LOCUS       sequence-357226-       15177 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Gene targeting vector for the mouse Rosa26 locus, including a CAG 
            promoter, loxP flanked stop cassette and GFP, for cloning into AscI.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15177)
  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 15177)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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/GFP Asc
FEATURES             Location/Qualifiers
     source          1..15177
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    6..1088
                     /label=Rosa26 left arm
                     /note="left homology arm from intron 1 of the mouse Rosa26 
                     gene (Soriano, 1999)"
     misc_feature    join(1095..1160,1161..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    complement(3388..3421)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     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    complement(5893..5926)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    5959..6006
                     /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    6045..6597
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(6189..6206)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     6416..6435
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(6609..6611,6612..6614,6615..7328)
                     /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             join(6609..6611,6612..6614,6615..7325)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(6654..6675)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6915..6934)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7262..7283
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    7353..7400
                     /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(7425..7441)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    7460..7684
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     misc_feature    7774..12114
                     /label=Rosa26 right arm
                     /note="right homology arm from intron 1 of the mouse Rosa26
                     gene (Soriano, 1999)"
     primer_bind     complement(12199..12222)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     complement(12332..12352)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(12332..12350)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(12371..12387)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12371..12387)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12384..12406)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12395..12411
                     /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(12419..12425,12426..12443,12444..12449))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12464..12485
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(12602..12619)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12773..13361)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12853..12872)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(13532..14323,14324..14392))
                     /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     14155..14174
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(14393..14497)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      14524..14979
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(14611..14630)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     14821..14842
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     15105..15127
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     15119..15136
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     15120..15136
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     15146..15165
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        15146..15164
                     /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 gggcggggga
      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 ggggsgcgcc
     2161 aggcggggcg gggcggggcg aggggcgggg cggggcgagg cggagaggtg 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 gcgccgcggc
     5941 cgcgatcaat tcggtaccga agttcctatt ccgaagttcc tattctctag aaagtatagg
     6001 aacttcctcg agggtacccc aattccgccc cccccccccc cctaacgtta ctggccgaag
     6061 ccgcttggaa taaggccggt gtgcgtttgt ctatatgtta ttttccacca tattgccgtc
     6121 ttttggcaat gtgagggccc ggaaacctgg ccctgtcttc ttgacgagca ttcctagggg
     6181 tctttcccct ctcgccaaag gaatgcaagg tctgttgaat gtcgtgaagg aagcagttcc
     6241 tctggaagct tcttgaagac aaacaacgtc tgtagcgacc ctttgcaggc agcggaaccc
     6301 cccacctggc gacaggtgcc tctgcggcca aaagccacgt gtataagata cacctgcaaa
     6361 ggcggcacaa ccccagtgcc acgttgtgag ttggatagtt gtggaaagag tcaaatggct
     6421 ctcctcaagc gtattcaaca aggggctgaa ggatgcccag aaggtacccc attgtatggg
     6481 atctgatctg gggcctcggt gcacatgctt tacatgtgtt tagtcgaggt taaaaaaacg
     6541 tctaggcccc ccgaaccacg gggacgtggt tttcctttga aaaacacgat gataatatgg
     6601 ccacaaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
