LOCUS       sequence-433397-       12296 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Lentiviral expression of Flag-tagged mouse Miwi in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12296)
  AUTHORS   Arif A, Bailey S, Izumi N, Anzelon TA, Ozata DM, Andersson C, 
            Gainetdinov I, MacRae IJ, Tomari Y, Zamore PD
  TITLE     GTSF1 accelerates target RNA cleavage by PIWI-clade Argonaute 
            proteins.
  JOURNAL   Nature. 2022 Aug;608(7923):618-625. doi: 10.1038/s41586-022-05009-0.
            Epub 2022 Jun 30.
  PUBMED    35772669
REFERENCE   2  (bases 1 to 12296)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/s41586-022-05009-0"; journalName: "Nature"; date: 
            "2022-08"; volume: "608"; issue: "7923"; pages: "618-625"
COMMENT     Sequence Label: pSCALPS-Flag-Miwi-EGFP
FEATURES             Location/Qualifiers
     source          1..12296
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     228..248
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     primer_bind     422..441
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        516..620
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(621..689,690..1481)
                     /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     complement(839..858)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1652..2240
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2141..2160
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     LTR             2311..2491
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    3009..3597
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             3115..3126
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     3524..3544
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(4707..5420,5421..5423,5424..5426))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4752..4773)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     5101..5120
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5360..5381
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     misc_feature    5438..5990
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5600..5619)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     5829..5846
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     CDS             complement(8628..8648)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(8667..9212)
                     /codon_start=1
                     /product="human O6-alkylguanine-DNA-alkyltransferase 
                     variant that forms covalent bonds with benzylguanine 
                     derivatives"
                     /label=SNAP-tag(R)
                     /note="fast-reacting variant, optimized for mammalian 
                     expression"
                     /translation="MDKDCEMKRTTLDSPLGKLELSGCEQGLHRIIFLGKGTSAADAVE
                     VPAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLK
                     VVKFGEVISYSHLAALAGNPAATAAVKTALSGNPVPILIPCHRVVQGDLDVGGYEGGLA
                     VKEWLLAHEGHRLGKPGLG"
     CDS             complement(9225..9290)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             complement(9297..9314)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        complement(9332..9739)
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     misc_feature    9876..9993
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(10291..10335)
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    10519..10752
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    11249..11374
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             11421..11601
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(11619..11817)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(11644..11664)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        11819..12198
