LOCUS       Exported               16896 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Gene targeting vector for the mouse Rosa26 locus, including a CAG 
            promoter, loxP flanked stop cassette, GFP, for gateway cloning.
ACCESSION   .
VERSION     .
KEYWORDS    pR26 CAG/GFP Dest
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 16896)
  AUTHORS   Chu VT, Weber T, Graf R, Sommermann T, Petsch K, Sack U, Volchkov P,
            Rajewsky K, Kuhn R
  TITLE     Efficient generation of Rosa26 knock-in mice using CRISPR/Cas9 in 
            C57BL/6 zygotes.
  JOURNAL   BMC Biotechnol. 2016 Jan 16;16(1):4. doi: 10.1186/s12896-016-0234-4.
  PUBMED    26772810
REFERENCE   2  (bases 1 to 16896)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..16896
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    6..1088
                     /label=Rosa26 left arm
                     /note="left homology arm from intron 1 of the mouse Rosa26 
                     gene (Soriano, 1999)"
     misc_feature    1095..1198
                     /label=Ad SA
                     /note="adenovirus major late transcript splice acceptor 
                     (Friedrich and Soriano, 1991)"
     polyA_signal    1217..1424
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     polyA_signal    1438..1645
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     enhancer        1654..2033
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          2309..3325
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     3249..3271
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     3333..3352
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    3388..3421
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     polyA_signal    3428..3476
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     CDS             complement(3483..4286)
                     /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
                     EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3594..3613)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4204..4223
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     intron          5099..5164
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     polyA_signal    5739..5873
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5776..5795)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5830..5849
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    5893..5926
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     protein_bind    5939..6063
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        6088..6118
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             6172..6831
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(6238..6257)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             7173..7478
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     7357..7376
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(7519..7643)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    7678..7725
                     /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)."
     misc_feature    7764..8316
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(7908..7925)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     8135..8154
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             8328..9047
                     /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             8328..9044
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(8373..8394)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(8634..8653)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     8981..9002
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    9072..9119
