LOCUS       sequence-362149-       13100 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Calcium biosensor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13100)
  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 13100)
  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: pCSIIhyg-iRFP670-P2Av3-RGECO1.0
FEATURES             Location/Qualifiers
     source          1..13100
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(57..76)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        248..627
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        629..827
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     782..802
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             845..1025
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1072..1197
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1689..1922
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2107..2151
                     /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    2425..2542
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2598..3760
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2829..3751
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3708..3728
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      3786..3795
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4728..4757
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4764..4820
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             4836..4892
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKAGHAVRAIGRLSS"
     primer_bind     5588..5607
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             5631..6074
                     /codon_start=1
                     /product="calcium-binding protein"
                     /label=calmodulin
                     /translation="DQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTEA
                     ELQDMINEVDADGDGTFDFPEFLTMMARKMNDTDSEEEIREAFRVFDKDGNGYIGAAEL
                     RHVMTDLGEKLTDEEVDEMIRVADIDGDGQVNYEEFVQMMTAK"
     misc_feature    6147..6714
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(6308..6325)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     6535..6554
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             6753..7772
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGYV
                     LRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPET
                     ELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQTV
                     MDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFGD
                     SQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGNF
                     DDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE"
     misc_feature    7808..8396
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7861..7881)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8279..8290)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8915..9095
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(9121..9138)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9127..9351
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      9397..9825
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9484..9503)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9694..9715
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9834..9854)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        9839..10168
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      10019..10154
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     10081..10100
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        10216..10263
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             10282..10656
                     /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    10786..10907
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10823..10842)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10877..10896
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(10956..10972)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10956..10972)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10969..10991)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10980..10996
                     /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(11004..11010,11011..11028,11029..11034))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11049..11070
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11187..11204)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11358..11946)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11438..11457)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12117..12908,12909..12977))
                     /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     12740..12759
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12978..13082)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca
       61 atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc
      121 gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc
      181 atgaagaatc tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat
