LOCUS       sequence-41505-e       18025 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cre-dependent expression of hChR2(H134R)-tdTomato, for optogenetic 
            excitation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 18025)
  AUTHORS   Madisen L, Mao T, Koch H, Zhuo JM, Berenyi A, Fujisawa S, Hsu YW, 
            Garcia AJ 3rd, Gu X, Zanella S, Kidney J, Gu H, Mao Y, Hooks BM, 
            Boyden ES, Buzsaki G, Ramirez JM, Jones AR, Svoboda K, Han X, Turner
            EE, Zeng H
  TITLE     A toolbox of Cre-dependent optogenetic transgenic mice for 
            light-induced activation and silencing.
  JOURNAL   Nat Neurosci. 2012 Mar 25;15(5):793-802. doi: 10.1038/nn.3078.
  PUBMED    22446880
REFERENCE   2  (bases 1 to 18025)
  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"; doi: "10.1038/nn"; 
            journalName: "Nat Neurosci"; date: "2012-03-25- 25"; volume: "15"; 
            issue: "5"; pages: "793-802"
COMMENT     Sequence Label: Ai27
FEATURES             Location/Qualifiers
     source          1..18025
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    3..1087
                     /label=Rosa26 left arm
                     /note="left homology arm from intron 1 of the mouse Rosa26 
                     gene (Soriano, 1999)"
     enhancer        1181..1484
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1486..1763
                     /label=chicken beta-actin promoter
     intron          1765..2782
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     2706..2728
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     2790..2809
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    2858..2891
                     /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)"
     protein_bind    2915..2948
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    3116..3250
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    3370..3504
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    3624..3758
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    3787..3820
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      3945..3954
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3951..4877
                     /codon_start=1
                     /product="H134R mutant of an N-terminal fragment of 
                     channelrhodopsin-2 from the alga Chlamydomonas reinhardtii 
                     (Nagel et al., 2005)"
                     /label=ChR2(H134R)
                     /note="blue light-activated cation channel"
                     /translation="MDYGGALSAVGRELLFVTNPVVVNGSVLVPEDQCYCAGWIESRGT
                     NGAQTASNVLQWLAAGFSILLLMFYAYQTWKSTCGWEEIYVCAIEMVKVILEFFFEFKN
                     PSMLYLATGHRVQWLRYAEWLLTCPVILIRLSNLTGLSNDYSRRTMGLLVSDIGTIVWG
                     ATSAMATGYVKVIFFCLGLCYGANTFFHAAKAYIEGYHTVPKGRCRQVVTGMAWLFFVS
                     WGMFPILFILGPEGFGVLSVYGSTVGHTIIDLMSKNCWGLLGHYLRVLIHEHILIHGDI
                     RKTTKLNIGGTEIEVETLVEDEAEAGAV"
     CDS             4890..6317
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGL
                     VTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIHQ
                     ALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRHH
                     LFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYE
                     GTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMN
                     FEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVL
                     KGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYER
                     SEGRHHLFLYGMDELYK"
     misc_feature    6486..7074
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6539..6559)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6957..6968)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(7077..7093)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    7095..7302
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    7310..7343
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     promoter        7365..7864
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     protein_bind    7884..7917
