LOCUS       sequence-199304-       17785 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Rosa26 gene targeting vector to induce a Cre-dependent mosaic of 
            cells expressing different membrane localized fluorescent proteins 
            and Notch signalling genes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 17785)
  AUTHORS   Pontes-Quero S, Heredia L, Casquero-Garcia V, Fernandez-Chacon M, 
            Luo W, Hermoso A, Bansal M, Garcia-Gonzalez I, Sanchez-Munoz MS, 
            Perea JR, Galiana-Simal A, Rodriguez-Arabaolaza I, Del Olmo-Cabrera 
            S, Rocha SF, Criado-Rodriguez LM, Giovinazzo G, Benedito R
  TITLE     Dual ifgMosaic: A Versatile Method for Multispectral and 
            Combinatorial Mosaic Gene-Function Analysis.
  JOURNAL   Cell. 2017 Aug 10;170(4):800-814.e18. doi: 
            10.1016/j.cell.2017.07.031.
  PUBMED    28802047
REFERENCE   2  (bases 1 to 17785)
  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: 
            "10.1016/j.cell.2017.07"; journalName: "Cell"; date: "2017-08-10- 
            10"; volume: "170"; issue: "4"; pages: "800-814.e18"
COMMENT     Sequence Label: iMb-Notch-Mosaic (IR99.40)
FEATURES             Location/Qualifiers
     source          1..17785
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     332..354
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     416..435
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    complement(469..502)
                     /label=loxN
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AGGTATAC) (Shaw et al., 2021). loxN sites are 
                     compatible with each other, but incompatible with loxP or 
                     lox2272 sites."
     protein_bind    complement(510..543)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(552..585)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      599..608
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        659..1158
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(694..717)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     1091..1110
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     promoter        1170..1217
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             1236..2039
                     /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(1299..1318)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1909..1928
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    2077..2301
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     polyA_signal    2458..2592
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2495..2514)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2549..2568
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(2612..2645)
                     /label=loxN
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AGGTATAC) (Shaw et al., 2021). loxN sites are 
                     compatible with each other, but incompatible with loxP or 
                     lox2272 sites."
     regulatory      2768..2777
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2774..2776,2777..2779,2780..3490)
                     /codon_start=1
                     /product="enhanced YFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2819..2840)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3080..3099)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3427..3448
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3500..3553
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             3584..3961
