LOCUS       sequence-362123-       11502 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 11502)
  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 11502)
  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: pCSIIbsr-GCaMP6s
FEATURES             Location/Qualifiers
     source          1..11502
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(165..182)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     392..411
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             585..983
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    1027..1615
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1080..1100)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1498..1509)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2134..2314
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(2340..2357)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2346..2570
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2616..3044
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2703..2722)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2913..2934
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3053..3073)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3058..3387
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3238..3373
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3300..3319
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3435..3482
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3501..3875
                     /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    4005..4126
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4042..4061)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4096..4115
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4175..4191)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4175..4191)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4188..4210)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4199..4215
                     /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(4223..4229,4230..4247,4248..4253))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4268..4289
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4406..4423)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4577..5165)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4657..4676)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5336..6127,6128..6196))
                     /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     5959..5978
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6197..6301)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6376..6395)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        6567..6946
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6948..7146
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7101..7121
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             7164..7344
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7391..7516
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    8008..8241
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8426..8470
                     /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    8744..8861
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        8917..10079
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          9148..10070
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     10027..10047
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      10132..10141
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(10138..10146,10147..10164,10165..10167,10168..10200,
                     10201..10203,10204..10227,10228..10254,10255..10311,
                     10312..10317,10318..10587,10588..10605,10606..11040,
                     11041..11046,11047..11490)
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=GCaMP6s
                     /translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK
                     TGHAVRAIGRLSSLENVYIKADKQKNGIKANFHIRHNIEDGGVQLAYHYQQNTPIGDGP
                     VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE
                     LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT
                     YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE
                     LKGIDFKEDGNILGHKLEYNLPDQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRS
                     LGQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKMKYRDTEEEIREAFGVFDKDGN
                     GYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     regulatory      10234..10243
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     10524..10545
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(10651..10672)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(10912..10931)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    join(11500..11502,1..572,573..575,576..584)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
ORIGIN
        1 ctctccctcc ccccccccta acgttactgg ccgaagccgc ttggaataag gccggtgtgc
       61 gtttgtctat atgttatttt ccaccatatt gccgtctttt ggcaatgtga gggcccggaa
      121 acctggccct gtcttcttga cgagcattcc taggggtctt tcccctctcg ccaaaggaat
      181 gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt gaagacaaac
      241 aacgtctgta gcgacccttt gcaggcagcg gaacccccca cctggcgaca ggtgcctctg
      301 cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc agtgccacgt
      361 tgtgagttgg atagttgtgg aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg
      421 gctgaaggat gcccagaagg taccccattg tatgggatct gatctggggc ctcggtacac
      481 atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga accacgggga
      541 cgtggttttc ctttgaaaaa cacgatgata atatggccac aaccatggcc aagcctttgt
      601 ctcaagaaga atccaccctc attgaaagag caacggctac aatcaacagc atccccatct
      661 ctgaagacta cagcgtcgcc agcgcagctc tctctagcga cggccgcatc ttcactggtg
      721 tcaatgtata tcattttact gggggacctt gtgcagaact cgtggtgctg ggcactgctg
      781 ctgctgcggc agctggcaac ctgacttgta tcgtcgcgat cggaaatgag aacaggggca
      841 tcttgagccc ctgcggacgg tgccgacagg tgcttctcga tctgcatcct gggatcaaag
      901 ccatagtgaa ggacagtgat ggacagccga cggcagttgg gattcgtgaa ttgctgccct
