pR26 iBFP Dest

LOCUS        pR26 iBFP Dest               14523 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Gene targeting vector for the mouse Rosa26 locus, including a loxP 
            flanked stop cassette and BFP, for gateway cloning.
ACCESSION   .
VERSION     .
KEYWORDS    pR26 iBFP Dest
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14523)
  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 14523)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..14523
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    3..1085
                     /label=Rosa26 left arm
                     /note="left homology arm from intron 1 of the mouse Rosa26 
                     gene (Soriano, 1999)"
     misc_feature    1092..1195
                     /label=Ad SA
                     /note="adenovirus major late transcript splice acceptor 
                     (Friedrich and Soriano, 1991)"
     protein_bind    1206..1239
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     polyA_signal    1246..1294
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     CDS             complement(1301..2104)
                     /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(1412..1431)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2022..2041
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     intron          2918..2983
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             3113..3133
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    3558..3692
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3595..3614)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3649..3668
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    3712..3745
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     protein_bind    3758..3882
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        3907..3937
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             3991..4650
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(4057..4076)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             4992..5297
                     /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     5176..5195
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(5338..5462)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(5474..5507)
                     /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)"
     misc_feature    5544..6096
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5688..5705)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5915..5934
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             6117..6836
                     /codon_start=1
                     /product="enhanced blue variant of GFP (Ai et al., 2007)"
                     /label=EBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLSHGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGVDFKEDGNILGHKLEYNFNSHNIYIMAVKQKNGIK
                     VNFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDSHYLSTQSVLSKDPNEKRDHMVLL
                     EFRTAAGITLGMDELYK"
     primer_bind     complement(6162..6183)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     protein_bind    complement(6844..6877)
                     /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)"
     polyA_signal    6886..7093
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     misc_feature    7132..11471
                     /label=Rosa26 right arm
                     /note="right homology arm from intron 1 of the mouse Rosa26
                     gene (Soriano, 1999)"
     primer_bind     complement(11676..11696)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(11676..11694)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(11715..11731)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11715..11731)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11728..11750)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11739..11755
                     /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(11763..11793)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11808..11829
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11946..11963)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12117..12705)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12197..12216)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(12876..13736)