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     CDS             3986..4039
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     polyA_signal    4099..4323
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    complement(4358..4391)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     CDS             4435..5862
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGRYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     CDS             5872..5925
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             5956..6333
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     CDS             6358..6411
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             8776..8799
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    8825..9049
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    complement(9084..9117)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      9240..9249
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             9246..9941
                     /codon_start=1
                     /product="monomeric far-red fluorescent protein (Shcherbo 
                     et al., 2009)"
                     /label=mKate2
                     /note="mammalian codon-optimized"
                     /translation="MVSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK
                     AVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRADMAL
                     KLVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAVARY
                     CDLPSKLGHR"
     CDS             9960..10013
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             10044..10421
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     CDS             10446..10499
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     polyA_signal    12121..12345
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    12397..12444
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     primer_bind     complement(12623..12640)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     complement(12812..12828)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    12847..13071
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     misc_feature    13161..17501
                     /label=Rosa26 right arm
                     /note="right homology arm from intron 1 of the mouse Rosa26
                     gene (Soriano, 1999)"
     primer_bind     complement(17552..17568)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(17600..17619)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(17601..17619)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(17629..17646)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(17629..17645)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(17638..17660)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 cgggccgggg agggctcggg ggaggggcgc ggcggccccc ggagcgccgg cggctgtcga
       61 ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt gcgagagggc gcagggactt
      121 cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac cccctctagc
      181 gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga gggccttcgt
      241 gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc gcggggggac
      301 ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt gaccggcggc
      361 tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct cctgggcaac
      421 gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tcctcgagat aacttcgtat
      481 agtatacctt atacgaagtt atcgagctca taacttcgta taggatactt tatacgaagt
      541 tataactcga gataacttcg tatagcatac attatacgaa gttataccgg cagatctcgc
