LOCUS       sequence-309018-       16215 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Ubiquitous expression of LbCas12a in Drosophila.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 16215)
  AUTHORS   Port F, Starostecka M, Boutros M
  TITLE     Multiplexed conditional genome editing with Cas12a in Drosophila.
  JOURNAL   Proc Natl Acad Sci U S A. 2020 Aug 25. pii: 2004655117. doi: 
            10.1073/pnas.2004655117.
  PUBMED    32843348
REFERENCE   2  (bases 1 to 16215)
  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: "Proc Natl 
            Acad Sci U S A. 2020 Aug 25. pii: 2004655117. doi: 
            10.1073/pnas.2004655117."
COMMENT     Sequence Label: act5c-LbCas12a
FEATURES             Location/Qualifiers
     source          1..16215
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(30..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(140..161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     352..371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     599..616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     623..642
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        623..641
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(675..693)
                     /label=pCasper-F
                     /note="Drosophila mini-white gene, reverse primer"
     CDS             join(1145..1216,1618..1891,1966..2620,2682..2997,3201..3332,
                     3403..4017)
                     /codon_start=1
                     /gene="white"
                     /product="Drosophila white gene eye color pigment"
                     /label=mini-white
                     /note="This is a modified version of the white gene lacking
                     part of the first intron."
                     /translation="MGQEDQELLIRGGSKHPSAEHLNNGDSGAASQSCINQGFGQAKNY
                     GTLRPPSPPEDSGSGSGQLAENLTYAWHNMDIFGAVNQPGSGWRQLVNRTRGLFCNERH
                     IPAPRKHLLKNVCGVAYPGELLAVMGSSGAGKTTLLNALAFRSPQGIQVSPSGMRLLNG
                     QPVDAKEMQARCAYVQQDDLFIGSLTAREHLIFQAMVRMPRHLTYRQRVARVDQVIQEL
                     SLSKCQHTIIGVPGRVKGLSGGERKRLAFASEALTDPPLLICDEPTSGLDSFTAHSVVQ
                     VLKKLSQKGKTVILTIHQPSSELFELFDKILLMAEGRVAFLGTPSEAVDFFSYVGAQCP
                     TNYNPADFYVQVLAVVPGREIESRDRIAKICDNFAISKVARDMEQLLATKNLEKPLEQP
                     ENGYTYKATWFMQFRAVLWRSWLSVLKEPLLVKVRLIQTTMVAILIGLIFLGQQLTQVG
                     VMNINGAIFLFLTNMTFQNVFATINVFTSELPVFMREARSRLYRCDTYFLGKTIAELPL
                     FLTVPLVFTAIAYPMIGLRAGVLHFFNCLALVTLVANVSTSFGYLISCASSSTSMALSV
                     GPPVIIPFLLFGGFFLNSGSVPVYLKWLSYLSWFRYANEGLLINQWADVEPGEISCTSS
                     NTTCPSSGKVILETLNFSAADLPLDYVGLAILIVSFRVLAYLALRLRARRKE"
     protein_bind    4792..4825
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        4896..7406
                     /label=Ac5 promoter
                     /note="Drosophila melanogaster actin 5C promoter"
     CDS             9172..12855
                     /codon_start=1
                     /product="CRISPR-associated protein from Lachnospiraceae 
                     bacterium ND2006 (Zetsche et al., 2015)"
                     /label=LbCpf1
