LOCUS       sequence-137074-       16491 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression plasmid of AsCpf1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 16491)
  AUTHORS   Port F, Bullock SL
  TITLE     Augmenting CRISPR applications in Drosophila with tRNA-flanked 
            sgRNAs.
  JOURNAL   Nat Methods. 2016 Oct;13(10):852-4. doi: 10.1038/nmeth.3972. Epub 
            2016 Sep 5.
  PUBMED    27595403
REFERENCE   2  (bases 1 to 16491)
  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"; doi: 
            "10.1038/nmeth.3972"; journalName: "Nat Methods"; date: "2016-10"; 
            volume: "13"; issue: "10"; pages: "852-4"
COMMENT     Sequence Label: act-AsCpf1
FEATURES             Location/Qualifiers
     source          1..16491
                     /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"
     gene            666..4802
                     /gene="white"
                     /label=mini-white
                     /note="This modified version of the white gene lacks part 
                     of the first intron."
     primer_bind     complement(675..693)
                     /label=pCasper-F
                     /note="Drosophila mini-white gene, reverse primer"
     CDS             join(1145..1216,1618..1891,1966..2620,2682..2997,3218..3349,
                     3420..4034)
                     /codon_start=1
                     /gene="white"
                     /product="Drosophila white gene eye color pigment"
                     /label=mini-white
                     /note="This modified version of the white gene lacks part 
                     of the first intron."
                     /translation="MGQEDQELLIRGGSKHPSAEHLNNGDSGAASQSCINQGFGQAKNY
                     GTLRPPSPPEDSGSGSGQLAENLTYAWHNMDIFGAVNQPGSGWRQLVNRTRGLFCNERH
                     IPAPRKHLLKNVCGVAYPGELLAVMGSSGAGKTTLLNALAFRSPQGIQVSPSGMRLLNG
                     QPVDAKEMQARCAYVQQDDLFIGSLTAREHLIFQAMVRMPRHLTYRQRVARVDQVIQEL
                     SLSKCQHTIIGVPGRVKGLSGGERKRLAFASEALTDPPLLICDEPTSGLDSFTAHSVVQ
                     VLKKLSQKGKTVILTIHQPSSELFELFDKILLMAEGRVAFLGTPSEAVDFFSYVGAQCP
                     TNYNPADFYVQVLAVVPGREIESRDRIAKICDNFAISKVARDMEQLLATKNLEKPLEQP
                     ENGYTYKATWFMQFRAVLWRSWLSVLKEPLLVKVRLIQTTMVAILIGLIFLGQQLTQVG
                     VMNINGAIFLFLTNMTFQNVFATINVFTSELPVFMREARSRLYRCDTYFLGKTIAELPL
                     FLTVPLVFTAIAYPMIGLRAGVLHFFNCLALVTLVANVSTSFGYLISCASSSTSMALSV
                     GPPVIIPFLLFGGFFLNSGSVPVYLKWLSYLSWFRYANEGLLINQWADVEPGEISCTSS
                     NTTCPSSGKVILETLNFSAADLPLDYVGLAILIVSFRVLAYLALRLRARRKE"
     protein_bind    4809..4842
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        4990..7445
                     /label=Ac5 promoter
                     /note="Drosophila melanogaster actin 5c promoter"
     CDS             9211..13131
                     /codon_start=1
                     /product="CRISPR-associated protein from Acidaminococcus 
                     sp. BV3L6 (Zetsche et al., 2015)"
                     /label=AsCpf1
                     /translation="MTQFEGFTNLYQVSKTLRFELIPQGKTLKHIQEQGFIEEDKARND
                     HYKELKPIIDRIYKTYADQCLQLVQLDWENLSAAIDSYRKEKTEETRNALIEEQATYRN
                     AIHDYFIGRTDNLTDAINKRHAEIYKGLFKAELFNGKVLKQLGTVTTTEHENALLRSFD
                     KFTTYFSGFYENRKNVFSAEDISTAIPHRIVQDNFPKFKENCHIFTRLITAVPSLREHF
                     ENVKKAIGIFVSTSIEEVFSFPFYNQLLTQTQIDLYNQLLGGISREAGTEKIKGLNEVL
