LOCUS       sequence-309014-       12262 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gal4 dependent expression of LbCas12a in Drosophila.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12262)
  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 12262)
  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: UAST-LbCas12a
FEATURES             Location/Qualifiers
     source          1..12262
                     /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)."
     protein_bind    4858..4952
                     /label=5X UAS
                     /bound_moiety="GAL4"
                     /note="five tandem copies of the ""ScaI site"" 17-mer 
                     CGGAGTACTGTCCTCCG, an upstream activating sequence (UAS) 
                     that efficiently binds yeast Gal4 (Webster et al., 1988; 
                     Pfeiffer et al., 2010)"
     promoter        4971..5209
                     /label=hsp70 promoter
                     /note="Drosophila melanogaster hsp70Bb promoter"
     primer_bind     5156..5177
                     /label=pCasper-hs
                     /note="Drosophila Hsp70 promoter, forward primer"
     CDS             5232..8915
                     /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             8916..8963
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             8970..9050
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     intron          9136..9201
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             9331..9351
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    9623..9757
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9660..9679)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9714..9733
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    9786..9855
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     promoter        complement(10074..10092)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(10113..10129)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10113..10129)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10126..10148)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10137..10153
                     /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(10161..10167,10168..10185,10186..10191))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10206..10227
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10344..10361)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10515..11103)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10595..10614)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11274..12065,12066..12134))
                     /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     11897..11916
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12135..12239)
                     /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 cttgcatgcc tgcaggtcgg
     4861 agtactgtcc tccgagcgga gtactgtcct ccgagcggag tactgtcctc cgagcggagt
     4921 actgtcctcc gagcggagta ctgtcctccg agcggagact ctagcgagcg ccggagtata
     4981 aatagaggcg cttcgtctac ggagcgacaa ttcaattcaa acaagcaaag tgaacacgtc
     5041 gctaagcgaa agctaagcaa ataaacaagc gcagctgaac aagctaaaca atctgcagta
     5101 aagtgcaagt taaagtgaat caattaaaag taaccagcaa ccaagtaaat caactgcaac
     5161 tactgaaatc tgccaagaag taattattga atacaagaag agaactctga atagggaatt
     5221 gggaattcac catgagcaag ctggagaagt ttacaaactg ctactccctg tctaagaccc
     5281 tgaggttcaa ggccatccct gtgggcaaga cccaggagaa catcgacaat aagcggctgc
     5341 tggtggagga cgagaagaga gccgaggatt ataagggcgt gaagaagctg ctggatcgct
     5401 actatctgtc ttttatcaac gacgtgctgc acagcatcaa gctgaagaat ctgaacaatt
     5461 acatcagcct gttccggaag aaaaccagaa ccgagaagga gaataaggag ctggagaacc
     5521 tggagatcaa tctgcggaag gagatcgcca aggccttcaa gggcaacgag ggctacaagt
     5581 ccctgtttaa gaaggatatc atcgagacaa tcctgccaga gttcctggac gataaggacg
     5641 agatcgccct ggtgaacagc ttcaatggct ttaccacagc cttcaccggc ttctttgata