      601 caccatggtg agtgtgatta cagaggctag cttggctgca ggtcgtcgaa attctaccgg
      661 gtaggggagg cgcttttccc aaggcagtct ggagcatgcg ctttagcagc cccgctgggc
      721 acttggcgct acacaagtgg cctctggcct cgcacacatt ccacatccac cggtaggcgc
      781 caaccggctc cgttctttgg tggccccttc gcgccacctt ctactcctcc cctagtcagg
      841 aagttccccc ccgccccgca gctcgcgtcg tgcaggacgt gacaaatgga agtagcacgt
      901 ctcactagtc tcgtgcagat ggacagcacc gctgagcaat ggaagcgggt aggcctttgg
      961 ggcagcggcc aatagcagct ttgctccttc gctttctggg ctcagaggct gggaaggggt
     1021 gggtccgggg gcgggctcag gggcgggctc aggggcgggg cgggcgcccg aaggtcctcc
     1081 ggaggcccgg cattctgcac gcttcaaaag cgcacgtctg ccgcgctgtt ctcctcttcc
     1141 tcatctccgg gcctttcgac ctgcagcctg ttgacaatta atcatcggca tagtatatcg
     1201 gcatagtata atacgacaag gtgaggaact aaaccatggg atcggccatt gaacaagatg
     1261 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     1321 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     1381 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc
     1441 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     1501 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     1561 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     1621 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     1681 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     1741 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgat gatctcgtcg
     1801 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     1861 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     1921 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     1981 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     2041 gggatcaatt ctctagagct cgctgatcag cctcgactgt gccttctagt tgccagccat
     2101 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     2161 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     2221 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg
     2281 gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc tcgactagag
     2341 cttgcggaac ccttcgtaat cagccatacc acatttgtag aggttttact tgctttaaaa
     2401 aacctcccac acctccccct gaacctgaaa catagaatga atgcaattgt tgttgttaac
     2461 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     2521 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     2581 catgtctgga tctgacatgg taagtaagct tataacttcg tatagtatac cttatacgaa
     2641 gttatggcgc gcctaattca cagccaccat gctgtgctgc atcagaagaa ctaaaccggt
     2701 tgagaagaat gaagaggccg atcaggagct gcagtcgacg gtaccgcggg cccgggatcc
     2761 accggtcgcc accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
     2821 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg
     2881 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
     2941 gccctggccc accctcgtga ccaccttcgg ctacggcctg cagtgcttcg cccgctaccc
     3001 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
     3061 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
     3121 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
     3181 catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga
     3241 caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag
     3301 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     3361 gcccgacaac cactacctga gctaccagtc caagctgagc aaagacccca acgagaagcg
     3421 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
     3481 gctgtacaag ggcagtggag agggcagagg aagtctgcta acatgcggtg acgtcgagga
     3541 gaatcctggc ccagaattct gtgtggatac ctaccgctac atcatgccag agccagcgaa