                     NLAIQKNDETAHIIASLPHRFIPLFKQILSDRNTLSFILEEFKSDEEVIQSFCKYKTLL
                     RNENVLETAEALFNELNSIDLTHIFISHKKLETISSALCDHWDTLRNALYERRISELTG
                     KITKSAKEKVQRSLKHEDINLQEIISAAGKELSEAFKQKTSEILSHAHAALDQPLPTTL
                     KKQEEKEILKSQLDSLLGLYHLLDWFAVDESNEVDPEFSARLTGIKLEMEPSLSFYNKA
                     RNYATKKPYSVEKFKLNFQMPTLASGWDVNKEKNNGAILFVKNGLYYLGIMPKQKGRYK
                     ALSFEPTEKTSEGFDKMYYDYFPDAAKMIPKCSTQLKAVTAHFQTHTTPILLSNNFIEP
                     LEITKEIYDLNNPEKEPKKFQTAYAKKTGDQKGYREALCKWIDFTRDFLSKYTKTTSID
                     LSSLRPSSQYKDLGEYYAELNPLLYHISFQRIAEKEIMDAVETGKLYLFQIYNKDFAKG
                     HHGKPNLHTLYWTGLFSPENLAKTSIKLNGQAELFYRPKSRMKRMAHRLGEKMLNKKLK
                     DQKTPIPDTLYQELYDYVNHRLSHDLSDEARALLPNVITKEVSHEIIKDRRFTSDKFFF
                     HVPITLNYQAANSPSKFNQRVNAYLKEHPETPIIGIDRGERNLIYITVIDSTGKILEQR
                     SLNTIQQFDYQKKLDNREKERVAARQAWSVVGTIKDLKQGYLSQVIHEIVDLMIHYQAV
                     VVLENLNFGFKSKRTGIAEKAVYQQFEKMLIDKLNCLVLKDYPAEKVGGVLNPYQLTDQ
                     FTSFAKMGTQSGFLFYVPAPYTSKIDPLTGFVDPFVWKTIKNHESRKHFLEGFDFLHYD
                     VKTGDFILHFKMNRNLSFQRGLPGFMPAWDIVFEKNETQFDAKGTPFIAGKRIVPVIEN
                     HRFTGRYRDLYPANELIALLEEKGIVFRDGSNILPKLLENDDSHAIDTMVALIRSVLQM
                     RNSNAATGEDYINSPVRDLNGVCFDSRFQNPEWPMDADANGAYHIALKGQLLLNHLKES
                     KDLKLQNGISNQDWLAYIQELRN"
     regulatory      12851..12860
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             13132..13179
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             13186..13266
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     intron          13365..13430
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             13560..13580
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    13852..13986
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(13889..13908)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     13943..13962
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    14015..14084
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     promoter        complement(14303..14321)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(14342..14358)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(14342..14358)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(14355..14377)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    14366..14382
                     /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(14390..14396,14397..14414,14415..14420))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    14435..14456
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(14573..14590)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(14744..15332)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(14824..14843)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(15503..16294,16295..16363))
                     /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     16126..16145
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(16364..16468)
                     /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 gattttccag gccatggtgc
     2221 ggatgccacg acatctgacc tatcggcagc gagtggcccg cgtggatcag gtgatccagg
     2281 agctttcgct cagcaaatgt cagcacacga tcatcggtgt gcccggcagg gtgaaaggtc
     2341 tgtccggcgg agaaaggaag cgtctggcat tcgcctccga ggcactaacc gatccgccgc
