LOCUS       sequence-142357-       16705 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gene targeting vector for the mouse Rosa26 locus, including a CAG 
            promoter, loxP flanked stop cassette, BFP, for gateway cloning.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 16705)
  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 16705)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "BMC 
            Biotechnol."; date: "2016-01-16"; volume: "16(1):4. doi"; pages: " 
            10.1186/s12896-016-0234-4"
COMMENT     Sequence Label: pR26 CAG/BFP Dest
FEATURES             Location/Qualifiers
     source          1..16705
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    3..1085
                     /label=Rosa26 left arm
                     /note="left homology arm from intron 1 of the mouse Rosa26 
                     gene (Soriano, 1999)"
     misc_feature    join(1092..1157,1158..1195)
                     /label=Ad SA
                     /note="adenovirus major late transcript splice acceptor 
                     (Friedrich and Soriano, 1991)"
     polyA_signal    1214..1421
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     polyA_signal    1435..1642
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     enhancer        1651..2030
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2032..2305
                     /label=chicken beta-actin promoter
     intron          2306..3322
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     3246..3268
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     3330..3349
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    complement(3385..3418)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    3425..3473
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     CDS             complement(3480..4283)
                     /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(3591..3610)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4201..4220
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     intron          5097..5162
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             5292..5312
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    5737..5871
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5774..5793)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5828..5847
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(5891..5924)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    5937..6061
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(6086..6091,6092..6109,6110..6116)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             6170..6829
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(6236..6255)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             7171..7476
                     /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     7355..7374
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(7517..7641)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(7653..7686)
                     /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    7706..8273
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    7723..8275
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(7867..7884)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     8094..8113
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(8291..8293,8294..8296,8297..9010)
                     /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(8336..8357)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     protein_bind    complement(9018..9051)
                     /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    9060..9267
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     misc_feature    9306..13646
                     /label=Rosa26 right arm
                     /note="right homology arm from intron 1 of the mouse Rosa26
                     gene (Soriano, 1999)"