     3601 gtctgctccc gccccgaaaa agggctccaa gaaggcggtg actaaggcgc agaagaaagg
     3661 cggcaagaag cgcaagcgca gccgcaagga gagctattcc atctatgtgt acaaggttct
     3721 gaagcaggtc caccctgaca ccggcatttc gtccaaggcc atgggcatca tgaactcgtt
     3781 tgtgaacgac attttcgagc gcatcgcagg tgaggcttcc cgcctggcgc attacaacaa
     3841 gcgctcgacc atcacctcca gggagatcca gacggccgtg cgcctgctgc tgcctgggga
     3901 gttggccaag cacgccgtgt ccgagggcac taaggccatc accaagtaca ccagcgctaa
     3961 ggatccaccg gtcgcctgaa gtggagaggg cagaggaagt ctgctaacat gcggtgacgt
     4021 cgaggagaat cctggcccag aattctgcag atatccagca cagtggcggc cgctcgagtc
     4081 taggcgcgcc agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     4141 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     4201 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     4261 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta
     4321 tggcttctga ggcggaaaga accagctggg gagatctata acttcgtata ggatacttta
     4381 tacgaagtta tggtaccggc gcgccgatcc atgggttgct gtttctccaa gaccatggtg
     4441 agcaagggag aggaggtcat caaagagttc atgcgcttca aggtgcgcat ggagggctcc
     4501 atgaacggcc acgagttcga gatcgagggc gagggcgagg gccgccccta cgagggcacc
     4561 cagaccgcca agctgaaggt gaccaagggc ggccccctgc ccttcgcctg ggacatcctg
     4621 tccccccagt tcatgtacgg ctccaaggcg tacgtgaagc accccgccga catccccgat
     4681 tacaagaagc tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac
     4741 ggcggtctgg tgaccgtgac ccaggactcc tccctgcagg acggcacgct gatctacaag
     4801 gtgaagatgc gcggcaccaa cttccccccc gacggccccg taatgcagaa gaagaccatg
     4861 ggctgggagg cctccaccga gcgcctgtac ccccgcgacg gcgtgctgaa gggcgagatc
     4921 caccaggccc tgaagctgaa ggacggcggc cactacctgg tggagttcaa gaccatctac
     4981 atggccaaga agcccgtgca actgcccggc tactactacg tggacaccaa gctggacatc
     5041 acctcccaca acgaggacta caccatcgtg gaacagtacg agcgctccga gggccgccac
     5101 cacctgttcc tggggcatgg caccggcagc accggcagcg gcagctccgg caccgcctcc
     5161 tccgaggaca acaacatggc cgtcatcaaa gagttcatgc gcttcaaggt gcgcatggag
     5221 ggctccatga acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag
     5281 ggcacccaga ccgccaagct gaaggtgacc aagggcggcc ccctgccctt cgcctgggac
     5341 atcctgtccc cccagttcat gtacggctcc aaggcgtacg tgaagcaccc cgccgacatc
     5401 cccgattaca agaagctgtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc
     5461 gaggacggcg gtctggtgac cgtgacccag gactcctccc tgcaggacgg cacgctgatc
     5521 tacaaggtga agatgcgcgg caccaacttc ccccccgacg gccccgtaat gcagaagaag
     5581 accatgggct gggaggcctc caccgagcgc ctgtaccccc gcgacggcgt gctgaagggc
     5641 gagatccacc aggccctgaa gctgaaggac ggcggccgct acctggtgga gttcaagacc
     5701 atctacatgg ccaagaagcc cgtgcaactg cccggctact actacgtgga caccaagctg
     5761 gacatcacct cccacaacga ggactacacc atcgtggaac agtacgagcg ctccgagggc
     5821 cgccaccacc tgttcctgta cggcatggac gagctgtaca agggcagtgg agagggcaga
     5881 ggaagtctgc taacatgcgg tgacgtcgag gagaatcctg gcccagaatt ctgtgtggat
     5941 acctaccgct acatcatgcc agagccagcg aagtctgctc ccgccccgaa aaagggctcc
     6001 aagaaggcgg tgactaaggc gcagaagaaa ggcggcaaga agcgcaagcg cagccgcaag
     6061 gagagctatt ccatctatgt gtacaaggtt ctgaagcagg tccaccctga caccggcatt
     6121 tcgtccaagg ccatgggcat catgaactcg tttgtgaacg acattttcga gcgcatcgca
     6181 ggtgaggctt cccgcctggc gcattacaac aagcgctcga ccatcacctc cagggagatc
     6241 cagacggccg tgcgcctgct gctgcctggg gagttggcca agcacgccgt gtccgagggc
     6301 actaaggcca tcaccaagta caccagcgct aaggatccac cggtcgccgg cagtggagag
     6361 ggcagaggaa gtctgctaac atgcggtgac gtcgaggaga atcctggccc agaattccct
     6421 gagggcttca aagtgtctga ggccagcaag aagaagcggc gggagcccct cggcgaggac