     2401 ttctgatctg cgatgagccc acctccggac tggactcatt 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 agcattacat tacatctcaa ctcctatcca gcgtgggtgc ccagtgtcct
     2701 accaactaca atccggcgga cttttacgta caggtgttgg ccgttgtgcc cggacgggag
     2761 atcgagtccc gtgatcggat cgccaagata tgcgacaatt ttgctattag caaagtagcc
     2821 cgggatatgg agcagttgtt ggccaccaaa aatttggaga agccactgga gcagccggag
     2881 aatgggtaca cctacaaggc cacctggttc atgcagttcc gggcggtcct gtggcgatcc
     2941 tggctgtcgg tgctcaagga accactcctc gtaaaagtgc gacttattca gacaacggtg
     3001 agtggttcca gtggaaacaa atgatataac gcttacaatt cttggaaaca aattcgctag
     3061 attttagtta gaattgcctg attccacacc cttcttagtt tttttcaatg agatgtatag
     3121 tttatagttt tgcagaaaat aaataaattt catttaactc gcgaacatgt tgaagatatg
     3181 aatattaatg agatgcgagt aacattttaa tttgcagatg gttgccatct tgattggcct
     3241 catctttttg ggccaacaac tcacgcaagt gggcgtgatg aatatcaacg gagccatctt
     3301 cctcttcctg accaacatga cctttcaaaa cgtctttgcc acgataaatg taagtcttgt
     3361 ttagaataca tttgcatatt aataatttac taactttcta atgaatcgat tcgatttagg
     3421 tgttcacctc agagctgcca gtttttatga gggaggcccg aagtcgactt tatcgctgtg
     3481 acacatactt tctgggcaaa acgattgccg aattaccgct ttttctcaca gtgccactgg
     3541 tcttcacggc gattgcctat ccgatgatcg gactgcgggc cggagtgctg cacttcttca
     3601 actgcctggc gctggtcact ctggtggcca atgtgtcaac gtccttcgga tatctaatat
     3661 cctgcgccag ctcctcgacc tcgatggcgc tgtctgtggg tccgccggtt atcataccat
     3721 tcctgctctt tggcggcttc ttcttgaact cgggctcggt gccagtatac ctcaaatggt
     3781 tgtcgtacct ctcatggttc cgttacgcca acgagggtct gctgattaac caatgggcgg
     3841 acgtggagcc gggcgaaatt agctgcacat cgtcgaacac cacgtgcccc agttcgggca
     3901 aggtcatcct ggagacgctt aacttctccg ccgccgatct gccgctggac tacgtgggtc
     3961 tggccattct catcgtgagc ttccgggtgc tcgcatatct ggctctaaga cttcgggccc
     4021 gacgcaagga gtagccgaca tatatccgaa ataactgctt gttttttttt tttaccatta
     4081 ttaccatcgt gtttactgtt tattgccccc tcaaaaagct aatgtaatta tatttgtgcc
     4141 aataaaaaca agatatgacc tatagaatac aagtatttcc ccttcgaaca tccccacaag
     4201 tagactttgg atttgtcttc taaccaaaag acttacacac ctgcatacct tacatcaaaa
     4261 actcgtttat cgctacataa aacaccggga tatatttttt atatacatac ttttcaaatc
     4321 gcgcgccctc ttcataattc acctccacca caccacgttt cgtagttgct ctttcgctgt
     4381 ctcccacccg ctctccgcaa cacattcacc ttttgttcga cgaccttgga gcgactgtcg
     4441 ttagttccgc gcgattcggt tcgctcaaat ggttccgagt ggttcatttc gtctcaatag
     4501 aaattagtaa taaatatttg tatgtacaat ttatttgctc caatatattt gtatatattt
     4561 ccctcacagc tatatttatt ctaatttaat attatgactt tttaaggtaa ttttttgtga
     4621 cctgttcgga gtgattagcg ttacaatttg aactgaaagt gacatccagt gtttgttcct
     4681 tgtgtagatg catctcaaaa aaatggtggg cataatagtg ttgtttatat atatcaaaaa
     4741 taacaactat aataataaga atacatttaa tttagaaaat gcttggattt cactggaact
     4801 aggctagcat aacttcgtat aatgtatgct atacgaagtt atgctagcgg atccaagctt
     4861 tgcggccgca attctatatt ctaaaaacac aaatgatact tctaaaaaaa aatcatgaat
     4921 ggcatcaact ctgaatcaaa tctttgcaga tgcacctact tctcatttcc actgtcacat
     4981 catttttcca gatctcgctg cctgttatgt ggcccacaaa ccaagacacg ttttatggcc
     5041 attaaagctg gctgatcgtc gccaaacacc aaatacataa tgaatatgta cacattcgag
     5101 aaagaagcga tcaaagaagc gtcttcgggc ggagtaggag aatgcggagg agaaggagaa
     5161 cgagctgatc tagtatctct ccacaatcca atgccaactg accaactggc catattcgga
     5221 gcaatttgaa gccaatttcc atcgcctggc gatcgctcca ttcttggcta tatgtttttc