                     /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)"
     regulatory      7918..7927
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             7924..8727
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(7987..8006)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     8597..8616
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    8737..9198
                     /label=PGK poly(A) signal
                     /note="mouse phosphoglycerate kinase 1 polyadenylation 
                     signal"
     misc_feature    9262..13531
                     /label=Rosa26 right arm
                     /note="right homology arm from intron 1 of the mouse Rosa26
                     gene (Soriano, 1999)"
     promoter        13580..14073
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     CDS             14100..14669
                     /codon_start=1
                     /gene="Corynebacterium diphtheriae tox"
                     /product="diphteria toxin Fragment A"
                     /label=DTA
                     /translation="DDVVILLIFCMENFSSYHGTKPGYVDSIQKGIQKPKSGTQGNYDD
                     DWKGFYSTDNKYDAAGYSVDNENPLSGKAGGVVKVTYPGLTKVLALKVDNAETIKKELG
                     LSLTEPLMEQVGTEEFIKRFGDGASRVVLSLPFAEGSSSVEYINNWEQAKALSVELEIN
                     FETRGKRGQDAMYEYMAQACAGNRVRR"
     intron          14746..14811
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     polyA_signal    14885..15109
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(15177..15197)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(15177..15195)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(15216..15232)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(15216..15232)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(15229..15251)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    15240..15256
                     /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(15264..15270,15271..15288,15289..15294))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    15309..15330
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(15447..15464)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(15618..16206)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(15698..15717)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(16377..17168,17169..17237))
                     /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     17000..17019
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(17238..17342)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(17368..17823)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(17505..17526)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     17717..17736
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     17950..17972
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     17964..17981
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     17965..17981
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     17991..18010
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        17991..18009
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ccgcggcagg ccctccgagc gtggtggagc cgttctgtga gacagccggg tacgagtcgt
       61 gacgctggaa ggggcaagcg ggtggtgggc aggaatgcgg tccgccctgc agcaaccgga
      121 gggggaggga gaagggagcg gaaaagtctc caccggacgc ggccatggct cggggggggg
      181 ggggcagcgg aggascgctt ccggccgacg tctcgtcgct gattggctty ttttcctccc
      241 gccgtgtgtg aaaacacaaa tggcgtgttt tggttggcgt aaggcgcctg tcagttaacg
      301 gcagccggag tgcgcagccg ccggcagcct cgctctgccc actgggtggg gcgggaggta
      361 ggtggggtga ggcgagctgn acgtgcgggc gcggtcggcc tctggcgggg cgggggaggg
      421 gagggagggt cagcgaaagt agctcgcgcg cgagcggccg cccaccctcc ccttcctctg
      481 ggggagtcgt tttacccgcc gccggccggg cctcgtcgtc tgattggctc tcggggccca
      541 gaaaactggc ccttgccatt ggctcgtgtt cgtgcaagtt gagtccatcc gccggccagc
      601 gggggcggcg aggaggcgct cccaggttcc ggccctcccc tcggccccgc gccgcagagt
      661 ctggccgcgc gcccctgcgc aacgtggcag gaagcgcgcg ctgggggcgg ggacgggcag
      721 tagggctgag cggctgcggg gcgggtgcaa gcacgtttcc gacttgagtt gcctcaagag
      781 gggcgtgctg agccagacct ccatcgcgca ctccggggag tggagggaag gagcgagggc
