LOCUS       sequence-362349-        9015 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PKA biosensor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9015)
  AUTHORS   Watabe T, Yamahira S, Takakura K, Thumkeo D, Narumiya S, Matsuda M, 
            Terai K
  TITLE     Calcium transients trigger switch-like discharge of prostaglandin 
            E(2) in an extracellular signal-regulated kinase-dependent manner.
  JOURNAL   eLife. 2024 Jan 26;12:RP86727. doi: 10.7554/eLife.86727.
  PUBMED    38276879
REFERENCE   2  (bases 1 to 9015)
  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: "eLife."; 
            date: "2024-01-26"; volume: "12:RP86727. doi"; pages: " 
            10.7554/eLife.86727"
COMMENT     Sequence Label: pPBpuro-UbC-4493NES(Booster-PKA)
FEATURES             Location/Qualifiers
     source          1..9015
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(30..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(140..161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     352..371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     599..616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   674..708
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        1004..2215
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     2052..2071
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     regulatory      2233..2242
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2239..2934
                     /codon_start=1
                     /product="monomeric far-red fluorescent protein (Shcherbo 
                     et al., 2009)"
                     /label=mKate2
                     /note="mammalian codon-optimized"
                     /translation="MVSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK
                     AVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRADMAL
                     KLVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAVARY
                     CDLPSKLGHR"
     CDS             3370..3717
                     /codon_start=1
                     /product="long flexible linker consisting of SAGG repeats 
                     (Komatsu et al., 2011)"
                     /label=EV linker
                     /translation="SAGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSAGGSAGGS
                     AGGSAGGSGSAGGSAGGSTSAGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSAGG
                     SAGGSAGGSAGG"
     CDS             3466..3813
                     /codon_start=1
                     /product="long flexible linker consisting of SAGG repeats 
                     (Komatsu et al., 2011)"
                     /label=EV linker
                     /translation="SAGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSAGGSAGGS
                     AGGSAGGSGSAGGSAGGSTSAGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSAGG
                     SAGGSAGGSAGG"
     CDS             3562..3909
                     /codon_start=1
                     /product="long flexible linker consisting of SAGG repeats 
                     (Komatsu et al., 2011)"
                     /label=EV linker
                     /translation="SAGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSAGGSAGGS
                     AGGSAGGSGSAGGSAGGSTSAGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSAGG
                     SAGGSAGGSAGG"
     CDS             3658..4005
                     /codon_start=1
                     /product="long flexible linker consisting of SAGG repeats 
                     (Komatsu et al., 2011)"
                     /label=EV linker
                     /translation="SAGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSAGGSAGGS
                     AGGSAGGSGSAGGSAGGSTSAGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSAGG
                     SAGGSAGGSAGG"
     CDS             3754..4101
                     /codon_start=1
                     /product="long flexible linker consisting of SAGG repeats 
                     (Komatsu et al., 2011)"
                     /label=EV linker
                     /translation="SAGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSAGGSAGGS
                     AGGSAGGSGSAGGSAGGSTSAGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSAGG
                     SAGGSAGGSAGG"
     CDS             4108..4761
                     /codon_start=1
                     /product="monomeric Kusabira-Orange 2 fluorescent protein"
                     /label=mKO2
                     /translation="MVSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRV
                     TMAEGGPMPFAFDLVSHVFCYGHRVFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSA
                     SVSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMY
                     LKLEGGGNHKCQFKTTYKAAKEILEMPGDHYIGHRLVRKTEGNITEQVEDAVAHS"
     CDS             4807..4833
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     misc_feature    4921..5470
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5065..5082)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5292..5311
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5473..6072
                     /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(5473..5492)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5969..5989
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(6099..6116)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6105..6329
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6376..6392)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6400..6416