                     /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(9144..9160)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9144..9160)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(9173..9190)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9179..9403
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     misc_feature    9493..13833
                     /label=Rosa26 right arm
                     /note="right homology arm from intron 1 of the mouse Rosa26
                     gene (Soriano, 1999)"
     primer_bind     complement(13918..13941)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     complement(14051..14071)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(14051..14069)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(14090..14106)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(14090..14106)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(14103..14125)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    14114..14130
                     /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(14138..14168)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    14183..14204
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(14321..14338)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(14492..15080)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(14572..14591)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(15251..16111)
                     /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     15874..15893
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(16112..16216)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(16269..16697)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(16379..16400)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     16591..16610
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     16824..16846
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     16838..16855
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     16839..16855
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     16865..16884
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        16865..16883
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ccaccgcggc aggccctccg agcgtggtgg agccgttctg tgagacagcc gggtacgagt
       61 cgtgacgctg gaaggggcaa gcgggtggtg ggcaggaatg cggtccgccc tgcagcaacc
      121 ggagggggag ggagaaggga gcggaaaagt ctccaccgga cgcggccatg gctcgggggg
      181 gggggggcag cggaggagcg cttccggccg acgtctcgtc gctgattggc ttcttttcct
      241 cccgccgtgt gtgaaaacac aaatggcgtg ttttggttgg cgtaaggcgc ctgtcagtta
      301 acggcagccg gagtgcgcag ccgccggcag cctcgctctg cccactgggt ggggcgggag
      361 gtaggtgggg tgaggcgagc tggacgtgcg ggcgcggtcg gcctctggcg gggcgggggg
      421 ggggagggag ggtcagcgaa agtagctcgc gcgcgagcgg ccgcccaccc tccccttcct
      481 ctgggggagt cgttttaccc gccgccggcc gggcctcgtc gtctgattgg ctctcggggc
      541 ccagaaaact ggcccttgcc attggctcgt gttcgtgcaa gttgagtcca tccgccggcc
      601 agcgggggcg gcgaggaggc gctcccaggt tccggccctc ccctcggccc cgcgccgcag
      661 agtctggccg cgcgcccctg cgcaacgtgg caggaagcgc gcgctggggg cggggacggg
      721 cagtagggct gagcggctgc ggggcgggtg caagcacgtt tccgacttga gttgcctcaa
      781 gaggggcgtg ctgagccaga cctccatcgc gcactccggg gagtggaggg aaggagcgag
      841 ggctcagttg ggctgttttg gaggcaggaa gcacttgctc tcccaaagtc gctctgagtt
      901 gttatcagta agggagctgc agtggagtag gcggggagaa ggccgcaccc ttctccggag
      961 gggggagggg agtgttgcaa tacctttctg ggagttctct gctgcctcct ggcttctgag
     1021 gaccgccctg ggcctgggag aatcccttcc ccctcttccc tcgtgatctg caactccagt
     1081 ctttctagca tctgtagggc gcagtagtcc agggtttcct tgatgatgtc atacttatcc
     1141 tgtccctttt ttttccacag ctcgcggttg aggacaaact cttcgcggtc tttccagtgg