                     /translation="MSKLEKFTNCYSLSKTLRFKAIPVGKTQENIDNKRLLVEDEKRAE
                     DYKGVKKLLDRYYLSFINDVLHSIKLKNLNNYISLFRKKTRTEKENKELENLEINLRKE
                     IAKAFKGNEGYKSLFKKDIIETILPEFLDDKDEIALVNSFNGFTTAFTGFFDNRENMFS
                     EEAKSTSIAFRCINENLTRYISNMDIFEKVDAIFDKHEVQEIKEKILNSDYDVEDFFEG
                     EFFNFVLTQEGIDVYNAIIGGFVTESGEKIKGLNEYINLYNQKTKQKLPKFKPLYKQVL
                     SDRESLSFYGEGYTSDEEVLEVFRNTLNKNSEIFSSIKKLEKLFKNFDEYSSAGIFVKN
                     GPAISTISKDIFGEWNVIRDKWNAEYDDIHLKKKAVVTEKYEDDRRKSFKKIGSFSLEQ
                     LQEYADADLSVVEKLKEIIIQKVDEIYKVYGSSEKLFDADFVLEKSLKKNDAVVAIMKD
                     LLDSVKSFENYIKAFFGEGKETNRDESFYGDFVLAYDILLKVDHIYDAIRNYVTQKPYS
                     KDKFKLYFQNPQFMGGWDKDKETDYRATILRYGSKYYLAIMDKKYAKCLQKIDKDDVNG
                     NYEKINYKLLPGPNKMLPKVFFSKKWMAYYNPSEDIQKIYKNGTFKKGDMFNLNDCHKL
                     IDFFKDSISRYPKWSNAYDFNFSETEKYKDIAGFYREVEEQGYKVSFESASKKEVDKLV
                     EEGKLYMFQIYNKDFSDKSHGTPNLHTMYFKLLFDENNHGQIRLSGGAELFMRRASLKK
                     EELVVHPANSPIANKNPDNPKKTTTLSYDVYKDKRFSEDQYELHIPIAINKCPKNIFKI
                     NTEVRVLLKHDDNPYVIGIDRGERNLLYIVVVDGKGNIVEQYSLNEIINNFNGIRIKTD
                     YHSLLDKKEKERFEARQNWTSIENIKELKAGYISQVVHKICELVEKYDAVIALEDLNSG
                     FKNSRVKVEKQVYQKFEKMLIDKLNYMVDKKSNPCATGGALKGYQITNKFESFKSMSTQ
                     NGFIFYIPAWLTSKIDPSTGFVNLLKTKYTSIADSKKFISSFDRIMYVPEEDLFEFALD
                     YKNFSRTDADYIKKWKLYSYGNRIRIFRNPKKNNVFDWEEVCLTSAYKELFNKYGINYQ
                     QGDIRALLCEQSDKAFYSSFMALMSLMLQMRNSITGRTDVDFLISPVKNSDGIFYDSRN
                     YEAQENAILPKNADANGAYNIARKVLWAIGQFKKAEDEKLDKVKIAISNKEWLEYAQTS
                     VKH"
     CDS             12856..12903
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             12910..12990
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     intron          13089..13154
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             13284..13304
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    13576..13710
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(13613..13632)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     13667..13686
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    13739..13808
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     promoter        complement(14027..14045)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(14066..14082)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(14066..14082)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(14079..14101)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    14090..14106
                     /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(14114..14120,14121..14138,14139..14144))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    14159..14180
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(14297..14314)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(14468..15056)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(14548..14567)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(15227..16018,16019..16087))
                     /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     15850..15869