     primer_bind     complement(13731..13754)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     complement(13858..13878)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(13858..13876)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(13897..13913)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(13897..13913)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(13910..13932)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    13921..13937
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(13945..13951,13952..13969,13970..13975))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    13990..14011
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(14128..14145)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(14299..14887)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(14379..14398)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(15058..15849,15850..15918))
                     /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     15681..15700
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(15919..16023)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(16049..16504)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(16186..16207)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     16398..16417
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     16631..16653
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     16645..16662
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     16646..16662
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     16672..16691
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        16672..16690
                     /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 attaactcga ggactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     1261 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     1321 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     1381 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtcta gcgactgtgc
     1441 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
     1501 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
     1561 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
     1621 acaatagcag gcatgctggg gatgcgggtc gacattgatt attgactagt tattaatagt
     1681 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
     1741 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
     1801 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
     1861 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
     1921 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
     1981 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtcgaggtga
     2041 gccccacgtt ctgcttcact ctccccatct cccccccctc cccaccccca attttgtatt
     2101 tatttatttt ttaattattt tgtgcagcga tgggggcggg gggggggggc gcgcgccagg
     2161 cggggcgggg cggggcgagg ggcggggcgg ggcgaggcgg agaggtgcgg cggcagccaa
     2221 tcagagcggc gcgctccgaa agtttccttt tatggcgagg cggcggcggc ggcggcccta
     2281 taaaaagcga agcgcgcggc gggcgggagt cgctgcgcgc tgccttcgcc ccgtgccccg
     2341 ctccgccgcc gcctcgcgcc gcccgccccg gctctgactg accgcgttac tcccacaggt
     2401 gagcgggcgg gacggccctt ctcctccggg ctgtaattag cgcttggttt aatgacggct
     2461 tgtttctttt ctgtggctgc gtgaaagcct tgaggggctc cgggagggcc ctttgtgcgg
     2521 ggggagcggc tcggggggtg cgtgcgtgtg tgtgtgcgtg gggagcgccg cgtgcggctc
     2581 cgcgctgccc ggcggctgtg agcgctgcgg gcgcggcgcg gggctttgtg cgctccgcag
     2641 tgtgcgcgag gggagcgcgg ccgggggcgg tgccccgcgg tgcggggggg gctgcgaggg
     2701 gaacaaaggc tgcgtgcggg gtgtgtgcgt gggggggtga gcagggggtg tgggcgcgtc
     2761 ggtcgggctg caaccccccc tgcacccccc tccccgagtt gctgagcacg gcccggcttc
     2821 gggtgcgggg ctccgtacgg ggcgtggcgc ggggctcgcc gtgccgggcg gggggtggcg
     2881 gcaggtgggg gtgccgggcg gggcggggcc gcctcgggcc ggggagggct cgggggaggg
     2941 gcgcggcggc ccccggagcg ccggcggctg tcgaggcgcg gcgagccgca gccattgcct
     3001 tttatggtaa tcgtgcgaga gggcgcaggg acttcctttg tcccaaatct gtgcggagcc
     3061 gaaatctggg aggcgccgcc gcaccccctc tagcgggcgc ggggcgaagc ggtgcggcgc
     3121 cggcaggaag gaaatgggcg gggagggcct tcgtgcgtcg ccgcgccgcc gtccccttct
     3181 ccctctccag cctcggggct gtccgcgggg ggacggctgc cttcgggggg gacggggcag
     3241 ggcggggttc ggcttctggc gtgtgaccgg cggctctaga gcctctgcta accatgttca
     3301 tgccttcttc tttttcctac agctcctggg caacgtgctg gttattgtgc tgtctcatca
     3361 ttttggcaaa gaattcttaa ttaaataact tcgtatagca tacattatac gaagttatgg
     3421 atccgaacaa acgacccaac acccgtgcgt tttattctgt ctttttattg ccgatcccct
     3481 cagaagaact cgtcaagaag gcgatagaag gcgatgcgct gcgaatcggg agcggcgata
     3541 ccgtaaagca cgaggaagcg gtcagcccat tcgccgccaa gctcttcagc aatatcacgg
     3601 gtagccaacg ctatgtcctg atagcggtcc gccacaccca gccggccaca gtcgatgaat
     3661 ccagaaaagc ggccattttc caccatgata ttcggcaagc aggcatcgcc atgggtcacg
     3721 acgagatcct cgccgtcggg catgcgcgcc ttgagcctgg cgaacagttc ggctggcgcg
     3781 agcccctgat gctcttcgtc cagatcatcc tgatcgacaa gaccggcttc catccgagta
     3841 cgtgctcgct cgatgcgatg tttcgcttgg tggtcgaatg ggcaggtagc cggatcaagc
     3901 gtatgcagcc gccgcattgc atcagccatg atggatactt tctcggcagg agcaaggtga
     3961 gatgacagga gatcctgccc cggcacttcg cccaatagca gccagtccct tcccgcttca
     4021 gtgacaacgt cgagcacagc tgcgcaagga acgcccgtcg tggccagcca cgatagccgc
     4081 gctgcctcgt cctgcagttc attcagggca ccggacaggt cggtcttgac aaaaagaacc
     4141 gggcgcccct gcgctgacag ccggaacacg gcggcatcag agcagccgat tgtctgttgt
     4201 gcccagtcat agccgaatag cctctccacc caagcggccg gagaacctgc gtgcaatcca
     4261 tcttgttcaa tggccgatcc catattggct gcagggtcgc tcggtgttcg aggccacacg
     4321 cgtcacctta atatgcgaag tggacctggg accgcgccgc cccgactgca tctgcgtgtt
     4381 cgaattcgcc aatgacaaga cgctgggcgg ggtttgctcg acattgggtg gaaacattcc
     4441 aggcctgggt ggagaggctt tttgcttcct cttgcaaaac cacactgctc gacattgggt
     4501 ggaaacattc caggcctggg tggagaggct ttttgcttcc tcttgcaaaa ccacactgct
     4561 cgactagtga ttaaagtcga ctcggggaca ccaaatatgg cgatctcggc cttttcgttt
     4621 cttggagctg ggacatgttt gccatcgatc catctaccac cagaacggcc gttagatctg
     4681 ctgccaccgt tgtttccacc gaagaaacca ccgttgccgt aaccaccacg acggttgttg
     4741 ctaaagaagc tgccaccgcc acggccaccg ttgtagccgc cgttgttgtt attgtagttg
     4801 ctcatgttat ttctggcact tcttggtttt cctcttaagt gaggaggaac ataaccattc
     4861 tcgttgttgt cgttgatgct taaattttgc acttgttcgc tcagttcagc cataatatga
     4921 aatgcttttc ttgttgttct tacggaatac cacttgccac ctatcaccac aactaacttt
     4981 ttcccgttcc tccatctctt ttatattttt tttctcgagg gatctttgtg aaggaacctt
     5041 acttctgtgg tgtgacataa ttggacaaac tacctacaga gatttaaagc tctaaggtaa
     5101 atataaaatt tttaagtgta taatgtgtta aactactgat tctaattgtt tgtgtatttt
     5161 agattccaac ctatggaact gatgaatggg agcagtggtg gaatgccttt aatgaggaaa
     5221 acctgttttg ctcagaagaa atgccatcta gtgatgatga ggctactgct gactctcaac
     5281 attctactcc tccaaaaaag aagagaaagg tagaagaccc caaggacttt ccttcagaat
     5341 tgctaagttt tttgagtcat gctgtgttta gtaatagaac tcttgcttgc tttgctattt
     5401 acaccacaaa ggaaaaagct gcactgctat acaagaaaat tatggaaaaa tattctgtaa
     5461 cctttataag taggcataac agttataatc ataacatact gttttttctt actccacaca
     5521 ggcatagagt gtctgctatt aataactatg ctcaaaaatt gtgtaccttt agctttttaa
     5581 tttgtaaagg ggttaataag gaatatttga tgtatagtgc cttgactaga gatcataatc
     5641 agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg
     5701 aacctgaaac atagaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat
     5761 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     5821 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggat ctgacatggt
     5881 aagtaagctt ataacttcgt atagcataca ttatacgaag ttatcggcgc gccatcacaa
     5941 gtttgtacaa aaaagctgaa cgagaaacgt aaaatgatat aaatatcaat atattaaatt
     6001 agattttgca taaaaaacag actacataat actgtaaaac acaacatatc cagtcatatt
     6061 ggcggccgca ttaggcaccc caggctttac actttatgct tccggctcgt ataatgtgtg
     6121 gattttgagt taggatccgt cgagattttc aggagctaag gaagctaaaa tggagaaaaa
     6181 aatcactgga tataccaccg ttgatatatc ccaatggcat cgtaaagaac attttgaggc
     6241 atttcagtca gttgctcaat gtacctataa ccagaccgtt cagctggata ttacggcctt
     6301 tttaaagacc gtaaagaaaa ataagcacaa gttttatccg gcctttattc acattcttgc
     6361 ccgcctgatg aatgctcatc cggaattccg tatggcaatg aaagacggtg agctggtgat
     6421 atgggatagt gttcaccctt gttacaccgt tttccatgag caaactgaaa cgttttcatc
     6481 gctctggagt gaataccacg acgatttccg gcagtttcta cacatatatt cgcaagatgt
     6541 ggcgtgttac ggtgaaaacc tggcctattt ccctaaaggg tttattgaga atatgttttt