      241 acgcgttgac attgattatt gactagttat taatagtaat caattacggg gtcattagtt
      301 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
      361 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
      421 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
      481 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
      541 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
      601 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt
      661 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
      721 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
      781 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg
      841 tactgggtct ctctggttag accagatctg agcctgggag ctctctggct aactagggaa
      901 cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct
      961 gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc
     1021 tagcagtggc gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga
     1081 cgcaggactc ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt
     1141 acgccaaaaa ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt
     1201 attaagcggg ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag
     1261 aaaaaatata aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt
     1321 aatcctggcc tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca
     1381 tcccttcaga caggatcaga agaacttaga tcattatata atacagtagc aaccctctat
     1441 tgtgtgcatc aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa
     1501 gagcaaaaca aaagtaagac caccgcacag caagggccgc tgatcttcag acctggagga
     1561 ggagatatga gggacaattg gagaagtgaa ttatataaat ataaagtagt aaaaattgaa
     1621 ccattaggag tagcacccac caaggcaaag agaagagtgg tgcagagaga aaaaagagca
     1681 gtgggaatag gagctttgtt ccttgggttc ttgggagcag caggaagcac tatgggcgca
     1741 gcgtcaatga cgctgacggt acaggccaga caattattgt ctggtatagt gcagcagcag
     1801 aacaatttgc tgagggctat tgaggcgcaa cagcatctgt tgcaactcac agtctggggc
     1861 atcaagcagc tccaggcaag aatcctggct gtggaaagat acctaaagga tcaacagctc
     1921 ctggggattt ggggttgctc tggaaaactc atttgcacca ctgctgtgcc ttggaatgct
     1981 agttggagta ataaatctct ggaacagatt tggaatcaca cgacctggat ggagtgggac
     2041 agagaaatta acaattacac aagcttaata cactccttaa ttgaagaatc gcaaaaccag
     2101 caagaaaaga atgaacaaga attattggaa ttagataaat gggcaagttt gtggaattgg
     2161 tttaacataa caaattggct gtggtatata aaattattca taatgatagt aggaggcttg
     2221 gtaggtttaa gaatagtttt tgctgtactt tctatagtga atagagttag gcagggatat
     2281 tcaccattat cgtttcagac ccacctccca accccgaggg gacccgacag gcccgaagga
     2341 atagaagaag aaggtggaga gagagacaga gacagatcca ttcgattagt gaacggatct
     2401 acaaatggca gtattcatcc acaattttaa aagaaaaggg gggattgggg ggtacagtgc
     2461 aggggaaaga atagtagaca taatagcaac agacatacaa actaaagaat tacaaaaaca
     2521 aattacaaaa attcaaaatt ttcgggttta ttacagggac agcagaaatt cactttgaat
     2581 taattcaagc ttcgtgaggc tccggtgccc gtcagtgggc agagcgcaca tcgcccacag
     2641 tccccgagaa gttgggggga ggggtcggca attgaaccgg tgcctagaga aggtggcgcg
     2701 gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag ggtgggggag
     2761 aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgcaacggg tttgccgcca
     2821 gaacacaggt aagtgccgtg tgtggttccc gcgggcctgg cctctttacg ggttatggcc
     2881 cttgcgtgcc ttgaattact tccacctggc tccagtacgt gattcttgat cccgagctgg
     2941 agccaggggc gggccttgcg ctttaggagc cccttcgcct cgtgcttgag ttgaggcctg
     3001 gcctgggcgc tggggccgcc gcgtgcgaat ctggtggcac cttcgcgcct gtctcgctgc
     3061 tttcgataag tctctagcca tttaaaattt ttgatgacct gctgcgacgc tttttttctg
     3121 gcaagatagt cttgtaaatg cgggccagga tctgcacact ggtatttcgg tttttggggc
     3181 cgcgggcggc gacggggccc gtgcgtccca gcgcacatgt tcggcgaggc ggggcctgcg
     3241 agcgcggcca ccgagaatcg gacgggggta gtctcaagct ggccggcctg ctctggtgcc
     3301 tggcctcgcg ccgccgtgta tcgccccgcc ctgggcggca aggctggccc ggtcggcacc
     3361 agttgcgtga gcggaaagat ggccgcttcc cggccctgct ccagggggct caaaatggag
     3421 gacgcggcgc tcgggagagc gggcgggtga gtcacccaca caaaggaaag gggcctttcc
     3481 gtcctcagcc gtcgcttcat gtgactccac ggagtaccgg gcgccgtcca ggcacctcga
     3541 ttagttctgg agcttttgga gtacgtcgtc tttaggttgg ggggaggggt tttatgcgat
     3601 ggagtttccc cacactgagt gggtggagac tgaagttagg ccagcttggc acttgatgta
     3661 attctccttg gaatttgccc tttttgagtt tggatcttgg ttcattctca agcctcagac
     3721 agtggttcaa agtttttttc ttccatttca ggtgtcgtga acacgctacc ggtctcgacg
     3781 aattcgccac catggcccgc aaggtggacc tgacctcctg cgaccgcgag cccatccaca
     3841 tccccggctc catccagccc tgcggctgcc tgctggcctg cgacgcccag gccgtgcgca
     3901 tcacccgcat caccgagaac gccggcgcct tcttcgggcg cgagaccccc cgcgtgggcg
     3961 agctgctggc cgactacttc ggcgagaccg aggcccacgc cctgcgcaac gccctggccc
     4021 agtcctccga ccccaagcgc cccgccctga tcttcggctg gcgcgacggc ctgaccgggc
     4081 gcaccttcga catctccctg caccgccacg acggcacctc catcatcgag ttcgagcccg
     4141 ccgcggccga gcaggctgac aaccccctgc gcctgacccg ccagatcatc gcccgcacca
     4201 aggagctgaa gtccctggag gagatggccg cccgcgtgcc ccgctacctg caggccatgc
     4261 tgggctacca tcgcgtgatg ctgtaccgct tcgccgacga cggctccggc atggtgatcg
     4321 gcgaggccaa gcgctccgac ctggagtcct tcctgggcca gcacttcccc gcctccctgg