     6481 tccgtgggcc tcaagcccct gaagaacgct tcagacggtg ccctcatgga cgacaaccag
     6541 aatgagtggg gggacgagga cctggagacc aagaagttcc ggttcgagga gcccgtggtt
     6601 ctgcctgacc tggacgacca gacagaccac cggcagtgga ctcagcagca cctggatgcc
     6661 gctgacctgc gcatgtctgc catggccccc acaccgcccc agggtgaggt tgacgccgac
     6721 tgcatggacg tcaatgtccg cgggcctgat ggcttcaccc cgctcatgat cgcctcctgc
     6781 agcgggggcg gcctggagac gggcaacagc gaggaagagg aggacgcgcc ggccgtcatc
     6841 tccgacttca tctaccaggg cgccagcctg cacaaccaga cagaccgcac gggcgagacc
     6901 gccttgcacc tggccgcccg ctactcacgc tctgatgccg ccaagcgcct gctggaggcc
     6961 agcgcagatg ccaacatcca ggacaacatg ggccgcaccc cgctgcatgc ggctgtgtct
     7021 gccgacgcac aaggtgtctt ccagatcctg atccggaacc gagccacaga cctggatgcc
     7081 cgcatgcatg atggcacgac gccactgatc ctggctgccc gcctggccgt ggagggcatg
     7141 ctggaggacc tcatcaactc acacgccgac gtcaacgccg tagatgacct gggcaagtcc
     7201 gccctgcact gggccgccgc cgtgaacaat gtggatgccg cagttgtgct cctgaagaac
     7261 ggggctaaca aagatatgca gaacaacagg gaggagacac ccctgtttct ggccgcccgg
     7321 gagggcagct acgagaccgc caaggtgctg ctggaccact ttgccaaccg ggacatcacg
     7381 gatcatatgg accgcctgcc gcgcgacatc gcacaggagc gcatgcatca cgacatcgtg
     7441 aggctgctgg acgagtacaa cctggtgcgc agcccgcagc tgcacggagc cccgctgggg
     7501 ggcacgccca ccctgtcgcc cccgctctgc tcgcccaacg gctacctggg cagcctcaag
     7561 cccggcgtgc agggcaagaa ggtccgcaag cccagcagca aaggcctggc ctgtggaagc
     7621 aaggaggcca aggacctcaa ggcacggagg aagaagtccc aggatggcaa gggctgcctg
     7681 ctggacagct ccggcatgct ctcgcccgtg gactccctgg agtcacccca tggctacctg
     7741 tcagacgtgg cctcgccgcc actgctgccc tccccgttcc agcagtctcc gtccgtgccc
     7801 ctcaaccacc tgcctgggat gcccgacacc cacctgggca tcgggcacct gaacgtggcg
     7861 gccaagcccg agatggcggc gctgggtggg ggcggccggc tggcctttga gactggccca
     7921 cctcgtctct cccacctgcc tgtggcctct ggcaccagca ccgtcctggg ctccagcagc
     7981 ggaggggccc tgaatttcac tgtgggcggg tccaccagtt tgaatggtca atgcgagtgg
     8041 ctgtcccggc tgcagagcgg catggtgccg aaccaataca accctctgcg ggggagtgtg
     8101 gcaccaggcc ccctgagcac acaggccccc tccctgcagc atggcatggt aggcccgctg
     8161 cacagtagcc ttgctgccag cgccctgtcc cagatgatga gctaccaggg cctgcccagc
     8221 acccggctgg ccacccagcc tcacctggtg cagacccagc aggtgcagcc acaaaactta
     8281 cagatgcagc agcagaacct gcagccagca aacatccagc agcagcaaag cctgcagccg
     8341 ccaccaccac caccacagcc gcaccttggc gtgagctcag cagccagcgg ccacctgggc
     8401 cggagcttcc tgagtggaga gccgagccag gcagacgtgc agccactggg ccccagcagc
     8461 ctggcggtgc acactattct gccccaggag agccccgccc tgcccacgtc gctgccatcc
     8521 tcgctggtcc cacccgtgac cgcagcccag ttcctgacgc ccccctcgca gcacagctac
     8581 tcctcgcctg tggacaacac ccccagccac cagctacagg tgcctgagca ccccttcctg
     8641 accccttcgc cggagtcgcc cgaccaatgg tcgtcctcgt cgccgcactc taatgtgtct
     8701 gactggtctg agggcgtgtc gtcgcccccg ccctccatgc agtcccagat cgcgcgcatc
     8761 ccggaggcct tcaaggacta caaggatgac gatgacaaat aaggcgcgcc ctcgagagcc
     8821 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     8881 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     8941 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     9001 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg
     9061 gaaagaacca gctggggact agtataactt cgtatagcat acattatacg aagttatggc
     9121 gcgcctaatt cacagccacc atgctgtgct gcatcagaag aactaagccg gttgagaaga
     9181 atgaagaggc cgatcaggag ctgcagtcga cggtaccgcg ggcccgggat ccaccggtcg
     9241 ccaccatggt gagcgagctg attaaggaga acatgcacat gaagctctac atggagggca
