LOCUS       sequence-195763-       11039 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Used to sense miRNA activity using fluorescence based tools. 
            hsa-miR-218-5p 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 11039)
  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 11039)
  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-218-5p
FEATURES             Location/Qualifiers
     source          1..11039
                     /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    5975..6030
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     misc_feature    6414..6439
                     /label=SA
                     /note="splice acceptor site"
     protein_bind    7111..7131
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     promoter        7180..8358
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          7411..8349
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     protein_bind    8375..8399
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      8401..8410
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(8407..8409,8410..8412,8413..9126)
                     /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(8452..8473)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     protein_bind    complement(9131..9155)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    9310..9365
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(9726..9742)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9726..9742)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9739..9761)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9750..9766
                     /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(9774..9780,9781..9798,9799..9804))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9819..9840
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10054..10073)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             10065..10874
                     /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(10132..10151)
                     /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 tctgcttcat aaatcacatg gttagatcaa gcacaaacat ggttagatca
     5761 agcacaaaca tggttagatc aagcacaaac atggttagat caagcacaac aaccagaaga
     5821 gacctgtgct tcactcctca ggtgcaggct gcctatcaga aggtggtggc tggtgtggcc
     5881 aatgccctgg ctcacaaata ccactgagat ctttttccct ctgccaaaaa ttatggggac
     5941 atcatgaagc cccttgagca tctgacttct ggctaataaa ggaaatttat tttcattgca
     6001 atagtgtgtt ggaatttttt gtgtctctca ctcggaagga catatgggag ggcaaatcat
     6061 ttaaaacatc agaatgagta tttggtttag agtttggcaa catatgccca tatgctggct
     6121 gccatgaaca aaggttggct ataaagaggt catcagtata tgaaacagcc ccctgctgtc
     6181 cattccttat tccatagaaa agccttgact tgaggttaga ttttttttat attttgtttt
     6241 gtgttatttt tttctttaac atccctaaaa ttttccttac atgttttact agccagattt
     6301 ttcctcctct cctgactact cccagtcata gctgtccctc ttctcttatg gagatcccga
     6361 gcggtgcgtt tttatgcttg tagtattgta taatgttttt aagatccaag cttctgacct
     6421 cttctcttcc tcccacaggg cctcgagaca attgcatcga tggtaccagt gaaagtctgc
     6481 aatgaattcg agttggcgcg cctgtcattc taaatctctc tttcagccta aagctttttc
     6541 cccgtatccc cccaggtgtc tgcaggctca aagagcagcg agaagcgttc agaggaaagc
     6601 gatcccgtgc caccttcccc gtgcccgggc tgtccccgca cgctgccggc tcggggatgc
     6661 ggggggagcg ccggaccgga gcggagcccc gggcggctcg ctgctgcccc ctagcggggg
     6721 agggacgtaa ttacatccct gggggctttg ggagggggct gtccccgtga gctcccaggc
     6781 gcgcctgtca ttctaaatct ctctttcagc ctaaagcttt ttccccgtat ccccccaggt
     6841 gtctgcaggc tcaaagagca gcgagaagcg ttcagaggaa agcgatcccg tgccaccttc
     6901 cccgtgcccg ggctgtcccc gcacgctgcc ggctcgggga tgcgggggga gcgccggacc
     6961 ggagcggagc cccgggcggc tcgctgctgc cccctagcgg gggagggacg taattacatc
     7021 cctgggggct ttgggggggg gctgtccccg tgagctccca ggcgcgccaa ctcggtaccg
     7081 tatcgatgtc gacgttaacg ctagtgatat caactttgta tagaaaagtt ggctccgaat
     7141 tcgcccttga cagcagagat ccagtttggt taattaattg gctccggtgc ccgtcagtgg
     7201 gcagagcgca catcgcccac agtccccgag aagttgtggg gaggggtcgg caattgaacc
     7261 ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc
     7321 ctttttcccg agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt
     7381 tttcgcaacg ggtttgccgc cagaacacag gtaagtgccg tgtgtggttc ccgcgggcct
     7441 ggcctcttta cgggttatgg cccttgcgtg ccttgaatta cttccacctg gctgcagtac