     1201 ttaattaact cgaggactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     1261 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     1321 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     1381 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt ctagcgactg
     1441 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     1501 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     1561 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     1621 aagacaatag caggcatgct ggggatgcgg gtcgacattg attattgact agttattaat
     1681 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
     1741 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
     1801 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
     1861 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
     1921 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat
     1981 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtcgagg
     2041 tgagccccac gttctgcttc actctcccca tctccccccc ctccccaccc ccaattttgt
     2101 atttatttat tttttaatta ttttgtgcag cgatgggggc gggggggggg ggcgcgcgcc
     2161 aggcgggggg gggggggggg gggggggggg gggggggggg gggggggggg cggcggcagc
     2221 caatcagagc ggcgcgctcc gaaagtttcc ttttatggcg aggcggcggc ggcggcggcc
     2281 ctataaaaag cgaagcgcgc ggcgggcggg agtcgctgcg cgctgccttc gccccgtgcc
     2341 ccgctccgcc gccgcctcgc gccgcccgcc ccggctctga ctgaccgcgt tactcccaca
     2401 ggtgagcggg cgggacggcc cttctcctcc gggctgtaat tagcgcttgg tttaatgacg
     2461 gcttgtttct tttctgtggc tgcgtgaaag ccttgagggg ctccgggagg gccctttgtg
     2521 cggggggagc ggctcggggg gtgcgtgcgt gtgtgtgtgc gtggggagcg ccgcgtgcgg
     2581 ctccgcgctg cccggcggct gtgagcgctg cgggcgcggc gcggggcttt gtgcgctccg
     2641 cagtgtgcgc gaggggagcg cggccggggg cggtgccccg cggtgcgggg ggggctgcga
     2701 ggggaacaaa ggctgcgtgc ggggtgtgtg cgtggggggg tgagcagggg gtgtgggcgc
     2761 gtcggtcggg ctgcaacccc ccctgcaccc ccctccccga gttgctgagc acggcccggc
     2821 ttcgggtgcg gggctccgta cggggcgtgg cgcggggctc gccgtgccgg gcggggggtg
     2881 gcggcaggtg ggggtgccgg gcggggcggg gccgcctcgg gccggggagg gctcggggga
     2941 ggggcgcggc ggcccccgga gcgccggcgg ctgtcgaggc gcggcgagcc gcagccattg
     3001 ccttttatgg taatcgtgcg agagggcgca gggacttcct ttgtcccaaa tctgtgcgga
     3061 gccgaaatct gggaggcgcc gccgcacccc ctctagcggg cgcggggcga agcggtgcgg
     3121 cgccggcagg aaggaaatgg gcggggaggg ccttcgtgcg tcgccgcgcc gccgtcccct
     3181 tctccctctc cagcctcggg gctgtccgcg gggggacggc tgccttcggg ggggacgggg
     3241 cagggcgggg ttcggcttct ggcgtgtgac cggcggctct agagcctctg ctaaccatgt
     3301 tcatgccttc ttctttttcc tacagctcct gggcaacgtg ctggttattg tgctgtctca
     3361 tcattttggc aaagaattct taattaaata acttcgtata gcatacatta tacgaagtta
     3421 tggatccgaa caaacgaccc aacacccgtg cgttttattc tgtcttttta ttgccgatcc
     3481 cctcagaaga actcgtcaag aaggcgatag aaggcgatgc gctgcgaatc gggagcggcg
     3541 ataccgtaaa gcacgaggaa gcggtcagcc cattcgccgc caagctcttc agcaatatca
     3601 cgggtagcca acgctatgtc ctgatagcgg tccgccacac ccagccggcc acagtcgatg
     3661 aatccagaaa agcggccatt ttccaccatg atattcggca agcaggcatc gccatgggtc
     3721 acgacgagat cctcgccgtc gggcatgcgc gccttgagcc tggcgaacag ttcggctggc
     3781 gcgagcccct gatgctcttc gtccagatca tcctgatcga caagaccggc ttccatccga
     3841 gtacgtgctc gctcgatgcg atgtttcgct tggtggtcga atgggcaggt agccggatca
     3901 agcgtatgca gccgccgcat tgcatcagcc atgatggata ctttctcggc aggagcaagg
     3961 tgagatgaca ggagatcctg ccccggcact tcgcccaata gcagccagtc ccttcccgct
     4021 tcagtgacaa cgtcgagcac agctgcgcaa ggaacgcccg tcgtggccag ccacgatagc
     4081 cgcgctgcct cgtcctgcag ttcattcagg gcaccggaca ggtcggtctt gacaaaaaga
     4141 accgggcgcc cctgcgctga cagccggaac acggcggcat cagagcagcc gattgtctgt
     4201 tgtgcccagt catagccgaa tagcctctcc acccaagcgg ccggagaacc tgcgtgcaat
     4261 ccatcttgtt caatggccga tcccatattg gctgcagggt cgctcggtgt tcgaggccac
     4321 acgcgtcacc ttaatatgcg aagtggacct gggaccgcgc cgccccgact gcatctgcgt