      841 tcagttgggc tgttttggag gcaggaagca cttgctctcc caaagtcgct ctgagttgtt
      901 atcagtaagg gagctgcagt ggagtaggcg gggagaaggc cgcacccttc tccggagggg
      961 ggaggggagt gttgcaatac ctttctggga gttctctgct gcctcctggc ttctgaggac
     1021 cgccctgggc ctgggagaat cccttgcccc ctcttcccct cgtgatctgc aactccagtc
     1081 tttctagtta attaaccgtt taaacaattc tgcaggaatc tagttattaa tagtaatcaa
     1141 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
     1201 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
     1261 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     1321 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
     1381 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
     1441 ctacttggca gtacatctac gtattagtca tcgctattac catggtcgag gtgagcccca
     1501 cgttctgctt cactctcccc atctcccccc cctccccacc cccaattttg tatttattta
     1561 ttttttaatt attttgtgca gcgatggggg cggggggggg gggggggcgc gcgccaggcg
     1621 gggcggggcg gggcgagggg cggggcgggg cgaggcggag aggtgcggcg gcagccaatc
     1681 agagcggcgc gctccgaaag tttcctttta tggcgaggcg gcggcggcgg cggccctata
     1741 aaaagcgaag cgcgcggcgg gcggggagtc gctgcgacgc tgccttcgcc ccgtgccccg
     1801 ctccgccgcc gcctcgcgcc gcccgccccg gctctgactg accgcgttac tcccacaggt
     1861 gagcgggcgg gacggccctt ctcctccggg ctgtaattag cgcttggttt aatgacggct
     1921 tgtttctttt ctgtggctgc gtgaaagcct tgaggggctc cgggagggcc ctttgtgcgg
     1981 ggggagcggc tcggggggtg cgtgcgtgtg tgtgtgcgtg gggagcgccg cgtgcggctc
     2041 cgcgctgccc ggcggctgtg agcgctgcgg gcgcggcgcg gggctttgtg cgctccgcag
     2101 tgtgcgcgag gggagcgcgg ccgggggcgg tgccccgcgg tgcggggggg gctgcgaggg
     2161 gaacaaaggc tgcgtgcggg gtgtgtgcgt gggggggtga gcagggggtg tgggcgcgtc
     2221 ggtcgggctg caaccccccc tgcacccccc tccccgagtt gctgagcacg gcccggcttc
     2281 gggtgcgggg ctccgtacgg ggcgtggcgc ggggctcgcc gtgccgggcg gggggtggcg
     2341 gcaggtgggg gtgccgggcg gggcggggcc gcctcgggcc ggggagggct cgggggaggg
     2401 gcgcggcggc ccccggagcg ccggcggctg tcgaggcgcg gcgagccgca gccattgcct
     2461 tttatggtaa tcgtgcgaga gggcgcaggg acttcctttg tcccaaatct gtgcggagcc
     2521 gaaatctggg aggcgccgcc gcaccccctc tagcgggcgc ggggcgaagc ggtgcggcgc
     2581 cggcaggaag gaaatgggcg gggagggcct tcgtgcgtcg ccgcgccgcc gtccccttct
     2641 ccctctccag cctcggggct gtccgcgggg ggacggctgc cttcgggggg gacggggcag
     2701 ggcggggttc ggcttctggc gtgtgaccgg cggctctaga gcctctgcta accatgttca
     2761 tgccttcttc tttttcctac agctcctggg caacgtgctg gttattgtgc tgtctcatca
     2821 ttttggcaaa gaattgattt gataccgcgg gccctaagaa gttcctattc tctagaaagt
     2881 ataggaactt cgtcgacatt taaatcattt aaatataact tcgtataatg tatgctatac
     2941 gaagttattc gcgatgaata aatgaaagct tgcagatctg cgactctaga ggatctgcga
     3001 ctctagagga tcataatcag ccntaccaca ttttgtagag gttttactng ctttaaaaaa
     3061 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
     3121 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     3181 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     3241 tgtctggatc tgcgactcta gaggatcata atcagccata ccacatttgt agaggtttta
     3301 cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt
     3361 gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     3421 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     3481 aatgtatctt atcatgtctg gatctgcgac tctagaggat cataatcagc cataccacat
     3541 ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata
     3601 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa
     3661 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     3721 tgtccaaact catcaatgta tcttatcatg tctggatccc catcaagctg atccggaacc
     3781 cttaatataa cttcgtataa tgtatgctat acgaagttat taggtccctc gacctgcagc
     3841 ccaagctaga tcgaattcgg ccggcccctt cacgtgttgc acttaacgcg taagcggtac
     3901 tagtccaggc tagcttatcc ctggattgtt taattcgatc tagcgccacc atggactatg
     3961 gcggcgcttt gtctgccgtc ggacgcgaac ttttgttcgt tactaatcct gtggtggtga
     4021 acgggtccgt cctggtccct gaggatcaat gttactgtgc cggatggatt gaatctcgcg
     4081 gcacgaacgg cgctcagacc gcgtcaaatg tcctgcagtg gcttgcagca ggattcagca
     4141 ttttgctgct gatgttctat gcctaccaaa cctggaaatc tacatgcggc tgggaggaga
     4201 tctatgtgtg cgccattgaa atggttaagg tgattctcga gttctttttt gagtttaaga
