LOCUS       sequence-362301-       10477 bp    DNA     circular SYN 15-APR-2025
DEFINITION  OptoSTIM1(CRY2clust).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10477)
  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 10477)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: pPBbsr2-OptoSTIM1(CRY2clust)
FEATURES             Location/Qualifiers
     source          1..10477
                     /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"
     intron          1668..2684
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     2608..2630
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     2692..2711
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      2751..2760
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2757..2759,2760..2762,2763..3473)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2757..2759,2760..2762,2763..3473)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2802..2823)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3063..3082)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3410..3431
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             complement(5044..5061)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    6536..7085
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(6680..6697)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     6907..6926
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     complement(7556..7573)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7562..7786
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(7833..7849)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7857..7873
                     /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(7881..7887,7888..7905,7906..7911))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7926..7947
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8328..8344)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    8352..8368
                     /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(8376..8382,8383..8400,8401..8406))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8421..8442
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(8730..9318)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8810..8829)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9489..10280,10281..10349))
                     /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     10112..10131
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10350..10454)
                     /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 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
     1261 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
     1321 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
     1381 tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct cccccccctc
     1441 cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga tgggggcggg
     1501 gggggggggg gggcgcgccr gggggggggg gggggggggg ggggsggggs ggggsgrggc
     1561 ggagaggtgc ggcggcagcc aatcagagcg gcgcgctccg aaagtttcct tttatggcga
     1621 ggcggcggcg gcggcggccc tataaaaagc gaagcgcgcg gcgggcggga gtcgctgcgc
     1681 gctgccttcg ccccgtgccc cgctccgccg ccgcctcgcg ccgcccgccc cggctctgac
     1741 tgaccgcgtt actcccacag gtgagcgggc gggacggccc ttctcctccg ggctgtaatt
     1801 agcgcttggt ttaatgacgg cttgtttctt ttctgtggct gcgtgaaagc cttgaggggc
     1861 tccgggaggg ccctttgtgc ggggggagcg gctcgggggg tgcgtgcgtg tgtgtgtgcg
     1921 tggggagcgc cgcgtgcggc tccgcgctgc ccggcggctg tgagcgctgc gggcgcggcg
     1981 cggggctttg tgcgctccgc agtgtgcgcg aggggagcgc ggccgggggc ggtgccccgc
     2041 ggtgcggggg gggctgcgag gggaacaaag gctgcgtgcg gggtgtgtgc gtgggggggt
     2101 gagcaggggg tgtgggcgcg tcggtcgggc tgcaaccccc cctgcacccc cctccccgag
     2161 ttgctgagca cggcccggct tcgggtgcgg ggctccgtac ggggcgtggc gcggggctcg
     2221 ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg ccgcctcggg
     2281 ccggggaggg ctcgggggag gggcgcggcg gcccccggag cgccggcggc tgtcgaggcg
     2341 cggcgagccg cagccattgc cttttatggt aatcgtgcga gagggcgcag ggacttcctt
     2401 tgtcccaaat ctgtgcggag ccgaaatctg ggaggcgccg ccgcaccccc tctagcgggc
     2461 gcggggcgaa gcggtgcggc gccggcagga aggaaatggg cggggagggc cttcgtgcgt
     2521 cgccgcgccg ccgtcccctt ctccctctcc agcctcgggg ctgtccgcgg ggggacggct
     2581 gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc ggcggctcta
     2641 gagcctctgc taaccatgtt catgccttct tctttttcct acagctcctg ggcaacgtgc
     2701 tggttattgt gctgtctcat cattttggca aagaattggg atccgaattc gccaccatgg
     2761 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