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        complement(join(12210..12296,1..243))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(12224..12296,1..63)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(join(12278..12296,1))
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
ORIGIN
        1 aaaaaaaatt agtcagccat ggggcggaga atgggcggaa ctgggcggag ttaggggcgg
       61 gatgggcgga gttaggggcg ggactatggt tgctgactaa ttgagatgca tgctttgcat
      121 acttctgcct gctggggagc ctggggactt tccacacctg gttgctgact aattgagatg
      181 catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc ctaactgaca
      241 cacattccac agcgcgtata tctggcccgt acatcgcgaa gcagcgcaaa acgcctaacc
      301 ctaagcagat tcttcatgca attgtcggtc aagccttgcc ttgttgtagc ttaaattttg
      361 ctcgcgcact actcagcgac ctccaacaca caagcaggga gcagatactg gcttaactat
      421 gcggcatcag agcagattgt actgagagtg caccataggg gatcgggaga tctcccgatc
      481 cgtcgacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
      541 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
      601 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
      661 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
      721 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
      781 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
      841 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
      901 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
      961 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     1021 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     1081 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     1141 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     1201 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     1261 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     1321 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     1381 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     1441 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     1501 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     1561 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     1621 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     1681 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1741 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     1801 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     1861 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     1921 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     1981 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     2041 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     2101 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     2161 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     2221 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     2281 gctggccttt tgctcacatg aaacgggccc tgctagagat tttccacact gactaaaagg
     2341 gtctgaggga tctctagtta ccagagtcac acaacagacg ggcacacact acttgaagca
     2401 ctcaaggcaa gctttattga ggcttaagca gtgggttccc tagttagcca gagagctccc
     2461 aggctcagat ctggtctaac cagagagacc cagtacagtc cggatgcagc tctcgggcca
     2521 tgtgatgaaa tgctaggcgg ctgtcaaacc tccactctaa tacttctctc tccgggtcat
     2581 ccatcccatg caggctcaca gggtgtaaca agcgggtgtt ctctccttca ttggcttctt
     2641 ctaccttctc ttgctcaact ggtactagct tgtagcacca tccaaaggtc agtggatatc
     2701 tgatccctgg ccctggtgtg tagttctgcc aatcagggaa gtagccttgt gtgtggtaga
     2761 tccacagatc aaggatatct tgtcttcgtt gggagtgaat tagcccttcc agtcccccct
     2821 tttcttttaa aaagtggcta agatctacag ctgccttgta agtcattggt cttaaaggta
     2881 cctgaggtgt gactggaaaa cccacctcct cctcctcttg tgcttctagc caggcacaat
     2941 cagcattggt agctgctgta ttgctacttg tgattgctcc atgtttttct aggtctcgac
     3001 gcgattaagc ggggaggcgg cccaaaggga gatccgactc gtctgagggc gaaggcgaag
     3061 acgcggaaga ggccgcagag ccggcagcag gccgcgggaa ggaaggtccg ctggattgag
