
    15181 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
    15241 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
    15301 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
    15361 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
    15421 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaa
//