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(16088..16192)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgaat tgtaatacga ctcactatag ggcgaattgg gtacgtaccg
      661 ggcccctagt atgtatgtaa gttaataaaa cccatttttg cggaaagtag ataaaaaaaa
      721 catttttttt ttttactgca ctggatatca ttgaacttat ctgatcagtt ttaaatttac
      781 ttcgatccaa gggtatttga tgtaccaggt tctttcgatt acctctcact caaaatgaca
      841 ttccactcaa agtcagcgct gtttgcctcc ttctctgtcc acagaaatat cgccgtctct
      901 ttcgccgctg cgtccgctat ctctttcgcc accgtttgta gcgttacgta gcgtcaatgt
      961 ccgccttcag ttgcattttg tcagcggttt cgtgacgaag ctccaagcgg tttacgccat
     1021 caattaaaca caaagtgctg tgccaaaact cctctcgctt cttatttttg tttgtttttt
     1081 gagtgattgg ggtggtgatt ggttttgggt gggtaagcag gggaaagtgt gaaaaatccc
     1141 ggcaatgggc caagaggatc aggagctatt aattcgcgga ggcagcaaac acccatctgc
     1201 cgagcatctg aacaatgtga gtagtacatg tgcatacatc ttaagttcac ttgatctata
     1261 ggaactgcga ttgcaacatc aaattgtctg cggcgtgaga actgcgaccc acaaaaatcc
     1321 caaaccgcaa ttgcacaaac aaatagtgac acgaaacaga ttattctggt agctgttctc
     1381 gctatataag acaatttttg agatcatatc atgatcaaga catctaaagg cattcatttt
     1441 cgactatatt cttttttaca aaaaatataa caaccagata ttttaagctg atcctagatg
     1501 cacaaaaaat aaataaaagt ataaacctac ttcgtaggat acttcggggt actttttgtt
     1561 cggggttaga tgagcataac gcttgtagtt gatatttgag atcccctatc attgcagggt
     1621 gacagcggag cggcttcgca gagctgcatt aaccagggct tcgggcaggc caaaaactac
     1681 ggcacgctcc ggccacccag tccgccggag gactccggtt cagggagcgg ccaactagcc
     1741 gagaacctca cctatgcctg gcacaatatg gacatctttg gggcggtcaa tcagccgggc
     1801 tccggatggc ggcagctggt caaccggaca cgcggactat tctgcaacga gcgacacata
     1861 ccggcgccca ggaaacattt gctcaagaac ggtgagtttc tattcgcagt cggctgatct
     1921 gtgtgaaatc ttaataaagg gtccaattac caatttgaaa ctcagtttgc ggcgtggcct
     1981 atccgggcga acttttggcc gtgatgggca gttccggtgc cggaaagacg accctgctga
     2041 atgcccttgc ctttcgatcg ccgcagggca tccaagtatc gccatccggg atgcgactgc
     2101 tcaatggcca acctgtggac gccaaggaga tgcaggccag gtgcgcctat gtccagcagg
     2161 atgacctctt tatcggctcc ctaacggcca gggaacacct gattttccaa gccatggtgc
     2221 ggatgccacg acatctgacc tatcggcagc gagtggcccg cgtggatcag gtgatccagg
     2281 agctttcgct cagcaaatgt cagcacacga tcatcggtgt gcccggcagg gtgaaaggtc
     2341 tgtccggcgg agaaaggaag cgtctggcat tcgcctccga ggctctaacc gatccgccgc
     2401 ttctgatctg cgatgagccc acctccggac tggactcctt taccgcccac agcgtcgtcc
     2461 aggtgctgaa gaagctgtcg cagaagggca agaccgtcat cctgaccatt catcagccgt
     2521 cttccgagct gtttgagctc tttgacaaga tccttctgat ggccgagggc agggtagctt
     2581 tcttgggcac tcccagcgaa gccgtcgact tcttttccta gtgagttcga tgtgtttatt
     2641 aagggtatct agtattacat aacatctcaa ctcctatcca gcgtgggtgc ccagtgtcct
     2701 accaactaca atccggcgga cttttacgta caggtgttgg ccgttgtgcc cggacgggag
     2761 atcgagtccc gtgatcggat cgccaagata tgcgacaatt ttgccattag caaagtagcc
     2821 cgggatatgg agcagttgtt ggccaccaaa aatctggaga agccactgga gcagccggag
     2881 aatgggtaca cctacaaggc cacctggttc atgcagttcc gggcggtcct gtggcgatcc
     2941 tggctgtcgg tgctcaagga accactcctc gtaaaagtgc gacttattca gacaacggtg
     3001 agtggttcca gtggaaacaa atgatataac gcttacaatt cttggaaaca aattcgctag
     3061 attttagata gaattgcctg attccacacc cttcttagtt tttttcaatg agatgtatag