     3121 ggccgaaggg acgtagcaga aggacgtccc gcgcagaatc caggtggcaa cacaggcgag
     3181 cagccatgga aaggacgtca gcttccccga caacaccacg gaattgtcag tgcccaacag
     3241 ccgagcccct gtccagcagc gggcaaggca ggcggcgatg agttccgccg tggcaatagg
     3301 gagggggaaa gcgaaagtcc cggaaaggag ctgacaggtg gtggcaatgc cccaaccagt
     3361 gggggttgcg tcagcaaaca cagtgcacac cacgccacgt tgcctgacaa cgggccacaa
     3421 ctcctcataa agagacagca accaggattt atacaaggag gagaaaatga aagccatacg
     3481 ggaagcaata gcatgataca aaggcattaa agcagcgtat ccacatagcg taaaaggagc
     3541 aacatagtta agaataccag tcaatctttc acaaattttg taatccagag gttgattagc
     3601 gatcgctcag gcgccgggtt tgcgtgtcat gcaccaggtc ctaggtcctt cggggacctc
     3661 cacatctgcg gtcacagtaa agcccagccg ttcatagaag ggcaggtttc gtggagcaga
     3721 ggtctccaga aatgcaggga ctccagctcg ttctgcagcc tccactcctg gcaggaccac
     3781 agcacttccc agtcctttgc cctgatggtc aggagagacc cccacggtag ccaggaacca
     3841 tgctggttcc ttaggcctgt gtggggccag cagtccctcc atctgctgct gtgctgccag
     3901 cctggagccg ctcagttctg ccattctagg gccgatctct gcaaagacag ctccggcttc
     3961 cacgctctct ggtgtggtcc agacagccac agctgctcca tcgtctgcga cccagacctt
     4021 cccaatgtcc aggcccactc gggtcaggaa cagttcctgc agctcagtca ctctctcgat
     4081 atgccgatct gggtccacag tgtggcgtgt ggcagggtaa tcggcaaaag cggcggccag
     4141 tgttctcaca gcccttggga catcatctct tgtagccagc ctcacggtag gtttgtattc
     4201 ggtcatggcg gcacgcgtgg ctaatagtac acggttttcc tcggcggtgg cgtctgcaaa
     4261 acggcacgag cctggcctcc cgcccctttt ataccacgtt cggccccgcc cgcaacccgg
     4321 tctcgcgcct ccgtcccggc caatcgggtc tgcgacttcg ccagagtgac agtgcatctg
     4381 gcctatagac ggaagccggc gctgatgggg acgacacatg aggcgcgcgt gcgcgctaac
     4441 gcgcggaggc aggttcccgc ctgcgggctc cgtcgcggtg gctgcggcat gggactcggg
     4501 gcggggcggc agtacgggac gcggggcacg ggggagggtc agtgcggaac gggggtgggc
     4561 agcatggcgc gcgcacgcgg gagacggaag cgcggcggct cttcggccgt tgtcaagcga
     4621 tgaggcgcgt ctccccattg cacagagggg gcaactgagg cgaacgcagg cctcaaggtc
     4681 gcgccgccag tcaaggtcga gaattcttac ttgtacagct cgtccatgcc gagagtgatc
     4741 ccggcggcgg tcacgaactc cagcaggacc atgtgatcgc gcttctcgtt ggggtctttg
     4801 ctcagggcgg actgggtgct caggtagtgg ttgtcgggca gcagcacggg gccgtcgccg
     4861 atgggggtgt tctgctggta gtggtcggcg agctgcacgc tgccgtcctc gatgttgtgg
     4921 cggatcttga agttcgcctt gatgccgttc ttctgcttgt cggccatgat atagacgttg
     4981 tggctgttgt agttgtactc cagcttgtgc cccaggatgt tgccgtcctc cttgaagtcg
     5041 atgcccttca gctcgatgcg gttcaccagg gtgtcgccct cgaacttcac ctcggcgcgg
     5101 gtcttgtagt tgccgtcgtc cttgaagaag atggtgcgct cctggacgta gccttcgggc
     5161 atggcggact tgaagaagtc gtgctgcttc atgtggtcgg ggtagcggct gaagcactgc
     5221 acgccgtagg tcagggtggt cacgagggtg ggccagggca cgggcagctt gccggtggtg
     5281 cagatgaact tcagggtcag cttgccgtag gtggcatcgc cctcgccctc gccagacacg
     5341 ctgaacttgt ggccgtttac gtcgccgtcc agctcgacca ggatgggcac caccccggtg
     5401 aacagctcct cgcccttgct caccatggtt gtggccatat tatcatcgtg tttttcaaag
     5461 gaaaaccacg tccccgtggt tcggggggcc tagacgtttt tttaacctcg actaaacaca
     5521 tgtaaagcat gtgcaccgag gccccagatc agatcccata caatggggta ccttctgggc
     5581 atccttcagc cccttgttga atacgcttga ggagagccat ttgactcttt ccacaactat
     5641 ccaactcaca acgtggcact ggggttgtgc cgcctttgca ggtgtatctt atacacgtgg
     5701 cttttggccg cagaggcacc tgtcgccagg tggggggttc cgctgcctgc aaagggtcgc
     5761 tacagacgtt gtttgtcttc aagaagcttc cagaggaact gcttccttca cgacattcaa
     5821 cagaccttgc attcctttgg cgagagggga aagaccccta ggaatgctcg tcaagaagac
     5881 agggccaggt ttccgggccc tcacattgcc aaaagacggc aatatggtgg aaaataacat
     5941 atagacaaac gcacaccggc cttattccaa gcggcttcgg ccagtaacgt gttaacccgc
     6001 ggctcgagtt agaggtagta gaggcggttg gacagggaga ggtttggctc tctgtggatg
     6061 ctctggccca cgaggaaggc cagcttgtgt gcatactggc aaggtgcagg gactcggatg
     6121 actccaggcc aattatagta cacgtggcag agcttgtatg tcagccgctg gatgtggtcg