     7501 gtgattcttg atcccgagct tcgggttgga agtgggtggg agagttcgag gccttgcgct
     7561 taaggagccc cttcgcctcg tgcttgagtt gaggcctggc ctgggcgctg gggccgccgc
     7621 gtgcgaatct ggtggcacct tcgcgcctgt ctcgctgctt tcgataagtc tctagccatt
     7681 taaaattttt gatgacctgc tgcgacgctt tttttctggc aagatagtct tgtaaatgcg
     7741 ggccaagatc tgcacactgg tatttcggtt tttggggccg cgggcggcga cggggcccgt
     7801 gcgtcccagc gcacatgttc ggcgaggcgg ggcctgcgag cgcggccacc gagaatcgga
     7861 cgggggtagt ctcaagctgg ccggcctgct ctggtgcctg gcctcgcgcc gccgtgtatc
     7921 gccccgccct gggcggcaag gctggcccgg tcggcaccag ttgcgtgagc ggaaagatgg
     7981 ccgcttcccg gccctgctgc agggagctca aaatggagga cgcggcgctc gggagagcgg
     8041 gcgggtgagt cacccacaca aaggaaaagg gcctttccgt cctcagccgt cgcttcatgt
     8101 gactccacgg agtaccgggc gccgtccagg cacctcgatt agttctcgag cttttggagt
     8161 acgtcgtctt taggttgggg ggaggggttt tatgcgatgg agtttcccca cactgagtgg
     8221 gtggagactg aagttaggcc agcttggcac ttgatgtaat tctccttgga atttgccctt
     8281 tttgagtttg gatcttggtt cattctcaag cctcagacag tggttcaaag tttttttctt
     8341 ccatttcagg tgtcgtgaga agggcgaatt cgacccaagt ttgtacaaaa aagcaggctc
     8401 gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
     8461 ctggacggcg acgtaaacgg ccacaagttc agcgtgaggg gcgagggcga gggcgatgcc
     8521 accaacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg
     8581 cccaccctcg tgaccaccct gagccacggc gtgcagtgct tcgcccgcta ccccgaccac
     8641 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
     8701 atcttcttca aggacgacgg cacctacaag acccgcgccg aggtgaagtt cgagggcgac
     8761 accctagtga accgcatcga gctgaagggc gtcgacttca aggaggacgg caacatcctg
     8821 gggcacaagc tggagtacaa cttcaacagc cacaacatct atatcatggc cgtcaagcag
     8881 aagaacggca tcaaggtgaa cttcaagatc cgccacaacg tggaggacgg cagcgtgcag
     8941 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     9001 agccactacc tgagcaccca gtccgtgctg agcaaagacc ccaacgagaa gcgcgatcac
     9061 atggtcctgc tggagttccg caccgccgcc gggatcactc tcggcatgga cgagctgtac
     9121 aagtaatacc acccagcttt cttgtacaaa gtggtacgcg tgaattcact cctcaggtgc
     9181 aggctgccta tcagaaggtg gtggctggtg tggccaatgc cctggctcac aaataccact
     9241 gagatctttt tccctctgcc aaaaattatg gggacatcat gaagcccctt gagcatctga
     9301 cttctggcta ataaaggaaa tttattttca ttgcaatagt gtgttggaat tttttgtgtc
     9361 tctcactcgg aaggacatat gggagggcaa atcatttaaa acatcagaat gagtatttgg
     9421 tttagagttt ggcaacatat gcccatatgc tggctgccat gaacaaaggt tggctataaa
     9481 gaggtcatca gtatatgaaa cagccccctg ctgtccattc cttattccat agaaaagcct
     9541 tgacttgagg ttagattttt tttatatttt gttttgtgtt atttttttct ttaacatccc
     9601 taaaattttc cttacatgtt ttactagcca gatttttcct cctctcctga ctactcccag
     9661 tcatagctgt ccctcttctc ttatggagat ccctcgacct gcagcccaag cttggcgtaa
     9721 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     9781 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     9841 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gcggatcgac
     9901 agtactaagc ttggtctaat acccaatctc tcgtcttatc cagatgtttt atacgcctct
     9961 ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga taaaataata tcatcatgaa
    10021 caataaaact gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac
    10081 gggaaacgtc gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat
    10141 gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgcttgtat gggaagcccg
    10201 atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg
    10261 agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta
    10321 tccgtactcc tgatgatgca tggttactca ccactgcgat ccccggaaaa acagcattcc
    10381 aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc
    10441 tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc
    10501 gtctcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg
    10561 acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataaa cttttgccat
    10621 tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg
    10681 aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg
    10741 atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt
    10801 ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg
    10861 atgagttttt ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc attacgctga
    10921 cttgacggga cggcgcaagc tcatgaccaa aatcccttaa cgtgagttac gcgtcgttcc
    10981 actgagcgtc agaggtatca ctagtattac agaggtaagt tataacagtc gcctaaccc
//