                     /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(6424..6430,6431..6448,6449..6454))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6469..6490
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     repeat_region   6724..6786
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(6827..6847)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(6827..6845)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6866..6882)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6890..6906
                     /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(6914..6920,6921..6938,6939..6944))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6959..6980
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7097..7114)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7268..7856)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7348..7367)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8027..8818,8819..8887))
                     /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     8650..8669
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8888..8992)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tggggcgcgc
      661 cattctagat taaccctaga aagatagtct gcgtaaaatt gacgcatgca ttcttgaaat
      721 attgctctct ctttctaaat agcgcgaatc cgtcgctgtg catttaggac atctcagtcg
      781 ccgcttggag ctcccgtgag gcgtgcttgt caatgcggta agtgtcactg attttgaact
      841 ataacgaccg cgtgagtcaa aatgacgcat gattatcttt tacgtgactt ttaagattta
      901 actcatacga taattatatt gttatttcat gttctactta cgtgataact tattatatat
      961 atattttctt gttatagata tcaactagaa tgctagcctc gagggcctcc gcgccgggtt
     1021 ttggcgcctc ccgcgggcgc ccccctcctc acggcgagcg ctgccacgtc agacgaaggg
     1081 cgcagcgagc gtcctgatcc ttccgcccgg acgctcagga cagcggcccg ctgctcataa
     1141 gactcggcct tagaacccca gtatcagcag aaggacattt taggacggga cttgggtgac
     1201 tctagggcac tggttttctt tccagagagc ggaacaggcg aggaaaagta gtcccttctc
     1261 ggcgattctg cggagggatc tccgtggggc ggtgaacgcc gatgattata taaggacgcg
     1321 ccgggtgtgg cacagctagt tccgtcgcag ccgggatttg ggtcgcagtt cttgtttgtg
     1381 gatcgctgtg atcgtcactt ggtgagtagc gggctgctgg gctggccggg gctttcgtgg
     1441 ccgccgggcc gctcggtggg acggaggcgt gtggagagac cgccaagggc tgtagtctgg
     1501 gtccgcgagc aaggttgccc tgaactgggg gttgggggga gcgcagcaaa atggcggctg
     1561 ttcccgagtc ttgaatggaa gacgcttgtg aggcgggctg tgaggtcgtt gaaacaaggt
     1621 ggggggcatg gtgggcggca agaacccaag gtcttgaggc cttcgctaat gcgggaaagc
     1681 tcttattcgg gtgagatggg ctggggcacc atctggggac cctgacgtga agtttgtcac
     1741 tgactggaga actcggtttg tcgtctgttg cgggggcggc agttatggcg gtgccgttgg
     1801 gcagtgcacc cgtacctttg ggagcgcgcg ccctcgtcgt gtcgtgacgt cacccgttct
     1861 gttggcttat aatgcagggt ggggccacct gccggtaggt gtgcggtagg cttttctccg
     1921 tcgcaggacg cagggttcgg gcctagggta ggctctcctg aatcgacagg cgccggacct
     1981 ctggtgaggg gagggataag tgaggcgtca gtttctctgg tcggttttat gtacctatct
     2041 tcttaagtag ctgaagctcc ggttttgaac tatgcgctcg gggttggcga gtgtgttttg
     2101 tgaagttttt taggcacctt ttgaaatgta atcatttggg tcaatatgta attttcagtg
     2161 ttagactagt aaattgtccg ctaaattctg gccgtttttg gcttttttgt tagacggatc
     2221 catgaattcg ccgccaccat ggtgagcgag ctgattaagg agaacatgca catgaagctg
     2281 tacatggagg gcaccgtgaa caaccaccac ttcaagtgca catccgaggg cgaaggcaag
     2341 ccctacgagg gcacccagac catgagaatc aaggcggtcg agggcggccc tctccccttc
     2401 gccttcgaca tcctggctac cagcttcatg tacggcagca aaaccttcat caaccacacc
     2461 cagggcatcc ccgacttctt taagcagtcc ttccccgagg gcttcacatg ggagagagtc
     2521 accacatacg aagacggggg cgtgctgacc gctacccagg acaccagcct ccaggacggc
     2581 tgcctcatct acaacgtcaa gatcagaggg gtgaacttcc catccaacgg ccctgtgatg
     2641 cagaagaaaa cactcggctg ggaggcctcc accgagaccc tgtaccccgc tgacggcggc
     2701 ctggaaggca gagccgacat ggccctgaag ctcgtgggcg ggggccacct gatctgcaac
     2761 ttgaagacca catacagatc caagaaaccc gctaagaacc tcaagatgcc cggcgtctac
     2821 tatgtggaca gaagactgga aagaatcaag gaggccgaca aagagaccta cgtcgagcag
     2881 cacgaggtgg ctgtggccag atactgcgac ctccctagca aactggggca cagactcgag
     2941 aagttttctc aagaacagat cggcgaaaac attgtgtgca gggtcatttg taccacgggt
     3001 caaattccca tccgagattt gtcagctgat atttcacaag tgcttaagga aaaacgatcc
     3061 ataaagaaag tttggacatt tggtagaaac ccagcctgtg actatcattt aggaaacatt
     3121 tcaagactgt caaataagca tttccaaata ctactaggag aagacggtaa ccttttattg
     3181 aatgacattt ccactaatgg gacctggtta aatgggcaaa aagtcgagaa gaacagcaat
     3241 cagttactgt cccaaggtga tgaaataacc gttggtgtag gcgtggaatc agatatttta
     3301 tctctggtca ttttcataaa cgacaaattt aagcagtgcc tggagcagaa caaagttgat
     3361 cgcggtacca gtgctggtgg tagtgctggt ggtagtgctg gtggtagtgc tggtggtagt
     3421 gctggtggtt ccggcagtgc tggtggtagt gctggtggta gtaccagtgc tggtggtagt
     3481 gctggtggta gtgctggtgg tagtgctggt ggtagtgctg gtggttccgg cagtgctggt
     3541 ggtagtgctg gtggtagtac cagtgctggt ggtagtgctg gtggtagtgc tggtggtagt
     3601 gctggtggta gtgctggtgg ttccggcagt gctggtggta gtgctggtgg tagtaccagt
     3661 gctggtggta gtgctggtgg tagtgctggt ggtagtgctg gtggtagtgc tggtggttcc
     3721 ggcagtgctg gtggtagtgc tggtggtagt accagtgctg gtggtagtgc tggtggtagt
     3781 gctggtggta gtgctggtgg tagtgctggt ggttccggca gtgctggtgg tagtgctggt
     3841 ggtagtacca gtgctggtgg tagtgctggt ggtagtgctg gtggtagtgc tggtggtagt
     3901 gctggtggtt ccggyagtgc tggtggtagt gctggtggta gtaccagtgc tggtggtagt
     3961 gctggtggta gtgctggtgg tagtgctggt ggtagtgctg gtggttccgg cagtgctggt
     4021 ggtagtgctg gtggtagtac cagtgctggt ggtagtgctg gtggtagtgc tggtggtagt
     4081 gctggtggta gtgctggtgg ttccggaatg gtgagtgtga ttaaaccaga gatgaagatg