     5281 accgttaccc ggggccattt tcaaagactc gtcggcaaga taagattgtg tcactcgctg
     5341 tctctcttca tttgtcgaag aatgctgagg aatttcgcga tgacgtcggc gagtattttg
     5401 aagaatgaga ataatttgta tttatacgaa aatcagttag tggaattttc tacaaaaaca
     5461 tgttatctat agataatttt gttgcaaaat atgttgacta tgacaaagat tgtatgtata
     5521 tacctttaat gtattctcat tttcttatgt atttataatg gcaatgatga tactgatgat
     5581 attttaagat gatgccagac cacaggctga tttctgcgtc ttttgccgaa cgcagtgcat
     5641 gtgcggttgt tgttttttgg aatagtttca attttcggac tgtccgcttt gatttcagtt
     5701 tcttggctta ttcaaaaagc aaagtaaagc caaaaaagcg agatggcaat accaaatgcg
     5761 gcaaaacggt agtggaagga aaggggtgcg gggcagcgga aggaagggtg gggcggggcg
     5821 tggcggggtc tgtggctggg cgcgacgtca ccgacgttgg agccactcct ttgaccatgt
     5881 gtgcgtgtgt gtattattcg tgtctcgcca ctcgccggtt gtttttttct ttttatctcg
     5941 ctctctctag cgccatctcg tacgcatgct caacgcaccg catgttgccg tgtcctttat
     6001 gcgtcatttt ggctcgaaat aggcaattat ttaaacaaag attagtcaac gaaaacgcta
     6061 aaataaataa gtctacaata tggttactta ttgccatgtg tgtgcagcca acgatagcaa
     6121 caaaagcaac aacacagtgg ctttccctct ttcacttttt gtttgcaagc gcgtgcgagc
     6181 aagacggcac gaccggcaaa cgcaattacg ctgacaaaga gcagacgaag ttttggccga
     6241 aaaacatcaa ggcgcctgat acgaatgcat ttgcaataac aattgcgata tttaatattg
     6301 tttatgaagc tgtttgactt caaaacacac aaaaaaaaaa ataaaacaaa ttatttgaaa
     6361 gagaattagg aatcggacag cttatcgtta cgggctaaca gcacaccgag acgaaatagc
     6421 ttacctgacg tcacagcctc tggaagaact gccgccaagc agacgatgca gaggacgaca
     6481 catagagtag cggagtaggc cagcgtagta cgcatgtgct tgtgtgtgag gcgtctctct
     6541 cttcgtctcc tgtttgcgca aacgcataga ctgcactgag aaaatcgatt acctattttt
     6601 tatgaatgaa tatttgcact attactattc aaaactatta agatagcaat cacattcaat
     6661 agccaaatac tataccacct gagcgatgca acgaaatgat caatttgagc aaaaatgctg
     6721 catatttagg acggcatcat tatagaaatg cttcttgctg tgtacttttc tctcgtctgg
     6781 cagctgtttc gccgttattg ttaaaaccgg cttaagttag gtgtgttttc tacgactagt
     6841 gatgccccta ctagaagatg tgtgttgcac aaatgtccct gaataaccaa tttgaagtgc
     6901 agatagcagt aaacgtaagc taatatgaat attatttaac tgtaatgttt taatatcgct
     6961 ggacattact aataaaccca ctataaacac atgtacatat gtatgttttg gcatacaatg
     7021 agtagttggg gaaaaaatgt gtaaaagcac cgtgaccatc acagcataaa gataaccagc
     7081 tgaagtatcg aatatgagta acccccaaat tgaatcacat gccgcaactg ataggaccca
     7141 tggaagtaca ctcttcatgg cgatatacaa gacacacaca agcacgaaca cccagttgcg
     7201 gaggaaattc tccgtaaatg aaaacccaat cggcgaacaa ttcataccca tatatggtaa
     7261 aagttttgaa cgcgacttga gagcggagag cattgcggct gataaggttt tagcgctaag
     7321 cgggctttat aaaacgggct gcgggaccag ttttcatatc actaccgttt gagttcttgt
     7381 gctgtgtgga tactcctccc gacacaaagc cgctccatca gccagcagtc gtctaatcca
     7441 gagacaccaa accgaaagac ttaatttata tttatttaat taattttaat aaaacacacc
     7501 aaatgtaagt agctttcccc ttcccaacaa caaaacacca tcgaaccact cccaccaaga
     7561 aaaagcaata atcgagaaaa gccgcggaaa atgtgtgatt ttttttgtaa acaaaacatt
     7621 tttttatgtg ccagtgctga aagtgatcaa aaaatactag ccacgagcta aagagttatt
     7681 gtattgacca aaactccaaa aatacccaag tttggcccta aattgtcaat caaaatacca
     7741 ataggtcgaa agacatcaaa attaacaaaa ccagggtttc aaataccata actcaagaat
     7801 caggattaca actgcagatt tcaggatata tacatacaaa ttatacgaaa ttataaaaac
     7861 caaagcaatt caatagcccc aactcaaatg ttaggatcta atatagtgtt taaagccaag
     7921 ctcgctgatg tgggcgtgtc acgatttcac ccaaagatat gccaaattac gaattgcaaa