      961 ctggttatgt gtgggagggc taagtcgagc ctctagagtt aacatcgagg gatcaagctt
     1021 atcgataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
     1081 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
     1141 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag
     1201 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc
     1261 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc
     1321 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
     1381 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg
     1441 ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg
     1501 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg
     1561 cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcatcga
     1621 taccgtcgag acctagaaaa acatggagca atcacaagta gcaatacagc agctaccaat
     1681 gctgattgtg cctggctaga agcacaagag gaggaggagg tgggttttcc agtcacacct
     1741 caggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa
     1801 gaaaaggggg gactggaagg gctaattcac tcccaacgaa gacaagatat ccttgatctg
     1861 tggatctacc acacacaagg ctacttccct gattggcaga actacacacc agggccaggg
     1921 atcagatatc cactgacctt tggatggtgc tacaagctag taccagttga gcaagagaag
     1981 gtagaagaag ccaatgaagg agagaacacc cgcttgttac accctgtgag cctgcatggg
     2041 atggatgacc cggagagaga agtattagag tggaggtttg acagccgcct agcatttcat
     2101 cacatggccc gagagctgca tccggactgt actgggtctc tctggttaga ccagatctga
     2161 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
     2221 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
     2281 agaccctttt agtcagtgtg gaaaatctct agcagggccc gtttaaaccc gctgatcagc
     2341 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     2401 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     2461 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     2521 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc
     2581 ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag
     2641 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     2701 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     2761 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     2821 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     2881 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     2941 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     3001 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg
     3061 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3121 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     3181 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     3241 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     3301 atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc
     3361 ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga
     3421 tctgatcagc acgtgttgac aattaatcat cggcatagta tatcggcata gtataatacg
     3481 acaaggtgag gaactaaacc atggccaagt tgaccagtgc cgttccggtg ctcaccgcgc
     3541 gcgacgtcgc cggagcggtc gagttctgga ccgaccggct cgggttctcc cgggacttcg
     3601 tggaggacga cttcgccggt gtggtccggg acgacgtgac cctgttcatc agcgcggtcc
     3661 aggaccaggt ggtgccggac aacaccctgg cctgggtgtg ggtgcgcggc ctggacgagc
     3721 tgtacgccga gtggtcggag gtcgtgtcca cgaacttccg ggacgcctcc gggccggcca
     3781 tgaccgagat cggcgagcag ccgtgggggc gggagttcgc cctgcgcgac ccggccggca
     3841 actgcgtgca cttcgtggcc gaggagcagg actgacacgt gctacgagat ttcgattcca
     3901 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     3961 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag
     4021 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     4081 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac
     4141 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     4201 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     4261 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     4321 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     4381 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     4441 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     4501 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     4561 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     4621 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     4681 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     4741 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     4801 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     4861 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     4921 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     4981 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     5041 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     5101 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     5161 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     5221 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     5281 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     5341 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     5401 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     5461 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     5521 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     5581 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     5641 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     5701 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     5761 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     5821 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     5881 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     5941 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     6001 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     6061 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     6121 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     6181 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     6241 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     6301 gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct
     6361 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc
     6421 tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca
     6481 aggcttgacc gacaattgca tgaagaatct gcttagggtt aggcgttttg cgctgcttcg
