LOCUS       sequence-195625-       11043 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Used to sense miRNA activity using fluorescence based tools. 
            hsa-miR-186-3p target in 3' UTR of of mKate. miRNA activity can be 
            measured by the repression of mKate2 relative to EBFP2 fluorescent 
            proteins . Plasmid is inherently unstable; please see Depositor 
            Comments.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11043)
  AUTHORS   Gam JJ, Babb J, Weiss R
  TITLE     A mixed antagonistic/synergistic miRNA repression model enables 
            accurate predictions of multi-input miRNA sensor activity.
  JOURNAL   Nat Commun. 2018 Jun 22;9(1):2430. doi: 10.1038/s41467-018-04575-0.
  PUBMED    29934631
REFERENCE   2  (bases 1 to 11043)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2018-06-22"; volume: "9(1):2430. doi"; pages: " 
            10.1038/s41467-018-04575-0"
COMMENT     Sequence Label: LSB-hsa-miR-186-3p
FEATURES             Location/Qualifiers
     source          1..11043
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(87..675)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(167..186)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(846..1637,1638..1706))
                     /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     1469..1488
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1707..1778)
                     /gene="bla"
                     /label=AmpR promoter
     protein_bind    1798..1835
                     /label=Bxb1 attB
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="Minimal attB site for the Mycopbacteriophage Bxb1 
                     integrase (Voutev and Mann, 2017)"
     CDS             1857..2456
                     /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(1857..1876)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2353..2373
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    2612..2667
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     protein_bind    3646..3666
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     promoter        3715..4893
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          3946..4884
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     protein_bind    4910..4934
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      4945..4954
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4951..5643
                     /codon_start=1
                     /product="monomeric far-red fluorescent protein (Shcherbo 
                     et al., 2009)"
                     /label=mKate2
                     /note="mammalian codon-optimized"
                     /translation="MVSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK
                     AVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRADMAL
                     KLVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAVARY
                     CDLPSKLGH"
     protein_bind    complement(5666..5690)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    5979..6034
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     misc_feature    6418..6443
                     /label=SA
                     /note="splice acceptor site"
     protein_bind    7115..7135
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     promoter        7184..8362
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          7415..8353
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     protein_bind    8379..8403
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      8405..8414
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(8411..8413,8414..8416,8417..9130)
                     /codon_start=1
                     /product="enhanced blue variant of GFP (Ai et al., 2007)"
                     /label=EBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLSHGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGVDFKEDGNILGHKLEYNFNSHNIYIMAVKQKNGIK
                     VNFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDSHYLSTQSVLSKDPNEKRDHMVLL
                     EFRTAAGITLGMDELYK"
     primer_bind     complement(8456..8477)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     protein_bind    complement(9135..9159)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    9314..9369
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(9730..9746)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9730..9746)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9743..9765)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9754..9770
                     /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)."
