LOCUS       sequence-204269-       15006 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For expressing Trpa1b-2A-EGFP in zebrafish cardiomyocytes..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15006)
  AUTHORS   Lam PY, Mendu SK, Mills RW, Zheng B, Padilla H, Milan DJ, Desai BN, 
            Peterson RT
  TITLE     A high-conductance chemo-optogenetic system based on the vertebrate 
            channel Trpa1b.
  JOURNAL   Sci Rep. 2017 Sep 19;7(1):11839. doi: 10.1038/s41598-017-11791-z.
  PUBMED    28928472
REFERENCE   2  (bases 1 to 15006)
  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: "Sci Rep."; 
            date: "2017-09-19"; volume: "7(1):11839. doi"; pages: " 
            10.1038/s41598-017-11791-z"
COMMENT     Sequence Label: Tol2-cmlc2-Trpa1b-2A-EGFP
FEATURES             Location/Qualifiers
     source          1..15006
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(493..1284,1285..1353))
                     /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"
     promoter        complement(1354..1445)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1487..1506
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1512..1531
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     protein_bind    1738..1771
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     primer_bind     3239..3257
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     protein_bind    complement(4990..5023)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     primer_bind     complement(5493..5510)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5493..5509)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(5779..6367)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5859..5878)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6538..7329,7330..7398))
                     /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"
     promoter        complement(7399..7503)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7666..7687)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    join(8036..8047,8048..8210)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     primer_bind     8583..8599
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    8619..8639
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     protein_bind    9483..9507
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             12845..12901
                     /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             join(12902..12904,12905..12907,12908..13621)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(12902..12904,12905..12907,12908..13618)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(13208..13227)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     13555..13576
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(13623..13647)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    13649..13783
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(13673..13692)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     13727..13746
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     misc_feature    join(14222..14409,14410..14421)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(14455..14471)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(14455..14471)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(14508..14528)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(14508..14526)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(14547..14563)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(14560..14582)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    14571..14587
                     /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(14595..14601,14602..14619,14620..14625))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    14640..14661
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
       61 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
      121 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
      181 cagagttctt gaagtggtgg gctaactacg gctacactag aagaacagta tttggtatct
      241 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
      301 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
      361 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgacg
      421 cgcgcgtaac tcacgttaag ggattttggt catgagcttg cgccgtcccg tcaagtcagc
      481 gtaatgctct gcttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