     9301 ccgtgaacaa ccaccacttc aagtgcacat ccgagggcga aggcaagccc tacgagggca
     9361 cccagaccat gagaatcaag gcggtcgagg gcggccctct ccccttcgcc ttcgacatcc
     9421 tggctaccag cttcatgtac ggcagcaaaa ccttcatcaa ccacacccag ggcatccccg
     9481 acttctttaa gcagtccttc cccgagggct tcacatggga gagagtcacc acatacgaag
     9541 acgggggcgt gctgaccgct acccaggaca ccagcctcca ggacggctgc ctcatctaca
     9601 acgtcaagat cagaggggtg aacttcccat ccaacggccc tgtgatgcag aagaaaacac
     9661 tcggctggga ggcctccacc gagaccctgt accccgctga cggcggcctg gaaggcagag
     9721 ccgacatggc cctgaagctc gtgggcgggg gccacctgat ctgcaacttg aagaccacat
     9781 acagatccaa gaaacccgct aagaacctca agatgcccgg cgtctactat gtggacagaa
     9841 gactggaaag aatcaaggag gccgacaaag agacctacgt cgagcagcac gaggtggctg
     9901 tggccagata ctgcgacctc cctagcaaac tggggcacag aatgtacaag ggcagtggag
     9961 agggcagagg aagtctgcta acatgcggtg acgtcgagga gaatcctggc ccagaattct
    10021 gtgtggatac ctaccgctac atcatgccag agccagcgaa gtctgctccc gccccgaaaa
    10081 agggctccaa gaaggcggtg actaaggcgc agaagaaagg cggcaagaag cgcaagcgca
    10141 gccgcaagga gagctattcc atctatgtgt acaaggttct gaagcaggtc caccctgaca
    10201 ccggcatttc gtccaaggcc atgggcatca tgaactcgtt tgtgaacgac attttcgagc
    10261 gcatcgcagg tgaggcttcc cgcctggcgc attacaacaa gcgctcgacc atcacctcca
    10321 gggagatcca gacggccgtg cgcctgctgc tgcctgggga gttggccaag cacgccgtgt
    10381 ccgagggcac taaggccatc accaagtaca ccagcgctaa ggatccaccg gtcgccggca
    10441 gtggagaggg cagaggaagt ctgctaacat gcggtgacgt cgaggagaat cctggcccag
    10501 aattcccagt aatgccctcc ggttttcctc agtctccacg tacgtccccg agggcgcgtc
    10561 ccaaaacccg gataaccgga gcgctcccca tggactactc ggagggcttg tccgcggagg
    10621 agcggcctgc gcatgctcca tcggcgggga agtttggtga gcggcctcca cccaaacgac
    10681 tcactaggga agctatgcga aactatttaa aagaacgagg ggatcaaaca gtgctcattc
    10741 ttcatgcaaa agtcgcacag aagtcttacg gaaatgaaaa acgatttttt tgccctcctc
    10801 cttgtgtgta tcttatgggc agtggttgga agaaaaaaaa agagcaaatg gaacgagatg
    10861 gatgttctga acaagagtct caaccctgtg cgtttattgg aataggaaac agtgaccaag
    10921 aaatgcagca gctgaacttg gaagggaaga actactgtac agccaaaaca ttgtacatat
    10981 ctgattcaga caagagaaaa catttcatgt tgtctgtaaa gatgttctat ggcaacagcg
    11041 atgacattgg tgtgttcctc agcaagcgga taaaggtcat ctccaaaccc tccaaaaaga
    11101 agcagtcact gaaaaatgct gacttgtgca ttgcttcagg aacgaaggtg gcactgttca
    11161 atcaccttcg gtcccagaca gttagtacca ggtacctgca tgtagaagga gggaatttcc
    11221 acgccagttc acaacagtgg ggagcatttt acatccatct cttggacgac gacgagtcgg
    11281 aaggagagga gttcacagtt agagatggct acatccatta cgggcagact gtcaagcttg
    11341 tgtgctcagt gactggcatg gcactcccaa gattgataat taggaaagtt gataagcaga
    11401 cggcattact ggatgcagac gaccctgtat cacaactcca caaatgtgca ttttacctta
    11461 aggatacaga aagaatgtac ttgtgccttt ctcaagaaag aataatccaa tttcaggcca
    11521 ctccatgtcc aaaagaacaa aataaggaaa tgataaacga tggagcttcc tggacaatca
    11581 ttagcacaga caaggccgag tacacgttct atgagggaat gggccccgtc cttgccccag
    11641 tcacccctgt gcctgtcgta gaaagtcttc agttgaatgg cggcggggac gtagcaatgc
    11701 ttgaacttac aggacaaaac tttactccaa atttaagagt gtggtttggg gatgtagaag
    11761 ccgaaacaat gtacagatgt ggagagagca tgctctgtgt ggtcccagac atttctgcat
    11821 tccgggaagg ttggagatgg gtccgccagc cagtccaggt tccagtgact ttggtccgaa
    11881 atgacggggt catttactcc accagcctta ccttcaccta cacaccagag ccagggccga
    11941 ggccacactg cagcgctgca ggagcgattc tcagagccaa ctccagccaa gtgccctcca
    12001 atgagtcaaa cacaaacagc gaggggaatt acacaaatgc cagcacaaat tctaccagtg
    12061 tcacatcgtc cacagcaacc gtggtgtcct aactctcgag tctaggcgcg ccagcctcga