     4261 atccctctat gctctacctt gccacaggac accgggtgca gtggctgcgc tatgcagagt
     4321 ggctgctcac ttgtcctgtc atccttatcc gcctgagcaa cctcaccggc ctgagcaacg
     4381 actacagcag gagaaccatg ggactccttg tctcagacat cgggactatc gtgtgggggg
     4441 ctaccagcgc catggcaacc ggctatgtta aagtcatctt cttttgtctt ggattgtgct
     4501 atggcgcgaa cacatttttt cacgccgcca aagcatatat cgagggttat catactgtgc
     4561 caaagggtcg gtgccgccag gtcgtgaccg gcatggcatg gctgtttttc gtgagctggg
     4621 gtatgttccc aattctcttc attttggggc ccgaaggttt tggcgtcctg agcgtctatg
     4681 gctccaccgt aggtcacacg attattgatc tgatgagtaa aaattgttgg gggttgttgg
     4741 gacactacct gcgcgtcctg atccacgagc acatattgat tcacggagat atccgcaaaa
     4801 ccaccaaact gaacatcggc ggaacggaga tcgaggtcga gactctcgtc gaagacgaag
     4861 ccgaggccgg agccgtgcca gccgcggccg tgagcaaggg cgaggaggtc atcaaagagt
     4921 tcatgcgctt caaggtgcgc atggagggct ccatgaacgg ccacgagttc gagatcgagg
     4981 gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag gtgaccaagg
     5041 gcggccccct gcccttcgcc tgggacatcc tgtcccccca gttcatgtac ggctccaagg
     5101 cgtacgtgaa gcaccccgcc gacatccccg attacaagaa gctgtccttc cccgagggct
     5161 tcaagtggga gcgcgtgatg aacttcgagg acggcggtct ggtgaccgtg acccaggact
     5221 cctccctgca ggacggcacg ctgatctaca aggtgaagat gcgcggcacc aacttccccc
     5281 ccgacggccc cgtaatgcag aagaagacca tgggctggga ggcctccacc gagcgcctgt
     5341 acccccgcga cggcgtgctg aagggcgaga tccaccaggc cctgaagctg aaggacggcg
     5401 gccactacct ggtggagttc aagaccatct acatggccaa gaagcccgtg caactgcccg
     5461 gctactacta cgtggacacc aagctggaca tcacctccca caacgaggac tacaccatcg
     5521 tggaacagta cgagcgctcc gagggccgcc accacctgtt cctggggcat ggcaccggca
     5581 gcaccggcag cggcagctcc ggcaccgcct cctccgagga caacaacatg gccgtcatca
     5641 aagagttcat gcgcttcaag gtgcgcatgg agggctccat gaacggccac gagttcgaga
     5701 tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag ctgaaggtga
     5761 ccaagggcgg ccccctgccc ttcgcctggg acatcctgtc cccccagttc atgtacggct
     5821 ccaaggcgta cgtgaagcac cccgccgaca tccccgatta caagaagctg tccttccccg
     5881 agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggtctggtg accgtgaccc
     5941 aggactcctc cctgcaggac ggcacgctga tctacaaggt gaagatgcgc ggcaccaact
     6001 tcccccccga cggccccgta atgcagaaga agaccatggg ctgggaggcc tccaccgagc
     6061 gcctgtaccc ccgcgacggc gtgctgaagg gcgagatcca ccaggccctg aagctgaagg
     6121 acggcggcca ctacctggtg gagttcaaga ccatctacat ggccaagaag cccgtgcaac
     6181 tgcccggcta ctactacgtg gacaccaagc tggacatcac ctcccacaac gaggactaca
     6241 ccatcgtgga acagtacgag cgctccgagg gccgccacca cctgttcctg tacggcatgg
     6301 acgagctgta caagtaagat caagcttgca aagcggttca atttgcggcc gcctttaaat
     6361 aggccggccc tttctgcagc ttccactaaa ttgtcgactt tcgaaattta gaaacttaat
     6421 tttggctatc gattaaaacg cgtacaaggc cggccctgca ggaattcgat atcaagctta
     6481 tcgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     6541 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     6601 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     6661 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     6721 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
     6781 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     6841 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc
     6901 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
     6961 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
     7021 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcatcgat
     7081 accgtcgacc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     7141 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     7201 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     7261 caagggggag gattgggaag acaatagcag gcatgctggg gaactagtgg tgccagggcg
     7321 tgcccttggg ctccccgggc gcggcggccg catcgaattc taccgggtag gggaggcgct
     7381 tttcccaagg cagtctggag catgcgcttt agcagccccg ctgggcactt ggcgctacac
     7441 aagtggcctc tggcctcgca cacattccac atccaccggt aggcgccaac cggctccgtt
     7501 ctttggtggc cccttcgcgc caccttctac tcctccccta gtcaggaagt tcccccccgc
     7561 cccgcagctc gcgtcgtgca ggacgtgaca aatggaagta gcacgtctca ctagtctcgt