     4381 tgccccagca ggcccgcctg ctgtacctga agaacgccat ccgcgtggtg tccgactccc
     4441 gcggcatctc ctcccgcatc gtgcccgagc acgacgcctc cggcgccgcc ctggacctgt
     4501 ccttcgccca cctgcgctcc atctccccct gccacctgga gttcctgcgc aacatgggcg
     4561 tgtccgcctc catgtccctg tccatcatca tcgacggcac cctgtggggc ctgatcatct
     4621 gccaccacta cgagccccgc gccgtgccta tggcccagcg cgtggccgcc gagatgttcg
     4681 ccgacttcct gtccctgcac ttcaccgccg cccaccagag aacgtacgag caaaagctga
     4741 tctcggagga ggacctgctc gaggcgacga attttagtct actgaaacaa gcgggagacg
     4801 tggaggaaaa ccctggacct atggtcgact catcacgtcg taagtggaat aaggcaggtc
     4861 acgcagtcag agctataggt cggctgagct cacccgtggt ttccgagcgg atgtaccccg
     4921 aggacggcgc cctgaagagc gagatcaaga aggggctgag gctgaaggac ggcggccact
     4981 acgccgccga ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg cccggcgcct
     5041 acatcgtcga catcaagttg gacatcgtgt cccacaacga ggactacacc atcgtggaac
     5101 agtgcgaacg cgccgagggc cgccactcca ccggcggcat ggacgagctg tacaagggag
     5161 gtacaggcgg gagtctggtg agcaagggcg aggaggataa catggccatc atcaaggagt
     5221 tcatgcgctt caaggtgcac atggagggct ccgtgaacgg ccacgagttc gagatcgagg
     5281 gcgagggcga gggccgcccc tacgaggcct ttcagaccgc taagctgaag gtgaccaagg
     5341 gtggccccct gcccttcgcc tgggacatcc tgtcccctca gttcatgtac ggctccaagg
     5401 cctacattaa gcacccagcc gacatccccg actacttcaa gctgtccttc cccgagggct
     5461 tcaggtggga gcgcgtgatg aacttcgagg acggcggcat tattcacgtt aaccaggact
     5521 cctccctgca ggacggcgta ttcatctaca aggtgaagct gcgcggcacc aacttccccc
     5581 ccgacggccc cgtaatgcag aagaagacca tgggctggga ggctacgcgt gaccaactga
     5641 ctgaagagca gatcgcagaa tttaaagagg ctttctccct atttgacaag gacggggatg
     5701 ggacgataac aaccaaggag ctggggacgg tgatgcggtc tctggggcag aaccccacag
     5761 aagcagagct gcaggacatg atcaatgaag tagatgccga cggtgacggc acattcgact
     5821 tccctgagtt cctgacgatg atggcaagaa aaatgaatga cacagacagt gaagaggaaa
     5881 ttagagaagc gttccgcgtg tttgataagg acggcaatgg ctacatcggc gcagcagagc
     5941 ttcgccacgt gatgacagac cttggagaga agttaacaga tgaggaggtt gatgaaatga
     6001 tcagggtagc agacatcgat ggggatggtc aggtaaacta cgaagagttt gtacaaatga
     6061 tgacagcgaa gtaatcggcg gccgcggctc tagaggatcc gttaactaac ttaagctagc
     6121 aacggtttcc ctctagcggg atcaattccg cccccccccc ctaacgttac tggccgaagc
     6181 cgcttggaat aaggccggtg tgcgtttgtc tatatgttat tttccaccat attgccgtct
     6241 tttggcaatg tgagggcccg gaaacctggc cctgtcttct tgacgagcat tcctaggggt
     6301 ctttcccctc tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct
     6361 ctggaagctt cttgaagaca aacaacgtct gtagcgaccc tttgcaggca gcggaacccc
     6421 ccacctggcg acaggtgcct ctgcggccaa aagccacgtg tataagatac acctgcaaag
     6481 gcggcacaac cccagtgcca cgttgtgagt tggatagttg tggaaagagt caaatggctc
     6541 tcctcaagcg tattcaacaa ggggctgaag gatgcccaga aggtacccca ttgtatggga
     6601 tctgatctgg ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt
     6661 ctaggccccc cgaaccacgg ggacgtggtt ttcctttgaa aaacacgatg ataagcttgc
     6721 cacaacccgt accaaagatg gatagatccg gaaagcctga actcaccgcg acgtctgtcg
     6781 agaagtttct gatcgaaaag ttcgacagcg tctccgacct gatgcagctc tcggagggcg
     6841 aagaatctcg tgctttcagc ttcgatgtag gagggcgtgg atatgtcctg cgggtaaata
     6901 gctgcgccga tggtttctac aaagatcgtt atgtttatcg gcactttgca tcggccgcgc
     6961 tcccgattcc ggaagtgctt gacattgggg agttcagcga gagcctgacc tattgcatct
     7021 cccgccgtgc acagggtgtc acgttgcaag acctgcctga aaccgaactg cccgctgttc
     7081 tgcagccggt cgcggaggcc atggatgcga tcgctgcggc cgatcttagc cagacgagcg
     7141 ggttcggccc attcggaccg caaggaatcg gtcaatacac tacatggcgt gatttcatat
     7201 gcgcgattgc tgatccccat gtgtatcact ggcaaactgt gatggacgac accgtcagtg
     7261 cgtccgtcgc gcaggctctc gatgagctga tgctttgggc cgaggactgc cccgaagtcc
     7321 ggcacctcgt gcacgcggat ttcggctcca acaatgtcct gacggacaat ggccgcataa
     7381 cagcggtcat tgactggagc gaggcgatgt tcggggattc ccaatacgag gtcgccaaca
     7441 tcttcttctg gaggccgtgg ttggcttgta tggagcagca gacgcgctac ttcgagcgga
     7501 ggcatccgga gcttgcagga tcgccgcggc tccgggcgta tatgctccgc attggtcttg
     7561 accaactcta tcagagcttg gttgacggca atttcgatga tgcagcttgg gcgcagggtc
     7621 gatgcgacgc aatcgtccga tccggagccg ggactgtcgg gcgtacacaa atcgcccgca
     7681 gaagcgcggc cgtctggacc gatggctgtg tagaagtact cgccgatagt ggaaaccgac
     7741 gccccagcac tcgtccgagg gcaaaggaat agactagagt taacatcgag ggatcaagct
     7801 tatcgataat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     7861 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     7921 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     7981 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     8041 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     8101 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     8161 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg
     8221 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     8281 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     8341 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcatcg
     8401 ataccgtcga gacctagaaa aacatggagc aatcacaagt agcaatacag cagctaccaa
     8461 tgctgattgt gcctggctag aagcacaaga ggaggaggag gtgggttttc cagtcacacc