                     /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     13499..13518
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13737..13841)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(13867..14322)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(14004..14025)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     14216..14235
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     14449..14471
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     14463..14480
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     14464..14480
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     14490..14509
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        14490..14508
                     /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 aggagcgctt ccggccgacg tctcgtcgct gattggcttc ttttcctccc
      241 gccgtgtgtg aaaacacaaa tggcgtgttt tggttggcgt aaggcgcctg tcagttaacg
      301 gcagccggag tgcgcagccg ccggcagcct cgctctgccc actgggtggg gcgggaggta
      361 ggtggggtga ggcgagctgg 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 cccttccccc tcttccctcg tgatctgcaa ctccagtctt
     1081 tctagcatct gtagggcgca gtagtccagg gtttccttga tgatgtcata cttatcctgt
     1141 cccttttttt tccacagctc gcggttgagg acaaactctt cgcggtcttt ccagtggtta
     1201 attaaataac ttcgtatagc atacattata cgaagttatg gatccgaaca aacgacccaa
     1261 cacccgtgcg ttttattctg tctttttatt gccgatcccc tcagaagaac tcgtcaagaa
     1321 ggcgatagaa ggcgatgcgc tgcgaatcgg gagcggcgat accgtaaagc acgaggaagc
     1381 ggtcagccca ttcgccgcca agctcttcag caatatcacg ggtagccaac gctatgtcct
     1441 gatagcggtc cgccacaccc agccggccac agtcgatgaa tccagaaaag cggccatttt
     1501 ccaccatgat attcggcaag caggcatcgc catgggtcac gacgagatcc tcgccgtcgg
     1561 gcatgcgcgc cttgagcctg gcgaacagtt cggctggcgc gagcccctga tgctcttcgt
     1621 ccagatcatc ctgatcgaca agaccggctt ccatccgagt acgtgctcgc tcgatgcgat
     1681 gtttcgcttg gtggtcgaat gggcaggtag ccggatcaag cgtatgcagc cgccgcattg
     1741 catcagccat gatggatact ttctcggcag gagcaaggtg agatgacagg agatcctgcc
     1801 ccggcacttc gcccaatagc agccagtccc ttcccgcttc agtgacaacg tcgagcacag
     1861 ctgcgcaagg aacgcccgtc gtggccagcc acgatagccg cgctgcctcg tcctgcagtt
     1921 cattcagggc accggacagg tcggtcttga caaaaagaac cgggcgcccc tgcgctgaca
     1981 gccggaacac ggcggcatca gagcagccga ttgtctgttg tgcccagtca tagccgaata
     2041 gcctctccac ccaagcggcc ggagaacctg cgtgcaatcc atcttgttca atggccgatc
     2101 ccatattggc tgcagggtcg ctcggtgttc gaggccacac gcgtcacctt aatatgcgaa
     2161 gtggacctgg gaccgcgccg ccccgactgc atctgcgtgt tcgaattcgc caatgacaag
     2221 acgctgggcg gggtttgctc gacattgggt ggaaacattc caggcctggg tggagaggct
     2281 ttttgcttcc tcttgcaaaa ccacactgct cgacattggg tggaaacatt ccaggcctgg
     2341 gtggagaggc tttttgcttc ctcttgcaaa accacactgc tcgactagtg attaaagtcg
     2401 actcggggac accaaatatg gcgatctcgg ccttttcgtt tcttggagct gggacatgtt
     2461 tgccatcgat ccatctacca ccagaacggc cgttagatct gctgccaccg ttgtttccac
     2521 cgaagaaacc accgttgccg taaccaccac gacggttgtt gctaaagaag ctgccaccgc
     2581 cacggccacc gttgtagccg ccgttgttgt tattgtagtt gctcatgtta tttctggcac
     2641 ttcttggttt tcctcttaag tgaggaggaa cataaccatt ctcgttgttg tcgttgatgc
     2701 ttaaattttg cacttgttcg ctcagttcag ccataatatg aaatgctttt cttgttgttc
     2761 ttacggaata ccacttgcca cctatcacca caactaactt tttcccgttc ctccatctct
     2821 tttatatttt ttttctcgag ggatctttgt gaaggaacct tacttctgtg gtgtgacata
     2881 attggacaaa ctacctacag agatttaaag ctctaaggta aatataaaat ttttaagtgt
     2941 ataatgtgtt aaactactga ttctaattgt ttgtgtattt tagattccaa cctatggaac
     3001 tgatgaatgg gagcagtggt ggaatgcctt taatgaggaa aacctgtttt gctcagaaga
     3061 aatgccatct agtgatgatg aggctactgc tgactctcaa cattctactc ctccaaaaaa
     3121 gaagagaaag gtagaagacc ccaaggactt tccttcagaa ttgctaagtt ttttgagtca
     3181 tgctgtgttt agtaatagaa ctcttgcttg ctttgctatt tacaccacaa aggaaaaagc
     3241 tgcactgcta tacaagaaaa ttatggaaaa atattctgta acctttataa gtaggcataa
     3301 cagttataat cataacatac tgttttttct tactccacac aggcatagag tgtctgctat
     3361 taataactat gctcaaaaat tgtgtacctt tagcttttta atttgtaaag gggttaataa
     3421 ggaatatttg atgtatagtg ccttgactag agatcataat cagccatacc acatttgtag
     3481 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa catagaatga
     3541 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     3601 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     3661 aactcatcaa tgtatcttat catgtctgga tctgacatgg taagtaagct tataacttcg
