LOCUS       sequence-362356-       12390 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 12390)
  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 12390)
  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: pCSIIpuro-4493NES(Booster-PKA)
FEATURES             Location/Qualifiers
     source          1..12390
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             160..813
                     /codon_start=1
                     /product="monomeric Kusabira-Orange 2 fluorescent protein"
                     /label=mKO2
                     /translation="MVSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRV
                     TMAEGGPMPFAFDLVSHVFCYGHRVFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSA
                     SVSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMY
                     LKLEGGGNHKCQFKTTYKAAKEILEMPGDHYIGHRLVRKTEGNITEQVEDAVAHS"
     CDS             859..885
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     misc_feature    988..1537
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1132..1149)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1359..1378
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             1540..2139
                     /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(1540..1559)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2036..2056
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    2174..2762
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2227..2247)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2645..2656)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3281..3461
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(3487..3504)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3493..3717
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3763..4191
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3850..3869)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4060..4081
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4200..4220)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4205..4534
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4385..4520
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4447..4466
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4582..4629
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4648..5022
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    5152..5273
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5189..5208)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5243..5262
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5322..5338)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5322..5338)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5335..5357)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5346..5362
                     /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(5370..5376,5377..5394,5395..5400))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5415..5436
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5553..5570)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5724..6312)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5804..5823)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6483..7274,7275..7343))
                     /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     7106..7125
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7344..7448)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7523..7542)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        7714..8093
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8095..8293
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8248..8268
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             8311..8491
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    8538..8663
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    9155..9388
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             9573..9617
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    9891..10008
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        10064..11226
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          10295..11217
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     11174..11194
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      11251..11260
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             11257..11952
                     /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"
ORIGIN
        1 gctggtggta gtgctggtgg tagtgctggt ggtagtgctg gtggtagtgc tggtggttcc
       61 ggcagtgctg gtggtagtgc tggtggtagt accagtgctg gtggtagtgc tggtggtagt
      121 gctggtggta gtgctggtgg tagtgctggt ggttccggaa tggtgagtgt gattaaacca
      181 gagatgaaga tgaggtacta catggacggc tccgtcaatg ggcatgagtt cacaattgaa
