LOCUS       sequence-196586-       12672 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Used to make miRNA sensors with targets in 5' and 3/ UTRs (with BsaI
            cloning) . Plasmid is inherently unstable; please see Depositor 
            Comments.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12672)
  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 12672)
  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: LSBr5and3
FEATURES             Location/Qualifiers
     source          1..12672
                     /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)"
     promoter        3791..4002
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     intron          4003..4941
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     protein_bind    5019..5040
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5055..5060,5061..5078,5079..5085)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5093..5109
                     /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     5117..5133
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             5129..5452
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    5146..5202
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(5203..5220)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5203..5219)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5212..5234)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5428..5447)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     regulatory      5565..5574
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5571..6263
                     /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"
     RBS             6378..6389
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             6397..7107
                     /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(6542..6560)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     polyA_signal    7342..7397
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     misc_feature    7789..7814
                     /label=SA
                     /note="splice acceptor site"
     protein_bind    8486..8506
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     promoter        8555..9733
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          8786..9724
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     protein_bind    9750..9774
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      9776..9785
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(9782..9784,9785..9787,9788..10501)
                     /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(9827..9848)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     protein_bind    complement(10506..10530)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    10685..10740
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(11101..11117)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    11125..11141
                     /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(11149..11155,11156..11173,11174..11179))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11194..11215
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        11368..11472
                     /gene="bla"
                     /label=AmpR promoter
     CDS             11692..12483
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
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 ccctaggttt accgagctct
     3061 tattggtttt caaacttcat tgactgtgcc ggagcctcga gacaattgat taacatcgat
     3121 acggtaccga gttggcgcgc ctgggagctc acggggacag ccccccccca aagcccccag
     3181 ggatgtaatt acgtccctcc cccgctaggg ggcagcagcg agccgcccgg ggctccgctc
     3241 cggtccggcg ctccccccgc atccccgagc cggcagcgtg cggggacagc ccgggcacgg
     3301 ggaaggtggc acgggatcgc tttcctctga acgcttctcg ctgctctttg agcctgcaga
     3361 cacctggggg gatacgggga aaaagcttta ggctgaaaga gagatttaga atgacaggcg
