LOCUS       sequence-358257-        9528 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 9528)
  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 9528)
  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-4493NES(Booster-PKA)
FEATURES             Location/Qualifiers
     source          1..9528
                     /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)"
     enhancer        1011..1390
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1393..1668
                     /label=chicken beta-actin promoter
     intron          1669..2677
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     2601..2623
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     regulatory      2747..2756
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2753..3448
                     /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             3884..4231
                     /codon_start=1
                     /product="long flexible linker consisting of SAGG repeats 
                     (Komatsu et al., 2011)"
                     /label=EV linker
                     /translation="SAGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSAGGSAGGS
                     AGGSAGGSGSAGGSAGGSTSAGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSAGG
                     SAGGSAGGSAGG"
     CDS             3980..4327
                     /codon_start=1
                     /product="long flexible linker consisting of SAGG repeats 
                     (Komatsu et al., 2011)"
                     /label=EV linker
                     /translation="SAGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSAGGSAGGS
                     AGGSAGGSGSAGGSAGGSTSAGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSAGG
                     SAGGSAGGSAGG"
     CDS             4076..4423
                     /codon_start=1
                     /product="long flexible linker consisting of SAGG repeats 
                     (Komatsu et al., 2011)"
                     /label=EV linker
                     /translation="SAGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSAGGSAGGS
                     AGGSAGGSGSAGGSAGGSTSAGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSAGG
                     SAGGSAGGSAGG"
     CDS             4172..4519
                     /codon_start=1
                     /product="long flexible linker consisting of SAGG repeats 
                     (Komatsu et al., 2011)"
                     /label=EV linker
                     /translation="SAGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSAGGSAGGS
                     AGGSAGGSGSAGGSAGGSTSAGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSAGG
                     SAGGSAGGSAGG"
     CDS             4268..4615
                     /codon_start=1
                     /product="long flexible linker consisting of SAGG repeats 
                     (Komatsu et al., 2011)"
                     /label=EV linker
                     /translation="SAGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSAGGSAGGS
                     AGGSAGGSGSAGGSAGGSTSAGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSAGG
                     SAGGSAGGSAGG"
     CDS             4622..5275
                     /codon_start=1
                     /product="monomeric Kusabira-Orange 2 fluorescent protein"
                     /label=mKO2
                     /translation="MVSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRV
                     TMAEGGPMPFAFDLVSHVFCYGHRVFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSA
                     SVSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMY
                     LKLEGGGNHKCQFKTTYKAAKEILEMPGDHYIGHRLVRKTEGNITEQVEDAVAHS"
     CDS             5321..5347
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     misc_feature    5435..5983
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5579..5596)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5806..5825
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5986..6585
                     /codon_start=1
                     /gene="pac from Streptomyces"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVECPKDRATWCMTRKPGA"
     primer_bind     complement(5986..6005)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6482..6502
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(6612..6629)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6618..6842
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6889..6905)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6913..6929
                     /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(6937..6943,6944..6961,6962..6967))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6982..7003
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     repeat_region   7237..7299
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(7340..7360)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(7340..7358)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7379..7395)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7403..7419
                     /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(7427..7433,7434..7451,7452..7457))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7472..7493