     6661 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
     6721 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
     6781 ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc
     6841 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
     6901 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
     6961 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
     7021 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc
     7081 agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc
     7141 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
     7201 acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc
     7261 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
     7321 acaagtaatc cgggatccgg agagctccca acgaagttcc tattccgaag ttcctattct
     7381 ctagaaagta taggaacttc ctcgaggttg gatgcagccc gggggatcca ctagttctag
     7441 agcggccgat cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc
     7501 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag
     7561 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag
     7621 gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct
     7681 atggcttctg aggcggaaag aaccagctgg ggctcgagat ccactagttc tagcctcgag
     7741 gctagagcgg ccgccgcgcc gatatcgaat tcgctagaag atgggcggga gtcttctggg
     7801 caggcttaaa ggctaacctg gtgtgtgggc gttgtcctgc aggggaattg aacaggtgta
     7861 aaattggagg gacaagactt cccacagatt ttcggttttg tcgggaagtt ttttaatagg
     7921 ggcaaataag gaaaatggga ggataggtag tcatctgggg ttttatgcag caaaactaca
     7981 ggttattatt gcttgtgatc cgcctcggag tattttccat cgaggtagat taaagacatg
     8041 ctcacccgag ttttatactc tcctgcttga gatccttact acagtatgaa attacagtgt
     8101 cgcgagttag actatgtaag cagaatttta atcattttta aagagcccag tacttcatat
     8161 ccatttctcc cgctccttct gcagccttat caaaaggtat tttagaacac tcattttagc
     8221 cccattttca tttattatac tggcttatcc aacccctaga cagagcattg gcattttccc
     8281 tttcctgatc ttagaagtct gatgactcat gaaaccagac agattagtta catacaccac
     8341 aaatcgaggc tgtagctggg gcctcaacac tgcagttctt ttataactcc ttagtacact
     8401 ttttgttgat cctttgcctt gatccttaat tttcagtgtc tatcacctct cccgtcaggt
     8461 ggtgttccac atttgggcct attctcagtc cagggagttt tacaacaata gatgtattga
     8521 gaatccaacc taaagcttaa ctttccactc ccatgaatgc ctctctcctt tttctccatt
     8581 tataaactga gctattaacc attaatggtt tccaggtgga tgtctcctcc cccaatatta
     8641 cctgatgtat cttacatatt gccaggctga tattttaaga cattaaaagg tatatttcat
     8701 tattgagcca catggtattg attactgctt actaaaattt tgtcattgta cacatctgta
     8761 aaaggtggtt ccttttggaa tgcaaagttc aggtgtttgt tgtctttcct gacctaaggt
     8821 cttgtgagct tgtatttttt ctatttaagc agtgctttct cttggactgg cttgactcat
     8881 ggcattctac acgttattgc tggtctaaat gtgattttgc caagcttctt caggacctat
     8941 aattttgctt gacttgtagc caaacacaag taaaatgatt aagcaacaaa tgtatttgtg
     9001 aagcttggtt tttaggttgt tgtgttgtgt gtgcttgtgc tctataataa tactatccag
     9061 gggctggaga ggtggctcgg agttcaagag cacagactgc tcttccagaa gtcctgagtt
     9121 caattcccag caaccacatg gtggctcaca accatctgta atgggatctg atgccctctt
     9181 ctggtgtgtc tgaagaccac aagtgtattc acattaaata aataaatcct ccttcttctt
     9241 cttttttttt tttttaaaga gaatactgtc tccagtagaa tttactgaag taatgaaata
     9301 ctttgtgttt gttccaatat ggtagccaat aatcaaatta ctctttaagc actggaaatg
     9361 ttaccaagga actaattttt atttgaagtg taactgtgga cagaggagcc ataactgcag
     9421 acttgtggga tacagaagac caatgcagac tttaatgtct tttctcttac actaagcaat
     9481 aaagaaataa aaattgaact tctagtatcc tatttgttta aactgctagc tttacttaac
     9541 ttttgtgctt catctataca aagctgaaag ctaagtctgc agccattact aaacatgaaa
     9601 gcaagtaatg ataattttgg atttcaaaaa tgtagggcca gagtttagcc agccagtggt
     9661 ggtgcttgcc tttatgcctt taatcccagc actctggagg cagagacagg cagatctctg
     9721 agtttgagcc cagcctggtc tacacatcaa gttctatcta ggatagccag gaatacacac
     9781 agaaaccctg ttggggaggg gggctctgag atttcataaa attataattg aagcattccc
     9841 taatgagcca ctatggatgt ggctaaatcc gtctaccttt ctgatgagat ttgggtatta
     9901 ttttttctgt ctctgctgtt ggttgggtct tttgacactg tgggctttct ttaaagcctc
     9961 cttcctgcca tgtggtctct tgtttgctac taacttccca tggcttaaat ggcatggctt
    10021 tttgccttct aagggcagct gctgagattt gcagcctgat ttccagggtg gggttgggaa
    10081 atctttcaaa cactaaaatt gtcctttaat ttttttttta aaaaatgggt tatataataa
    10141 acctcataaa atagttatga ggagtgaggt ggactaatat taaatgagtc cctcccctat
    10201 aaaagagcta ttaaggcttt ttgtcttata cttaactttt tttttaaatg tggtatcttt
    10261 agaaccaagg gtcttagagt tttagtatac agaaactgtt gcatcgctta atcagatttt
    10321 ctagtttcaa atccagagaa tccaaattct tcacagccaa agtcaaatta agaatttctg
    10381 acttttaatg ttaatttgct tactgtgaat ataaaaatga tagcttttcc tgaggcaggg
    10441 tctcactatg tatctctgcc tgatctgcaa caagatatgt agactaaagt tctgcctgct
    10501 tttgtctcct gaatactaag gttaaaatgt agtaatactt ttggaacttg caggtcagat
    10561 tcttttatag gggacacact aagggagctt gggtgatagt tggtaaaatg tgtttcaagt
    10621 gatgaaaact tgaattatta tcaccgcaac ctacttttta aaaaaaaaag ccaggcctgt
    10681 tagagcatgc ttaagggatc cctaggactt gctgagcaca caagagtagt tacttggcag