     4381 gttcgaattc gccaatgaca agacgctggg cggggtttgc tcgacattgg gtggaaacat
     4441 tccaggcctg ggtggagagg ctttttgctt cctcttgcaa aaccacactg ctcgacattg
     4501 ggtggaaaca ttccaggcct gggtggagag gctttttgct tcctcttgaa aaccacactg
     4561 ctcgactagt gattaaagtc gactcgggga caccaaatat ggcgatctcg gccttttcgt
     4621 ttcttggagc tgggacatgt ttgccatcga tccatctacc accagaacgg ccgttagatc
     4681 tgctgccacc gttgtttcca ccgaagaaac caccgttgcc gtaaccacca cgacggttgt
     4741 tgctaaagaa gctgccaccg ccacggccac cgttgtagcc gccgttgttg ttattgtagt
     4801 tgctactgtt atttctggca cttcttggtt ttcctcttaa gtgaggagga acataaccat
     4861 tctcgttgtt gtcgttgatg cttaaatttt gcacttgttc gctcagttca gccataatat
     4921 gaaatgcttt tcttgttgtt cttacggaat accacttgcc acctatcacc acaactaact
     4981 ttttcccgtt cctccatctc ttttatattt tttttctcga gggatctttg tgaaggaacc
     5041 ttacttctgt ggtgtgacat aattggacaa actacctaca gagatttaaa gctctaaggt
     5101 aaatataaaa tttttaagtg tataatgtgt taaactactg attctaattg tttgtgtatt
     5161 ttagattcca acctatggaa ctgatgaatg ggagcagtgg tggaatgcct ttaatgagga
     5221 aaacctgttt tgctcagaag aaatgccatc tagtgatgat gaggctactg ctgactctca
     5281 acattctact cctccaaaaa agaagagaag ggtagaagac cccaaggact ttccttcaga
     5341 attgctaagt tttttgagtc atgctgtgtt tagtaataga actcttgctt gctttgctat
     5401 ttacaccaca aaggaaaaag ctgcactgct atacaagaaa attatggaaa aatattctgt
     5461 aacctttata agtaggcata acagttataa tcataacata ctgttttttc ttactccaca
     5521 caggcataga gtgtctgcta ttaataacta tgctcaaaaa ttgtgtacct ttagcttttt
     5581 aatttgtaaa ggggttaata aggaatattt gatgtatagt gccttgacta gagatcataa
     5641 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
     5701 tgaacctgaa acatagaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
     5761 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     5821 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg atctgacatg
     5881 gtaagtaagc ttataacttc gtatagcata cattatacga agttatcggc gcgccatcac
     5941 aagtttgtac aaaaaagctg aacgagaaac gtaaaatgat ataaatatca atatattaaa
     6001 ttagattttg cataaaaaac agactacata atactgtaaa acacaacata tccagtcact
     6061 atggcggccg cattaggcac cccaggcttt acactttatg cttccggctc gtataatgtg
     6121 tggattttga gttaggatcc gtcgagattt tcaggagcta aggaagctaa aatggagaaa
     6181 aaaatcactg gatataccac cgttgatata tcccaatggc atcgtaaaga acattttgag
     6241 gcatttcagt cagttgctca atgtacctat aaccagaccg ttcagctgga tattacggcc
     6301 tttttaaaga ccgtaaagaa aaataagcac aagttttatc cggcctttat tcacattctt
     6361 gcccgcctga tgaatgctca tccggaattc cgtatggcaa tgaaagacgg tgagctggtg
     6421 atatgggata gtgttcaccc ttgttacacc gttttccatg agcaaactga aacgttttca
     6481 tcgctctgga gtgaatacca cgacgatttc cggcagtttc tacacatata ttcgcaagat
     6541 gtggcgtgtt acggtgaaaa cctggcctat ttccctaaag ggtttattga gaatatgttt
     6601 ttcgtctcag ccaatccctg ggtgagtttc accagttttg atttaaacgt ggccaatatg
     6661 gacaacttct tcgcccccgt tttcaccatg ggcaaatatt atacgcaagg cgacaaggtg
     6721 ctgatgccgc tggcgattca ggttcatcat gccgtttgtg atggcttcca tgtcggcaga
     6781 atgcttaatg aattacaaca gtactgcgat gagtggcagg gcggggcgta aacgcgtgga
     6841 tccggcttac taaaagccag ataacagtat gcgtatttgc gcgctgattt ttgcggtata
     6901 agaatatata ctgatatgta tacccgaagt atgtcaaaaa gaggtatgct atgaagcagc
     6961 gtattacagt gacagttgac agcgacagct atcagttgct caaggcatat atgatgtcaa
     7021 tatctccggt ctggtaagca caaccatgca gaatgaagcc cgtcgtctgc gtgccgaacg
     7081 ctggaaagcg gaaaatcagg aagggatggc tgaggtcgcc cggtttattg aaatgaacgg
     7141 ctcttttgct gacgagaaca ggggctggtg aaatgcagtt taaggtttac acctataaaa
     7201 gagagagccg ttatcgtctg tttgtggatg tacagagtga tattattgac acgcccgggc
     7261 gacggatggt gatccccctg gccagtgcac gtctgctgtc agataaagtc tcccgtgaac
     7321 tttacccggt ggtgcatatc ggggatgaaa gctggcgcat gatgaccacc gatatggcca