     3121 tttatagttt tgcagaagat aaataaattt catttaactc gcgaatatta atgagatgcg
     3181 agtaacattt taatttgcag atggttgcca tcttgattgg cctcatcttt ttgggccaac
     3241 aactcacgca agtgggtgtg atgaatatca acggagccat cttcctcttc ctgaccaaca
     3301 tgacctttca aaacgtcttt gccacgataa atgtaagtca tgtttagaat acatttgcat
     3361 ttcaataatt tactaacttt ctaatgaatc gattcgattt aggtgttcac ctcagagctg
     3421 ccagttttta tgagggaggc ccgaagtcga ctttatcgct gtgacacata ctttctgggc
     3481 aaaacgattg ccgaattgcc gctttttctc acagtgccac tggtcttcac ggcgattgcc
     3541 tatccgatga tcggactgcg ggccggagtg ctgcacttct tcaactgcct ggcgctggtc
     3601 actctggtgg ccaatgtgtc aacgtccttc ggatatctaa tatcctgcgc cagctcctcg
     3661 acctcgatgg cgctgtctgt gggtccgccg gttatcatac cattcctgct ctttggcggc
     3721 ttcttcttga actcgggctc ggtgccagta tacctcaaat ggttgtcgta cctctcatgg
     3781 ttccgttacg ccaacgaggg tctgctgatt aaccaatggg cggacgtgga gccgggcgaa
     3841 attagctgca catcgtcgaa caccacgtgc cccagttcgg gcaaggtcat cctggagacg
     3901 cttaacttct ccgccgccga tctgccgctg gactacgtgg gtctggccat tctcatcgtg
     3961 agcttccggg tgctcgcata tctggctcta agacttcggg cccgacgcaa ggagtagccg
     4021 acatatatcc gaaataactg cttgtttttt ttttttacca ttattaccat cgtgtttact
     4081 gtttattgcc ccctcaaaaa gctaatgtaa ttatatttgt gccaataaaa acaagatatg
     4141 acctatagaa tacaagtatt tccccttcga acatccccac aagtagactt tggatttgtc
     4201 ttctaaccaa aagacttaca cacctgcata ccttacatca aaaactcgtt tatcgctaca
     4261 taaaacaccg ggatatattt tttatataca tacttttcaa atcgcgcgcc ctcttcataa
     4321 ttcacctcca ccacaccacg tttcgtagtt gctctttcgc tgtctcccac ccgctctccg
     4381 caacacattc accttttgtt cgacgacctt ggagcgactg tcgttagttc cgcgcgattc
     4441 ggttcgctca aatggttccg agtggttcat ttcgtctcaa tagaaattag taataaatat
     4501 ttgtatgtac aatttatttg ctccaatata tttgtatata tttccctcac agctatattt
     4561 attctaattt aatattatga ctttttaagg taattttttg tgacctgttc ggagtgatta
     4621 gcgttacaat ttgaactgaa agtgacatcc agtgtttgtt ccttgtgtag atgcatctca
     4681 aaaaaatggt gggcataata gtgttgttta tatatatcaa aaataacaac tataataata
     4741 agaatacatt taatttagaa aatgcttgga tttcactgga actaggctag cataacttcg
     4801 tataatgtat gctatacgaa gttatgctag cggatccaag cttgcggccg caattctata
     4861 ttctaaaaac acaaatgata cttctaaaaa aaaatcatga atggcatcaa ctctgaatca
     4921 aatctttgca gatgcaccta cttctcattt ccactgtcac atcatttttc cagatctcgc
     4981 tgcctgttat gtggcccaca aaccaagaca cgttttatgg ccattaaagc tggctgatcg
     5041 tcgccaaaca ccaaatacat aatgaatatg tacacattcg agaaagaagc gatcaaagaa
     5101 gcgtcttcgg gcggagtagg agaatgcgga ggagaaggag aacgagctga tctagtatct
     5161 ctccacaatc caatgccaac tgaccaactg gccatattcg gagcaatttg aagccaattt
     5221 ccatcgcctg gcgatcgctc cattcttggc tatatgtttt tcaccgttac ccggggccat
     5281 tttcaaagac tcgtcggcaa gataagattg tgtcactcgc tgtctctctt catttgtcga
     5341 agaatgctga ggaatttcgc gatgacgtcg gcgagtattt tgaagaatga gaataatttg
     5401 tatttatacg aaaatcagtt agtggaattt tctacaaaaa catgttatct atagataatt
     5461 ttgttgcaaa atatgttgac tatgacaaag attgtatgta tataccttta atgtattctc
     5521 attttcttat gtatttataa tggcaatgat gatactgatg atattttaag atgatgccag
     5581 accaaaaggc ttgaatttct gcgtcttttg ccgaacgcag tgcatgtgca attgttgttt