     7621 gcagatggac agcaccgctg agcaatggaa gcgggtaggc ctttggggca gcggccaata
     7681 gcagctttgc tccttcgctt tctgggctca gaggctggga aggggtgggt ccgggggcgg
     7741 gctcaggggc gggctcaggg gcggggcggg cgcccgaagg tcctccggag gcccggcatt
     7801 ctgcacgctt caaaagcgca cgtctgccgc gctgttctcc tcttcctcat ctccgggcct
     7861 ttcgacctgc aatcgccgct agcgaagttc ctattctcta gaaagtatag gaacttcgcc
     7921 accatgggat cggccattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     7981 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     8041 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     8101 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     8161 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     8221 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     8281 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     8341 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     8401 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     8461 cgcatgcccg acggcgatga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     8521 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     8581 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     8641 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     8701 tatcgccttc ttgacgagtt cttctgaggg gatccgctgt aagtctgcag aaattgatga
     8761 tctattaaac aataaagatg tccactaaaa tggaagtttt tcctgtcata ctttgttaag
     8821 aagggtgaga acagagtacc tacattttga atggaaggat tggagctacg ggggtggggg
     8881 tggggtggga ttagataaat gcctgctctt tactgaaggc tctttactat tgctttatga
     8941 taatgtttca tagttggata tcataattta aacaagcaaa accaaattaa gggccagctc
     9001 attcctccca ctcatgatct atagatctat agatctctcg tgggatcatt gtttttctct
     9061 tgattcccac tttgtggttc taagtactgt ggtttccaaa tgtgtcagtt tcatagcctg
     9121 aagaacgaga tcagcagcct ctgttccaca tacacttcat tctcagtatt gttttgccaa
     9181 gttctaattc catcagaaag cttgcagaag atctccccaa ctggggtaac ctttgagttc
     9241 tctcagttgg gggggcgcgc cctagaagat gggcgggagt cttctgggca ggcttaaagg
     9301 ctaacctggt gtgtgggcgt tgtcctgcag gggaattgaa caggtgtaaa attggaggga
     9361 caagacttcc cacagatttt cggttttgtc gggaagtttt ttaatagggg caaataggaa
     9421 aatggaggat aggagtcatc tggggtttat gcagcaaaac tacaggtata ttgcttgtat
     9481 ccgcctcgga gatttccatg aggagataaa gacatgtcac ccgagtttat actctcctgc
     9541 ttagatccta ctacagtatg aaatacagtg tygcgaggta gactatgtaa gcagatttaa
     9601 tcattttaaa gagcccagta cttcatatcc atttctcccg ctccttctgc agccttatca
     9661 aaaggtattt agaacactca ttttagcccc attttcattt attatactgg cttatccaac
     9721 ccctagacag agcattggca ttttcccttt cctgatctta gaagtctgat gactcatgaa
     9781 accagacaga ttagttacat acaccacaaa tcgaggctgt agctggggcc tcaacactgc
     9841 agttctttta taactcctta gtacactttt tgttgatcct ttgccttgat ccttaatttt
     9901 cagtgtctat cacctctccc gtcaggtggt gttccacatt tgggcctatt ctcagtccag
     9961 ggagttttac aacaatagat gtattgagaa tccaacctaa agcttaactt tccactccca
    10021 tgaatgcctc tctccttttt ctccattata actgagctat waccattaat ggtttcaggt
    10081 ggatgtctcc tcccccaata tacctgatgt atctacatat tgccaggctg atattttaag
    10141 acatwaaagg tatatttcat tattgagcca catggtattg attactgcta ctaaaatttt
    10201 gtcattgtac acatctgtaa aaggtggttc cttttggaat gcaaagttca ggtgtttgtt
    10261 gtctttcctg acctaaggtc ttgtgagctt gtattttttc tatttaagca gtgctttctc
    10321 ttggactggc ttgactcatg gcattctaca cgttattgct ggtctaaatg tgattttgcc
    10381 aagcttcttc aggacctata attttgcttg acttgtagcc aaacacaagt aaaatgatta
    10441 agcaacaaat gtatttgtga agcttggttt ttaggttgtt gtgttgtgtg tgcttgtgct
    10501 ctataataat actatccagg ggctggagag gtggctcgga gttcaagagc acagactgct
    10561 cttccagaag tcctgagttc aattcccagc aaccacatgg tggctcacaa ccatctgtaa
    10621 tgggatctga tgccctcttc tggtgtgtct gaagaccaca agtgtattca cattaaataa
    10681 ataatcctcc ttcttcttct tttttttttt ttaaagagaa twctgtctcc agtagaatta
    10741 ctgaagtaat gaaatacttt gtgtttgttc caatatggwa gccaataatc aaatactctt
    10801 wagcactgga aatgtaccaa ggaactattt tatttaagtg wactgtggac agaggagcca
    10861 taactgcaga cttgtgggat acagaagacc aatgcagact taatgtcttt tctcttacac
    10921 taagcaataa agaaataaaa attgaacttc tagtatccta tttgttaaac tgctagcttt
    10981 actaactttt gtgcttcatc tatacaaagc tgaaagctaa gtctgcagcc attactaaac