     7981 tcaattcgcc aacactttct ttttttccca cgccctaaaa caccagatca tccataaatg
     8041 tacatacata cagtatatgc atattataat ctgtaaaact agatcaggtt cttgaaaata
     8101 gtgacgtagg cagccgtttt ggctgaagca gaaatttttg ccggtttttc aaagttgtag
     8161 ttgcaaaaat ggagaaaacc ttcgagcatt cgttcatata cacacactca cgcgcaaaat
     8221 aacgagagag agtgtatgtg tgtgtgagag agcgaaagcc agacgacggt ttgcttttcg
     8281 cctcgaaaca tgaccatata tggtcacaaa acttggccgc cgcaattcaa cacaccagcg
     8341 ctctccttcg cacccatagc gaccatggcg cggagcgagc gagatggcga gagcgagcga
     8401 cgcctatggc gacgtcgacg caggcagcga ttgaaaaacg cagttaactg gcattcaaca
     8461 ttcaccagcc actttcagtc ggtttattcc agtcattcct ttcaaaccgt gcggtcgctt
     8521 agctcagcct cgccacttgc gtttacagta gttttcacgc cttgaatttg ttaaatcgaa
     8581 caaaaaggta aagtttaact agctttgaaa agtttcgtgg ctcttaattg ttaaattttc
     8641 tagagtgcgt ttagtgtttt tttttttttt tattttgtaa tgttaatttc gggttccaat
     8701 tcgagtttta ggcagccgcc attttaaggg cgcatacaca caggcaactg tgctctcttt
     8761 gcggctttct tttgcaccgg cattcgttaa gtgtcgtcta gaagcttctc ccctcccttt
     8821 tcggcatatt cgtattgtgg ttttaatttt tcggggcggg gccttctatt ttgtaactgt
     8881 tcttttaatt tcttattaca attcgatcgc aagtgaaaat cagttttcaa tcggaaaagt
     8941 atttttttat gaaatttttt tttgtccaag attaaaattt tgtactaaaa aaacgtacat
     9001 tgcattgagt gatttttaat tgtacacgaa aaacaagtta gtttgttatg acaattgtac
     9061 tttggtagac cagcgcagtc caaggaaacc acgcaaattc tcagtttttt ttttgccatt
     9121 tctacattac caaataaggt aaccaaaaac taatgggaaa tccgcattct ttccattgca
     9181 gcttacagga tcgatccccg ggaattcacc atgacacagt tcgagggctt taccaacctg
     9241 tatcaggtga gcaagacact gcggtttgag ctgatcccac agggcaagac cctgaagcac
     9301 atccaggagc agggcttcat cgaggaggac aaggcccgca atgatcacta caaggagctg
     9361 aagcccatca tcgatcggat ctacaagacc tatgccgacc agtgcctgca gctggtgcag
     9421 ctggattggg agaacctgag cgccgccatc gactcctata gaaaggagaa aaccgaggag
     9481 acaaggaacg ccctgatcga ggagcaggcc acatatcgca atgccatcca cgactacttc
     9541 atcggccgga cagacaacct gaccgatgcc atcaataaga gacacgccga gatctacaag
     9601 ggcctgttca aggccgagct gtttaatggc aaggtgctga agcagctggg caccgtgacc
     9661 acaaccgagc acgagaacgc cctgctgcgg agcttcgaca agtttacaac ctacttctcc
     9721 ggcttttatg agaacaggaa gaacgtgttc agcgccgagg atatcagcac agccatccca
     9781 caccgcatcg tgcaggacaa cttccccaag tttaaggaga attgtcacat cttcacacgc
     9841 ctgatcaccg ccgtgcccag cctgcgggag cactttgaga acgtgaagaa ggccatcggc
     9901 atcttcgtga gcacctccat cgaggaggtg ttttccttcc ctttttataa ccagctgctg
     9961 acacagaccc agatcgacct gtataaccag ctgctgggag gaatctctcg ggaggcaggc
    10021 accgagaaga tcaagggcct gaacgaggtg ctgaatctgg ccatccagaa gaatgatgag
    10081 acagcccaca tcatcgcctc cctgccacac agattcatcc ccctgtttaa gcagatcctg
    10141 tccgatagga acaccctgtc tttcatcctg gaggagttta agagcgacga ggaagtgatc
    10201 cagtccttct gcaagtacaa gacactgctg agaaacgaga acgtgctgga gacagccgag
    10261 gccctgttta acgagctgaa cagcatcgac ctgacacaca tcttcatcag ccacaagaag
    10321 ctggagacaa tcagcagcgc cctgtgcgac cactgggata cactgaggaa tgccctgtat
    10381 gagcggagaa tctccgagct gacaggcaag atcaccaagt ctgccaagga gaaggtgcag
    10441 cgcagcctga agcacgagga tatcaacctg caggagatca tctctgccgc aggcaaggag
    10501 ctgagcgagg ccttcaagca gaaaaccagc gagatcctgt cccacgcaca cgccgccctg
    10561 gatcagccac tgcctacaac cctgaagaag caggaggaga aggagatcct gaagtctcag
    10621 ctggacagcc tgctgggcct gtaccacctg ctggactggt ttgccgtgga tgagtccaac
    10681 gaggtggacc ccgagttctc tgcccggctg accggcatca agctggagat ggagccttct