     5641 tttggaatat tcaattttcg gactgtccgc tttgatttca gtttcttggc ttattcaaaa
     5701 agcaaagtaa agccaaaaaa gcgagatggc aataccaaat gcggcaaaac ggtagtggaa
     5761 ggaaaggggt gcggggcagc ggaaggaagg gtggggcggg gcgtggcggg gtctgtggct
     5821 gggcgcgacg tcaccgacgt tggagccact cctttgacca tgtgtgcgtg tgtgtattat
     5881 tcgtgtctcg ccactcgccg gttgtttttt tctttttatg ctgcgctctc tctagcgcca
     5941 tctcgcttac gcatgctcaa cgcaccgcat gttgccgttt ccttttatgc gtcattttgg
     6001 ctcgaaatag gcaattattt aaacaaagat tagtcaacga aaacgctaaa ataaataagt
     6061 ctacaatatg gttacttatt gccatgtgtg tgcagccaac gatagcaaca aaagcaacaa
     6121 cacaggtggc tttccctctt tcactttttg tttgcaagcc gcgtgcgagc aagacggcac
     6181 gaccggcaaa cgcaattacg ctgacaaaga gcagacgaag ttttggcgaa aaacatcaag
     6241 gcgcctgata cgaatgcatt tgcaataaca attgcgatat ttaatattgt ttatgaagct
     6301 gtttgacttc aaaacacaca aaaaaaaaaa ataaaacaaa ttatttgaaa gagaattagg
     6361 aatcggacgc ttatcgttag ggtaacaaca agaaatgctt actgagtcac agcctctgga
     6421 aaactgccgc aagccagaga gagagagaaa aagagggaga gcagcttaga ccgcatgtgc
     6481 ttgtgtgtga ggcgtctctc tcttcgtctc tgttgcgcaa acgcatagac tgcactgaaa
     6541 aaatcgatta cctatttttt atgaatgaat atttgcacta ttactattca aaactattaa
     6601 gatagcaatc acattcaata gccaaatact ataccacctg agcgatgcaa cgaaatgatc
     6661 aatttgagca aaaatgctgc atatttagga cggcatcatt atagaaatgc ttcttgctgt
     6721 gtacttttct ctcgtctggc agctgtttcg ccgttattgt taaaaccggc ttaagttagg
     6781 tgtgttttct acgactagtg aatgccctac tagaagatgt gtgttgcaca aaatgtccct
     6841 ggaataacca atttgaagtg cagatagcag taaacgtaag ctaatatgaa tattatttaa
     6901 ctgtaatgtt ttaatatcgc tggacattac taataaaccc actataaaca catgtacata
     6961 tgtatgtttt ggcatacaat gagtagttgg ggaaaaaatg tgtaaaagca ccgtgaccat
     7021 cacagcataa agataaccag ctgaagtatc gaatatgagt aacccccaaa ttgaatcaca
     7081 tgccgcaact gataggaccc atggaagtac actcttcatg gcgatataca agacacacac
     7141 aagcacgaac acccagttgc ggaggaaatt ctccgtaaat gaaaacccaa tcggcgaaca
     7201 attcataccc atatatggta aaagttttga acgcgacttg agagcggaga gcattgcggc
     7261 tgataaggtt ttagcgctaa gcgggcttta taaaacgggc tgcgggacca gttttcatat
     7321 cactaccgtt tgagttcttg tgctgtgtgg atactcctcc cgacacaaag ccgctccatc
     7381 agccagcagt cgtctaatcc agagacacca aaccgaaaga cttaatttat atttatttaa
     7441 ttaattttaa taaaacacac caaatgtaag tagctttccc cttcccaaca acaaaacacc
     7501 atcgaaccac tcccaccaag aaaaagcaat aatcgagaaa agccgcggaa aatgtgtgat
     7561 tttttttgta aacaaaacat ttttttatgt gccagtgctg aaagtgatca aaaaatacta
     7621 gccacgagct aaagagttat tgtattgacc aaaactccaa aaatacccaa gtttggccct
     7681 aaattgtcaa tcaaaatacc aataggtcga aagacatcaa aattaacaaa accagggttt
     7741 caaataccat aactcaagaa tcaggattac aactgcagat ttcaggatat atacatacaa
     7801 attatacgaa attataaaaa ccaaagcaat tcaatagccc caactcaaat gttaggatct
     7861 aatatagtgt ttaaagccaa gctcgctgat gtgggcgtgt cacgatttca cccaaagata
     7921 tgccaaatta cgaattgcaa atcaattcgc caacactttc tttttttccc acgccctaaa
     7981 acaccagatc atccataaat gtacatacat acagtatatg catattataa tctgtaaaac
     8041 tagatcaggt tcttgaaaat agtgacgtag gcagccgttt tggctgaagc agaaattttt
     8101 gccggttttt caaagttgta gttgcaaaaa tggagaaaac cttcgagcat tcgttcatat
     8161 acacacactc acgcgcaaaa taacgagaga gagtgtatgt gtgtgtgaga gagcgaaagc
