LOCUS       sequence-468999-       16663 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Empty Cas12a vector for multiplex gene editing in fungi.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 16663)
  AUTHORS   Yuan G, Deng S, Czajka JJ, Dai Z, Hofstad BA, