LOCUS       sequence-483324-        9085 bp    DNA     circular SYN 14-AUG-2025
DEFINITION  mCherry reporter plasmid for live cell visualization of RNA m6A 
            modification.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9085)
  AUTHORS   Li Q, Liu J, Guo L, Zhang Y, Chen Y, Liu H, Cheng H, Deng L, Qiu J, 
            Zhang K, Goh WSS, Wang Y, Peng Q
  TITLE     Decoding the interplay between m(6)A modification and stress granule
            stability by live-cell imaging.
  JOURNAL   Sci Adv. 2024 Nov 15;10(46):eadp5689. doi: 10.1126/sciadv.adp5689. 
            Epub 2024 Nov 15.
  PUBMED    39546601
REFERENCE   2  (bases 1 to 9085)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Sci Adv."; 
            date: "2024-11-15"; volume: "10(46):eadp5689. doi"; pages: " 
            10.1126/sciadv.adp5689. Epub 2024 Nov 15"
COMMENT     Sequence Label: pLD1112 TRE-mCherry-SKL-22Xm6A MS2
FEATURES             Location/Qualifiers
     source          1..9085
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    41..158
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        212..526
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    220..238
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    256..274
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    291..309
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    327..345
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    363..381
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    398..416
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    434..452
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     499..523
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      543..552
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             549..1256
                     /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(694..712)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     962..981
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_RNA        1338..1356
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        1366..1384
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        1394..1412
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        1422..1440
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        1450..1468
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        1478..1496
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        1506..1524
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        1534..1552
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        1562..1580
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        1590..1608
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        1618..1636
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        1646..1664
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        1674..1692
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        1702..1720
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        1730..1748
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        1758..1776
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        1786..1804
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        1814..1832
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        1842..1860
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        1870..1888
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        1898..1916
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        1926..1944
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        1954..1972
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        1982..2000
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     promoter        2027..2537
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     2456..2474
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             2547..3143
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(2547..2566)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3043..3063
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             3144..3197
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             join(3198..3815,3816..3821,3822..3857,3858..3860,3861..3896,
                     3897..3899,3900..3935,3936..3944)
                     /codon_start=1
                     /product="improved tetracycline-controlled transactivator"
                     /label=rtTA-Advanced
                     /note="In the Tet-On(R) system, rtTA-Advanced binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the presence of tetracycline or doxycycline."
                     /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     primer_bind     complement(3984..4004)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4402..4413)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4618..4851
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4923..5044
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4960..4979)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5014..5033
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5084..5219
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        5219..5290
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5291..5359,5360..6151)
                     /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(5509..5528)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6322..6910
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6811..6830
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7064..7081
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7198..7219
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7234..7239,7240..7257,7258..7264)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7272..7288