     8221 cagacgacgg tttgcttttc gcctcgaaac atgaccatat atggtcacaa aacttggccg
     8281 ccgcaattca acacaccagc gctctccttc gcacccatag cgaccatggc gcggagcgag
     8341 cgagatggcg agagcgagcg acgcctatgg cgacgtcgac gcaggcagcg attgaaaaac
     8401 gcagttaact ggcattcaac attcaccagc cactttcagt cggtttattc cagtcattcc
     8461 tttcaaaccg tgcggtcgct tagctcagcc tcgccacttg cgtttacagt agttttcacg
     8521 ccttgaattt gttaaatcga acaaaaaggt aaagtttaac tagctttgaa aagtttcgtg
     8581 gctcttaatt gttaaatttt ctagagtgcg tttagtgttt tttttttttt ttattttgta
     8641 atgttaattt cgggttccaa ttcgagtttt aggcagccgc cattttaagg gcgcatacac
     8701 acaggcaact gtgctctctt tgcggctttc ttttgcaccg gcattcgtta agtgtcgtct
     8761 agaagcttct cccctccctt ttcggcatat tcgtattgtg gttttaattt ttcggggcgg
     8821 ggccttctat tttgtaactg ttcttttaat ttcttattac aattcgatcg caagtgaaaa
     8881 tcagttttca atcggaaaag tattttttta tgaaattttt ttttgtccaa gattaaaatt
     8941 ttgtactaaa aaaacgtaca ttgcattgag tgatttttaa ttgtacacga aaaacaagtt
     9001 agtttgttat gacaattgta ctttggtaga ccagcgcagt ccaaggaaac cacgcaaatt
     9061 ctcagttttt tttttgccat ttctacatta ccaaataagg taaccaaaaa ctaatgggaa
     9121 atccgcattc tttccattgc agcttacagg atcgatcccc gggaattcac catgagcaag
     9181 ctggagaagt ttacaaactg ctactccctg tctaagaccc tgaggttcaa ggccatccct
     9241 gtgggcaaga cccaggagaa catcgacaat aagcggctgc tggtggagga cgagaagaga
     9301 gccgaggatt ataagggcgt gaagaagctg ctggatcgct actatctgtc ttttatcaac
     9361 gacgtgctgc acagcatcaa gctgaagaat ctgaacaatt acatcagcct gttccggaag
     9421 aaaaccagaa ccgagaagga gaataaggag ctggagaacc tggagatcaa tctgcggaag
     9481 gagatcgcca aggccttcaa gggcaacgag ggctacaagt ccctgtttaa gaaggatatc
     9541 atcgagacaa tcctgccaga gttcctggac gataaggacg agatcgccct ggtgaacagc
     9601 ttcaatggct ttaccacagc cttcaccggc ttctttgata acagagagaa tatgttttcc
     9661 gaggaggcca agagcacatc catcgccttc aggtgtatca acgagaatct gacccgctac
     9721 atctctaata tggacatctt cgagaaggtg gacgccatct ttgataagca cgaggtgcag
     9781 gagatcaagg agaagatcct gaacagcgac tatgatgtgg aggatttctt tgagggcgag
     9841 ttctttaact ttgtgctgac acaggagggc atcgacgtgt ataacgccat catcggcggc
     9901 ttcgtgaccg agagcggcga gaagatcaag ggcctgaacg agtacatcaa cctgtataat
     9961 cagaaaacca agcagaagct gcctaagttt aagccactgt ataagcaggt gctgagcgat
    10021 cgggagtctc tgagcttcta cggcgagggc tatacatccg atgaggaggt gctggaggtg
    10081 tttagaaaca ccctgaacaa gaacagcgag atcttcagct ccatcaagaa gctggagaag
    10141 ctgttcaaga attttgacga gtactctagc gccggcatct ttgtgaagaa cggccccgcc
    10201 atcagcacaa tctccaagga tatcttcggc gagtggaacg tgatccggga caagtggaat
    10261 gccgagtatg acgatatcca cctgaagaag aaggccgtgg tgaccgagaa gtacgaggac
    10321 gatcggagaa agtccttcaa gaagatcggc tccttttctc tggagcagct gcaggagtac
    10381 gccgacgccg atctgtctgt ggtggagaag ctgaaggaga tcatcatcca gaaggtggat
    10441 gagatctaca aggtgtatgg ctcctctgag aagctgttcg acgccgattt tgtgctggag
    10501 aagagcctga agaagaacga cgccgtggtg gccatcatga aggacctgct ggattctgtg
    10561 aagagcttcg agaattacat caaggccttc tttggcgagg gcaaggagac aaacagggac
    10621 gagtccttct atggcgattt tgtgctggcc tacgacatcc tgctgaaggt ggaccacatc
    10681 tacgatgcca tccgcaatta tgtgacccag aagccctact ctaaggataa gttcaagctg