    10741 ctgagcttct acaacaaggc cagaaattat gccaccaaga agccctactc cgtggagaag
    10801 ttcaagctga actttcagat gcctacactg gcctctggct gggacgtgaa taaggagaag
    10861 aacaatggcg ccatcctgtt tgtgaagaac ggcctgtact atctgggcat catgccaaag
    10921 cagaagggca ggtataaggc cctgagcttc gagcccacag agaaaaccag cgagggcttt
    10981 gataagatgt actatgacta cttccctgat gccgccaaga tgatcccaaa gtgcagcacc
    11041 cagctgaagg ccgtgacagc ccactttcag acccacacaa cccccatcct gctgtccaac
    11101 aatttcatcg agcctctgga gatcacaaag gagatctacg acctgaacaa tcctgagaag
    11161 gagccaaaga agtttcagac agcctacgcc aagaaaaccg gcgaccagaa gggctacaga
    11221 gaggccctgt gcaagtggat cgacttcaca agggattttc tgtccaagta taccaagaca
    11281 acctctatcg atctgtctag cctgcggcca tcctctcagt ataaggacct gggcgagtac
    11341 tatgccgagc tgaatcccct gctgtaccac atcagcttcc agagaatcgc cgagaaggag
    11401 atcatggatg ccgtggagac aggcaagctg tacctgttcc agatctataa caaggacttt
    11461 gccaagggcc accacggcaa gcctaatctg cacacactgt attggaccgg cctgttttct
    11521 ccagagaacc tggccaagac aagcatcaag ctgaatggcc aggccgagct gttctaccgc
    11581 cctaagtcca ggatgaagag gatggcacac cggctgggag agaagatgct gaacaagaag
    11641 ctgaaggatc agaaaacccc aatccccgac accctgtacc aggagctgta cgactatgtg
    11701 aatcacagac tgtcccacga cctgtctgat gaggccaggg ccctgctgcc caacgtgatc
    11761 accaaggagg tgtctcacga gatcatcaag gataggcgct ttaccagcga caagttcttt
    11821 ttccacgtgc ctatcacact gaactatcag gccgccaatt ccccatctaa gttcaaccag
    11881 agggtgaatg cctacctgaa ggagcacccc gagacaccta tcatcggcat cgatcggggc
    11941 gagagaaacc tgatctatat cacagtgatc gactccaccg gcaagatcct ggagcagcgg
    12001 agcctgaaca ccatccagca gtttgattac cagaagaagc tggacaacag ggagaaggag
    12061 agggtggcag caaggcaggc ctggtctgtg gtgggcacaa tcaaggatct gaagcagggc
    12121 tatctgagcc aggtcatcca cgagatcgtg gacctgatga tccactacca ggccgtggtg
    12181 gtgctggaga acctgaattt cggctttaag agcaagagga ccggcatcgc cgagaaggcc
    12241 gtgtaccagc agttcgagaa gatgctgatc gataagctga attgcctggt gctgaaggac
    12301 tatccagcag agaaagtggg aggcgtgctg aacccatacc agctgacaga ccagttcacc
    12361 tcctttgcca agatgggcac ccagtctggc ttcctgtttt acgtgcctgc cccatataca
    12421 tctaagatcg atcccctgac cggcttcgtg gaccccttcg tgtggaaaac catcaagaat
    12481 cacgagagcc gcaagcactt cctggagggc ttcgactttc tgcactacga cgtgaaaacc
    12541 ggcgacttca tcctgcactt taagatgaac agaaatctgt ccttccagag gggcctgccc
    12601 ggctttatgc ctgcatggga tatcgtgttc gagaagaacg agacacagtt tgacgccaag
    12661 ggcacccctt tcatcgccgg caagagaatc gtgccagtga tcgagaatca cagattcacc
    12721 ggcagatacc gggacctgta tcctgccaac gagctgatcg ccctgctgga ggagaagggc
    12781 atcgtgttca gggatggctc caacatcctg ccaaagctgc tggagaatga cgattctcac
    12841 gccatcgaca ccatggtggc cctgatccgc agcgtgctgc agatgcggaa ctccaatgcc
    12901 gccacaggcg aggactatat caacagcccc gtgcgcgatc tgaatggcgt gtgcttcgac
    12961 tcccggtttc agaacccaga gtggcccatg gacgccgatg ccaatggcgc ctaccacatc
    13021 gccctgaagg gccagctgct gctgaatcac ctgaaggaga gcaaggatct gaagctgcag
    13081 aacggcatct ccaatcagga ctggctggcc tacatccagg agctgcgcaa caaaaggccg
    13141 gcggccacga aaaaggccgg ccaggcaaaa aagaaaaagg gatcctaccc atacgatgtt
    13201 ccagattacg cttatcccta cgacgtgcct gattatgcat acccatatga tgtccccgac
    13261 tatgcctaac tcgagggtac ctctagagga tctttgtgaa ggaaccttac ttctgtggtg
    13321 tgacataatt ggacaaacta cctacagaga tttaaagctc taaggtaaat ataaaatttt
    13381 taagtgtata atgtgttaaa ctactgattc taattgtttg tgtattttag attccaacct