     6181 ggcttcaggc cactgctgtc atagatgaca ttgtagtgtg ttggggacac actcccgctt
     6241 ctcactgcct ggctcacgat gaaaaagtca taccactctg gtctggtgac ttccacatcg
     6301 atgactgtcc ctggaagagg gttctgaagt cttcccccag actgagcaaa aaacctggca
     6361 ttgacacgct tcttcaccac gattacagtc agtcttgggt tgtaacctct cccgactgac
     6421 ttgaggcaat ctaggaactg tgggacctca taattgacca gggtcttcag ctgcccgtcc
     6481 cccacaccgt ctcggtacac gatgacacgg ctgggcatgt attcattgca gccactccaa
     6541 gccctcagag cagcttgcaa gcacaccttg agaccatcca ccagctcctg cccgcggtcc
     6601 tgaaagacgc agcgggagaa ccagcgggtc atcccttcat tgatgctggc gacgaatcct
     6661 gcgatggacc tccgcccagc tgtggtgtca tggtaacagt cgatgcccac gatcattgcc
     6721 agtttcaggg gcatgtccac ccgccagagc tcgcctccca tcttgcagtt catctgcagg
     6781 gcgatcttgg tggcaatggc catgactgtt tgctgcttgc ccagggtccg ggccaccaca
     6841 cactgacttg gggtggggca gtctgtacac aagtacttct tgatggcatc atatttgtcc
     6901 ttccgattac ttgacaagag acagacaact atctgagtgt ctgacgtcac cttctgctgc
     6961 aaggctctca gataagcttc tgttctgtca tccacctcga tcatgattgc ctttttcatc
     7021 tggatgccca tggctggagt cactttgaac aggttctgta tcagtgagtt ggctgcttca
     7081 taattcctgc gggtatagat cagcagccag ttatccaatg gcttcacgct gatcagcggc
     7141 gcgcctcttg tttctttgga ccagtctgca aattgtgggt tgtaatcaaa cgtctttccg
     7201 ccctggtgga tcttctcaga ttgtaagatt cttccagaga aggacagcaa gtttgagtcg
     7261 aagctcaggc cccagtctcg aagctctctc tgcacattgt catccttgtg gatgtagtcg
     7321 atgaggcggc ccacctcccg ctgccgctgc tcaggggtca gccgcgtgtg cactgccagg
     7381 tccttcatca cattgaaatc attgcgcatt ttatcagtca ggcctgtgag atagcagagt
     7441 tcagggatga gcatagctgg gccaggcagg gtgccgccgg ggcctctcct ccgcttgggt
     7501 tggctcacca gcaccggctg cttcaggtcc gtgatctcct ggttgtattg cttcctgtag
     7561 tactccagga agctgacctc cgagccatcc gccttcttga aggtgctctt tggattctgg
     7621 tcccagtcaa tgtcatccac ccggtaggtc ttgttattgt acttggtgag aacgatgagg
     7681 cctatgagct ccttcgacac ttgctcctgg aacttgtgct cctctgtctg ctggtataga
     7741 ttgaacatga agtctaggac agtctcgctg cggagcacct tgtggctgac gtctgtgcag
     7801 agcatgatgt tgttctcata ctgaaggatg gaggtggtga agccgggcca gatcaccaac
     7861 ctgtggtttg gaatatcaat cgggtcactt ggattgtaat aattccgtcc aatttgttgc
     7921 aaattcatga ttttcaagag cctcctgaag atgatgttat agaactgcag gcaggtgggc
     7981 gaggtgggcg gcagctcgtt ggtcagggtg atggtgatcc tcacgtgttc cccattccga
     8041 gtctgactga atacttctgt gaccttgtgc tgtagtctct taggtaaaaa caatattgtc
     8101 ccatcgaaag catgacacct tccaatgagg tcttcatgct ggaagagcag tgcggagcga
     8161 agccttcggg cctccatcag gggattgtag tcgatgtggt actgatacag ggcccactgt
     8221 gggcgcgagg tcagccggaa gtggttggtg ctcagcttca caatgatgcc agaggagcct
     8281 gtctttgact ctttgacatg gtcaaggttc tgtctggtgt tcacaccaag gtcatggaag
     8341 tctcggcgac gccctcctct ctctgccagt gacagctcct gaaacccagc tgagatctgc
     8401 agttcttgtg acttggatgt ggctccgact accatccccc tctgtcgtcc tcggccaacc
     8461 aagtccccct ctgtggggga ctgttgaggt ctcggtggga tgtacccagg ttgctggctc
     8521 gcagcagccc caacatgctg caccgtctcc tgacctcgtg ccctgccgcg ggcccgagct
     8581 cgggctcggc cagtcatgga tccagcatcg ccagagccgc cagagcctcc ctggaaatac
     8641 aggttttcgc cagagccgcc agagccaccc agcccaggct tgcccagtct gtggccctcg
     8701 tgggccagca gccactcttt cacggcgagc ccgccctcgt agccccccac gtccaggtcg
     8761 ccctgcacca cccggtggca ggggatcaga atgggcacgg gatttccgct cagggcggtt
     8821 ttcacggcgg cggtggcggc gggattgccg gccagggcgg ccaggtggct gtagctgatg
     8881 acctctccga acttcaccac tttcagcagt ttccacagca cctggcgggt aaagctctcc
     8941 tgctggaaca ctgggtggtg cagggctggc acagggaact cctcgatggc ctcaggctgg
     9001 tgaaagtagg cgttgagcca ggcggtggcc tgcatcagtg gctctggtcc gcccagcacg
     9061 gcggctgggg caggcacttc cacggcgtcg gcggcagatg ttcctttgcc caggaagatg
     9121 atacggtgca ggccctgttc gcacccagac agttccagct tgcccagagg gctatccagg
     9181 gtggtgcgct tcatttcgca gtctttgtcc atgccgccag agcccttgtc gtcgtcgtcc