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7610..7627)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7781..8369)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7861..7880)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8540..9331,9332..9400))
                     /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     9163..9182
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9401..9505)
                     /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 tgctagcatg ggcccatctc gacattgatt
     1021 attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
     1081 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
     1141 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
     1201 acgtcaatgg gtggactatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
     1261 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
     1321 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
     1381 tattaccatg ggtcgaggtg agccccacgt tctgcttcac tctccccatc tcccccccct
     1441 ccccaccccc aattttgtat ttatttattt tttaattatt ttgtgcagcg atgggggcgg
     1501 gggggggggg ggcgcgcgcc aggcggggcg gggcggggcg aggggcgggg cggggcgagg
     1561 cggagaggtg cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc ttttatggcg
     1621 aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg agtcgctgcg
     1681 ttgccttcgc cccgtgcccc gctccgcgcc gcctcgcgcc gcccgccccg gctctgactg
     1741 accgcgttac tcccacaggt gagcgggcgg gacggccctt ctcctccggg ctgtaattag
     1801 cgcttggttt aatgacggct cgtttctttt ctgtggctgc gtgaaagcct taaagggctc
     1861 cgggagggcc ctttgtgcgg gggggagcgg ctcggggggt gcgtgcgtgt gtgtgtgcgt
     1921 ggggagcgcc gcgtgcggcc cgcgctgccc ggcggctgtg agcgctgcgg gcgcggcgcg
     1981 gggctttgtg cgctccgcgt gtgcgcgagg ggagcgcggc cgggggcggt gccccgcggt
     2041 gcgggggggc tgcgagggga acaaaggctg cgtgcggggt gtgtgcgtgg gggggtgagc
     2101 agggggtgtg ggcgcggcgg tcgggctgta acccccccct gcacccccct ccccgagttg
     2161 ctgagcacgg cccggcttcg ggtgcggggc tccgtgcggg gcgtggcgcg gggctcgccg
     2221 tgccgggcgg ggggtggcgg caggtggggg tgccgggcgg ggcggggccg cctcgggccg
     2281 gggagggctc gggggagggg cgcggcggcc ccggagcgcc ggcggctgtc gaggcgcggc
     2341 gagccgcagc cattgccttt tatggtaatc gtgcgagagg gcgcagggac ttcctttgtc
     2401 ccaaatctgg cggagccgaa atctgggagg cgccgccgca ccccctctag cgggcgcggg
     2461 cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga gggccttcgt gcgtcgccgc
     2521 gccgccgtcc ccttctccat ctccagcctc ggggctgccg cagggggacg gctgccttcg
     2581 ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg accggcggct ctagagcctc
     2641 tgctaaccat gttcatgcct tcttcttttt cctacagctc ctgggcaacg tgctggttgt
     2701 tgtgctgtct catcattttg gcaaagaatt gggatccgaa ttcgccgcca ccatggtgag
     2761 cgagctgatt aaggagaaca tgcacatgaa gctgtacatg gagggcaccg tgaacaacca
     2821 ccacttcaag tgcacatccg agggcgaagg caagccctac gagggcaccc agaccatgag
     2881 aatcaaggcg gtcgagggcg gccctctccc cttcgccttc gacatcctgg ctaccagctt
     2941 catgtacggc agcaaaacct tcatcaacca cacccagggc atccccgact tctttaagca
     3001 gtccttcccc gagggcttca catgggagag agtcaccaca tacgaagacg ggggcgtgct
     3061 gaccgctacc caggacacca gcctccagga cggctgcctc atctacaacg tcaagatcag
     3121 aggggtgaac ttcccatcca acggccctgt gatgcagaag aaaacactcg gctgggaggc
     3181 ctccaccgag accctgtacc ccgctgacgg cggcctggaa ggcagagccg acatggccct
     3241 gaagctcgtg ggcgggggcc acctgatctg caacttgaag accacataca gatccaagaa
     3301 acccgctaag aacctcaaga tgcccggcgt ctactatgtg gacagaagac tggaaagaat
     3361 caaggaggcc gacaaagaga cctacgtcga gcagcacgag gtggctgtgg ccagatactg
     3421 cgacctccct agcaaactgg ggcacagact cgagaagttt tctcaagaac agatcggcga
     3481 aaacattgtg tgcagggtca tttgtaccac gggtcaaatt cccatccgag atttgtcagc
     3541 tgatatttca caagtgctta aggaaaaacg atccataaag aaagtttgga catttggtag
     3601 aaacccagcc tgtgactatc atttaggaaa catttcaaga ctgtcaaata agcatttcca
     3661 aatactacta ggagaagacg gtaacctttt attgaatgac atttccacta atgggacctg
     3721 gttaaatggg caaaaagtcg agaagaacag caatcagtta ctgtcccaag gtgatgaaat
     3781 aaccgttggt gtaggcgtgg aatcagatat tttatctctg gtcattttca taaacgacaa
     3841 atttaagcag tgcctggagc agaacaaagt tgatcgcggt accagtgctg gtggtagtgc
     3901 tggtggtagt gctggtggta gtgctggtgg tagtgctggt ggttccggca gtgctggtgg
     3961 tagtgctggt ggtagtacca gtgctggtgg tagtgctggt ggtagtgctg gtggtagtgc
     4021 tggtggtagt gctggtggtt ccggcagtgc tggtggtagt gctggtggta gtaccagtgc
     4081 tggtggtagt gctggtggta gtgctggtgg tagtgctggt ggtagtgctg gtggttccgg
     4141 cagtgctggt ggtagtgctg gtggtagtac cagtgctggt ggtagtgctg gtggtagtgc
     4201 tggtggtagt gctggtggta gtgctggtgg ttccggcagt gctggtggta gtgctggtgg
     4261 tagtaccagt gctggtggta gtgctggtgg tagtgctggt ggtagtgctg gtggtagtgc
     4321 tggtggttcc ggcagtgctg gtggtagtgc tggtggtagt accagtgctg gtggtagtgc
     4381 tggtggtagt gctggtggta gtgctggtgg tagtgctggt ggttccggca gtgctggtgg
     4441 tagtgctggt ggtagtacca gtgctggtgg tagtgctggt ggtagtgctg gtggtagtgc
     4501 tggtggtagt gctggtggtt ccggcagtgc tggtggtagt gctggtggta gtaccagtgc
     4561 tggtggtagt gctggtggta gtgctggtgg tagtgctggt ggtagtgctg gtggttccgg
     4621 aatggtgagt gtgattaaac cagagatgaa gatgaggtac tacatggacg gctccgtcaa
     4681 tgggcatgag ttcacaattg aaggtgaagg cacaggcaga ccttacgagg gacatcaaga
     4741 gatgacacta cgcgtcacaa tggccgaggg cgggccaatg cctttcgcgt ttgacttagt
     4801 gtcacacgtg ttctgttacg gccacagagt atttactaaa tatccagaag agataccaga