     4141 aggtactaca tggacggctc cgtcaatggg catgagttca caattgaagg tgaaggcaca
     4201 ggcagacctt acgagggaca tcaagagatg acactacgcg tcacaatggc cgagggcggg
     4261 ccaatgcctt tcgcgtttga cttagtgtca cacgtgttct gttacggcca cagagtattt
     4321 actaaatatc cagaagagat accagactat ttcaaacaag catttcctga aggcctgtca
     4381 tgggaaaggt cgttggagtt cgaagatggt gggtccgctt cagtcagtgc gcatataagc
     4441 cttagaggaa acaccttcta ccacaaatcc aaatttactg gggttaactt tcctgccgat
     4501 ggtcctatca tgcaaaacca aagtgttgat tgggagccat caaccgagaa aattactgcc
     4561 agcgacggag ttctgaaggg tgatgttacg atgtacctaa aacttgaagg aggcggcaat
     4621 cacaaatgcc aattcaagac tacttacaag gcggcaaaag agattcttga aatgccagga
     4681 gaccattaca tcggccatcg cctcgtcagg aaaaccgaag gcaacattac tgagcaggta
     4741 gaagatgcag tagctcattc cgcggccgca ttgaggcgcg cgacgctggt tgactctaga
     4801 ctgcagctgc ctcctctgga acgcctgact ctggattaag tcgacggccg ctctagagtc
     4861 aactaactta agctagcaac ggtttccctc tagcgggatc aattccgccc ccccccccta
     4921 acgttactgg ccgaagccgc ttggaataag gccggtgtgc gtttgtctat atgttatttt
     4981 ccaccatatt gccgtctttt ggcaatgtga gggcccggaa acctggccct gtcttcttga
     5041 cgagcattcc taggggtctt tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg
     5101 tgaaggaagc agttcctctg gaagcttctt gaagacaaac aacgtctgta gcgacccttt
     5161 gcaggcagcg gaacccccca cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat
     5221 aagatacacc tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg atagttgtgg
     5281 aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg
     5341 taccccattg tatgggatct gatctggggc ctcggtgcac atgctttaca tgtgtttagt
     5401 cgaggttaaa aaaacgtcta ggccccccga accacgggga cgtggttttc ctttgaaaaa
     5461 cacgataata ccatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc
     5521 cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc
     5581 gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc
     5641 gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg
     5701 accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc
     5761 gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac
     5821 cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc
     5881 aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg
     5941 cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc
     6001 accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag
     6061 cccggtgcct gatcgatgat ctagagctcg ctgatcagcc tcgactgtgc cttctagttg
     6121 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
     6181 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
     6241 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
     6301 gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacct gcagcccaag
     6361 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     6421 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     6481 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     6541 gcggatctta aaagttttgt tactttatag aagaaatttt gagtttttgt ttttttttaa
     6601 taaataaata aacataaata aattgtttgt tgaatttatt attagtatgt aagtgtaaat
     6661 ataataaaac ttaatatcta ttcaaattaa taaataaacc tcgatataca gaccgataaa
     6721 acacatgcgt caattttacg catgattatc tttaacgtac gtcacaatat gattatcttt
     6781 ctagggttaa tctagtatac gcgtttaatt aatccagctt ttgttccctt tagtgagggt
     6841 taattgcgcg cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     6901 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     6961 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     7021 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     7081 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     7141 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     7201 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     7261 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     7321 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     7381 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     7441 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     7501 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     7561 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     7621 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     7681 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     7741 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     7801 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     7861 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     7921 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     7981 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     8041 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     8101 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     8161 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     8221 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     8281 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     8341 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     8401 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     8461 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     8521 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     8581 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     8641 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     8701 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     8761 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     8821 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     8881 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     8941 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     9001 cccgaaaagt gccac
//