     7381 gtgtgccggt ctccgttatc ggggaagaag tggctgatct cagccaccgc gaaaatgaca
     7441 tcaaaaacgc cattaacctg atgttctggg gaatataaat gtcaggctcc cttatacaca
     7501 gccagtctgc aggtcgacca tagtgactgg atatgttgtg ttttacagta ttatgtagtc
     7561 tgttttttat gcaaaatcta atttaatata ttgatattta tatcatttta cgtttctcgt
     7621 tcagctttct tgtacaaagt ggtgatggcg cgccgcggcc gcgatcaatt cggtaccgaa
     7681 gttcctattc cgaagttcct attctctaga aagtatagga acttcctcga gggtacccca
     7741 attccgcccc cccccccccc ctaacgttac tggccgaagc cgcttggaat aaggccggtg
     7801 tgcgtttgtc tatatgttat tttccaccat attgccgtct tttggcaatg tgagggcccg
     7861 gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc tcgccaaagg
     7921 aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct ctggaagctt cttgaagaca
     7981 aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg acaggtgcct
     8041 ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac cccagtgcca
     8101 cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg tattcaacaa
     8161 ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg ggcctcggtg
     8221 cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg
     8281 ggacgtggtt ttcctttgaa aaacacgatg ataatatggc cacaaccatg gtgagcaagg
     8341 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
     8401 gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc
     8461 tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc
     8521 tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct
     8581 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg
     8641 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
     8701 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
     8761 actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga
     8821 acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc
     8881 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc
     8941 agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
     9001 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaatcc gggatccgga
     9061 gagctcccaa cgaagttcct attccgaagt tcctattctc tagaaagtat aggaacttcc
     9121 tcgaggttgg atgcagcccg ggggatccac tagttctaga gcggccgatc agcctcgact
     9181 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     9241 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     9301 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     9361 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
     9421 accagctggg gctcgagatc cactagttct agcctcgagg ctagagcggc cgccgcgccg
     9481 atatcgaatt cgctagaaga tgggcgggag tcttctgggc aggcttaaag gctaacctgg
     9541 tgtgtgggcg ttgtcctgca ggggaattga acaggtgtaa aattggaggg acaagacttc
     9601 ccacagattt tcggttttgt cgggaagttt tttaataggg gcaaataagg aaaatgggag
     9661 gataggtagt catctggggt tttatgcagc aaaactacag gttattattg cttgtgatcc
     9721 gcctcggagt attttccatc gaggtagatt aaagacatgc tcacccgagt tttatactct
     9781 cctgcttgag atccttacta cagtatgaaa ttacagtgtc gcgagttaga ctatgtaagc
     9841 agaattttaa tcatttttaa agagcccagt acttcatatc catttctccc gctccttctg
     9901 cagccttatc aaaaggtatt ttagaacact cattttagcc ccattttcat ttattatact
     9961 ggcttatcca acccctagac agagcattgg cattttccct ttcctgatct tagaagtctg
    10021 atgactcatg aaaccagaca gattagttac atacaccaca aatcgaggct gtagctgggg
    10081 cctcaacact gcagttcttt tataactcct tagtacactt tttgttgatc ctttgccttg
    10141 atccttaatt ttcagtgtct atcacctctc ccgtcaggtg gtgttccaca tttgggccta
    10201 ttctcagtcc agggagtttt acaacaatag atgtattgag aatccaacct aaagcttaac
    10261 tttccactcc catgaatgcc tctctccttt ttctccattt ataaactgag ctattaacca
    10321 ttaatggttt ccaggtggat gtctcctccc ccaatattac ctgatgtatc ttacatattg
    10381 ccaggctgat attttaagac attaaaaggt atatttcatt attgagccac atggtattga
    10441 ttactgctta ctaaaatttt gtcattgtac acatctgtaa aaggtggttc cttttggaat