                     /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     7277..7299
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7296..7312
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7296..7312
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7331..7351
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7333..7351
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        7379..7605
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             7606..7786
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7833..7958
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    8451..8684
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8869..8913
                     /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"
ORIGIN
        1 aattctcgac ctcgagacaa atggcagtat tcatccacaa ttttaaaaga aaagggggga
       61 ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac atacaaacta
      121 aagaattaca aaaacaaatt acaaaaattc aaaattttcg ggtttattac agggacagca
      181 gagatccact ttggccgcga atcgatatgt cgagtttact ccctatcagt gatagagaac
      241 gtatgtcgag tttactccct atcagtgata gagaacgatg tcgagtttac tccctatcag
      301 tgatagagaa cgtatgtcga gtttactccc tatcagtgat agagaacgta tgtcgagttt
      361 actccctatc agtgatagag aacgtatgtc gagtttatcc ctatcagtga tagagaacgt
      421 atgtcgagtt tactccctat cagtgataga gaacgtatgt cgaggtaggc gtgtacggtg
      481 ggaggcctat ataagcagag ctcgtttagt gaaccgtcag atcgcctgga gaattggcta
      541 gcgccaccat ggtgagcaag ggcgaggagg ataacatggc catcatcaag gagttcatgc
      601 gcttcaaggt gcacatggag ggctccgtga acggccacga gttcgagatc gagggcgagg
      661 gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggtggcc
      721 ccctgccctt cgcctgggac atcctgtccc ctcagttcat gtacggctcc aaggcctacg
      781 tgaagcaccc cgccgacatc cccgactact tgaagctgtc cttccccgag ggcttcaagt
      841 gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac cgtgacccag gactcctccc
      901 tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg caccaacttc ccctccgacg
      961 gccccgtaat gcagaagaag accatgggct gggaggcctc ctccgagcgg atgtaccccg
     1021 aggacggcgc cctgaagggc gagatcaagc agaggctgaa gctgaaggac ggcggccact
     1081 acgacgctga ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg cccggcgcct
     1141 acaacgtcaa catcaagttg gacatcacct cccacaacga ggactacacc atcgtggaac
     1201 agtacgaacg cgccgagggc cgccactcca ccggcggcat ggacgagctg tacaagaccg
     1261 gtggcgctcc aggattctcc tcaatttccg cttccaaact gtaactggta ccatgcaatt
     1321 cgaagcttcc ggactgtaca tgaggatcac ccatgtccgg actgtacatg aggatcaccc
     1381 atgtccggac tgtacatgag gatcacccat gtccggactg tacatgagga tcacccatgt
     1441 ccggactgta catgaggatc acccatgtcc ggactgtaca tgaggatcac ccatgtccgg
     1501 actgtacatg aggatcaccc atgtccggac tgtacatgag gatcacccat gtccggactg
     1561 tacatgagga tcacccatgt ccggactgta catgaggatc acccatgtcc ggactgtaca
     1621 tgaggatcac ccatgtccgg actgtacatg aggatcaccc atgtccggac tgtacatgag
     1681 gatcacccat gtccggactg tacatgagga tcacccatgt ccggactgta catgaggatc
     1741 acccatgtcc ggactgtaca tgaggatcac ccatgtccgg actgtacatg aggatcaccc
     1801 atgtccggac tgtacatgag gatcacccat gtccggactg tacatgagga tcacccatgt
     1861 ccggactgta catgaggatc acccatgtcc ggactgtaca tgaggatcac ccatgtccgg
     1921 actgtacatg aggatcaccc atgtccggac tgtacatgag gatcacccat gtccggactg
     1981 tacatgagga tcacccatgt gcggccgcga tcgtgaattc ggatccgggg ttggggttgc
     2041 gccttttcca aggcagccct gggtttgcgc agggacgcgg ctgctctggg cgtggttccg
     2101 ggaaacgcag cggcgccgac cctgggtctc gcacattctt cacgtccgtt cgcagcgtca
     2161 cccggatctt cgccgctacc cttgtgggcc ccccggcgac gcttcctgct ccgcccctaa
     2221 gtcgggaagg ttccttgcgg ttcgcggcgt gccggacgtg acaaacggaa gccgcacgtc
     2281 tcactagtac cctcgcagac ggacagcgcc agggagcaat ggcagcgcgc cgaccgcgat
     2341 gggctgtggc caatagcggc tgctcagcag ggcgcgccga gagcagcggc cgggaagggg
     2401 cggtgcggga ggcggggtgt ggggcggtag tgtgggccct gttcctgccc gcgcggtgtt
     2461 ccgcattctg caagcctccg gagcgcacgt cggcagtcgg ctccctcgtt gaccgaatca
     2521 ccgacctctc tccccagcaa ttcaccatga ccgagtacaa gcccacggtg cgcctcgcca
     2581 cccgcgacga cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg
     2641 ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac
     2701 tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg
     2761 cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg
     2821 gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc
     2881 tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc
     2941 ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg
     3001 agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg
     3061 agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt
     3121 gcatgacccg caagcccggt gccgaaggta gaggttctct cctcacttgt ggtgatgttg
     3181 aagaaaaccc tggtccaatg tctagactgg acaagagcaa agtcataaac ggagctctgg
     3241 aattactcaa tggtgtcggt atcgaaggcc tgacgacaag gaaactcgct caaaagctgg
     3301 gagttgagca gcctaccctg tactggcacg tgaagaacaa gcgggccctg ctcgatgccc
     3361 tgccaatcga gatgctggac aggcatcata cccacttctg ccccctggaa ggcgagtcat
     3421 ggcaagactt tctgcggaac aacgccaagt cataccgctg tgctctcctc tcacatcgcg
     3481 acggggctaa agtgcatctc ggcacccgcc caacagagaa acagtacgaa accctggaaa
     3541 atcagctcgc gttcctgtgt cagcaaggct tctccctgga gaacgcactg tacgctctgt
     3601 ccgccgtggg ccactttaca ctgggctgcg tattggagga acaggagcat caagtagcaa
     3661 aagaggaaag agagacacct accaccgatt ctatgccccc acttctgaga caagcaattg
     3721 agctgttcga ccggcaggga gccgaacctg ccttcctttt cggcctggaa ctaatcatat
     3781 gtggcctgga gaaacagcta aagtgcgaaa gcggcgggcc gaccgacgcc cttgacgatt
     3841 ttgacttaga catgctccca gccgatgccc ttgacgactt tgaccttgat atgctgcctg
     3901 ctgacgctct tgacgatttt gaccttgaca tgctccccgg gtaactaagt aaggatcgat