     3721 tatagcatac attatacgaa gttatcggcg cgccatcaca agtttgtaca aaaaagctga
     3781 acgagaaacg taaaatgata taaatatcaa tatattaaat tagattttgc ataaaaaaca
     3841 gactacataa tactgtaaaa cacaacatat ccagtcacta tggcggccgc attaggcacc
     3901 ccaggcttta cactttatgc ttccggctcg tataatgtgt ggattttgag ttaggatccg
     3961 tcgagatttt caggagctaa ggaagctaaa atggagaaaa aaatcactgg atataccacc
     4021 gttgatatat cccaatggca tcgtaaagaa cattttgagg catttcagtc agttgctcaa
     4081 tgtacctata accagaccgt tcagctggat attacggcct ttttaaagac cgtaaagaaa
     4141 aataagcaca agttttatcc ggcctttatt cacattcttg cccgcctgat gaatgctcat
     4201 ccggaattcc gtatggcaat gaaagacggt gagctggtga tatgggatag tgttcaccct
     4261 tgttacaccg ttttccatga gcaaactgaa acgttttcat cgctctggag tgaataccac
     4321 gacgatttcc ggcagtttct acacatatat tcgcaagatg tggcgtgtta cggtgaaaac
     4381 ctggcctatt tccctaaagg gtttattgag aatatgtttt tcgtctcagc caatccctgg
     4441 gtgagtttca ccagttttga tttaaacgtg gccaatatgg acaacttctt cgcccccgtt
     4501 ttcaccatgg gcaaatatta tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag
     4561 gttcatcatg ccgtttgtga tggcttccat gtcggcagaa tgcttaatga attacaacag
     4621 tactgcgatg agtggcaggg cggggcgtaa acgcgtggat ccggcttact aaaagccaga
     4681 taacagtatg cgtatttgcg cgctgatttt tgcggtataa gaatatatac tgatatgtat
     4741 acccgaagta tgtcaaaaag aggtatgcta tgaagcagcg tattacagtg acagttgaca
     4801 gcgacagcta tcagttgctc aaggcatata tgatgtcaat atctccggtc tggtaagcac
     4861 aaccatgcag aatgaagccc gtcgtctgcg tgccgaacgc tggaaagcgg aaaatcagga
     4921 agggatggct gaggtcgccc ggtttattga aatgaacggc tcttttgctg acgagaacag
     4981 gggctggtga aatgcagttt aaggtttaca cctataaaag agagagccgt tatcgtctgt
     5041 ttgtggatgt acagagtgat attattgaca cgcccgggcg acggatggtg atccccctgg
     5101 ccagtgcacg tctgctgtca gataaagtct cccgtgaact ttacccggtg gtgcatatcg
     5161 gggatgaaag ctggcgcatg atgaccaccg atatggccag tgtgccggtc tccgttatcg
     5221 gggaagaagt ggctgatctc agccaccgcg aaaatgacat caaaaacgcc attaacctga
     5281 tgttctgggg aatataaatg tcaggctccc ttatacacag ccagtctgca ggtcgaccat
     5341 agtgactgga tatgttgtgt tttacagcat tatgtagtct gttttttatg caaaatctaa
     5401 tttaatatat tgatatttat atcattttac gtttctcgtt cagctttctt gtacaaagtg
     5461 gtgatggcgc gccgaagttc ctatactttc tagagaatag gaacttctct ctcgaggtta
     5521 acgaattccg cccccccccc ctaacgttac tggccgaagc cgcttggaat aaggccggtg
     5581 tgcgtttgtc tatatgttat tttccaccat attgccgtct tttggcaatg tgagggcccg
     5641 gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc tcgccaaagg
     5701 aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct ctggaagctt cttgaagaca
     5761 aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg acaggtgcct
     5821 ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac cccagtgcca
     5881 cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg tattcaacaa
     5941 ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg ggcctcggtg
     6001 cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg
     6061 ggacgtggtt ttcctttgaa aaacacgatg ataatatggc cacaacccgg gacaccatgg
     6121 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
     6181 acgtaaacgg ccacaagttc agcgtgaggg gcgagggcga gggcgatgcc accaacggca
     6241 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
     6301 tgaccaccct gagccacggc gtgcagtgct tcgcccgcta ccccgaccac atgaagcagc
     6361 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
     6421 aggacgacgg cacctacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
     6481 accgcatcga gctgaagggc gtcgacttca aggaggacgg caacatcctg gggcacaagc
     6541 tggagtacaa cttcaacagc cacaacatct atatcatggc cgtcaagcag aagaacggca
     6601 tcaaggtgaa cttcaagatc cgccacaacg tggaggacgg cagcgtgcag ctcgccgacc
     6661 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac agccactacc
     6721 tgagcaccca gtccgtgctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     6781 tggagttccg caccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaacccc
     6841 ggggaagttc ctatactttc tagagaatag gaacttcgct agagactgtg ccttctagtt
     6901 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     6961 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     7021 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     7081 ggcatgctgg ggatgcggtc tagcgcggcc gccgcgccga tatcgaattc gctagaagat
     7141 gggcgggagt cttctgggca ggcttaaagg ctaacctggt gtgtgggcgt tgtcctgcag