     6601 cgtctcagcc aatccctggg tgagtttcac cagttttgat ttaaacgtgg ccaatatgga
     6661 caacttcttc gcccccgttt tcaccatggg caaatattat acgcaaggcg acaaggtgct
     6721 gatgccgctg gcgattcagg ttcatcatgc cgtttgtgat ggcttccatg tcggcagaat
     6781 gcttaatgaa ttacaacagt actgcgatga gtggcagggc ggggcgtaaa cgcgtggatc
     6841 cggcttacta aaagccagat aacagtatgc gtatttgcgc gctgattttt gcggtataag
     6901 aatatatact gatatgtata cccgaagtat gtcaaaaaga ggtatgctat gaagcagcgt
     6961 attacagtga cagttgacag cgacagctat cagttgctca aggcatatat gatgtcaata
     7021 tctccggtct ggtaagcaca accatgcaga atgaagcccg tcgtctgcgt gccgaacgct
     7081 ggaaagcgga aaatcaggaa gggatggctg aggtcgcccg gtttattgaa atgaacggct
     7141 cttttgctga cgagaacagg ggctggtgaa atgcagttta aggtttacac ctataaaaga
     7201 gagagccgtt atcgtctgtt tgtggatgta cagagtgata ttattgacac gcccgggcga
     7261 cggatggtga tccccctggc cagtgcacgt ctgctgtcag ataaagtctc ccgtgaactt
     7321 tacccggtgg tgcatatcgg ggatgaaagc tggcgcatga tgaccaccga tatggccagt
     7381 gtgccggtct ccgttatcgg ggaagaagtg gctgatctca gccaccgcga aaatgacatc
     7441 aaaaacgcca ttaacctgat gttctgggga atataaatgt caggctccct tatacacagc
     7501 cagtctgcag gtcgaccata gtgactggat atgttgtgtt ttacagcatt atgtagtctg
     7561 ttttttatgc aaaatctaat ttaatatatt gatatttata tcattttacg tttctcgttc
     7621 agctttcttg tacaaagtgg tgatggcgcg ccgaagttcc tatactttct agagaatagg
     7681 aacttctctc tcgaggttaa cgaattccgc cccccccccc taacgttact ggccgaagcc
     7741 gcttggaata aggccggtgt gcgtttgtct atatgttatt ttccaccata ttgccgtctt
     7801 ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt gacgagcatt cctaggggtc
     7861 tttcccctct cgccaaagga atgcaaggtc tgttgaatgt cgtgaaggaa gcagttcctc
     7921 tggaagcttc ttgaagacaa acaacgtctg tagcgaccct ttgcaggcag cggaaccccc
     7981 cacctggcga caggtgcctc tgcggccaaa agccacgtgt ataagataca cctgcaaagg
     8041 cggcacaacc ccagtgccac gttgtgagtt ggatagttgt ggaaagagtc aaatggctct
     8101 cctcaagcgt attcaacaag gggctgaagg atgcccagaa ggtaccccat tgtatgggat
     8161 ctgatctggg gcctcggtgc acatgcttta catgtgttta gtcgaggtta aaaaaacgtc
     8221 taggcccccc gaaccacggg gacgtggttt tcctttgaaa aacacgatga taatatggcc
     8281 acaacccggg atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
     8341 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg aggggcgagg gcgagggcga
     8401 tgccaccaac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
     8461 ctggcccacc ctcgtgacca ccctgagcca cggcgtgcag tgcttcgccc gctaccccga
     8521 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
     8581 caccatcttc ttcaaggacg acggcaccta caagacccgc gccgaggtga agttcgaggg
     8641 cgacaccctg gtgaaccgca tcgagctgaa gggcgtcgac ttcaaggagg acggcaacat
     8701 cctggggcac aagctggagt acaacttcaa cagccacaac atctatatca tggccgtcaa
     8761 gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacgtggagg acggcagcgt
     8821 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     8881 cgacagccac tacctgagca cccagtccgt gctgagcaaa gaccccaacg agaagcgcga
     8941 tcacatggtc ctgctggagt tccgcaccgc cgccgggatc actctcggca tggacgagct
     9001 gtacaagtaa ccccggggaa gttcctatac tttctagaga ataggaactt cgctagagac
     9061 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     9121 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     9181 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     9241 ggaagacaat agcaggcatg ctggggatgc ggtctagcgc ggccgccgcg ccgatatcga
     9301 attcgctaga agatgggcgg gagtcttctg ggcaggctta aaggctaacc tggtgtgtgg
     9361 gcgttgtcct gcaggggaat tgaacaggtg taaaattgga gggacaagac ttcccacaga
     9421 ttttcggttt tgtcgggaag ttttttaata ggggcaaata aggaaaatgg gaggataggt
     9481 agtcatctgg ggttttatgc agcaaaacta caggttatta ttgcttgtga tccgcctcgg
     9541 agtattttcc atcgaggtag attaaagaca tgctcacccg agttttatac tctcctgctt
     9601 gagatcctta ctacagtatg aaattacagt gtcgcgagtt agactatgta agcagaattt
     9661 taatcatttt taaagagccc agtacttcat atccatttct cccgctcctt ctgcagcctt
     9721 atcaaaaggt attttagaac actcatttta gccccatttt catttattat actggcttat
     9781 ccaaccccta gacagagcat tggcattttc cctttcctga tcttagaagt ctgatgactc