     promoter        complement(join(9778..9784,9785..9802,9803..9808))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9823..9844
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10058..10077)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             10069..10878
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(10136..10155)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 aattgagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
       61 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
      121 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
      181 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
      241 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
      301 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
      361 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
      421 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
      481 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
      541 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
      601 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
      661 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
      721 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
      781 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
      841 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
      901 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
      961 ggccccagtg ctgcaatgat accgcgagaa ccacgctcac cggctccaga tttatcagca
     1021 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     1081 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     1141 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     1201 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     1261 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     1321 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     1381 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     1441 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     1501 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     1561 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     1621 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     1681 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     1741 cagggttatt gtctcatgag cggatacata tttgaatgta gctccgaatt tctcgagggc
     1801 ttgtcgacga cggcggtctc cgtcgtcagg atcatgaatt cgacccgcct gccaccatga
     1861 ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtcccccgg gccgtacgca
     1921 ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgac ccggaccgcc
     1981 acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg
     2041 gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg
     2101 tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc
     2161 ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg
     2221 cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg
     2281 ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga
     2341 catccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg
     2401 tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctgatacg
     2461 cgtgaattca ctcctcaggt gcaggctgcc tatcagaagg tggtggctgg tgtggccaat
     2521 gccctggctc acaaatacca ctgagatctt tttccctctg ccaaaaatta tggggacatc
     2581 atgaagcccc ttgagcatct gacttctggc taataaagga aatttatttt cattgcaata
     2641 gtgtgttgga attttttgtg tctctcactc ggaaggacat atgggagggc aaatcattta
     2701 aaacatcaga atgagtattt ggtttagagt ttggcaacat atgcccatat gctggctgcc
     2761 atgaacaaag gttggctata aagaggtcat cagtatatga aacagccccc tgctgtccat
     2821 tccttattcc atagaaaagc cttgacttga ggttagattt tttttatatt ttgttttgtg
     2881 ttattttttt ctttaacatc cctaaaattt tccttacatg ttttactagc cagatttttc
     2941 ctcctctcct gactactccc agtcatagct gtccctcttc tcttatggag atccctcgac
     3001 ctgcagccca agcttgaatt ctctagaatt accctgttat ccctatgctt ggtttaccga
     3061 gctcttattg gttttcaaac ttcattgact gtgcccaacg cgcctgggag ctcacgggga
     3121 cagccccccc ccaaagcccc cagggatgta attacgtccc tcccccgcta gggggcagca
     3181 gcgagccgcc cggggctccg ctccggtccg gcgctccccc cgcatccccg agccggcagc
     3241 gtgcggggac agcccgggca cggggaaggt ggcacgggat cgctttcctc tgaacgcttc
     3301 tcgctgctct ttgagcctgc agacacctgg ggggatacgg ggaaaaagct aggctgaaag
     3361 accggttctt agaatgacac gcgcctggga gctcacgggg acagcccccc cccaaagccc
     3421 ccagggatgt aattacgtcc ctcccccgct agggggcagc agcgagccgc ccggggctcc
     3481 gctccggtcc ggcgctcccc ccgcatcccc gagccggcag cgtgcgggga cagcccgggc
     3541 acggggaagg tggcacggga tcgctttcct ctgaacgctt ctcgctgctc tttgagcctg
     3601 cagacacctg gggggatacg gggaaaaagc taacttctga gacgacaact ttgtatagaa
     3661 aagttggctc cgaattcgcc cttgacagca gagatccagt ttggttaatt aattggctcc