      541 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
      601 accatctggc cccagcgctg cgatgatacc gcgagaacca cgctcaccgg ctccggattt
      661 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
      721 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
      781 tagtttgcgc aacgttgttg ccatcgctac aggcatcgtg gtgtcacgct cgtcgtttgg
      841 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
      901 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
      961 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     1021 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     1081 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     1141 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     1201 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     1261 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     1321 aataagggcg acacggaaat gttgaatact catattcttc ctttttcaat attattgaag
     1381 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     1441 acaaataggg gtcagtgtta caaccaatta accaattctg aacattatcg cgagcccatt
     1501 tatacctgaa tatggctcat aacacccctt gtttgcctgg cggcagtagc gcggtggtcc
     1561 cacctgaccc catgccgaac tcagaagtga aacgccgtag cgccgatggt agtgtgggga
     1621 ctccccatgc gagagtaggg aactgccagg catcaaataa aacgaaaggc tcagtcgaaa
     1681 gactgggcct ttcgcccggg ctaattatgg ggtgtcgccc ttattcgact ctatagtgaa
     1741 gttcctattc tctagaaagt ataggaactt ctgaagtggg gatttaaatg cggccgcgaa
     1801 tgcacgcgat cgcacgcatt ccgttgcatc aaatttaggg atgccatcaa ctttcagttc
     1861 ataattaata tcttaccaaa ttaggtaatc tgcaaaagtt cagactgtga aatgtaacat
     1921 tttatatatc aagctctatt taatgcctca cagtagttaa cataaagaga tacagaattg
     1981 tcgtgtcagt gtatactatc catgtgtata ctctggatat ccatttgtat tccattatct
     2041 acgaattggc ggggttctgc tcgagctggg tggcgaggtt gttaagtcgg tccatttttg
     2101 tgtgtcttgg ttggatgagg tccaaagtgg ggggagtcta ccaggcttta tatacacctg
     2161 gtagttgcaa ctttcggtat gatccccacg aaagataaag ttcagcatgt ggagagccga
     2221 ggtagtatgc gtgtgacaga gacccaaact gggggtcacc gcgtgcacac tggcgacgcc
     2281 gctctggaag gtcacatgac cgcagatact ccgttctccc ctcacgtcga ttcggctcgg
     2341 ggacacggtc agcgccattg ccacgtgtcc acggttctct ccactctgta gaaagtcact
     2401 tccaccaagt tccatgaagc ttattaggtt tttggagaaa tccggggcgg catttctggg
     2461 gtaacttggg caagccagta gtggatcaat tgggacccga attgggtgga aaaaggtgat
     2521 aacgagccct ttgggatccc cacaattgat ctaagacctc tgcttgcccc gatcttggcc
     2581 ccctcagccc caacactgtt gctcttcaaa ctcccccata cctcggttca gatgcagcct
     2641 tcaaaacttt aacgtacctc cgagcctgtg aaaacgtggt catgaaaaaa cgagagatgt
     2701 aagtcccgtc acataccgct gcgagaccac cagctgtaca tacacttggc gttccagcat
     2761 gcacggtcgc tcaacaggcg ctcccccatt ggcggaaaga caagccccca ctagattcct
     2821 gttcaagggc aaagtctaga agcccgcccc ccccagcgcc cgtttccttg ttagtgcggc
     2881 ctgcacatgt cacctgtctg gcaggtatcg tcgctacttt gccagtcgta tcgtcgtcaa
     2941 cggattgcgc tgcccattca cgtgtaaact ctcgacggat atcccggagt ttcccgccga
     3001 tcctcccgcg agccactgcc acgtcgcctc caagcctgct cgagcgtccc aaaaccttct
     3061 caagcaaggt tttcagtata atgttacatg cgtacacgcg tctgtacaga aaaaaaagaa
     3121 aaatttgaaa tataaataac gttcttaata ctaacataac tataaaaaaa taaataggga
     3181 cctagacttc aggttgtcta actccttcct tttcggttag agcggatgtg gggggagggc
     3241 gtgaatgtaa gcgtgacata actaattaca tgatcatgga catggcatag acatatacaa
     3301 ggcttgttca ccatctgatg cagtaccatc gtataatgca gtatccaaac cacacaaggt
     3361 aaaacccatt cttctatagg catgaatagc tggagcgtta acattagtaa cttccaacca
     3421 caaatgacca gcacctcttt ctctagcaaa ttcagtagcc aaacccatca aagctctacc
     3481 aacaccatga cctctatgtt ctggagcaac ttcaatatct tcgacggtca atcttctatt
     3541 ccaaccagaa taagaaacaa caacaaaacc agccaagtca ccgtcatcac cataagcaac
     3601 aaaagttcta gaatctggat caccatcttc accatcatca gattcgtcat cggattcatc
     3661 atctggaaaa accttagtca atggtggatc aactggaact tctctcaaag tgaaaccatc
     3721 accagtagca gtaactctga aaacagtgtc agtagtgaaa gaaccatcca aagcttcaat
     3781 agcttcagca tcacctggaa cagaagttct gtatctgtaa gcagtatcat ccaaagtagt
     3841 acccattgct gtagatatgt cttgtgtgta agggggttgg ggtggttgtt tgtgttcttg
     3901 acttttgtgt tagcaaggga agacgggcaa aaaagtgagt gtggttggga gggagagacg
     3961 agccttatat ataatgcttg tttgtgtttg tgcaagtgga cgccgaaacg ggcaggagcc
     4021 aaactaaaca aggcagacaa tgcgagctta attggattgc ctgatgggca ggggttaggg