    10501 gcaaagttca ggtgtttgtt gtctttcctg acctaaggtc ttgtgagctt gtattttttc
    10561 tatttaagca gtgctttctc ttggactggc ttgactcatg gcattctaca cgttattgct
    10621 ggtctaaatg tgattttgcc aagcttcttc aggacctata attttgcttg acttgtagcc
    10681 aaacacaagt aaaatgatta agcaacaaat gtatttgtga agcttggttt ttaggttgtt
    10741 gtgttgtgtg tgcttgtgct ctataataat actatccagg ggctggagag gtggctcgga
    10801 gttcaagagc acagactgct cttccagaag tcctgagttc aattcccagc aaccacatgg
    10861 tggctcacaa ccatctgtaa tgggatctga tgccctcttc tggtgtgtct gaagaccaca
    10921 agtgtattca cattaaataa ataaatcctc cttcttcttc tttttttttt ttttaaagag
    10981 aatactgtct ccagtagaat ttactgaagt aatgaaatac tttgtgtttg ttccaatatg
    11041 gtagccaata atcaaattac tctttaagca ctggaaatgt taccaaggaa ctaattttta
    11101 tttgaagtgt aactgtggac agaggagcca taactgcaga cttgtgggat acagaagacc
    11161 aatgcagact ttaatgtctt ttctcttaca ctaagcaata aagaaataaa aattgaactt
    11221 ctagtatcct atttgtttaa actgctagct ttacttaact tttgtgcttc atctatacaa
    11281 agctgaaagc taagtctgca gccattacta aacatgaaag caagtaatga taattttgga
    11341 tttcaaaaat gtagggccag agtttagcca gccagtggtg gtgcttgcct ttatgccttt
    11401 aatcccagca ctctggaggc agagacaggc agatctctga gtttgagccc agcctggtct
    11461 acacatcaag ttctatctag gatagccagg aatacacaca gaaaccctgt tggggagggg
    11521 ggctctgaga tttcataaaa ttataattga agcattccct aatgagccac tatggatgtg
    11581 gctaaatccg tctacctttc tgatgagatt tgggtattat tttttctgtc tctgctgttg
    11641 gttgggtctt ttgacactgt gggctttctt taaagcctcc ttcctgccat gtggtctctt
    11701 gtttgctact aacttcccat ggcttaaatg gcatggcttt ttgccttcta agggcagctg
    11761 ctgagatttg cagcctgatt tccagggtgg ggttgggaaa tctttcaaac actaaaattg
    11821 tcctttaatt ttttttttaa aaaatgggtt atataataaa cctcataaaa tagttatgag
    11881 gagtgaggtg gactaatatt aaatgagtcc ctcccctata aaagagctat taaggctttt
    11941 tgtcttatac ttaacttttt ttttaaatgt ggtatcttta gaaccaaggg tcttagagtt
    12001 ttagtataca gaaactgttg catcgcttaa tcagattttc tagtttcaaa tccagagaat
    12061 ccaaattctt cacagccaaa gtcaaattaa gaatttctga cttttaatgt taatttgctt
    12121 actgtgaata taaaaatgat agcttttcct gaggcagggt ctcactatgt atctctgcct
    12181 gatctgcaac aagatatgta gactaaagtt ctgcctgctt ttgtctcctg aatactaagg
    12241 ttaaaatgta gtaatacttt tggaacttgc aggtcagatt cttttatagg ggacacacta
    12301 agggagcttg ggtgatagtt ggtaaaatgt gtttcaagtg atgaaaactt gaattattat
    12361 caccgcaacc tactttttaa aaaaaaaagc caggcctgtt agagcatgct taagggatcc
    12421 ctaggacttg ctgagcacac aagagtagtt acttggcagg ctcctggtga gagcatattt
    12481 caaaaaacaa ggcagacaac caagaaacta cagttaaggt tacctgtctt taaaccatct
    12541 gcatatacac agggatatta aaatattcca aataatattt cattcaagtt ttcccccatc
    12601 aaattgggac atggatttct ccggtgaata ggcagagttg gaaactaaac aaatgttggt
    12661 tttgtgattt gtgaaattgt tttcaagtga tagttaaagc ccatgagata cagaacaaag
    12721 ctgctatttc gaggtctctt ggtttatact cagaagcact tctttgggtt tccctgcact
    12781 atcctgatca tgtgctaggc ctaccttagg ctgattgttg ttcaaataaa cttaagtttc
    12841 ctgtcaggtg atgtcatatg atttcatata tcaaggcaaa acatgttata tatgttaaac
    12901 atttgtactt aatgtgaaag ttaggtcttt gtgggtttga tttttaattt tcaaaacctg
    12961 agctaaataa gtcattttta catgtcttac atttggtgga attgtataat tgtggtttgc
    13021 aggcaagact ctctgaccta gtaaccctac ctatagagca ctttgctggg tcacaagtct
    13081 aggagtcaag catttcacct tgaagttgag acgttttgtt agtgtatact agtttatatg
    13141 ttggaggaca tgtttatcca gaagatattc aggactattt ttgactgggc taaggaattg
    13201 attctgatta gcactgttag tgagcattga gtggccttta ggcttgaatt ggagtcactt
    13261 gtatatctca aataatgctg gcctttttta aaaagccctt gttctttatc accctgtttt
    13321 ctacataatt tttgttcaaa gaaatacttg tttggatctc cttttgacaa caatagcatg
    13381 ttttcaagcc atattttttt tccttttttt tttttttttt ggtttttcga gacagggttt
    13441 ctctgtatag ccctggctgt cctggaactc actttgtaga ccaggctggc ctcgaactca