     7201 gggaattgaa caggtgtaaa attggaggga caagacttcc cacagatttt cggttttgtc
     7261 gggaagtttt ttaatagggg caaataagga aaatgggagg ataggtagtc atctggggtt
     7321 ttatgcagca aaactacagg ttattattgc ttgtgatccg cctcggagta ttttccatcg
     7381 aggtagatta aagacatgct cacccgagtt ttatactctc ctgcttgaga tccttactac
     7441 agtatgaaat tacagtgtcg cgagttagac tatgtaagca gaattttaat catttttaaa
     7501 gagcccagta cttcatatcc atttctcccg ctccttctgc agccttatca aaaggtattt
     7561 tagaacactc attttagccc cattttcatt tattatactg gcttatccaa cccctagaca
     7621 gagcattggc attttccctt tcctgatctt agaagtctga tgactcatga aaccagacag
     7681 attagttaca tacaccacaa atcgaggctg tagctggggc ctcaacactg cagttctttt
     7741 ataactcctt agtacacttt ttgttgatcc tttgccttga tccttaattt tcagtgtcta
     7801 tcacctctcc cgtcaggtgg tgttccacat ttgggcctat tctcagtcca gggagtttta
     7861 caacaataga tgtattgaga atccaaccta aagcttaact ttccactccc atgaatgcct
     7921 ctctcctttt tctccattta taaactgagc tattaaccat taatggtttc caggtggatg
     7981 tctcctcccc caatattacc tgatgtatct tacatattgc caggctgata ttttaagaca
     8041 ttaaaaggta tatttcatta ttgagccaca tggtattgat tactgcttac taaaattttg
     8101 tcattgtaca catctgtaaa aggtggttcc ttttggaatg caaagttcag gtgtttgttg
     8161 tctttcctga cctaaggtct tgtgagcttg tattttttct atttaagcag tgctttctct
     8221 tggactggct tgactcatgg cattctacac gttattgctg gtctaaatgt gattttgcca
     8281 agcttcttca ggacctataa ttttgcttga cttgtagcca aacacaagta aaatgattaa
     8341 gcaacaaatg tatttgtgaa gcttggtttt taggttgttg tgttgtgtgt gcttgtgctc
     8401 tataataata ctatccaggg gctggagagg tggctcggag ttcaagagca cagactgctc
     8461 ttccagaagt cctgagttca attcccagca accacatggt ggctcacaac catctgtaat
     8521 gggatctgat gccctcttct ggtgtgtctg aagaccacaa gtgtattcac attaaataaa
     8581 taaatcctcc ttcttcttct tttttttttt tttaaagaga atactgtctc cagtagaatt
     8641 tactgaagta atgaaatact ttgtgtttgt tccaatatgg tagccaataa tcaaattact
     8701 ctttaagcac tggaaatgtt accaaggaac taatttttat ttgaagtgta actgtggaca
     8761 gaggagccat aactgcagac ttgtgggata cagaagacca atgcagactt taatgtcttt
     8821 tctcttacac taagcaataa agaaataaaa attgaacttc tagtatccta tttgtttaaa
     8881 ctgctagctt tacttaactt ttgtgcttca tctatacaaa gctgaaagct aagtctgcag
     8941 ccattactaa acatgaaagc aagtaatgat aattttggat ttcaaaaatg tagggccaga
     9001 gtttagccag ccagtggtgg tgcttgcctt tatgccttta atcccagcac tctggaggca
     9061 gagacaggca gatctctgag tttgagccca gcctggtcta cacatcaagt tctatctagg
     9121 atagccagga atacacacag aaaccctgtt ggggaggggg gctctgagat ttcataaaat
     9181 tataattgaa gcattcccta atgagccact atggatgtgg ctaaatccgt ctacctttct
     9241 gatgagattt gggtattatt ttttctgtct ctgctgttgg ttgggtcttt tgacactgtg
     9301 ggctttcttt aaagcctcct tcctgccatg tggtctcttg tttgctacta acttcccatg
     9361 gcttaaatgg catggctttt tgccttctaa gggcagctgc tgagatttgc agcctgattt
     9421 ccagggtggg gttgggaaat ctttcaaaca ctaaaattgt cctttaattt tttttttaaa
     9481 aaatgggtta tataataaac ctcataaaat agttatgagg agtgaggtgg actaatatta
     9541 aatgagtccc tcccctataa aagagctatt aaggcttttt gtcttatact taactttttt
     9601 tttaaatgtg gtatctttag aaccaagggt cttagagttt tagtatacag aaactgttgc
     9661 atcgcttaat cagattttct agtttcaaat ccagagaatc caaattcttc acagccaaag
     9721 tcaaattaag aatttctgac ttttaatgtt aatttgctta ctgtgaatat aaaaatgata
     9781 gcttttcctg aggcagggtc tcactatgta tctctgcctg atctgcaaca agatatgtag
     9841 actaaagttc tgcctgcttt tgtctcctga atactaaggt taaaatgtag taatactttt
     9901 ggaacttgca ggtcagattc ttttataggg gacacactaa gggagcttgg gtgatagttg
     9961 gtaaaatgtg tttcaagtga tgaaaacttg aattattatc accgcaacct actttttaaa
    10021 aaaaaaagcc aggcctgtta gagcatgctt aagggatccc taggacttgc tgagcacaca
    10081 agagtagtta cttggcaggc tcctggtgag agcatatttc aaaaaacaag gcagacaacc
    10141 aagaaactac agttaaggtt acctgtcttt aaaccatctg catatacaca gggatattaa
    10201 aatattccaa ataatatttc attcaagttt tcccccatca aattgggaca tggatttctc
    10261 cggtgaatag gcagagttgg aaactaaaca aatgttggtt ttgtgatttg tgaaattgtt
    10321 ttcaagtgat agttaaagcc catgagatac agaacaaagc tgctatttcg aggtctcttg
    10381 gtttatactc agaagcactt ctttgggttt ccctgcacta tcctgatcat gtgctaggcc
    10441 taccttaggc tgattgttgt tcaaataaac ttaagtttcc tgtcaggtga tgtcatatga
    10501 tttcatatat caaggcaaaa catgttatat atgttaaaca tttgtactta atgtgaaagt
    10561 taggtctttg tgggtttgat ttttaatttt caaaacctga gctaaataag tcatttttac