     3421 cgcctggcca tacatcgata cggtaccgag ttggcgcgcc tgggagctca cggggacagc
     3481 ccccccccaa agcccccagg gatgtaatta cgtccctccc ccgctagggg gcagcagcga
     3541 gccgcccggg gctccgctcc ggtccggcgc tccccccgca tccccgagcc ggcagcgtgc
     3601 ggggacagcc cgggcacggg gaaggtggca cgggatcgct ttcctctgaa cgcttctcgc
     3661 tgctctttga gcctgcagac acctgggggg atacggggaa aaagctttag gctgaaagag
     3721 agatttagaa tgacaggcgc gcctggtggc gaacgatcaa ctactttaat tggctccggt
     3781 gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttgtg gggaggggtc
     3841 ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg
     3901 tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc
     3961 gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc cgtgtgtggt
     4021 tcccgcgggc ctggcctctt tacgggttat ggcccttgcg tgccttgaat tacttccacc
     4081 tggctgcagt acgtgattct tgatcccgag cttcgggttg gaagtgggtg ggagagttcg
     4141 aggccttgcg cttaaggagc cccttcgcct cgtgcttgag ttgaggcctg gcctgggcgc
     4201 tggggccgcc gcgtgcgaat ctggtggcac cttcgcgcct gtctcgctgc tttcgataag
     4261 tctctagcca tttaaaattt ttgatgacct gctgcgacgc tttttttctg gcaagatagt
     4321 cttgtaaatg cgggccaaga tctgcacact ggtatttcgg tttttggggc cgcgggcggc
     4381 gacggggccc gtgcgtccca gcgcacatgt tcggcgaggc ggggcctgcg agcgcggcca
     4441 ccgagaatcg gacgggggta gtctcaagct ggccggcctg ctctggtgcc tggcctcgcg
     4501 ccgccgtgta tcgccccgcc ctgggcggca aggctggccc ggtcggcacc agttgcgtga
     4561 gcggaaagat ggccgcttcc cggccctgct gcagggagct caaaatggag gacgcggcgc
     4621 tcgggagagc gggcgggtga gtcacccaca caaaggaaaa gggcctttcc gtcctcagcc
     4681 gtcgcttcat gtgactccac ggagtaccgg gcgccgtcca ggcacctcga ttagttctcg
     4741 agcttttgga gtacgtcgtc tttaggttgg ggggaggggt tttatgcgat ggagtttccc
     4801 cacactgagt gggtggagac tgaagttagg ccagcttggc acttgatgta attctccttg
     4861 gaatttgccc tttttgagtt tggatcttgg ttcattctca agcctcagac agtggttcaa
     4921 agtttttttc ttccatttca ggtggaagac aacagatgag accgcagctg gcacgacagg
     4981 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
     5041 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
     5101 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagct tgcatgcctg
     5161 caggtcgact ctagaggatc cccgggtacc gagctcgaat tcactggccg tcgttttaca
     5221 acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc
     5281 tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg
     5341 cagcctgaat ggcgaatggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     5401 ttcacaccgc atatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     5461 gccccgacac tcgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     5521 cgcttacaga caagctgtga cggtctcaag gtttgtcttc aggtgccacc atggtgagcg
     5581 agctgattaa ggagaacatg cacatgaagc tgtacatgga gggcaccgtg aacaaccacc
     5641 acttcaagtg cacatccgag ggcgaaggca agccctacga gggcacccag accatgagaa
     5701 tcaaggcggt cgagggcggc cctctcccct tcgccttcga catcctggct accagcttca
     5761 tgtacggcag caaaaccttc atcaaccaca cccagggcat ccccgacttc tttaagcagt
     5821 ccttccccga gggcttcaca tgggagagag tcaccacata cgaagatggg ggcgtgctga
     5881 ccgctaccca ggacaccagc ctccaggacg gctgcctcat ctacaacgtc aagatcagag
     5941 gggtgaactt cccatccaac ggccctgtga tgcagaagaa aacactcggc tgggaggcct
     6001 ccaccgagac actgtacccc gctgacggcg gcctggaagg cagagccgac atggccctga
     6061 agctcgtggg cgggggccac ctgatctgca accttaagac cacatacaga tccaagaaac
     6121 ccgctaagaa cctcaagatg cccggcgtct actatgtgga caggagactg gaaagaatca
     6181 aggaggccga caaagagaca tacgtcgagc agcacgaggt ggctgtggcc agatactgcg
     6241 acctccctag caaactgggg cacaaactta attgagaaga caagctttga gaccgaattc
     6301 aagcttggag cgttgacacc atcgaatggt gcaaaacctt tcgcggtatg gcatgatagc
     6361 gcccggtact tctagagaaa gaggagaaat actagaatgg tgagcaaggg cgaggaggat
     6421 aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg ctccgtgaac