     9841 atgaaaccag acagattagt tacatacacc acaaatcgag gctgtagctg gggcctcaac
     9901 actgcagttc ttttataact ccttagtaca ctttttgttg atcctttgcc ttgatcctta
     9961 attttcagtg tctatcacct ctcccgtcag gtggtgttcc acatttgggc ctattctcag
    10021 tccagggagt tttacaacaa tagatgtatt gagaatccaa cctaaagctt aactttccac
    10081 tcccatgaat gcctctctcc tttttctcca tttataaact gagctattaa ccattaatgg
    10141 tttccaggtg gatgtctcct cccccaatat tacctgatgt atcttacata ttgccaggct
    10201 gatattttaa gacattaaaa ggtatatttc attattgagc cacatggtat tgattactgc
    10261 ttactaaaat tttgtcattg tacacatctg taaaaggtgg ttccttttgg aatgcaaagt
    10321 tcaggtgttt gttgtctttc ctgacctaag gtcttgtgag cttgtatttt ttctatttaa
    10381 gcagtgcttt ctcttggact ggcttgactc atggcattct acacgttatt gctggtctaa
    10441 atgtgatttt gccaagcttc ttcaggacct ataattttgc ttgacttgta gccaaacaca
    10501 agtaaaatga ttaagcaaca aatgtatttg tgaagcttgg tttttaggtt gttgtgttgt
    10561 gtgtgcttgt gctctataat aatactatcc aggggctgga gaggtggctc ggagttcaag
    10621 agcacagact gctcttccag aagtcctgag ttcaattccc agcaaccaca tggtggctca
    10681 caaccatctg taatgggatc tgatgccctc ttctggtgtg tctgaagacc acaagtgtat
    10741 tcacattaaa taaataaatc ctccttcttc ttcttttttt tttttttaaa gagaatactg
    10801 tctccagtag aatttactga agtaatgaaa tactttgtgt ttgttccaat atggtagcca
    10861 ataatcaaat tactctttaa gcactggaaa tgttaccaag gaactaattt ttatttgaag
    10921 tgtaactgtg gacagaggag ccataactgc agacttgtgg gatacagaag accaatgcag
    10981 actttaatgt cttttctctt acactaagca ataaagaaat aaaaattgaa cttctagtat
    11041 cctatttgtt taaactgcta gctttactta acttttgtgc ttcatctata caaagctgaa
    11101 agctaagtct gcagccatta ctaaacatga aagcaagtaa tgataatttt ggatttcaaa
    11161 aatgtagggc cagagtttag ccagccagtg gtggtgcttg cctttatgcc tttaatccca
    11221 gcactctgga ggcagagaca ggcagatctc tgagtttgag cccagcctgg tctacacatc
    11281 aagttctatc taggatagcc aggaatacac acagaaaccc tgttggggag gggggctctg
    11341 agatttcata aaattataat tgaagcattc cctaatgagc cactatggat gtggctaaat
    11401 ccgtctacct ttctgatgag atttgggtat tattttttct gtctctgctg ttggttgggt
    11461 cttttgacac tgtgggcttt ctttaaagcc tccttcctgc catgtggtct cttgtttgct
    11521 actaacttcc catggcttaa atggcatggc tttttgcctt ctaagggcag ctgctgagat
    11581 ttgcagcctg atttccaggg tggggttggg aaatctttca aacactaaaa ttgtccttta
    11641 attttttttt taaaaaatgg gttatataat aaacctcata aaatagttat gaggagtgag
    11701 gtggactaat attaaatgag tccctcccct ataaaagagc tattaaggct ttttgtctta
    11761 tacttaactt tttttttaaa tgtggtatct ttagaaccaa gggtcttaga gttttagtat
    11821 acagaaactg ttgcatcgct taatcagatt ttctagtttc aaatccagag aatccaaatt
    11881 cttcacagcc aaagtcaaat taagaatttc tgacttttaa tgttaatttg cttactgtga
    11941 atataaaaat gatagctttt cctgaggcag ggtctcacta tgtatctctg cctgatctgc
    12001 aacaagatat gtagactaaa gttctgcctg cttttgtctc ctgaatacta aggttaaaat
    12061 gtagtaatac ttttggaact tgcaggtcag attcttttat aggggacaca ctaagggagc
    12121 ttgggtgata gttggtaaaa tgtgtttcaa gtgatgaaaa cttgaattat tatcaccgca
    12181 acctactttt taaaaaaaaa agccaggcct gttagagcat gcttaaggga tccctaggac
    12241 ttgctgagca cacaagagta gttacttggc aggctcctgg tgagagcata tttcaaaaaa
    12301 caaggcagac aaccaagaaa ctacagttaa ggttacctgt ctttaaacca tctgcatata
    12361 cacagggata ttaaaatatt ccaaataata tttcattcaa gttttccccc atcaaattgg
    12421 gacatggatt tctccggtga ataggcagag ttggaaacta aacaaatgtt ggttttgtga
    12481 tttgtgaaat tgttttcaag tgatagttaa agcccatgag atacagaaca aagctgctat
    12541 ttcgaggtct cttggtttat actcagaagc acttctttgg gtttccctgc actatcctga
    12601 tcatgtgcta ggcctacctt aggctgattg ttgttcaaat aaacttaagt ttcctgtcag
    12661 gtgatgtcat atgatttcat atatcaaggc aaaacatgtt atatatgtta aacatttgta
    12721 cttaatgtga aagttaggtc tttgtgggtt tgatttttaa ttttcaaaac ctgagctaaa
    12781 taagtcattt ttacatgtct tacatttggt ggaattgtat aattgtggtt tgcaggcaag
    12841 actctctgac ctagtaaccc tacctataga gcactttgct gggtcacaag tctaggagtc
    12901 aagcatttca ccttgaagtt gagacgtttt gttagtgtat actagtttat atgttggagg
    12961 acatgtttat ccagaagata ttcaggacta tttttgactg ggctaaggaa ttgattctga
    13021 ttagcactgt tagtgagcat tgagtggcct ttaggcttga attggagtca cttgtatatc