      241 ggtgaaggca caggcagacc ttacgaggga catcaagaga tgacactacg cgtcacaatg
      301 gccgagggcg ggccaatgcc tttcgcgttt gacttagtgt cacacgtgtt ctgttacggc
      361 cacagagtat ttactaaata tccagaagag ataccagact atttcaaaca agcatttcct
      421 gaaggcctgt catgggaaag gtcgttggag ttcgaagatg gtgggtccgc ttcagtcagt
      481 gcgcatataa gccttagagg aaacaccttc taccacaaat ccaaatttac tggggttaac
      541 tttcctgccg atggtcctat catgcaaaac caaagtgttg attgggagcc atcaaccgag
      601 aaaattactg ccagcgacgg agttctgaag ggtgatgtta cgatgtacct aaaacttgaa
      661 ggaggcggca atcacaaatg ccaattcaag actacttaca aggcggcaaa agagattctt
      721 gaaatgccag gagaccatta catcggccat cgcctcgtca ggaaaaccga aggcaacatt
      781 actgagcagg tagaagatgc agtagctcat tccgcggccg cattgaggcg cgcgacgctg
      841 gttgactcta gactgcagct gcctcctctg gaacgcctga ctctggatta agtcgagggc
      901 ggcggccgcg gctctagagg atccgttaac taacttaagc tagcaacggt ttccctctag
      961 cgggatcaat tccgcccccc ccccctaacg ttactggccg aagccgcttg gaataaggcc
     1021 ggtgtgcgtt tgtctatatg ttattttcca ccatattgcc gtcttttggc aatgtgaggg
     1081 cccggaaacc tggccctgtc ttcttgacga gcattcctag gggtctttcc cctctcgcca
     1141 aaggaatgca aggtctgttg aatgtcgtga aggaagcagt tcctctggaa gcttcttgaa
     1201 gacaaacaac gtctgtagcg accctttgca ggcagcggaa ccccccacct ggcgacaggt
     1261 gcctctgcgg ccaaaagcca cgtgtataag atacacctgc aaaggcggca caaccccagt
     1321 gccacgttgt gagttggata gttgtggaaa gagtcaaatg gctctcctca agcgtattca
     1381 acaaggggct gaaggatgcc cagaaggtac cccattgtat gggatctgat ctggggcctc
     1441 ggtgcacatg ctttacatgt gtttagtcga ggttaaaaaa acgtctaggc cccccgaacc
     1501 acggggacgt ggttttcctt tgaaaaacac gataatacca tgaccgagta caagcccacg
     1561 gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc
     1621 gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc
     1681 gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg
     1741 gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg
     1801 ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa
     1861 cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc
     1921 gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga
     1981 gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac
     2041 ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga
     2101 ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgac tagagttaac atcgagggat
     2161 caagcttatc gataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     2221 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     2281 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
     2341 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
     2401 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc
     2461 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     2521 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt
     2581 tccatggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
     2641 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
     2701 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc
     2761 gcatcgatac cgtcgagacc tagaaaaaca tggagcaatc acaagtagca atacagcagc
     2821 taccaatgct gattgtgcct ggctagaagc acaagaggag gaggaggtgg gttttccagt
     2881 cacacctcag gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt
     2941 tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaagac aagatatcct
     3001 tgatctgtgg atctaccaca cacaaggcta cttccctgat tggcagaact acacaccagg
     3061 gccagggatc agatatccac tgacctttgg atggtgctac aagctagtac cagttgagca
     3121 agagaaggta gaagaagcca atgaaggaga gaacacccgc ttgttacacc ctgtgagcct
     3181 gcatgggatg gatgacccgg agagagaagt attagagtgg aggtttgaca gccgcctagc
     3241 atttcatcac atggcccgag agctgcatcc ggactgtact gggtctctct ggttagacca
     3301 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag
     3361 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag
     3421 atccctcaga cccttttagt cagtgtggaa aatctctagc agggcccgtt taaacccgct
     3481 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     3541 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     3601 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     3661 agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt
     3721 ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg
     3781 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     3841 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     3901 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     3961 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     4021 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     4081 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     4141 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg
     4201 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     4261 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     4321 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     4381 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     4441 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg
     4501 aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat
     4561 tttcggatct gatcagcacg tgttgacaat taatcatcgg catagtatat cggcatagta
     4621 taatacgaca aggtgaggaa ctaaaccatg gccaagttga ccagtgccgt tccggtgctc
     4681 accgcgcgcg acgtcgccgg agcggtcgag ttctggaccg accggctcgg gttctcccgg
     4741 gacttcgtgg aggacgactt cgccggtgtg gtccgggacg acgtgaccct gttcatcagc
     4801 gcggtccagg accaggtggt gccggacaac accctggcct gggtgtgggt gcgcggcctg
     4861 gacgagctgt acgccgagtg gtcggaggtc gtgtccacga acttccggga cgcctccggg
     4921 ccggccatga ccgagatcgg cgagcagccg tgggggcggg agttcgccct gcgcgacccg
     4981 gccggcaact gcgtgcactt cgtggccgag gagcaggact gacacgtgct acgagatttc
     5041 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     5101 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt
     5161 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     5221 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     5281 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg
     5341 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     5401 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     5461 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     5521 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     5581 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     5641 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     5701 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     5761 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     5821 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     5881 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     5941 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     6001 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     6061 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     6121 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     6181 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     6241 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     6301 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     6361 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     6421 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     6481 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     6541 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     6601 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     6661 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     6721 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     6781 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     6841 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     6901 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     6961 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     7021 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     7081 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     7141 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     7201 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     7261 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     7321 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     7381 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     7441 gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg
     7501 atcccctatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtatc
     7561 tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa
     7621 caaggcaagg cttgaccgac aattgcatga agaatctgct tagggttagg cgttttgcgc
     7681 tgcttcgcga tgtacgggcc agatatacgc gttgacattg attattgact agttattaat
     7741 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
     7801 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
     7861 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
     7921 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
     7981 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat
     8041 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc
     8101 ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc
     8161 tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa
     8221 aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg
     8281 tctatataag cagcgcgttt tgcctgtact gggtctctct ggttagacca gatctgagcc
     8341 tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga
     8401 gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga
     8461 cccttttagt cagtgtggaa aatctctagc agtggcgccc gaacagggac ttgaaagcga
     8521 aagggaaacc agaggagctc tctcgacgca ggactcggct tgctgaagcg cgcacggcaa
     8581 gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt gactagcgga ggctagaagg