     4081 ctcgatcaat gggggtgcga agtgacaaaa ttgggaatta ggttcgcaag caaggctgac
     4141 aagactttgg cccaaacatt tgtacgcggt ggacaacagg agccacccat cgtctgtcac
     4201 gggctagccg gtcgtgcgtc ctgtcaggct ccacctaggc tccatgccac tccatacaat
     4261 cccactagtg taccgctagg ccgcttttag ctcccatcta agaccccccc aaaacctcca
     4321 ctgtacagtg cactgtactg tgtggcgatc aagggcaagg gaaaaaaggc gcaaacatgc
     4381 atgcatggaa tgacgtaggt aaggcgttac tagactgaaa agtggcacat ttcggcgtgc
     4441 caaagggtcc taggtgcgtt tcgcgagctg ggcgccaggc caagccgctc caaaacgcct
     4501 ctccgactcc ctccagcggc ctccatatcc ccatccctct ccacagcaat gttgttaagc
     4561 cttgcaaacg aaaaaataga aaggctaata agcttccaat attgtggtgt acgctgcata
     4621 acgcaacaat gagcgccaaa caacacacac acacagcaca cagcagcatt aaccacgatg
     4681 aacagcatga cattacaggt gggtgtgtaa tcagggccct gattgctggt ggtgggagcc
     4741 cccatcatgg gctctcgtcc agccgcggaa accggaaacg ctcgaatctg ccgccgctca
     4801 tactcaccga aacgtgtggg gctcgaaaaa acgggcgcca aactccttat actactgtat
     4861 attcaagcaa gtatatccgt gggtgcgggt gatttggatc taaggttcgt actcaacact
     4921 cacgagcagc ttgcctatgt tacatccttt tatcagacat agcggccgca aatttaaata
     4981 aaatgaagtg aagttcctat actttctaga gaataggaac ttctatagtg agtcgaataa
     5041 gggcgacaca aaatttattc taaatgcata ataaatactg ataacatctt atagtttgta
     5101 ttatattttg tattatcgtt gacatgtata attttgatat caaaaactga ttttcccttt
     5161 attattttcg agatttattt tcttaattct ctttaacaaa ctagaaatat tgtatataca
     5221 aaaaatcata aataatagat gaatagttta attataggtg ttcatcaatc gaaaaagcaa
     5281 cgtatcttat ttaaagtgcg ttgctttttt ctcatttata aggttaaata attctcatat
     5341 atcaagcaaa gtgacaggcg cccttaaata ttctgacaaa tgctctttcc ctaaactccc
     5401 cccataaaaa aacccgccga agcgggtttt tacgttattt gcggattaac gattactcgt
     5461 tatcagaacc gcccaggggg cccgagctta agactggccg tcgttttaca acacagaaag
     5521 agtttgtaga aacgcaaaaa ggccatccgt caggggcctt ctgcttagtt tgatgcctgg
     5581 cagttcccta ctctcgcctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     5641 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     5701 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     5761 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     5821 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     5881 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     5941 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     6001 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     6061 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     6121 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     6181 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     6241 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     6301 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     6361 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     6421 acgttaaggc attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     6481 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     6541 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     6601 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     6661 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     6721 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     6781 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     6841 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     6901 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     6961 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     7021 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     7081 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     7141 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     7201 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     7261 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     7321 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     7381 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     7441 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     7501 gcgcacattt ccccgaaaag tgccacctaa attgtaagcg ttaatatttt gttaaaattc
     7561 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     7621 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag
     7681 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     7741 gatggcccac gctggctaag aactcatcag cctccccggt ccatctaccc acgtaccaat
     7801 gcaccaattg gccacaatga cggctactac atggtgccat tccttcctct ttataggaat
     7861 ggagactacc tcctgtccaa caatgctctt ggatacgagt acgcctacct gttggaccca