    13081 tcaaataatg ctggcctttt ttaaaaagcc cttgttcttt atcaccctgt tttctacata
    13141 atttttgttc aaagaaatac ttgtttggat ctccttttga caacaatagc atgttttcaa
    13201 gccatatttt ttttcctttt tttttttttt tttggttttt cgagacaggg tttctctgta
    13261 tagccctggc tgtcctggaa ctcactttgt agaccaggct ggcctcgaac tcagaaatcc
    13321 gcctgcctct gcctcctgag tgccgggatt aaaggcgtgc accaccacgc ctggctaagt
    13381 tggatatttt gttatataac tataaccaat actaactcca ctgggtggat ttttaattca
    13441 gtcagtagtc ttaagtggtc tttattggcc cttcattaaa atctactgtt cactctaaca
    13501 gaggctgttg gtactagtgg cacttaagca acttcctacg gatatactag cagattaagg
    13561 gtcagggata gaaactagtc tagcgttttg tatacctacc agctttatac taccttgttc
    13621 tgatagaaat atttcaggac atctagctta tcgataccgt cgacggtatc gataagcttg
    13681 atatcgaatt ctaccgggta ggggaggcgc ttttcccaag gcagtctgga gcatgcgctt
    13741 tagcagcccc gctgggcact tggcgctaca caagtggcct ctggcctcgc acacattcca
    13801 catccaccgg taggcgccgc cgccctgcag ggcgatcgct acgtacagct tttgttccct
    13861 ttagtgaggg ttaattgcgc gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
    13921 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
    13981 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
    14041 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
    14101 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
    14161 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
    14221 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
    14281 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
    14341 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
    14401 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
    14461 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
    14521 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
    14581 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
    14641 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
    14701 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
    14761 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
    14821 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
    14881 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
    14941 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
    15001 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
    15061 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
    15121 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
    15181 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
    15241 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
    15301 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
    15361 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
    15421 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
    15481 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
    15541 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
    15601 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
    15661 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
    15721 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
    15781 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
    15841 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
    15901 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
    15961 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
    16021 gcgcacattt ccccgaaaag tgccacctaa attgtaagcg ttaatatttt gttaaaattc
    16081 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
    16141 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag
    16201 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
    16261 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
    16321 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
    16381 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
    16441 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
    16501 gcgtcccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
    16561 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg
    16621 ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgagcgcgc gtaatacgac
    16681 tcactatagg gcgaattgga gctcc
//