     8641 agagagatgg gtgcgagagc gtcagtatta agcgggggag aattagatcg cgatgggaaa
     8701 aaattcggtt aaggccaggg ggaaagaaaa aatataaatt aaaacatata gtatgggcaa
     8761 gcagggagct agaacgattc gcagttaatc ctggcctgtt agaaacatca gaaggctgta
     8821 gacaaatact gggacagcta caaccatccc ttcagacagg atcagaagaa cttagatcat
     8881 tatataatac agtagcaacc ctctattgtg tgcatcaaag gatagagata aaagacacca
     8941 aggaagcttt agacaagata gaggaagagc aaaacaaaag taagaccacc gcacagcaag
     9001 ggccgctgat cttcagacct ggaggaggag atatgaggga caattggaga agtgaattat
     9061 ataaatataa agtagtaaaa attgaaccat taggagtagc acccaccaag gcaaagagaa
     9121 gagtggtgca gagagaaaaa agagcagtgg gaataggagc tttgttcctt gggttcttgg
     9181 gagcagcagg aagcactatg ggcgcagcgt caatgacgct gacggtacag gccagacaat
     9241 tattgtctgg tatagtgcag cagcagaaca atttgctgag ggctattgag gcgcaacagc
     9301 atctgttgca actcacagtc tggggcatca agcagctcca ggcaagaatc ctggctgtgg
     9361 aaagatacct aaaggatcaa cagctcctgg ggatttgggg ttgctctgga aaactcattt
     9421 gcaccactgc tgtgccttgg aatgctagtt ggagtaataa atctctggaa cagatttgga
     9481 atcacacgac ctggatggag tgggacagag aaattaacaa ttacacaagc ttaatacact
     9541 ccttaattga agaatcgcaa aaccagcaag aaaagaatga acaagaatta ttggaattag
     9601 ataaatgggc aagtttgtgg aattggttta acataacaaa ttggctgtgg tatataaaat
     9661 tattcataat gatagtagga ggcttggtag gtttaagaat agtttttgct gtactttcta
     9721 tagtgaatag agttaggcag ggatattcac cattatcgtt tcagacccac ctcccaaccc
     9781 cgaggggacc cgacaggccc gaaggaatag aagaagaagg tggagagaga gacagagaca
     9841 gatccattcg attagtgaac ggatctacaa atggcagtat tcatccacaa ttttaaaaga
     9901 aaagggggga ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac
     9961 atacaaacta aagaattaca aaaacaaatt acaaaaattc aaaattttcg ggtttattac
    10021 agggacagca gaaattcact ttgaattaat tcaagcttcg tgaggctccg gtgcccgtca
    10081 gtgggcagag cgcacatcgc ccacagtccc cgagaagttg gggggagggg tcggcaattg
    10141 aaccggtgcc tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg tgtactggct
    10201 ccgccttttt cccgagggtg ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt
    10261 tctttttcgc aacgggtttg ccgccagaac acaggtaagt gccgtgtgtg gttcccgcgg
    10321 gcctggcctc tttacgggtt atggcccttg cgtgccttga attacttcca cctggctcca
    10381 gtacgtgatt cttgatcccg agctggagcc aggggcgggc cttgcgcttt aggagcccct
    10441 tcgcctcgtg cttgagttga ggcctggcct gggcgctggg gccgccgcgt gcgaatctgg
    10501 tggcaccttc gcgcctgtct cgctgctttc gataagtctc tagccattta aaatttttga
    10561 tgacctgctg cgacgctttt tttctggcaa gatagtcttg taaatgcggg ccaggatctg
    10621 cacactggta tttcggtttt tggggccgcg ggcggcgacg gggcccgtgc gtcccagcgc
    10681 acatgttcgg cgaggcgggg cctgcgagcg cggccaccga gaatcggacg ggggtagtct
    10741 caagctggcc ggcctgctct ggtgcctggc ctcgcgccgc cgtgtatcgc cccgccctgg
    10801 gcggcaaggc tggcccggtc ggcaccagtt gcgtgagcgg aaagatggcc gcttcccggc
    10861 cctgctccag ggggctcaaa atggaggacg cggcgctcgg gagagcgggc gggtgagtca
    10921 cccacacaaa ggaaaggggc ctttccgtcc tcagccgtcg cttcatgtga ctccacggag
    10981 taccgggcgc cgtccaggca cctcgattag ttctggagct tttggagtac gtcgtcttta
    11041 ggttgggggg aggggtttta tgcgatggag tttccccaca ctgagtgggt ggagactgaa
    11101 gttaggccag cttggcactt gatgtaattc tccttggaat ttgccctttt tgagtttgga
    11161 tcttggttca ttctcaagcc tcagacagtg gttcaaagtt tttttcttcc atttcaggtg
    11221 tcgtgaacac gctaccggtc cgaattcgcc gccaccatgg tgagcgagct gattaaggag
    11281 aacatgcaca tgaagctgta catggagggc accgtgaaca accaccactt caagtgcaca
    11341 tccgagggcg aaggcaagcc ctacgagggc acccagacca tgagaatcaa ggcggtcgag
    11401 ggcggccctc tccccttcgc cttcgacatc ctggctacca gcttcatgta cggcagcaaa
    11461 accttcatca accacaccca gggcatcccc gacttcttta agcagtcctt ccccgagggc
    11521 ttcacatggg agagagtcac cacatacgaa gacgggggcg tgctgaccgc tacccaggac
    11581 accagcctcc aggacggctg cctcatctac aacgtcaaga tcagaggggt gaacttccca
    11641 tccaacggcc ctgtgatgca gaagaaaaca ctcggctggg aggcctccac cgagaccctg
    11701 taccccgctg acggcggcct ggaaggcaga gccgacatgg ccctgaagct cgtgggcggg
    11761 ggccacctga tctgcaactt gaagaccaca tacagatcca agaaacccgc taagaacctc
    11821 aagatgcccg gcgtctacta tgtggacaga agactggaaa gaatcaagga ggccgacaaa
    11881 gagacctacg tcgagcagca cgaggtggct gtggccagat actgcgacct ccctagcaaa
    11941 ctggggcaca gactcgagaa gttttctcaa gaacagatcg gcgaaaacat tgtgtgcagg
    12001 gtcatttgta ccacgggtca aattcccatc cgagatttgt cagctgatat ttcacaagtg
    12061 cttaaggaaa aacgatccat aaagaaagtt tggacatttg gtagaaaccc agcctgtgac
    12121 tatcatttag gaaacatttc aagactgtca aataagcatt tccaaatact actaggagaa
    12181 gacggtaacc ttttattgaa tgacatttcc actaatggga cctggttaaa tgggcaaaaa
    12241 gtcgagaaga acagcaatca gttactgtcc caaggtgatg aaataaccgt tggtgtaggc
    12301 gtggaatcag atattttatc tctggtcatt ttcataaacg acaaatttaa gcagtgcctg
    12361 gagcagaaca aagttgatcg cggtaccagt
//