    11041 atgaaagcaa gtaatgataa ttttggattt caaaaatgta gggccagagt ttagccagcc
    11101 agtggtggtg cttgccttta tgccttaatc ccagcactct ggaggcagag acaggcagat
    11161 ctctgagttt gagcccagcc tggtctacac atcaagttct atctaggata gccaggaata
    11221 cacacagaaa ccctgttggg gaggggggct ctgagatttc ataaaattat aattgaagca
    11281 ttccctaatg agccactatg gatgtggcta aatccgtcta cctttctgat gagatttggg
    11341 tattattttt tctgtctctg ctgttggttg ggtcttttga cactgtgggc tttcttaaag
    11401 cctccttccc tgccatgtgg tctcttgttt gctactaact tcccatggct taaatggcat
    11461 ggctttttgc cttctaaggg cagctgctga gwtttgcagc ctgatttcca gggtggggtt
    11521 gggaaatctt tcaaacacta aaattgtcct ttaatttttt tttaaaaaat gggttatata
    11581 ataaacctca taaaatagtt atgaggagtg aggtggacta atattaatga gtccctcccc
    11641 tataaaagag ctattaaggc tttttgtctt atactaactt tttttttaaa tgtggtatct
    11701 ttagaaccaa gggtcttaga gttttagtat acagaaactg ttgcatcgct taatcagatt
    11761 ttctagtttc aaatccagag aatccaaatt cttcacagcc aaagtcaaat taagaatttc
    11821 tgactttaat gttatttgct actgtgaata taaaatgata gcttttcctg aggcagggtc
    11881 tcactatgta tctctgcctg atctgcaaca agatatgtag actaaagttc tgcctgcttt
    11941 tgtctcctga atactaaggt taaaatgtag taatactttt ggaacttgca ggtcagattc
    12001 ttttataggg gacacactaa gggagcttgg gtgatagttg gtaaatgtgt ttaagtgatg
    12061 aaaacttgaa ttattatcac cgcaacctac tttttaaaaa aaaaagccag gcctgttaga
    12121 gcatgctaag ggatccctag gacttgctga gcacacaaga gtagtacttg gcaggctcct
    12181 ggtgagagca tatttcaaaa aacaaggcag acaaccaaga aactacagta aggttacctg
    12241 tctttaacca tctgcatata cacagggata ttaaaatatt ccaaataata tttcattcaa
    12301 gttttccccc atcaaattgg gacatggatt tctccggtga ataggcagag ttggaaacta
    12361 aacaaatgtt ggttttgtga tttgtgaaat tgttttcaag tgatagttaa agcccatgag
    12421 atacagaaca aagctgctat ttcgaggtct cttggttata ctcagaagca cttctttggg
    12481 tttccctgca ctatcctgat catgtgctag gcctwcctta ggctgattgt tgttcaaata
    12541 acttaagttt cctgtcaggt gatgtcatat gatttcatat atcaaggcaa aacatgttat
    12601 atatgttaaa catttgkact taatgtgaaa gttaggtctt tgtgggtttt gattttaatt
    12661 tcaaaacctg agctaaataa gtcattttac atgtcttaca tttggtgaat tgtatattgt
    12721 ggtttgcagg caagactctc tgacctagta accctcctat agagcacttt gctgggtcac
    12781 aagtctagga gtcaagcatt tcaccttgaa gttgagacgt tttgttagtg tatactagtt
    12841 atatgttgga ggacatgttt atccagaaga tattcaggac tatttttgac tgggctaagg
    12901 aattgattct gattagcact gttagtgagc attgagtggc ctttaggctt gaattggagt
    12961 cacttgtata tctcaaataa tgctggcctt ttttwaaaag cccttgttct ttatcaccct
    13021 gttttctaca taatttttgt tcaaagaaat acttgtttgg atctcctttt gacaacaata
    13081 gcatgttttc aagccatatt ttttttcctt tttttttttt tttttggttt ttcgagacag
    13141 ggtttctctg tatagccctg gctgtcctgg aactcacttt gtagaccagg ctggcctcga
    13201 actcagaaat ccgcctgcct ctgcctcctg agtgccggga ttaaaggcgt gcaccaccac
    13261 gcctggctaa gttggatatt ttgtatataa ctataaccaa tactaactcc actgggtgga
    13321 tttttaattc agtcagtagt cttaagtggt ctttattggc ccttattaaa atctactgtt
    13381 cactctaaca gaggctgttg gactagtggs actaagcaac ttcctacgga tatactagca
    13441 gataagggtc agggatagaa actagtctag cgttttgtat acctaccagc ttatactacc
    13501 ttgttctgat agaaatattt aggacatcta gcttatcgat ccgtcgacgg tatcgataag
    13561 cttgatatcg aattctaccg ggtaggggag gcgcttttcc aaggcagtct gagcatgcgc
    13621 ttagcagccc cgctggcact tggcgctaca caagtggcct ytggcctcgc acacattcca
    13681 catccaccgg taggcgccaa ccggctccgt tctttggtgg ccccttcgcg ccaccttctw
    13741 ctcctcccct agtcaggaag ttcccccccg ccccgcagct cgcgtcgtsa ggacgtgaca
    13801 aatggaagta gcacgtctca ctagtctcgt cagatggaca gcaccgctga gcaatggaag
    13861 cgggtaggcc tttggggcag cggccaatag cagctttgct ccttcgcttt ctgggctcag
    13921 aggctgggaa ggggtgggtc cgggggcggg ctcaggggcg ggctcagggg cggggcgggc
    13981 gcccgaaggt cctccggagg cccggcattc tgcacgcttc aaaagcgcac gtctgccgcg
    14041 ctgttctcct cttcctcatc tccgggcctt tcgacctgca ggtcctcgcc atggatcctg
    14101 atgatgttgt tattcttcta atcttttgta tggaaaactt ttcttcgtac cacgggacta
    14161 aacctggtta tgtagattcc attcaaaaag gtatacaaaa gccaaaatct ggtacacaag
    14221 gaaattatga cgatgattgg aaagggtttt atagtaccga caataaatac gacgctgcgg