     7921 ggtcattgca caacaccaga aatgccctct gatctgcaaa agacgtgaat atctgttcag
     7981 acacccatat ccactctgtt ccacacaggt cagaggtttg tccaggagtt cttgacagag
     8041 gtgtaaaaag tactcaaaaa ttttactcaa gtgaaagtac aagtacttag ggaaaatttt
     8101 actcaattaa aagtaaaagt atctggctag aatcttactt gagtaaaagt aaaaaagtac
     8161 tccattaaaa ttgtacttga gtattaagga agtaaaagta aaagcaagaa agaaaactag
     8221 agattcttgt ttaagctttt aatctcaaaa aacattaaat gaaatgcata caaggtttta
     8281 tcctgcttta gaactgtttg tatttaatta tcaaactata agacagacaa tctaatgcca
     8341 gtacacgcta ctcaaagttg taaaacctca gatttaactt cagtagaagc tgattctcaa
     8401 aattgttagt gtcaagccta gctcttttgg ggctgaaaag caatcctgca gtgctgaaaa
     8461 gcctctcaca ggcagccgat gcgggaagag gtgtattagt cttgatagag aggctgcaaa
     8521 tagcaggaaa cgtgagcaga gactccctgg tgtctgaaac acaggccaga tgggccctcg
     8581 agcaggaaac agctatgacc atgattacgc caagctatca actttgtata gaaaagttgc
     8641 tccaaatcag cagacttaac attctttaaa atgattgatt caatagtgat aaaaatcagg
     8701 catagccagt tgtaacttta gataaattac agaaaatgtc aaatacagag aaccgattct
     8761 tttttatgat acatccaagc acacatttaa cacaatccag gcaaaccccg aatttcacag
     8821 tcacaagcac tgtttgtaca agagctttgc ctaaggacac acagtctcta taagtccagg
     8881 tcgttggttt cactcttatt ttaaacatgt gacatttttc ctgccatcct gtcttaggct
     8941 gctgtttgct tcattccatg tcacattaat ttcctcagta gcacctttta cacacacagc
     9001 caatcttttc cagaaaattc aattgctttg aaaagataat gtgtgaacaa atccatttag
     9061 aaaaggaaaa ttaagaattt gtaaaatcat ctgtaaattg ttggcattct tctgtatatg
     9121 aacatcacat catttacagg taaaggtctg gtcattaatt atatgacaat ttactggtat
     9181 tattttgtga aaggggctat tttcaatgca ttcatccatc cttttcatcc ctcaaatctc
     9241 tcattcacgt ccccctcccc atctgcacac tttatctcat tttccaccct gctggaatct
     9301 gagcacttgt gcagttatca gggctcctgt atttaggagg ctctgggtgt ccatgtaggg
     9361 gacgaacaga aacactgcag acctttatag aagaacaaat gataagagtc ctcatacata
     9421 aagactccat tagaaacgtc agtgacccag gagcccagac caacagcaaa gcagacagtg
     9481 aaccaagttt gtacaaaaaa gcggcttcgc caccatgcag tttggaaagg agttagtgag
     9541 gcgaaacagc ttctacaaat gtgtgataaa cgaggacgaa acggaggagt ctgcagatgt
     9601 ctttgagttg gctttcaaag gaaaggcttc tgccatcgat cgtctgatcc agaaaggtcc
     9661 agaacaccta agcctcaggg atgagaatgg agccagtctg ctgcactatg cttcagctgg
     9721 aggaaacctg gatatcattc ggctcattgt ttccattgtg ggccctgagg tgctgaatgt
     9781 gcaggatgag cagggccgaa ctcctctgca ctgggctgtg gagcaggatc aacaacagag
     9841 ctgcgctgta ctgctggatc tgggtgctga ccccaacatt ctcaacaatg ccctgatggg
     9901 ccctctgcac ctggctgtca ccaaacaata caatcatcta gcagaggtcc ttctgtcttg
     9961 tgacaaaacc aatagcaatc tggaggggga tctgggaaac acccctgtca tgctggcctg
    10021 ctccaataac aactgccaag ctctccagat cctgatcaag cgcggtgcca agatgtgtat
    10081 tcagaacaag ctgggtcact atcccattca tactgttgct ttcgcaggtg ccaaagaggc
    10141 catggagatg gtattgaaaa ttggagaaga acttggtgta tcatctacat tgcacattaa
    10201 ttatttggat aagtccaaaa gcacacctct tcatctggct gtacgcggag gcaatatcga
    10261 ggtgatcaaa ctctgcatct tgaaaggagc caaagtagaa caacatcaga gtggcaaatg
    10321 cacggccctc cactttgcct gcagtcaggg ttctctggag gctgtcaaaa tcatgctttc
    10381 ctcttacaac agaacagaag atatcgtcaa cattcgggat ggagccaatc ggacaccttt
    10441 gcatagagca acgctgtttg atcacgttga gttggctgaa tatcttattt caaagggagc
    10501 agaaattgac tctattgact gtaagggact gtctccgctt ctgctcgcct ccagctgcag
    10561 cgcttggaaa acagttgcat atctgttgtc cattggtgca gattttaaaa tcaaagaaaa
    10621 aacaggtcgc aacttcctcc actttgtcat tcttcaaccc aaaggcctga aaaatcttcc
    10681 tgagacggtt ttgcagagca ccgccgtgaa ggagatgttg tctgatgagg atgttgaagg
    10741 ctgcactcct ctgcactacg cctgcaagct tggcatccac gattcggtca aaaatatgct
    10801 gggtctcaat atctgcttgg gccaaaagtc acgggagaag aaatcagcgc ttcacttcgc
    10861 tgcagaatat gggcgtataa atacatgcca ccggctgctg gagaccctca cagactctaa
    10921 aatgctgaat gattgggatg aaaagggtct gacgcccctc catcttgctt caagggcagg
    10981 acacgctcaa gtagtggatc tgctgcttag gaagggagca ctgtttcaaa gtgattacaa
    11041 gggctggacc tgtttacatc atgcagcagc tgaaggatac acacagacga tgaaaatcct
    11101 gctagcagcc aatgtgaaac tactggatga aaaaaatgaa gatgggaaca cagcccttca
    11161 tatagcagca caagctggac atgtaagtgc tgttttgctg ttattggaca gaggagctga
    11221 aatcgctctc aacgatgctg acaactcttt tttgcatgaa gctgtgcgga atgagagaag
    11281 agaagtagta aacgccacca ttgaacatga gcggtgtgat gagtcaatga cgtcatttaa