    13501 gaaatccgcc tgcctctgcc tcctgagtgc cgggattaaa ggcgtgcacc accacgcctg
    13561 gctaagttgg atattttgtt atataactat aaccaatact aactccactg ggtggatttt
    13621 taattcagtc agtagtctta agtggtcttt attggccctt cattaaaatc tactgttcac
    13681 tctaacagag gctgttggta ctagtggcac ttaagcaact tcctacggat atactagcag
    13741 attaagggtc agggatagaa actagtctag cgttttgtat acctaccagc tttatactac
    13801 cttgttctga tagaaatatt tcaggacatc tagcttatcg ataccgtcga cggtatcgat
    13861 aagcttgata tcgaattcta ccgggtaggg gaggcgcttt tcccaaggca gtctggagca
    13921 tgcgctttag cagccccgct gggcacttgg cgctacacaa gtggcctctg gcctcgcaca
    13981 cattccacat ccaccggtag gcgccgccgc cctgcagggc gatcgcggcc gcggtaccca
    14041 gcttttgttc cctttagtga gggttaattg cgcgcttggc gtaatcatgg tcatagctgt
    14101 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
    14161 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
    14221 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
    14281 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
    14341 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
    14401 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
    14461 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
    14521 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
    14581 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
    14641 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
    14701 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
    14761 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
    14821 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
    14881 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
    14941 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
    15001 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
    15061 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
    15121 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
    15181 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
    15241 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
    15301 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
    15361 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
    15421 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
    15481 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
    15541 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
    15601 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
    15661 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
    15721 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
    15781 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
    15841 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
    15901 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
    15961 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
    16021 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
    16081 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
    16141 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
    16201 aaataggggt tccgcgcaca tttccccgaa aagtgccacc taaattgtaa gcgttaatat
    16261 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
    16321 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
    16381 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
    16441 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
    16501 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
    16561 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
    16621 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
    16681 gccgctacag ggcacgttcc attcgccatt caggctgcgc aactgttggg aagggcgatc
    16741 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt
    16801 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgagcg
    16861 cgcgtaatac gactcactat agggcgaatt ggagct
//