     5701 acagagagaa tatgttttcc gaggaggcca agagcacatc catcgccttc aggtgtatca
     5761 acgagaatct gacccgctac atctctaata tggacatctt cgagaaggtg gacgccatct
     5821 ttgataagca cgaggtgcag gagatcaagg agaagatcct gaacagcgac tatgatgtgg
     5881 aggatttctt tgagggcgag ttctttaact ttgtgctgac acaggagggc atcgacgtgt
     5941 ataacgccat catcggcggc ttcgtgaccg agagcggcga gaagatcaag ggcctgaacg
     6001 agtacatcaa cctgtataat cagaaaacca agcagaagct gcctaagttt aagccactgt
     6061 ataagcaggt gctgagcgat cgggagtctc tgagcttcta cggcgagggc tatacatccg
     6121 atgaggaggt gctggaggtg tttagaaaca ccctgaacaa gaacagcgag atcttcagct
     6181 ccatcaagaa gctggagaag ctgttcaaga attttgacga gtactctagc gccggcatct
     6241 ttgtgaagaa cggccccgcc atcagcacaa tctccaagga tatcttcggc gagtggaacg
     6301 tgatccggga caagtggaat gccgagtatg acgatatcca cctgaagaag aaggccgtgg
     6361 tgaccgagaa gtacgaggac gatcggagaa agtccttcaa gaagatcggc tccttttctc
     6421 tggagcagct gcaggagtac gccgacgccg atctgtctgt ggtggagaag ctgaaggaga
     6481 tcatcatcca gaaggtggat gagatctaca aggtgtatgg ctcctctgag aagctgttcg
     6541 acgccgattt tgtgctggag aagagcctga agaagaacga cgccgtggtg gccatcatga
     6601 aggacctgct ggattctgtg aagagcttcg agaattacat caaggccttc tttggcgagg
     6661 gcaaggagac aaacagggac gagtccttct atggcgattt tgtgctggcc tacgacatcc
     6721 tgctgaaggt ggaccacatc tacgatgcca tccgcaatta tgtgacccag aagccctact
     6781 ctaaggataa gttcaagctg tattttcaga accctcagtt catgggcggc tgggacaagg
     6841 ataaggagac agactatcgg gccaccatcc tgagatacgg ctccaagtac tatctggcca
     6901 tcatggataa gaagtacgcc aagtgcctgc agaagatcga caaggacgat gtgaacggca
     6961 attacgagaa gatcaactat aagctgctgc ccggccctaa taagatgctg ccaaaggtgt
     7021 tcttttctaa gaagtggatg gcctactata accccagcga ggacatccag aagatctaca
     7081 agaatggcac attcaagaag ggcgatatgt ttaacctgaa tgactgtcac aagctgatcg
     7141 acttctttaa ggatagcatc tcccggtatc caaagtggtc caatgcctac gatttcaact
     7201 tttctgagac agagaagtat aaggacatcg ccggctttta cagagaggtg gaggagcagg
     7261 gctataaggt gagcttcgag tctgccagca agaaggaggt ggataagctg gtggaggagg
     7321 gcaagctgta tatgttccag atctataaca aggacttttc cgataagtct cacggcacac
     7381 ccaatctgca caccatgtac ttcaagctgc tgtttgacga gaacaatcac ggacagatca
     7441 ggctgagcgg aggagcagag ctgttcatga ggcgcgcctc cctgaagaag gaggagctgg
     7501 tggtgcaccc agccaactcc cctatcgcca acaagaatcc agataatccc aagaaaacca
     7561 caaccctgtc ctacgacgtg tataaggata agaggttttc tgaggaccag tacgagctgc
     7621 acatcccaat cgccatcaat aagtgcccca agaacatctt caagatcaat acagaggtgc
     7681 gcgtgctgct gaagcacgac gataacccct atgtgatcgg catcgatagg ggcgagcgca
     7741 atctgctgta tatcgtggtg gtggacggca agggcaacat cgtggagcag tattccctga
     7801 acgagatcat caacaacttc aacggcatca ggatcaagac agattaccac tctctgctgg
     7861 acaagaagga gaaggagagg ttcgaggccc gccagaactg gacctccatc gagaatatca
     7921 aggagctgaa ggccggctat atctctcagg tggtgcacaa gatctgcgag ctggtggaga
     7981 agtacgatgc cgtgatcgcc ctggaggacc tgaactctgg ctttaagaat agccgcgtga
     8041 aggtggagaa gcaggtgtat cagaagttcg agaagatgct gatcgataag ctgaactaca
     8101 tggtggacaa gaagtctaat ccttgtgcaa caggcggcgc cctgaagggc tatcagatca
     8161 ccaataagtt cgagagcttt aagtccatgt ctacccagaa cggcttcatc ttttacatcc
     8221 ctgcctggct gacatccaag atcgatccat ctaccggctt tgtgaacctg ctgaaaacca
     8281 agtataccag catcgccgat tccaagaagt tcatcagctc ctttgacagg atcatgtacg
     8341 tgcccgagga ggatctgttc gagtttgccc tggactataa gaacttctct cgcacagacg
     8401 ccgattacat caagaagtgg aagctgtact cctacggcaa ccggatcaga atcttccgga
     8461 atcctaagaa gaacaacgtg ttcgactggg aggaggtgtg cctgaccagc gcctataagg
     8521 agctgttcaa caagtacggc atcaattatc agcagggcga tatcagagcc ctgctgtgcg
     8581 agcagtccga caaggccttc tactctagct ttatggccct gatgagcctg atgctgcaga
     8641 tgcggaacag catcacaggc cgcaccgacg tggattttct gatcagccct gtgaagaact
     8701 ccgacggcat cttctacgat agccggaact atgaggccca ggagaatgcc atcctgccaa
     8761 agaacgccga cgccaatggc gcctataaca tcgccagaaa ggtgctgtgg gccatcggcc