     8521 tcaggtacct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa
     8581 agaaaagggg ggactggaag ggctaattca ctcccaacga agacaagata tccttgatct
     8641 gtggatctac cacacacaag gctacttccc tgattggcag aactacacac cagggccagg
     8701 gatcagatat ccactgacct ttggatggtg ctacaagcta gtaccagttg agcaagagaa
     8761 ggtagaagaa gccaatgaag gagagaacac ccgcttgtta caccctgtga gcctgcatgg
     8821 gatggatgac ccggagagag aagtattaga gtggaggttt gacagccgcc tagcatttca
     8881 tcacatggcc cgagagctgc atccggactg tactgggtct ctctggttag accagatctg
     8941 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc
     9001 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct
     9061 cagacccttt tagtcagtgt ggaaaatctc tagcagggcc cgtttaaacc cgctgatcag
     9121 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
     9181 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc
     9241 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
     9301 aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg
     9361 cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa
     9421 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     9481 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     9541 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     9601 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     9661 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     9721 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     9781 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt
     9841 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     9901 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     9961 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
    10021 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
    10081 tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg
    10141 cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg
    10201 atctgatcag cacgtgttga caattaatca tcggcatagt atatcggcat agtataatac
    10261 gacaaggtga ggaactaaac catggccaag ttgaccagtg ccgttccggt gctcaccgcg
    10321 cgcgacgtcg ccggagcggt cgagttctgg accgaccggc tcgggttctc ccgggacttc
    10381 gtggaggacg acttcgccgg tgtggtccgg gacgacgtga ccctgttcat cagcgcggtc
    10441 caggaccagg tggtgccgga caacaccctg gcctgggtgt gggtgcgcgg cctggacgag
    10501 ctgtacgccg agtggtcgga ggtcgtgtcc acgaacttcc gggacgcctc cgggccggcc
    10561 atgaccgaga tcggcgagca gccgtggggg cgggagttcg ccctgcgcga cccggccggc
    10621 aactgcgtgc acttcgtggc cgaggagcag gactgacacg tgctacgaga tttcgattcc
    10681 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
    10741 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca
    10801 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
    10861 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
    10921 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
    10981 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
    11041 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
    11101 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
    11161 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
    11221 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
    11281 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
    11341 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
    11401 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
    11461 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
    11521 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
    11581 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
    11641 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
    11701 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
    11761 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
    11821 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
    11881 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
    11941 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
    12001 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
    12061 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
    12121 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
    12181 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
    12241 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
    12301 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
    12361 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
    12421 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
    12481 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
    12541 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
    12601 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
    12661 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
    12721 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
    12781 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
    12841 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
    12901 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
    12961 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
    13021 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
    13081 cgcacatttc cccgaaaagt
//