     6481 ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg cacccagacc
     6541 gccaagctga aggtgaccaa gggtggcccc ctgcccttcg cctgggacat cctgtcccct
     6601 cagttcatgt acggctccaa ggcctacgtg aagcaccccg ccgacatccc cgactacttg
     6661 aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc
     6721 gtggtgaccg tgacccagga ctcctccttg caggacggcg agttcatcta caaggtgaag
     6781 ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc agaagaaaac catgggctgg
     6841 gaggcctcct ccgagcggat gtaccccgag gacggcgccc tgaagggcga gatcaagcag
     6901 aggctgaagc tgaaggacgg cggccactac gacgctgagg tcaagaccac ctacaaggcc
     6961 aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca tcaagttgga catcacctcc
     7021 cacaacgagg actacaccat cgtggaacag tacgaacgcg ccgagggccg ccactccacc
     7081 ggcggcatgg acgagcttta caagtaagct tctcggtacc aaattccaga aaagaggcct
     7141 cccgaaaggg gggccttttt tcgttttggt cccgctggtc tcacaacttg tcttcattca
     7201 ctcctcaggt gcaggctgcc tatcagaagg tggtggctgg tgtggccaat gccctggctc
     7261 acaaatacca ctgagatctt tttccctctg ccaaaaatta tggggacatc atgaagcccc
     7321 ttgagcatct gacttctggc taataaagga aatttatttt cattgcaata gtgtgttgga
     7381 attttttgtg tctctcactc ggaaggacat atgggagggc aaatcattta aaacatcaga
     7441 atgagtattt ggtttagagt ttggcaacat atgcccatat gctggctgcc atgaacaaag
     7501 gttggctata aagaggtcat cagtatatga aacagccccc tgctgtccat tccttattcc
     7561 atagaaaagc cttgacttga ggttagattt tttttatatt ttgttttgtg ttattttttt
     7621 ctttaacatc cctaaaattt tccttacatg ttttactagc cagatttttc ctcctctcct
     7681 gactactccc agtcatagct gtccctcttc tcttatggag atccctcgac ctgcgctggt
     7741 gcgtttttat gcttgtagta ttgtataatg tttttaagat ccaagcttct gacctcttct
     7801 cttcctccca cagggcctcg agacaattgc atcgatggta ccagtgaaag tctgcaatga
     7861 attcgagttg gcgcgcctgt cattctaaat ctctctttca gcctaaagct ttttccccgt
     7921 atccccccag gtgtctgcag gctcaaagag cagcgagaag cgttcagagg aaagcgatcc
     7981 cgtgccacct tccccgtgcc cgggctgtcc ccgcacgctg ccggctcggg gatgcggggg
     8041 gagcgccgga ccggagcgga gccccgggcg gctcgctgct gccccctagc gggggaggga
     8101 cgtaattaca tccctggggg ctttgggagg gggctgtccc cgtgagctcc caggcgcgcc
     8161 tgtcattcta aatctctctt tcagcctaaa gctttttccc cgtatccccc caggtgtctg
     8221 caggctcaaa gagcagcgag aagcgttcag aggaaagcga tcccgtgcca ccttccccgt
     8281 gcccgggctg tccccgcacg ctgccggctc ggggatgcgg ggggagcgcc ggaccggagc
     8341 ggagccccgg gcggctcgct gctgccccct agcgggggag ggacgtaatt acatccctgg
     8401 gggctttggg ggggggctgt ccccgtgagc tcccaggcgc gccaactcgg taccgtatcg
     8461 atgtcgacgt taacgctagt gatatcaact ttgtatagaa aagttggctc cgaattcgcc
     8521 cttgacagca gagatccagt ttggttaatt aattggctcc ggtgcccgtc agtgggcaga
     8581 gcgcacatcg cccacagtcc ccgagaagtt gtggggaggg gtcggcaatt gaaccggtgc
     8641 ctagagaagg tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt
     8701 tcccgagggt gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg
     8761 caacgggttt gccgccagaa cacaggtaag tgccgtgtgt ggttcccgcg ggcctggcct
     8821 ctttacgggt tatggccctt gcgtgccttg aattacttcc acctggctgc agtacgtgat
     8881 tcttgatccc gagcttcggg ttggaagtgg gtgggagagt tcgaggcctt gcgcttaagg
     8941 agccccttcg cctcgtgctt gagttgaggc ctggcctggg cgctggggcc gccgcgtgcg
     9001 aatctggtgg caccttcgcg cctgtctcgc tgctttcgat aagtctctag ccatttaaaa
     9061 tttttgatga cctgctgcga cgcttttttt ctggcaagat agtcttgtaa atgcgggcca
     9121 agatctgcac actggtattt cggtttttgg ggccgcgggc ggcgacgggg cccgtgcgtc
     9181 ccagcgcaca tgttcggcga ggcggggcct gcgagcgcgg ccaccgagaa tcggacgggg
     9241 gtagtctcaa gctggccggc ctgctctggt gcctggcctc gcgccgccgt gtatcgcccc
     9301 gccctgggcg gcaaggctgg cccggtcggc accagttgcg tgagcggaaa gatggccgct
     9361 tcccggccct gctgcaggga gctcaaaatg gaggacgcgg cgctcgggag agcgggcggg
     9421 tgagtcaccc acacaaagga aaagggcctt tccgtcctca gccgtcgctt catgtgactc
     9481 cacggagtac cgggcgccgt ccaggcacct cgattagttc tcgagctttt ggagtacgtc