     6541 cgatgtacgg gccagatata cgcgttgaca ttgattattg actagttatt aatagtaatc
     6601 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
     6661 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
     6721 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
     6781 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
     6841 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
     6901 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg
     6961 gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
     7021 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
     7081 taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
     7141 aagcagcgcg ttttgcctgt actgggtctc tctggttaga ccagatctga gcctgggagc
     7201 tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc
     7261 aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt
     7321 agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag cgaaagggaa
     7381 accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg caagaggcga
     7441 ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga aggagagaga
     7501 tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg aaaaaattcg
     7561 gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg caagcaggga
     7621 gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct gtagacaaat
     7681 actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat cattatataa
     7741 tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca ccaaggaagc
     7801 tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc aagggccgct
     7861 gatcttcaga cctggaggag gagatatgag ggacaattgg agaagtgaat tatataaata
     7921 taaagtagta aaaattgaac cattaggagt agcacccacc aaggcaaaga gaagagtggt
     7981 gcagagagaa aaaagagcag tgggaatagg agctttgttc cttgggttct tgggagcagc
     8041 aggaagcact atgggcgcag cgtcaatgac gctgacggta caggccagac aattattgtc
     8101 tggtatagtg cagcagcaga acaatttgct gagggctatt gaggcgcaac agcatctgtt
     8161 gcaactcaca gtctggggca tcaagcagct ccaggcaaga atcctggctg tggaaagata
     8221 cctaaaggat caacagctcc tggggatttg gggttgctct ggaaaactca tttgcaccac
     8281 tgctgtgcct tggaatgcta gttggagtaa taaatctctg gaacagattt ggaatcacac
     8341 gacctggatg gagtgggaca gagaaattaa caattacaca agcttaatac actccttaat
     8401 tgaagaatcg caaaaccagc aagaaaagaa tgaacaagaa ttattggaat tagataaatg
     8461 ggcaagtttg tggaattggt ttaacataac aaattggctg tggtatataa aattattcat
     8521 aatgatagta ggaggcttgg taggtttaag aatagttttt gctgtacttt ctatagtgaa
     8581 tagagttagg cagggatatt caccattatc gtttcagacc cacctcccaa ccccgagggg
     8641 acccgacagg cccgaaggaa tagaagaaga aggtggagag agagacagag acagatccat
     8701 tcgattagtg aacggatcta caaatggcag tattcatcca caattttaaa agaaaagggg
     8761 ggattggggg gtacagtgca ggggaaagaa tagtagacat aatagcaaca gacatacaaa
     8821 ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat tacagggaca
     8881 gcagaaattc actttgaatt aattcaagct tcgtgaggct ccggtgcccg tcagtgggca
     8941 gagcgcacat cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgaaccggt
     9001 gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt
     9061 tttcccgagg gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt
     9121 cgcaacgggt ttgccgccag aacacaggta agtgccgtgt gtggttcccg cgggcctggc
     9181 ctctttacgg gttatggccc ttgcgtgcct tgaattactt ccacctggct ccagtacgtg
     9241 attcttgatc ccgagctgga gccaggggcg ggccttgcgc tttaggagcc ccttcgcctc
     9301 gtgcttgagt tgaggcctgg cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc
     9361 ttcgcgcctg tctcgctgct ttcgataagt ctctagccat ttaaaatttt tgatgacctg
     9421 ctgcgacgct ttttttctgg caagatagtc ttgtaaatgc gggccaggat ctgcacactg
     9481 gtatttcggt ttttggggcc gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt
     9541 cggcgaggcg gggcctgcga gcgcggccac cgagaatcgg acgggggtag tctcaagctg
     9601 gccggcctgc tctggtgcct ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa
     9661 ggctggcccg gtcggcacca gttgcgtgag cggaaagatg gccgcttccc ggccctgctc
     9721 cagggggctc aaaatggagg acgcggcgct cgggagagcg ggcgggtgag tcacccacac
     9781 aaaggaaagg ggcctttccg tcctcagccg tcgcttcatg tgactccacg gagtaccggg
     9841 cgccgtccag gcacctcgat tagttctgga gcttttggag tacgtcgtct ttaggttggg
     9901 gggaggggtt ttatgcgatg gagtttcccc acactgagtg ggtggagact gaagttaggc
     9961 cagcttggca cttgatgtaa ttctccttgg aatttgccct ttttgagttt ggatcttggt
    10021 tcattctcaa gcctcagaca gtggttcaaa gtttttttct tccatttcag gtgtcgtgaa
    10081 cacgctaccg gtctcgacga attcggcctc gagggcggcg gccgcgatct cgccaccatg
    10141 ggttctcatc atcatcatca tcatggtatg gctagcatga ctggtggaca gcaaatgggt
    10201 cgggatctgt acgacgatga cgataaggat ctcgccacca tggtcgactc atcacgtcgt
    10261 aagtggaata agacaggtca cgcagtcaga gctataggtc ggctgagctc actcgagaac
    10321 gtctatatca aggccgacaa gcagaagaac ggcatcaagg cgaacttcca catccgccac
    10381 aacatcgagg acggcggcgt gcagctcgcc taccactacc agcagaacac ccccatcggc
    10441 gacggccccg tgctgctgcc cgacaaccac tacctgagcg tgcagtccaa actttcgaaa
    10501 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
    10561 actctcggca tggacgagct gtacaagggc ggtaccggag ggagcatggt gagcaagggc
    10621 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
    10681 cacaagttca gcgtgtccgg cgagggtgag ggcgatgcca cctacggcaa gctgaccctg
    10741 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg
    10801 acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc
    10861 aagtccgcca tgcccgaagg ctacatccag gagcgcacca tcttcttcaa ggacgacggc
    10921 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
    10981 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
    11041 ctgccggacc aactgactga agagcagatc gcagaattta aagaggcttt ctccctattt
    11101 gacaaggacg gggatgggac aataacaacc aaggagctgg ggacggtgat gcggtctctg
    11161 gggcagaacc ccacagaagc agagctgcag gacatgatca atgaagtaga tgccgacggt
    11221 gacggcacaa tcgacttccc tgagttcctg acaatgatgg caagaaaaat gaaatacagg
    11281 gacacggaag aagaaattag agaagcgttc ggtgtgtttg ataaggatgg caatggctac
    11341 atcagtgcag cagagcttcg ccacgtgatg acaaaccttg gagagaagtt aacagatgaa
    11401 gaggttgatg aaatgatcag ggaagcagac atcgatgggg atggtcaggt aaactacgaa
    11461 gagtttgtac aaatgatgac agcgaagtga gcggatccgc cc
//