    14281 gatactctgt agataatgaa aacccgctct ctggaaaagc tggaggcgtg gtcaaagtga
    14341 cgtatccagg actgacgaag gttctcgcac taaaagtgga taatgccgaa actattaaga
    14401 aagagttagg tttaagtctc actgaaccgt tgatggagca agtcggaacg gaagagttta
    14461 tcaaaaggtt cggtgatggt gcttcgcgtg tagtgctcag ccttcccttc gctgagggga
    14521 gttctagcgt tgaatatatt aataactggg aacaggcgaa agcgttaagc gtagaacttg
    14581 agattaattt tgaaacccgt ggaaaacgtg gccaagatgc gatgtatgag tatatggctc
    14641 aagcctgtgc aggaaatcgt gtcaggcgat ctctttgtga aggaacctta cttctgtggt
    14701 gtgacataat tggacaaact acctacagag atttaaagct ctaaggtaaa tataaaattt
    14761 ttaagtgtat aatgtgttaa actactgatt ctaattgttt gtgtatttta gattccaacc
    14821 tatggaactg atgaatggga gcagtggtgg aatgcagatc ctagagctcg ctgatcagcc
    14881 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
    14941 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
    15001 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
    15061 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg
    15121 gaaagaacca gctggggctc gacctcgagg gggggcccgg tacccagctt ttgttccctt
    15181 tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
    15241 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
    15301 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
    15361 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
    15421 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
    15481 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
    15541 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
    15601 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
    15661 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
    15721 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
    15781 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
    15841 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
    15901 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
    15961 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
    16021 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
    16081 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
    16141 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
    16201 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
    16261 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
    16321 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
    16381 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
    16441 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
    16501 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
    16561 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
    16621 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
    16681 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
    16741 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
    16801 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
    16861 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
    16921 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
    16981 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
    17041 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
    17101 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
    17161 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
    17221 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
    17281 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
    17341 cgcacatttc cccgaaaagt gccacctaaa ttgtaagcgt taatattttg ttaaaattcg
    17401 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
    17461 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
    17521 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
    17581 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
    17641 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
    17701 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
    17761 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
    17821 cgtcccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt
    17881 cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc
    17941 cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgagcgcgcg taatacgact
    18001 cactataggg cgaattggag ctcca
//