    13441 atggaactga tgaatgggag cagtggtgga atgcctttaa tgaggaaaac ctgttttgct
    13501 cagaagaaat gccatctagt gatgatgagg ctactgctga ctctcaacat tctactcctc
    13561 caaaaaagaa gagaaaggta gaagacccca aggactttcc ttcagaattg ctaagttttt
    13621 tgagtcatgc tgtgtttagt aatagaactc ttgcttgctt tgctatttac accacaaagg
    13681 aaaaagctgc actgctatac aagaaaatta tggaaaaata tttgatgtat agtgccttga
    13741 ctagagatca taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc
    13801 ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt
    13861 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
    13921 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
    13981 tggatccact agtgtcgacg atgtaggtca cggtctcgaa gccgcggtgc gggtgccagg
    14041 gcgtgccctt gggctccccg ggcgcgtact ccacctcacc catctggtcc atcatgatga
    14101 acgggtcgag gtggcggtag ttgatcccgg cgaacgcgcg gcgcaccggg aagccctcgc
    14161 cctcgaaacc gctgggcgcg gtggtcacgg tgagcacggg acgtgcgacg gcgtcggcgg
    14221 gtgcggatac gcggggcagc gtcagcgggt tctcgacggt cacggcgggc atgtcgacac
    14281 tagttctagc cagcttttgt tccctttagt gagggttaat ttcgagcttg gcgtaatcat
    14341 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
    14401 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
    14461 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
    14521 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
    14581 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
    14641 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
    14701 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
    14761 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
    14821 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
    14881 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
    14941 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
    15001 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
    15061 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
    15121 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
    15181 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
    15241 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
    15301 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
    15361 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
    15421 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
    15481 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
    15541 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
    15601 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
    15661 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
    15721 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
    15781 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
    15841 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
    15901 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
    15961 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
    16021 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
    16081 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
    16141 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
    16201 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
    16261 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
    16321 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
    16381 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
    16441 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca c
//