    10741 gctcctggtg agagcatatt tcaaaaaaca aggcagacaa ccaagaaact acagttaagg
    10801 ttacctgtct ttaaaccatc tgcatataca cagggatatt aaaatattcc aaataatatt
    10861 tcattcaagt tttcccccat caaattggga catggatttc tccggtgaat aggcagagtt
    10921 ggaaactaaa caaatgttgg ttttgtgatt tgtgaaattg ttttcaagtg atagttaaag
    10981 cccatgagat acagaacaaa gctgctattt cgaggtctct tggtttatac tcagaagcac
    11041 ttctttgggt ttccctgcac tatcctgatc atgtgctagg cctaccttag gctgattgtt
    11101 gttcaaataa acttaagttt cctgtcaggt gatgtcatat gatttcatat atcaaggcaa
    11161 aacatgttat atatgttaaa catttgtact taatgtgaaa gttaggtctt tgtgggtttg
    11221 atttttaatt ttcaaaacct gagctaaata agtcattttt acatgtctta catttggtgg
    11281 aattgtataa ttgtggtttg caggcaagac tctctgacct agtaacccta cctatagagc
    11341 actttgctgg gtcacaagtc taggagtcaa gcatttcacc ttgaagttga gacgttttgt
    11401 tagtgtatac tagtttatat gttggaggac atgtttatcc agaagatatt caggactatt
    11461 tttgactggg ctaaggaatt gattctgatt agcactgtta gtgagcattg agtggccttt
    11521 aggcttgaat tggagtcact tgtatatctc aaataatgct ggcctttttt aaaaagccct
    11581 tgttctttat caccctgttt tctacataat ttttgttcaa agaaatactt gtttggatct
    11641 ccttttgaca acaatagcat gttttcaagc catatttttt ttcctttttt tttttttttt
    11701 tggtttttcg agacagggtt tctctgtata gccctggctg tcctggaact cactttgtag
    11761 accaggctgg cctcgaactc agaaatccgc ctgcctctgc ctcctgagtg ccgggattaa
    11821 aggcgtgcac caccacgcct ggctaagttg gatattttgt tatataacta taaccaatac
    11881 taactccact gggtggattt ttaattcagt cagtagtctt aagtggtctt tattggccct
    11941 tcattaaaat ctactgttca ctctaacaga ggctgttggt actagtggca cttaagcaac
    12001 ttcctacgga tatactagca gattaagggt cagggataga aactagtcta gcgttttgta
    12061 tacctaccag ctttatacta ccttgttctg atagaaatat ttcaggacat ctagcttatc
    12121 gataccgtcg acggtatcga taagcttgat atcgaattct accgggtagg ggaggcgctt
    12181 ttcccaaggc agtctggagc atgcgcttta gcagccccgc tgggcacttg gcgctacaca
    12241 agtggcctct ggcctcgcac acattccaca tccaccggta ggcgccgccg ccctgcaggg
    12301 cgatcgcggc cgcggtaccc agcttttgtt ccctttagtg agggttaatt gcgcgcttgg
    12361 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
    12421 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
    12481 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
    12541 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
    12601 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
    12661 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
    12721 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
    12781 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
    12841 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
    12901 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
    12961 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
    13021 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
    13081 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
    13141 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
    13201 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
    13261 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
    13321 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
    13381 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
    13441 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
    13501 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
    13561 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
    13621 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
    13681 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
    13741 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
    13801 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
    13861 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
    13921 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
    13981 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
    14041 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
    14101 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
    14161 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
    14221 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
    14281 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
    14341 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
    14401 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
    14461 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
    14521 ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
    14581 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
    14641 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
    14701 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
    14761 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
    14821 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
    14881 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
    14941 ttaacgcgaa ttttaacaaa atattaacgc ttacaatttc cattcgccat tcaggctgcg
    15001 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
    15061 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
    15121 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tggagct
//