     9241 ttgtagtcga tgtcgtggtc cttgtagtca ccgtcgtggt ccttgtagtc gccaccatga
     9301 tggtggtgat ggtgcatgat gcttctagac ccccgggcga ctcagtctgt cggaggactg
     9361 gcgcgccgag tgaggggttg tgagctcttt tatagagctc gggaagcaga agcgcgcgaa
     9421 cagaagcgag aagcaggctg attggttaat tcaaataagg cgcagggtca tttcaggtcc
     9481 ttgggggagc ctggaaacat ctgatgggtc ttaagaaact gctgagggtt gggccatatc
     9541 tggggaccat ctgttcttgg cctcgggccg gggccgaaac tgcggtgacc atctgttctt
     9601 ggccccgggc cggggccgaa actgctcacc gcagatatcc tgtttggccc aacgttagct
     9661 attttcatgt acccgccctt gatctgaact tctctattct tggtttggta tttttccatg
     9721 ccttgcaaaa tggcgttact gcagctagct tgccaaacct acaggtgggg tctttcattc
     9781 cccccttttt ctggagacta aataaaatct tttattttat cggggctgca gttaattaac
     9841 caaactggat ctctgctgtc cctgtaataa acccgaaaat tttgaatttt tgtaatttgt
     9901 ttttgtaatt ctttagtttg tatgtctgtt gctattatgt ctactattct ttcccctgca
     9961 ctgtaccccc caatcccccc ttttctttta aaattgtgga tgaatactgc catttgtctg
    10021 cagaattggc gcacgcagtg ccgatccgtt cactaatcga atggatctgt ctctgtctct
    10081 ctctccacct tcttcttcta ttccttcggg cctgtcgggt cccctcgggg ttgggaggtg
    10141 ggtctgaaac gataatggtg aatatccctg cctaactcta ttcactatag aaagtacagc
    10201 aaaaactatt cttaaaccta ccaagcctcc tactatcatt atgaataatt ttatatacca
    10261 cagccaattt gttatgttaa accaattcca caaacttgcc catttatcta attccaataa
    10321 ttcttgttca ttcttttctt gctggttttg cgattcttca attaaggagt gtattaagct
    10381 tgtgtaattg ttaattctct gtcccactcc atccaggtcg tgtgattcca aatctgttcc
    10441 agagatttat tactccaact agcattccaa ggcacagcag tggtgcaaat gagttttcca
    10501 gagcaacccc aaatccccag gagctgttga tcctttaggt atctttccac agccaggatt
    10561 cttgcctgga gctgcttgat gccccagact gtgagttgca acagatgctg ttgcgcctca
    10621 atagccctca gcaaattgtt ctgctgctgc actataccag acaataattg tctggcctgt
    10681 accgtcagcg tcattgacgc tgcgcccata gtgcttcctg ctgctcccaa gaacccaagg
    10741 aacaaagctc ctattcccac tgctcttttt tctctctgca ccactcttct ctttgccttg
    10801 gtgggtgcta ctcctaatgg ttcaattttt actactttat atttatataa ttcacttctc
    10861 caattgtccc tcatatctcc tcctccaggt ctgaagatca gcggccggcc gcttgctgtg
    10921 cggtggtctt acttttgttt tgctcttcct ctatcttgtc taaagcttcc ttggtgtctt
    10981 ttatctctat cctttgatgc acacaataga gggttgctac tgtattatat aatgatctaa
    11041 gttcttctga tcctgtctga agggatggtt gtagctgtcc cagtatttgt ctacagcctt
    11101 ctgatgtttc taacaggcca ggattaactg cgaatcgttc tagctccctg cttgcccata
    11161 ctatatgttt taatttatat tttttctttc cccctggcct taaccgaatt ttttcccatc
    11221 gcgatctaat tctcccccgc ttaatactga cgctctcgca cccatctctc tccttctagc
    11281 ctccgctagt caaaattttt ggcgtactca ccagtcgccg cccctcgcct cttgccgtgc
    11341 gcgcttcagc aagccgagtc ctgcgtcgag agagctcctc tggtttccct ttcgctttca
    11401 agtccctgtt cgggcgccac tgctagagat tttccacact gactaaaagg gtctgaggga
    11461 tctctagtta ccagagtcac acaacagacg ggcacacact acttgaagca ctcaaggcaa
    11521 gctttattga ggcttaagca gtgggttccc tagttagcca gagagctccc aggctcagat
    11581 ctggtctaac cagagagacc cagtacaggc aaaacgcgct gcttatatag acctcccacc
    11641 gtacacgcct accgcccatt tgcgtcaatg gggcggagtt gttacgacat tttggaaagt
    11701 cccgttgatt ttggtgccaa aacaaactcc cattgacgtc aatggggtgg agacttggaa
    11761 atccccgtga gtcaaaccgc tatccacgcc cattgatgta ctgccaaaac cgcatcacca
    11821 tggtaatagc gatgactaat acgtagatgt actgccaagt aggaaagtcc cataaggtca
    11881 tgtactgggc ataatgccag gcgggccatt taccgtcatt gacgtcaata gggggcgtac
    11941 ttggcatatg atacacttga tgtactgcca agtgggcagt ttaccgtaaa tactccaccc
    12001 attgacgtca atggaaagtc cctattggcg ttactatggg aacatacgtc attattgacg
    12061 tcaatgggcg ggggtcgttg ggcggtcagc caggcgggcc atttaccgta agttatgtaa
    12121 cgcggaactc catatatggg ctatgaacta atgaccccgt aattgattac tattaataac
    12181 tagtcaataa tcaatgtcaa cgcgagcttt ttgcaaaagc ctaggcctcc aaaaaagcct
    12241 cctcactact tctggaatag ctcagaggca gaggcggcct cggcctctgc ataaat
//