    10741 tattttcaga accctcagtt catgggcggc tgggacaagg ataaggagac agactatcgg
    10801 gccaccatcc tgagatacgg ctccaagtac tatctggcca tcatggataa gaagtacgcc
    10861 aagtgcctgc agaagatcga caaggacgat gtgaacggca attacgagaa gatcaactat
    10921 aagctgctgc ccggccctaa taagatgctg ccaaaggtgt tcttttctaa gaagtggatg
    10981 gcctactata accccagcga ggacatccag aagatctaca agaatggcac attcaagaag
    11041 ggcgatatgt ttaacctgaa tgactgtcac aagctgatcg acttctttaa ggatagcatc
    11101 tcccggtatc caaagtggtc caatgcctac gatttcaact tttctgagac agagaagtat
    11161 aaggacatcg ccggctttta cagagaggtg gaggagcagg gctataaggt gagcttcgag
    11221 tctgccagca agaaggaggt ggataagctg gtggaggagg gcaagctgta tatgttccag
    11281 atctataaca aggacttttc cgataagtct cacggcacac ccaatctgca caccatgtac
    11341 ttcaagctgc tgtttgacga gaacaatcac ggacagatca ggctgagcgg aggagcagag
    11401 ctgttcatga ggcgcgcctc cctgaagaag gaggagctgg tggtgcaccc agccaactcc
    11461 cctatcgcca acaagaatcc agataatccc aagaaaacca caaccctgtc ctacgacgtg
    11521 tataaggata agaggttttc tgaggaccag tacgagctgc acatcccaat cgccatcaat
    11581 aagtgcccca agaacatctt caagatcaat acagaggtgc gcgtgctgct gaagcacgac
    11641 gataacccct atgtgatcgg catcgatagg ggcgagcgca atctgctgta tatcgtggtg
    11701 gtggacggca agggcaacat cgtggagcag tattccctga acgagatcat caacaacttc
    11761 aacggcatca ggatcaagac agattaccac tctctgctgg acaagaagga gaaggagagg
    11821 ttcgaggccc gccagaactg gacctccatc gagaatatca aggagctgaa ggccggctat
    11881 atctctcagg tggtgcacaa gatctgcgag ctggtggaga agtacgatgc cgtgatcgcc
    11941 ctggaggacc tgaactctgg ctttaagaat agccgcgtga aggtggagaa gcaggtgtat
    12001 cagaagttcg agaagatgct gatcgataag ctgaactaca tggtggacaa gaagtctaat
    12061 ccttgtgcaa caggcggcgc cctgaagggc tatcagatca ccaataagtt cgagagcttt
    12121 aagtccatgt ctacccagaa cggcttcatc ttttacatcc ctgcctggct gacatccaag
    12181 atcgatccat ctaccggctt tgtgaacctg ctgaaaacca agtataccag catcgccgat
    12241 tccaagaagt tcatcagctc ctttgacagg atcatgtacg tgcccgagga ggatctgttc
    12301 gagtttgccc tggactataa gaacttctct cgcacagacg ccgattacat caagaagtgg
    12361 aagctgtact cctacggcaa ccggatcaga atcttccgga atcctaagaa gaacaacgtg
    12421 ttcgactggg aggaggtgtg cctgaccagc gcctataagg agctgttcaa caagtacggc
    12481 atcaattatc agcagggcga tatcagagcc ctgctgtgcg agcagtccga caaggccttc
    12541 tactctagct ttatggccct gatgagcctg atgctgcaga tgcggaacag catcacaggc
    12601 cgcaccgacg tggattttct gatcagccct gtgaagaact ccgacggcat cttctacgat
    12661 agccggaact atgaggccca ggagaatgcc atcctgccaa agaacgccga cgccaatggc
    12721 gcctataaca tcgccagaaa ggtgctgtgg gccatcggcc agttcaagaa ggccgaggac
    12781 gagaagctgg ataaggtgaa gatcgccatc tctaacaagg agtggctgga gtacgcccag
    12841 accagcgtga agcacaaaag gccggcggcc acgaaaaagg ccggccaggc aaaaaagaaa
    12901 aagggatcct acccatacga tgttccagat tacgcttatc cctacgacgt gcctgattat
    12961 gcatacccat atgatgtccc cgactatgcc taactcgagg gtacctctag aggatctttg
    13021 tgaaggaacc ttacttctgt ggtgtgacat aattggacaa actacctaca gagatttaaa
    13081 gctctaaggt aaatataaaa tttttaagtg tataatgtgt taaactactg attctaattg
    13141 tttgtgtatt ttagattcca acctatggaa ctgatgaatg ggagcagtgg tggaatgcct
    13201 ttaatgagga aaacctgttt tgctcagaag aaatgccatc tagtgatgat gaggctactg
    13261 ctgactctca acattctact cctccaaaaa agaagagaaa ggtagaagac cccaaggact