     3961 ccaagatatc gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     4021 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     4081 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
     4141 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
     4201 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
     4261 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
     4321 gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat tctgcgcggg
     4381 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
     4441 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
     4501 ctttgggccg cctccccgcc tgtttcgcct cggcgtccgg actagaggta cctttaagac
     4561 caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg
     4621 aagggctaat tcactcccaa cgaagacaag atctgctttt tgcttgtact gggtctctct
     4681 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
     4741 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
     4801 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt
     4861 catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga
     4921 ggaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     4981 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     5041 cttatcatgt ctggctctag ctatcccgcc cctaactccg cccatcccgc ccctaactcc
     5101 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     5161 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     5221 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     5281 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     5341 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     5401 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     5461 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     5521 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     5581 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     5641 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     5701 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     5761 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     5821 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     5881 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     5941 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     6001 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     6061 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     6121 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     6181 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     6241 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     6301 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     6361 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     6421 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     6481 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     6541 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     6601 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     6661 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     6721 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     6781 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     6841 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     6901 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     6961 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     7021 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     7081 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     7141 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
     7201 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
     7261 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta
     7321 cgccaagcgc gcaattaacc ctcactaaag ggaacaaaag ctggagctgc aagcttaatg
     7381 tagtcttatg caatactctt gtagtcttgc aacatggtaa cgatgagtta gcaacatgcc
     7441 ttacaaggag agaaaaagca ccgtgcatgc cgattggtgg aagtaaggtg gtacgatcgt
     7501 gccttattag gaaggcaaca gacgggtctg acatggattg gacgaaccac tgaattgccg
     7561 cattgcagag atattgtatt taagtgccta gctcgataca taaacgggtc tctctggtta
     7621 gaccagatct gagcctggga gctctctggc taactaggga acccactgct taagcctcaa
     7681 taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac
     7741 tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca
     7801 gggacttgaa agcgaaaggg aaaccagagg agctctctcg acgcaggact cggcttgctg
     7861 aagcgcgcac ggcaagaggc gaggggcggc gactggtgag tacgccaaaa attttgacta
     7921 gcggaggcta gaaggagaga gatgggtgcg agagcgtcag tattaagcgg gggagaatta
     7981 gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat aaattaaaac
     8041 atatagtatg ggcaagcagg gagctagaac gattcgcagt taatcctggc ctgttagaaa
     8101 catcagaagg ctgtagacaa atactgggac agctacaacc atcccttcag acaggatcag
     8161 aagaacttag atcattatat aatacagtag caaccctcta ttgtgtgcat caaaggatag
     8221 agataaaaga caccaaggaa gctttagaca agatagagga agagcaaaac aaaagtaaga
     8281 ccaccgcaca gcaagcggcc gctgatcttc agacctggag gaggagatat gagggacaat
     8341 tggagaagtg aattatataa atataaagta gtaaaaattg aaccattagg agtagcaccc
     8401 accaaggcaa agagaagagt ggtgcagaga gaaaaaagag cagtgggaat aggagctttg
     8461 ttccttgggt tcttgggagc agcaggaagc actatgggcg cagcgtcaat gacgctgacg
     8521 gtacaggcca gacaattatt gtctggtata gtgcagcagc agaacaattt gctgagggct
     8581 attgaggcgc aacagcatct gttgcaactc acagtctggg gcatcaagca gctccaggca
     8641 agaatcctgg ctgtggaaag atacctaaag gatcaacagc tcctggggat ttggggttgc
     8701 tctggaaaac tcatttgcac cactgctgtg ccttggaatg ctagttggag taataaatct
     8761 ctggaacaga tttggaatca cacgacctgg atggagtggg acagagaaat taacaattac
     8821 acaagcttaa tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa
     8881 gaattattgg aattagataa atgggcaagt ttgtggaatt ggtttaacat aacaaattgg
     8941 ctgtggtata taaaattatt cataatgata gtaggaggct tggtaggttt aagaatagtt
     9001 tttgctgtac tttctatagt gaatagagtt aggcagggat attcaccatt atcgtttcag
     9061 acccacctcc caaccccgag gggac
//