    12121 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
    12181 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
    12241 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
    12301 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa
    12361 gaaccagctg ggggcggccg cgatcaattc ggtaccgaag ttcctattcc gaagttccta
    12421 ttctctagaa agtataggaa cttcctcgag ggtaccccaa ttccgccccc cccccctaac
    12481 gttactggcc gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat gttattttcc
    12541 accatattgc cgtcttttgg caatgtgagg gcccggaaac ctggccctgt cttcttgacg
    12601 agcattccta ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt gaatgtcgtg
    12661 aaggaagcag ttcctctgga agcttcttga agacaaacaa cgtctgtagc gaccctttgc
    12721 aggcagcgga accccccacc tggcgacagg tgcctctgcg gccaaaagcc accgagctcg
    12781 gatccactag tccagtgtgg tggaattggg ggatccacta gttctagagc ggccgatcag
    12841 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
    12901 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc
    12961 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
    13021 aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg
    13081 cggaaagaac cagctggggc tcgagatcca ctagttctag cctcgaggct agagcggccg
    13141 ccgcgccgat atcgaattcg ctagaagatg ggcgggagtc ttctgggcag gcttaaaggc
    13201 taacctggtg tgtgggcgtt gtcctgcagg ggaattgaac aggtgtaaaa ttggagggac
    13261 aagacttccc acagattttc ggttttgtcg ggaagttttt taataggggc aaataaggaa
    13321 aatgggagga taggtagtca tctggggttt tatgcagcaa aactacaggt tattattgct
    13381 tgtgatccgc ctcggagtat tttccatcga ggtagattaa agacatgctc acccgagttt
    13441 tatactctcc tgcttgagat ccttactaca gtatgaaatt acagtgtcgc gagttagact
    13501 atgtaagcag aattttaatc atttttaaag agcccagtac ttcatatcca tttctcccgc
    13561 tccttctgca gccttatcaa aaggtatttt agaacactca ttttagcccc attttcattt
    13621 attatactgg cttatccaac ccctagacag agcattggca ttttcccttt cctgatctta
    13681 gaagtctgat gactcatgaa accagacaga ttagttacat acaccacaaa tcgaggctgt
    13741 agctggggcc tcaacactgc agttctttta taactcctta gtacactttt tgttgatcct
    13801 ttgccttgat ccttaatttt cagtgtctat cacctctccc gtcaggtggt gttccacatt
    13861 tgggcctatt ctcagtccag ggagttttac aacaatagat gtattgagaa tccaacctaa
    13921 agcttaactt tccactccca tgaatgcctc tctccttttt ctccatttat aaactgagct
    13981 attaaccatt aatggtttcc aggtggatgt ctcctccccc aatattacct gatgtatctt
    14041 acatattgcc aggctgatat tttaagacat taaaaggtat atttcattat tgagccacat
    14101 ggtattgatt actgcttact aaaattttgt cattgtacac atctgtaaaa ggtggttcct
    14161 tttggaatgc aaagttcagg tgtttgttgt ctttcctgac ctaaggtctt gtgagcttgt
    14221 attttttcta tttaagcagt gctttctctt ggactggctt gactcatggc attctacacg
    14281 ttattgctgg tctaaatgtg attttgccaa gcttcttcag gacctataat tttgcttgac
    14341 ttgtagccaa acacaagtaa aatgattaag caacaaatgt atttgtgaag cttggttttt
    14401 aggttgttgt gttgtgtgtg cttgtgctct ataataatac tatccagggg ctggagaggt
    14461 ggctcggagt tcaagagcac agactgctct tccagaagtc ctgagttcaa ttcccagcaa
    14521 ccacatggtg gctcacaacc atctgtaatg ggatctgatg ccctcttctg gtgtgtctga
    14581 agaccacaag tgtattcaca ttaaataaat aaatcctcct tcttcttctt tttttttttt
    14641 ttaaagagaa tactgtctcc agtagaattt actgaagtaa tgaaatactt tgtgtttgtt
    14701 ccaatatggt agccaataat caaattactc tttaagcact ggaaatgtta ccaaggaact
    14761 aatttttatt tgaagtgtaa ctgtggacag aggagccata actgcagact tgtgggatac
    14821 agaagaccaa tgcagacttt aatgtctttt ctcttacact aagcaataaa gaaataaaaa
    14881 ttgaacttct agtatcctat ttgtttaaac tgctagcttt acttaacttt tgtgcttcat