     3721 ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt gtggggaggg
     3781 gtcggcaatt gaaccggtgc ctagagaagg tggcgcgggg taaactggga aagtgatgtc
     3841 gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag tgcagtagtc
     3901 gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacaggtaag tgccgtgtgt
     3961 ggttcccgcg ggcctggcct ctttacgggt tatggccctt gcgtgccttg aattacttcc
     4021 acctggctgc agtacgtgat tcttgatccc gagcttcggg ttggaagtgg gtgggagagt
     4081 tcgaggcctt gcgcttaagg agccccttcg cctcgtgctt gagttgaggc ctggcctggg
     4141 cgctggggcc gccgcgtgcg aatctggtgg caccttcgcg cctgtctcgc tgctttcgat
     4201 aagtctctag ccatttaaaa tttttgatga cctgctgcga cgcttttttt ctggcaagat
     4261 agtcttgtaa atgcgggcca agatctgcac actggtattt cggtttttgg ggccgcgggc
     4321 ggcgacgggg cccgtgcgtc ccagcgcaca tgttcggcga ggcggggcct gcgagcgcgg
     4381 ccaccgagaa tcggacgggg gtagtctcaa gctggccggc ctgctctggt gcctggcctc
     4441 gcgccgccgt gtatcgcccc gccctgggcg gcaaggctgg cccggtcggc accagttgcg
     4501 tgagcggaaa gatggccgct tcccggccct gctgcaggga gctcaaaatg gaggacgcgg
     4561 cgctcgggag agcgggcggg tgagtcaccc acacaaagga aaagggcctt tccgtcctca
     4621 gccgtcgctt catgtgactc cacggagtac cgggcgccgt ccaggcacct cgattagttc
     4681 tcgagctttt ggagtacgtc gtctttaggt tggggggagg ggttttatgc gatggagttt
     4741 ccccacactg agtgggtgga gactgaagtt aggccagctt ggcacttgat gtaattctcc
     4801 ttggaatttg ccctttttga gtttggatct tggttcattc tcaagcctca gacagtggtt
     4861 caaagttttt ttcttccatt tcaggtgtcg tgagaagggc gaattcgacc caagtttgta
     4921 caaaaaagca ggctggatcc aggtgccacc atggtgagcg agctgattaa ggagaacatg
     4981 cacatgaagc tgtacatgga gggcaccgtg aacaaccacc acttcaagtg cacatccgag
     5041 ggcgaaggca agccctacga gggcacccag accatgagaa tcaaggcggt cgagggcggc
     5101 cctctcccct tcgccttcga catcctggct accagcttca tgtacggcag caaaaccttc
     5161 atcaaccaca cccagggcat ccccgacttc tttaagcagt ccttccccga gggcttcaca
     5221 tgggagagag tcaccacata cgaagatggg ggcgtgctga ccgctaccca ggacaccagc
     5281 ctccaggacg gctgcctcat ctacaacgtc aagatcagag gggtgaactt cccatccaac
     5341 ggccctgtga tgcagaagaa aacactcggc tgggaggcct ccaccgagac actgtacccc
     5401 gctgacggcg gcctggaagg cagagccgac atggccctga agctcgtggg cgggggccac
     5461 ctgatctgca accttaagac cacatacaga tccaagaaac ccgctaagaa cctcaagatg
     5521 cccggcgtct actatgtgga caggagactg gaaagaatca aggaggccga caaagagaca
     5581 tacgtcgagc agcacgaggt ggctgtggcc agatactgcg acctccctag caaactgggg
     5641 cacaaactta attgagaatt ctaccaccca gctttcttgt acaaagtggt acgtctcaag
     5701 accagaggtc tctgcttaac tggttcccaa aaaattcacc tttgggcccc aaaaaattca
     5761 cctttgggcc ccaaaaaatt cacctttggg ccccaaaaaa ttcacctttg ggccaaccag
     5821 aagagacctg tgcttcactc ctcaggtgca ggctgcctat cagaaggtgg tggctggtgt
     5881 ggccaatgcc ctggctcaca aataccactg agatcttttt ccctctgcca aaaattatgg
     5941 ggacatcatg aagccccttg agcatctgac ttctggctaa taaaggaaat ttattttcat
     6001 tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga aggacatatg ggagggcaaa
     6061 tcatttaaaa catcagaatg agtatttggt ttagagtttg gcaacatatg cccatatgct
     6121 ggctgccatg aacaaaggtt ggctataaag aggtcatcag tatatgaaac agccccctgc
     6181 tgtccattcc ttattccata gaaaagcctt gacttgaggt tagatttttt ttatattttg
     6241 ttttgtgtta tttttttctt taacatccct aaaattttcc ttacatgttt tactagccag
     6301 atttttcctc ctctcctgac tactcccagt catagctgtc cctcttctct tatggagatc
     6361 ccgagcggtg cgtttttatg cttgtagtat tgtataatgt ttttaagatc caagcttctg
     6421 acctcttctc ttcctcccac agggcctcga gacaattgca tcgatggtac cagtgaaagt
     6481 ctgcaatgaa ttcgagttgg cgcgcctgtc attctaaatc tctctttcag cctaaagctt
     6541 tttccccgta tccccccagg tgtctgcagg ctcaaagagc agcgagaagc gttcagagga
     6601 aagcgatccc gtgccacctt ccccgtgccc gggctgtccc cgcacgctgc cggctcgggg
     6661 atgcgggggg agcgccggac cggagcggag ccccgggcgg ctcgctgctg ccccctagcg
     6721 ggggagggac gtaattacat ccctgggggc tttgggaggg ggctgtcccc gtgagctccc
     6781 aggcgcgcct gtcattctaa atctctcttt cagcctaaag ctttttcccc gtatcccccc
     6841 aggtgtctgc aggctcaaag agcagcgaga agcgttcaga ggaaagcgat cccgtgccac
     6901 cttccccgtg cccgggctgt ccccgcacgc tgccggctcg gggatgcggg gggagcgccg
     6961 gaccggagcg gagccccggg cggctcgctg ctgcccccta gcgggggagg gacgtaatta
     7021 catccctggg ggctttgggg gggggctgtc cccgtgagct cccaggcgcg ccaactcggt
     7081 accgtatcga tgtcgacgtt aacgctagtg atatcaactt tgtatagaaa agttggctcc
     7141 gaattcgccc ttgacagcag agatccagtt tggttaatta attggctccg gtgcccgtca
     7201 gtgggcagag cgcacatcgc ccacagtccc cgagaagttg tggggagggg tcggcaattg
     7261 aaccggtgcc tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg tgtactggct
     7321 ccgccttttt cccgagggtg ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt
     7381 tctttttcgc aacgggtttg ccgccagaac acaggtaagt gccgtgtgtg gttcccgcgg
     7441 gcctggcctc tttacgggtt atggcccttg cgtgccttga attacttcca cctggctgca
     7501 gtacgtgatt cttgatcccg agcttcgggt tggaagtggg tgggagagtt cgaggccttg
     7561 cgcttaagga gccccttcgc ctcgtgcttg agttgaggcc tggcctgggc gctggggccg
     7621 ccgcgtgcga atctggtggc accttcgcgc ctgtctcgct gctttcgata agtctctagc
     7681 catttaaaat ttttgatgac ctgctgcgac gctttttttc tggcaagata gtcttgtaaa
     7741 tgcgggccaa gatctgcaca ctggtatttc ggtttttggg gccgcgggcg gcgacggggc
     7801 ccgtgcgtcc cagcgcacat gttcggcgag gcggggcctg cgagcgcggc caccgagaat
     7861 cggacggggg tagtctcaag ctggccggcc tgctctggtg cctggcctcg cgccgccgtg
     7921 tatcgccccg ccctgggcgg caaggctggc ccggtcggca ccagttgcgt gagcggaaag
     7981 atggccgctt cccggccctg ctgcagggag ctcaaaatgg aggacgcggc gctcgggaga
     8041 gcgggcgggt gagtcaccca cacaaaggaa aagggccttt ccgtcctcag ccgtcgcttc
     8101 atgtgactcc acggagtacc gggcgccgtc caggcacctc gattagttct cgagcttttg
     8161 gagtacgtcg tctttaggtt ggggggaggg gttttatgcg atggagtttc cccacactga
     8221 gtgggtggag actgaagtta ggccagcttg gcacttgatg taattctcct tggaatttgc
     8281 cctttttgag tttggatctt ggttcattct caagcctcag acagtggttc aaagtttttt
     8341 tcttccattt caggtgtcgt gagaagggcg aattcgaccc aagtttgtac aaaaaagcag
     8401 gctcgccacc atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
     8461 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg aggggcgagg gcgagggcga
     8521 tgccaccaac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
     8581 ctggcccacc ctcgtgacca ccctgagcca cggcgtgcag tgcttcgccc gctaccccga
     8641 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
     8701 caccatcttc ttcaaggacg acggcaccta caagacccgc gccgaggtga agttcgaggg
     8761 cgacacccta gtgaaccgca tcgagctgaa gggcgtcgac ttcaaggagg acggcaacat
     8821 cctggggcac aagctggagt acaacttcaa cagccacaac atctatatca tggccgtcaa
     8881 gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacgtggagg acggcagcgt
     8941 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     9001 cgacagccac tacctgagca cccagtccgt gctgagcaaa gaccccaacg agaagcgcga
     9061 tcacatggtc ctgctggagt tccgcaccgc cgccgggatc actctcggca tggacgagct
     9121 gtacaagtaa taccacccag ctttcttgta caaagtggta cgcgtgaatt cactcctcag
     9181 gtgcaggctg cctatcagaa ggtggtggct ggtgtggcca atgccctggc tcacaaatac
     9241 cactgagatc tttttccctc tgccaaaaat tatggggaca tcatgaagcc ccttgagcat
     9301 ctgacttctg gctaataaag gaaatttatt ttcattgcaa tagtgtgttg gaattttttg
     9361 tgtctctcac tcggaaggac atatgggagg gcaaatcatt taaaacatca gaatgagtat
     9421 ttggtttaga gtttggcaac atatgcccat atgctggctg ccatgaacaa aggttggcta
     9481 taaagaggtc atcagtatat gaaacagccc cctgctgtcc attccttatt ccatagaaaa
     9541 gccttgactt gaggttagat tttttttata ttttgttttg tgttattttt ttctttaaca
     9601 tccctaaaat tttccttaca tgttttacta gccagatttt tcctcctctc ctgactactc
     9661 ccagtcatag ctgtccctct tctcttatgg agatccctcg acctgcagcc caagcttggc
     9721 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     9781 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     9841 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagcggat
     9901 cgacagtact aagcttggtc taatacccaa tctctcgtct tatccagatg ttttatacgc
     9961 ctctggcccg tgtctcaaaa tctctgatgt tacattgcac aagataaaat aatatcatca
    10021 tgaacaataa aactgtctgc ttacataaac agtaatacaa ggggtgttat gagccatatt
    10081 caacgggaaa cgtcgaggcc gcgattaaat tccaacatgg atgctgattt atatgggtat
    10141 aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa tctatcgctt gtatgggaag
    10201 cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca
    10261 gatgagatgg tcagactaaa ctggctgacg gaatttatgc ctcttccgac catcaagcat
    10321 tttatccgta ctcctgatga tgcatggtta ctcaccactg cgatccccgg aaaaacagca
    10381 ttccaggtat tagaagaata tcctgattca ggtgaaaata ttgttgatgc gctggcagtg
    10441 ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc cttttaacag cgatcgcgta
    10501 tttcgtctcg ctcaggcgca atcacgaatg aataacggtt tggttgatgc gagtgatttt
    10561 gatgacgagc gtaatggctg gcctgttgaa caagtctgga aagaaatgca taaacttttg
    10621 ccattctcac cggattcagt cgtcactcat ggtgatttct cacttgataa ccttattttt
    10681 gacgagggga aattaatagg ttgtattgat gttggacgag tcggaatcgc agaccgatac
    10741 caggatcttg ccatcctatg gaactgcctc ggtgagtttt ctccttcatt acagaaacgg
    10801 ctttttcaaa aatatggtat tgataatcct gatatgaata aattgcagtt tcatttgatg
    10861 ctcgatgagt ttttctaatc agaattggtt aattggttgt aacactggca gagcattacg
    10921 ctgacttgac gggacggcgc aagctcatga ccaaaatccc ttaacgtgag ttacgcgtcg
    10981 ttccactgag cgtcagaggt atcactagta ttacagaggt aagttataac agtcgcctaa
    11041 ccc
//