    11341 agcgaaatcc cgctgtgttg tgctggacat catagagttc cttccagaat cttttcagca
    11401 ccttctggac cagtgtatta cagaatctga tcatgatgcc aatagtcagg attatcatat
    11461 catgtacaat ttccaatggc ttcaggctcc catacagttg aagaaatatg ctaaaacaga
    11521 caaaacaaaa gcatttcaac cactggcagc attaaatgcc atggtgcggt acaaccgtct
    11581 ggagctcctc attcacccct tgagccgaaa atacctggag atgaagtgga cggcatatgg
    11641 aagcaaagtt catttcctaa acttggcgat atacctgctt ggattgttgc ccctcacata
    11701 cctcatcctc aatctgagac ccagccagga cttctccaaa ggcaatggca catctgtcat
    11761 catggtgccc gtatctttta gcgagcaaca atatttaatc tccgtgtgta taatcatggt
    11821 gatcgtcatg aatgtgtatt cgatttgcaa agaggtggta cagctggctc agcagcgagt
    11881 aaattacttc acagatttct caaaccctgc ggactggagt gcagctatta gcgctttggt
    11941 ttttgttgta cccatgtgct gcagtgtgga ggccacatgg caatgggaag ctggagctta
    12001 tgctattctg acatcatgga ttggctttct gctttacttt cagagatttg agcggattgg
    12061 gatttacgtg gtgatgttta atggaatcgt acggaccctg gtgtgcatca tggttctttt
    12121 tgtcttcctt ttgctggctt tcggcttggc tttctacgct ctgatgctcc acaggccgga
    12181 gttcagttcc atcagcctag cgttagcgca aacgtttgtg atgacagtag gagagctaaa
    12241 ctaccaaagc acatttctta actcctatga agaaggccac atggctttcc cagccatcac
    12301 ttatttagtt tttgtgtttt ttgttcttct catgcccatt cttctcatga acctaatgat
    12361 tggtttggct gtaggagaca ttgcagaggt acagagaaac gcagagctga aaaggattgc
    12421 tatgcagata gatcttcaca ctgcccttga ggagaaacta ccttactggt tcctgaagcg
    12481 agtggacaaa ccctctaccg ttgtctatcc caataaatgc aagaaagtgc ttctggaggc
    12541 cttggtgcat ggtgagattc agaacacagt tcgaactcgt ctaaaccttt gctcacggaa
    12601 ggaagggctt ttggagagag agctgcacaa gcagaagaac aggttgaaag aaatgtcgtg
    12661 catgctggaa aaacagcata acctgttgaa gctgattatc cagaagatgg aaattacatc
    12721 tgaagctgat gaatatgatg gtccacagaa tcatggagct ctaaaacagc acacctcatc
    12781 cacctcacag aaatcaaagt gggtcccgtt actgcaagca atcaaggcca agaagggatc
    12841 cggagccacg aacttctctc tgttaaagca agcaggagac gtggaagaaa accccggtcc
    12901 tatggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
    12961 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta
    13021 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
    13081 cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa
    13141 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
    13201 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
    13261 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
    13321 caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa
    13381 cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc
    13441 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
    13501 ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt
    13561 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta
    13621 agacccagct ttcttgtaca aagtggggga tccagacatg ataagataca ttgatgagtt
    13681 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
    13741 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
    13801 tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct
    13861 ctacaaatgt ggtatggctg attatgatcc tctagatcag atctgcgaag atacggccac
    13921 gggtgctctt gatcctgtgg ctgattttgg actgtgctgc tcgcagctgc tgatgaatca
    13981 catacttcct ccattttctt ccactgattg actgttataa tttccctaat ttccaggtca
    14041 aggtgctgtg cattgtggta atagatgtga catgacgtca cttccaaagg accaatgaac
    14101 atgtctgacc aatttcatat aatgtgaaaa cgattttcat aggcagaata aataacattt
    14161 aaattaaact gggcatcagc gcaattcaat tggtttggta atagcaaggg aaaatagaat
    14221 gaagtgatct ccaaaaaata agtacttttt gactgtaaat aaaattgtaa ggagtaaaaa
    14281 gtactttttt ttctaaaaaa atgtaattaa gtaaaagtaa aagtattgat ttttaattgt
    14341 actcaagtaa agtaaaaatc cccaaaaata atacttaagt acagtaatca agtaaaatta
    14401 ctcaagtact ttacacctct ggttcttgac cccctacctt cagcaagccc agcagatcca
    14461 ctagttctag agcggccgcc accgcggtgg agctccagct tttgttccct ttagtgaggg
    14521 ttaattgcgc gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
    14581 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
    14641 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
    14701 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
    14761 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
    14821 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
    14881 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
    14941 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
    15001 cagagg
//