    14941 ctatacaaag ctgaaagcta agtctgcagc cattactaaa catgaaagca agtaatgata
    15001 attttggatt tcaaaaatgt agggccagag tttagccagc cagtggtggt gcttgccttt
    15061 atgcctttaa tcccagcact ctggaggcag agacaggcag atctctgagt ttgagcccag
    15121 cctggtctac acatcaagtt ctatctagga tagccaggaa tacacacaga aaccctgttg
    15181 gggagggggg ctctgagatt tcataaaatt ataattgaag cattccctaa tgagccacta
    15241 tggatgtggc taaatccgtc tacctttctg atgagatttg ggtattattt tttctgtctc
    15301 tgctgttggt tgggtctttt gacactgtgg gctttcttta aagcctcctt cctgccatgt
    15361 ggtctcttgt ttgctactaa cttcccatgg cttaaatggc atggcttttt gccttctaag
    15421 ggcagctgct gagatttgca gcctgatttc cagggtgggg ttgggaaatc tttcaaacac
    15481 taaaattgtc ctttaatttt ttttttaaaa aatgggttat ataataaacc tcataaaata
    15541 gttatgagga gtgaggtgga ctaatattaa atgagtccct cccctataaa agagctatta
    15601 aggctttttg tcttatactt aacttttttt ttaaatgtgg tatctttaga accaagggtc
    15661 ttagagtttt agtatacaga aactgttgca tcgcttaatc agattttcta gtttcaaatc
    15721 cagagaatcc aaattcttca cagccaaagt caaattaaga atttctgact tttaatgtta
    15781 atttgcttac tgtgaatata aaaatgatag cttttcctga ggcagggtct cactatgtat
    15841 ctctgcctga tctgcaacaa gatatgtaga ctaaagttct gcctgctttt gtctcctgaa
    15901 tactaaggtt aaaatgtagt aatacttttg gaacttgcag gtcagattct tttatagggg
    15961 acacactaag ggagcttggg tgatagttgg taaaatgtgt ttcaagtgat gaaaacttga
    16021 attattatca ccgcaaccta ctttttaaaa aaaaaagcca ggcctgttag agcatgctta
    16081 agggatccct aggacttgct gagcacacaa gagtagttac ttggcaggct cctggtgaga
    16141 gcatatttca aaaaacaagg cagacaacca agaaactaca gttaaggtta cctgtcttta
    16201 aaccatctgc atatacacag ggatattaaa atattccaaa taatatttca ttcaagtttt
    16261 cccccatcaa attgggacat ggatttctcc ggtgaatagg cagagttgga aactaaacaa
    16321 atgttggttt tgtgatttgt gaaattgttt tcaagtgata gttaaagccc atgagataca
    16381 gaacaaagct gctatttcga ggtctcttgg tttatactca gaagcacttc tttgggtttc
    16441 cctgcactat cctgatcatg tgctaggcct accttaggct gattgttgtt caaataaact
    16501 taagtttcct gtcaggtgat gtcatatgat ttcatatatc aaggcaaaac atgttatata
    16561 tgttaaacat ttgtacttaa tgtgaaagtt aggtctttgt gggtttgatt tttaattttc
    16621 aaaacctgag ctaaataagt catttttaca tgtcttacat ttggtggaat tgtataattg
    16681 tggtttgcag gcaagactct ctgacctagt aaccctacct atagagcact ttgctgggtc
    16741 acaagtctag gagtcaagca tttcaccttg aagttgagac gttttgttag tgtatactag
    16801 tttatatgtt ggaggacatg tttatccaga agatattcag gactattttt gactgggcta
    16861 aggaattgat tctgattagc actgttagtg agcattgagt ggcctttagg cttgaattgg
    16921 agtcacttgt atatctcaaa taatgctggc cttttttaaa aagcccttgt tctttatcac
    16981 cctgttttct acataatttt tgttcaaaga aatacttgtt tggatctcct tttgacaaca
    17041 atagcatgtt ttcaagccat attttttttc cttttttttt ttttttttgg tttttcgaga
    17101 cagggtttct ctgtatagcc ctggctgtcc tggaactcac tttgtagacc aggctggcct
    17161 cgaactcaga aatccgcctg cctctgcctc ctgagtgccg ggattaaagg cgtgcaccac
    17221 cacgcctggc taagttggat attttgttat ataactataa ccaatactaa ctccactggg
    17281 tggattttta attcagtcag tagtcttaag tggtctttat tggcccttca ttaaaatcta
    17341 ctgttcactc taacagaggc tgttggtact agtggcactt aagcaacttc ctacggatat
    17401 actagcagat taagggtcag ggatagaaac tagtctagcg ttttgtatac ctaccagctt
    17461 tatactacct tgttctgata gaaatatttc aggacatcta gcttatcgat accgtcgacg
    17521 ttcgaacacg tggaattcct gcagcccggg ggatccacta gttctagagc ggccgccacc
    17581 gcggtggagc tccaattcgc cctatagtga gtcgtattac gcgcgctcac tggccgtcgt
    17641 tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca
    17701 tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
    17761 gttgcgcagc ctgaatggcg aatgg
//