     2821 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
     2881 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
     2941 tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc
     3001 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
     3061 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
     3121 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
     3181 tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca
     3241 tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc
     3301 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     3361 tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     3421 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagccaccgg
     3481 tcgccaccat gaagatggac aaaaagacca tcgtctggtt tcggagagat ttgagaatag
     3541 aagataatcc cgcgctcgcc gccgcggccc acgagggttc cgtcttcccc gttttcattt
     3601 ggtgtcctga agaagaaggc cagttttatc ccggaagggc ctctaggtgg tggatgaagc
     3661 aaagtctggc ccatcttagc cagtcactga aagcactggg cagtgatctt accctgatca
     3721 agacacacaa taccatctct gccatcctcg actgcatcag ggtgaccggc gcaacgaaag
     3781 tcgtgtttaa ccacctgtac gatccagtta gtctggtgcg cgaccacact gtgaaggaga
     3841 agctggtgga acgggggatc agtgtgcaga gctacaacgg ggaccttctg tacgagccat
     3901 gggagatcta ttgcgagaaa gggaaaccgt tcacctcctt caacagttac tggaagaaat
     3961 gtttggatat gtcaatagag tccgttatgt tgccccctcc ctggagactg atgccgatta
     4021 ctgctgctgc agaggccatc tgggcctgct ccatcgagga actcggtctg gaaaatgaag
     4081 cagaaaagcc aagcaatgca cttctcacta gagcctggag ccccggctgg tctaatgccg
     4141 acaagctgct taacgagttc atcgaaaaac aactgattga ctacgcgaag aactccaaga
     4201 aagtggtagg taactcaact agcttgctct ctccatatct ccattttggc gagatttctg
     4261 tccgccatgt atttcagtgc gctcggatga aacagattat ctgggctcgc gataaaaaca
     4321 gcgaaggcga agaaagcgcc gatctgttcc tgcgagggat cggacttcgg gaatactccc
     4381 ggtatatatg tttcaacttt ccattcacac acgagcagag tctgttgtcc cacctcaggt
     4441 tcttcccctg ggacgccgat gtcgacaagt tcaaagcctg gcggcaggga agaacaggct
     4501 atcccctggt ggatgctgga atgagagagc tgtgggccac tggctggatg cacaatagga
     4561 tccgcgtgat tgtgagctcc tttgctgtga agttcctgct gctgccttgg aagtggggca
     4621 tgaaatattt ttgggataca ctgctggatg ccgacctgga gtgtgacatc ctggggtggc
     4681 agtacatctc cggatctatt ccagatggcc atgaactgga taggctggac aaccccgctc
     4741 tgcagggcgc taagtatgac cctgagggag aatacatcag gcagtggctg cctgagctgg
     4801 ctaggctgcc aactgaatgg attcaccatc cctgggacgc tcctctgaca gtgctgaagg
     4861 ccagtggggt ggagctggga actaattacg ctaaacctat cgtggatatt gacaccgcca
     4921 gagagctgct ggctaaggcc atctcccgga ccagagaagc ccagatcatg attggcgctg
     4981 ccgctcgaga tcctccagac ctcgataacg tgtacagtgg gggaggcgga gggggtggtt
     5041 caggtggtgg tggtggtggt ggttcgggtg gtggcggagg gggaggctcc ggactcagat
     5101 ctaaggagca catgaagaag atgatgaagg acttggaggg gttacaccga gctgagcaga
     5161 gtctgcatga ccttcaggaa aggctgcaca aggcccagga ggagcaccgc acagtggagg
     5221 tggagaaggt ccatctggaa aagaagctgc gcgatgagat caaccttgct aagcaggaag
     5281 cccagcggct gaaggagctg cgggagggta ctgagaatga gcggagccgc caaaaatatg
     5341 ctgaggagga gttggagcag gttcgggagg ccttgaggaa agcagagaag gagctagaat
     5401 ctcacagctc atggtatgct ccagaggccc ttcagaagtg gctgcagctg acacatgagg
     5461 tggaggtgca atattacaac atcaagaagc aaaatgctga gaagcagctg ctggtggcca
     5521 aggagggggc tgagaagata aaaaagaaga gaaacacact ctttggcacc ttccacgtgg
     5581 cccacagctc ttccctggat gatgtagatc ataaaattct aacagctaag caagcactga
     5641 gcgaggtgac agcagcattg cgggagcgcc tgcaccgctg gcaacagatc gagatcctct
     5701 gtggcttcca gattgtcaac aaccctggca tccactcact ggtggctgcc ctcaacatag
     5761 accccagctg gatgggcagt acacgcccca accctgctca cttcatcatg actgacgacg
     5821 tggatgacat ggatgaggag attgtgtctc ccttgtccat gcagtcccct agcctgcaga
     5881 gcagtgttcg gcagcgcctg acggagccac agcatggcct gggatctcag agagatttga
     5941 cccattccga ttcggagtcc tccctccaca tgagtgaccg ccagcgtgtg gcccccaaac
     6001 ctcctcagat gagccgtgct gcagacgagg ctctcaatgc catgacttcc aatggcagcc
     6061 accggctgat cgagggggtc cacccagggt ctctggtgga gaaactgcct gacagccctg
     6121 ccctggccaa gaaggcatta ctggcgctga accatgggct ggacaaggcc cacagcctga
     6181 tggagctgag cccctcagcc ccacctggtg gctctccaca tttggattct tcccgttctc
     6241 acagccccag ctccccagac ccagacacac catctccagt tggggacagc cgagccctgc
     6301 aagccagccg aaacacacgc attccccacc tggctggcaa gaaggctgtg gctgaggagg
     6361 ataatggctc tattggcgag gaaacagact ccagcccagg ccggaagaag tttcccctca
     6421 aaatctttaa gaagcctctt aagaagtagg tcgacgggcc gcggtaacaa ttgttaacta