    10621 atgtcttaca tttggtggaa ttgtataatt gtggtttgca ggcaagactc tctgacctag
    10681 taaccctacc tatagagcac tttgctgggt cacaagtcta ggagtcaagc atttcacctt
    10741 gaagttgaga cgttttgtta gtgtatacta gtttatatgt tggaggacat gtttatccag
    10801 aagatattca ggactatttt tgactgggct aaggaattga ttctgattag cactgttagt
    10861 gagcattgag tggcctttag gcttgaattg gagtcacttg tatatctcaa ataatgctgg
    10921 ccttttttaa aaagcccttg ttctttatca ccctgttttc tacataattt ttgttcaaag
    10981 aaatacttgt ttggatctcc ttttgacaac aatagcatgt tttcaagcca tatttttttt
    11041 cctttttttt tttttttttg gtttttcgag acagggtttc tctgtatagc cctggctgtc
    11101 ctggaactca ctttgtagac caggctggcc tcgaactcag aaatccgcct gcctctgcct
    11161 cctgagtgcc gggattaaag gcgtgcacca ccacgcctgg ctaagttgga tattttgtta
    11221 tataactata accaatacta actccactgg gtggattttt aattcagtca gtagtcttaa
    11281 gtggtcttta ttggcccttc attaaaatct actgttcact ctaacagagg ctgttggtac
    11341 tagtggcact taagcaactt cctacggata tactagcaga ttaagggtca gggatagaaa
    11401 ctagtctagc gttttgtata cctaccagct ttatactacc ttgttctgat agaaatattt
    11461 aggacatcta gcttatcgat ccgtcgacgg tatcgataag cttgatatcg aattctaccg
    11521 ggtaggggag gcgcttttcc aaggcagtct gagcatgcgc ttagcagccc cgctggcact
    11581 tggcgctaca caagtggcct tggcctcgca cacattccac atccaccggt aggcgccgcc
    11641 gccctgcagg cgatcgctac gtacagcttt tgttcccttt agtgagggtt aattgcgcgc
    11701 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
    11761 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
    11821 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
    11881 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
    11941 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
    12001 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
    12061 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
    12121 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
    12181 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
    12241 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
    12301 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
    12361 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
    12421 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
    12481 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
    12541 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
    12601 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
    12661 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
    12721 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
    12781 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
    12841 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
    12901 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
    12961 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
    13021 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
    13081 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
    13141 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
    13201 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
    13261 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
    13321 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
    13381 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
    13441 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
    13501 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
    13561 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
    13621 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
    13681 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
    13741 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
    13801 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
    13861 ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
    13921 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
    13981 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
    14041 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
    14101 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
    14161 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
    14221 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
    14281 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg ccattcaggc
    14341 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga
    14401 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac
    14461 gttgtaaaac gacggccagt gagcgcgcgt aatacgactc actatagggc gaattggagc
    14521 tcc
//