    13321 ttccttcaga attgctaagt tttttgagtc atgctgtgtt tagtaataga actcttgctt
    13381 gctttgctat ttacaccaca aaggaaaaag ctgcactgct atacaagaaa attatggaaa
    13441 aatatttgat gtatagtgcc ttgactagag atcataatca gccataccac atttgtagag
    13501 gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat
    13561 gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc
    13621 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
    13681 ctcatcaatg tatcttatca tgtctggatc cactagtgtc gacgatgtag gtcacggtct
    13741 cgaagccgcg gtgcgggtgc cagggcgtgc ccttgggctc cccgggcgcg tactccacct
    13801 cacccatctg gtccatcatg atgaacgggt cgaggtggcg gtagttgatc ccggcgaacg
    13861 cgcggcgcac cgggaagccc tcgccctcga aaccgctggg cgcggtggtc acggtgagca
    13921 cgggacgtgc gacggcgtcg gcgggtgcgg atacgcgggg cagcgtcagc gggttctcga
    13981 cggtcacggc gggcatgtcg acactagttc tagccagctt ttgttccctt tagtgagggt
    14041 taatttcgag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
    14101 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
    14161 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
    14221 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
    14281 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
    14341 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
    14401 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
    14461 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
    14521 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
    14581 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
    14641 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
    14701 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
    14761 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
    14821 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
    14881 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
    14941 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
    15001 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
    15061 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
    15121 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
    15181 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
    15241 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
    15301 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
    15361 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
    15421 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
    15481 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
    15541 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
    15601 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
    15661 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
    15721 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
    15781 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
    15841 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
    15901 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
    15961 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
    16021 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
    16081 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
    16141 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
    16201 cccgaaaagt gccac
//