     6481 acttaagcta gcaacggttt ccctctagcg ggatcaattc cgcccccccc ccctaacgtt
     6541 actggccgaa gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt attttccacc
     6601 atattgccgt cttttggcaa tgtgagggcc cggaaacctg gccctgtctt cttgacgagc
     6661 attcctaggg gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag
     6721 gaagcagttc ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg
     6781 cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg tgtataagat
     6841 acacctgcaa aggcggcaca accccagtgc cacgttgtga gttggatagt tgtggaaaga
     6901 gtcaaatggc tctcctcaag cgtattcaac aaggggctga aggatgccca gaaggtaccc
     6961 cattgtatgg gatctgatct ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg
     7021 ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg ttttcctttg aaaaacacga
     7081 taataccatg gtcatgaaaa catttaacat ttctcaacaa gatctagaat tagtagaagt
     7141 agcgacagag aagattacaa tgctttatga ggataataaa catcatgtgg gagcggcaat
     7201 tcgtacgaaa acaggagaaa tcatttcggc agtacatatt gaagcgtata taggacgagt
     7261 aactgtttgt gcagaagcca ttgcgattgg tagtgcagtt tcgaatggac aaaaggattt
     7321 tgacacgatt gtagctgtta gacaccctta ttctgacgaa gtagatagaa gtattcgagt
     7381 ggtaagtcct tgtggtatgt gtagggagtt gatttcagac tatgcaccag attgttttgt
     7441 gttaatagaa atgaatggca agttagtcaa aactacgatt gaagaactca ttccactcaa
     7501 atatacccga aattaaaagt cgagatgcat cgatgatcta gagctcgctg atcagcctcg
     7561 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     7621 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     7681 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     7741 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa
     7801 agaacctgca gcccaagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     7861 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     7921 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     7981 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     8041 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     8101 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     8161 aacgcaggaa agaacatgtg agcaamakgc gtcaatttta cgcatgatta tctttaacgt
     8221 acgtcacaat atgattatct tkmyartrky aggcwtgctg gggatgcggt gggctctatg
     8281 gcttctgagg cggaaagaac ctgcagccca agcttggcgt aatcatggtc atagctgttt
     8341 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     8401 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     8461 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     8521 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     8581 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     8641 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     8701 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     8761 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     8821 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     8881 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     8941 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     9001 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     9061 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     9121 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     9181 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     9241 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     9301 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     9361 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     9421 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     9481 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     9541 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     9601 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     9661 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     9721 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     9781 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     9841 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     9901 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     9961 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
    10021 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
    10081 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
    10141 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
    10201 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
    10261 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
    10321 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
    10381 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
    10441 ataggggttc cgcgcacatt tccccgaaaa gtgccac
//