     9541 gtctttaggt tggggggagg ggttttatgc gatggagttt ccccacactg agtgggtgga
     9601 gactgaagtt aggccagctt ggcacttgat gtaattctcc ttggaatttg ccctttttga
     9661 gtttggatct tggttcattc tcaagcctca gacagtggtt caaagttttt ttcttccatt
     9721 tcaggtgtcg tgagaagggc gaattcgacc caagtttgta caaaaaagca ggctcgccac
     9781 catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
     9841 cggcgacgta aacggccaca agttcagcgt gaggggcgag ggcgagggcg atgccaccaa
     9901 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
     9961 cctcgtgacc accctgagcc acggcgtgca gtgcttcgcc cgctaccccg accacatgaa
    10021 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
    10081 cttcaaggac gacggcacct acaagacccg cgccgaggtg aagttcgagg gcgacaccct
    10141 agtgaaccgc atcgagctga agggcgtcga cttcaaggag gacggcaaca tcctggggca
    10201 caagctggag tacaacttca acagccacaa catctatatc atggccgtca agcagaagaa
    10261 cggcatcaag gtgaacttca agatccgcca caacgtggag gacggcagcg tgcagctcgc
    10321 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacagcca
    10381 ctacctgagc acccagtccg tgctgagcaa agaccccaac gagaagcgcg atcacatggt
    10441 cctgctggag ttccgcaccg ccgccgggat cactctcggc atggacgagc tgtacaagta
    10501 ataccaccca gctttcttgt acaaagtggt acgcgtgaat tcactcctca ggtgcaggct
    10561 gcctatcaga aggtggtggc tggtgtggcc aatgccctgg ctcacaaata ccactgagat
    10621 ctttttccct ctgccaaaaa ttatggggac atcatgaagc cccttgagca tctgacttct
    10681 ggctaataaa ggaaatttat tttcattgca atagtgtgtt ggaatttttt gtgtctctca
    10741 ctcggaagga catatgggag ggcaaatcat ttaaaacatc agaatgagta tttggtttag
    10801 agtttggcaa catatgccca tatgctggct gccatgaaca aaggttggct ataaagaggt
    10861 catcagtata tgaaacagcc ccctgctgtc cattccttat tccatagaaa agccttgact
    10921 tgaggttaga ttttttttat attttgtttt gtgttatttt tttctttaac atccctaaaa
    10981 ttttccttac atgttttact agccagattt ttcctcctct cctgactact cccagtcata
    11041 gctgtccctc ttctcttatg gagatccctc gacctgcagc ccaagcttgg cgtaatcatg
    11101 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
    11161 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
    11221 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagcgga tcgacagtac
    11281 taagcttggt ctaataccca atctctcgtc ttatccagat gttttatacg cccttagacg
    11341 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
    11401 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
    11461 aaaaggaaga gtatgcgctc acgcaactgg tccagaacct tgaccgaacg cagcggtggt
    11521 aacggcgcag tggcggtttt catggcttgt tatgactgtt tttttggggt acagtctatg
    11581 cctcgggcat ccaagcagca agcgcgttac gccgtgggtc gatgtttgat gttatggagc
    11641 agcaacgatg ttacgcagca gggcagtcgc cctaaaacaa agttaaacat catgagggaa
    11701 gcggtgatcg ccgaagtatc gactcaacta tcagaggtag ttggcgtcat cgagcgccat
    11761 ctcgaaccga cgttgctggc cgtacatttg tacggctccg cagtggatgg cggcctgaag
    11821 ccacacagcg atattgattt gctggttacg gtgaccgtaa ggcttgatga aacaacgcgg
    11881 cgagctttga tcaacgacct tttggaaact tcggcttccc ctggagagag cgagattctc
    11941 cgcgctgtag aagtcaccat tgttgtgcac gacgacatca ttccgtggcg ttatccagct
    12001 aagcgcgaac tgcaatttgg agaatggcag cgcaatgaca ttcttgcagg tatcttcgag
    12061 ccagccacga tcgacattga tctggctatc ttgctgacaa aagcaagaga acatagcgtt
    12121 gccttggtag gtccagcggc ggaggaactc tttgatccgg ttcctgaaca ggatctattt
    12181 gaggcgctaa atgaaacctt aacgctatgg aactcgccgc ccgactgggc tggcgatgag
    12241 cgaaatgtag tgcttacgtt gtcccgcatt tggtacagcg cagtaaccgg caaaatcgcg
    12301 ccgaaggatg tcgctgccga ctgggcaatg gagcgcctgc cggcccagta tcagcccgtc
    12361 atacttgaag ctagacaggc ttatcttgga caagaagaag atcgcttggc ctcgcgcgca
    12421 gatcagttgg aagaatttgt ccattacgta aaaggcgaga tcaccaaggt agtcggcaaa
    12481 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
    12541 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
    12601 gagttttcgt tccactgagc gtcagaggta tcactagtat tacagaggta agttataaca
    12661 gtcgcctaac cc
//