     4861 ctatttcaaa caagcatttc ctgaaggcct gtcatgggaa aggtcgttgg agttcgaaga
     4921 tggtgggtcc gcttcagtca gtgcgcatat aagccttaga ggaaacacct tctaccacaa
     4981 atccaaattt actggggtta actttcctgc cgatggtcct atcatgcaaa accaaagtgt
     5041 tgattgggag ccatcaaccg agaaaattac tgccagcgac ggagttctga agggtgatgt
     5101 tacgatgtac ctaaaacttg aaggaggcgg caatcacaaa tgccaattca agactactta
     5161 caaggcggca aaagagattc ttgaaatgcc aggagaccat tacatcggcc atcgcctcgt
     5221 caggaaaacc gaaggcaaca ttactgagca ggtagaagat gcagtagctc attccgcggc
     5281 cgcattgagg cgcgcgacgc tggttgactc tagactgcag ctgcctcctc tggaacgcct
     5341 gactctggat taagtcgacg gccgctctag agtcaactaa cttaagctag caacggtttc
     5401 cctctagcgg gatcaattcc gccccccccc cctaacgtta ctggccgaag ccgcttggaa
     5461 taaggccggt gtgcgtttgt ctatatgtta ttttccacca tattgccgtc ttttggcaat
     5521 gtgagggccc ggaaacctgg ccctgtcttc ttgacgagca ttcctagggg tctttcccct
     5581 ctcgccaaag gaatgcaagg tctgttgaat gtcgtgaagg aagcagttcc tctggaagct
     5641 tcttgaagac aaacaacgtc tgtagcgacc ctttgcaggc agcggaaccc cccacctggc
     5701 gacaggtgcc tctgcggcca aaagccacgt gtataagata cacctgcaaa ggcggcacaa
     5761 ccccagtgcc acgttgtgag ttggatagtt gtggaaagag tcaaatggct ctcctcaagc
     5821 gtattcaaca aggggctgaa ggatgcccag aaggtacccc attgtatggg atctgatctg
     5881 gggcctcggt gcacatgctt tacatgtgtt tagtcgaggt taaaaaacgt ctaggccccc
     5941 cgaaccacgg ggacgtggtt ttcctttgaa aaacacgata ataccatgac cgagtacaag
     6001 cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc
     6061 gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg
     6121 gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg
     6181 gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg
     6241 gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg
     6301 cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg
     6361 gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc
     6421 cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc
     6481 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgagtgcccg
     6541 aaggaccgcg cgacctggtg catgacccgc aagcccggtg cctgatcgat gatctagagc
     6601 tcgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     6661 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     6721 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     6781 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     6841 ggcttctgag gcggaaagaa cctgcagccc aagcttggcg taatcatggt catagctgtt
     6901 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     6961 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     7021 gcccgctttc cagtcgggaa acctgtcgtg ccagcggatc ttaaaagttt tgttacttta
     7081 tagaagaaat tttgagtttt tgtttttttt taataaataa ataaacataa ataaattgtt
     7141 tgttgaattt attattagta tgtaagtgta aatataataa aacttaatat ctattcaaat
     7201 taataaataa acctcgatat acagaccgat aaaacacatg cgtcaatttt acgcatgatt
     7261 atctttaacg tacgtcacaa tatgattatc tttctagggt taatctagta tacgcgttta
     7321 attaatccag cttttgttcc ctttagtgag ggttaattgc gcgcttggcg taatcatggt
     7381 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     7441 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     7501 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     7561 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     7621 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     7681 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     7741 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     7801 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     7861 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     7921 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     7981 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     8041 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     8101 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     8161 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     8221 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     8281 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     8341 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     8401 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     8461 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     8521 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     8581 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     8641 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     8701 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     8761 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     8821 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     8881 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     8941 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     9001 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     9061 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     9121 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     9181 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     9241 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     9301 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     9361 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     9421 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     9481 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccac
//