     8821 agttcaagaa ggccgaggac gagaagctgg ataaggtgaa gatcgccatc tctaacaagg
     8881 agtggctgga gtacgcccag accagcgtga agcacaaaag gccggcggcc acgaaaaagg
     8941 ccggccaggc aaaaaagaaa aagggatcct acccatacga tgttccagat tacgcttatc
     9001 cctacgacgt gcctgattat gcatacccat atgatgtccc cgactatgcc taactagagg
     9061 atctttgtga aggaacctta cttctgtggt gtgacataat tggacaaact acctacagag
     9121 atttaaagct ctaaggtaaa tataaaattt ttaagtgtat aatgtgttaa actactgatt
     9181 ctaattgttt gtgtatttta gattccaacc tatggaactg atgaatggga gcagtggtgg
     9241 aatgccttta atgaggaaaa cctgttttgc tcagaagaaa tgccatctag tgatgatgag
     9301 gctactgctg actctcaaca ttctactcct ccaaaaaaga agagaaaggt agaagacccc
     9361 aaggactttc cttcagaatt gctaagtttt ttgagtcatg ctgtgtttag taatagaact
     9421 cttgcttgct ttgctattta caccacaaag gaaaaagctg cactgctata caagaaaatt
     9481 atggaaaaat atttgatgta tagtgccttg actagagatc ataatcagcc ataccacatt
     9541 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
     9601 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag
     9661 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     9721 gtccaaactc atcaatgtat cttatcatgt ctggatccac tagtgtcgac gatgtaggtc
     9781 acggtctcga agccgcggtg cgggtgccag ggcgtgccct tgggctcccc gggcgcgtac
     9841 tccacctcac ccatctggtc catcatgatg aacgggtcga ggtggcggta gttgatcccg
     9901 gcgaacgcgc ggcgcaccgg gaagccctcg ccctcgaaac cgctgggcgc ggtggtcacg
     9961 gtgagcacgg gacgtgcgac ggcgtcggcg ggtgcggata cgcggggcag cgtcagcggg
    10021 ttctcgacgg tcacggcggg catgtcgaca ctagttctag ccagcttttg ttccctttag
    10081 tgagggttaa tttcgagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
    10141 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
    10201 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
    10261 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
    10321 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
    10381 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
    10441 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
    10501 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
    10561 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
    10621 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
    10681 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
    10741 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
    10801 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
    10861 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
    10921 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
    10981 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
    11041 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
    11101 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
    11161 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
    11221 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
    11281 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
    11341 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
    11401 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
    11461 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
    11521 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
    11581 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
    11641 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
    11701 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
    11761 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
    11821 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
    11881 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
    11941 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
    12001 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
    12061 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
    12121 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
    12181 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
    12241 acatttcccc gaaaagtgcc ac
//
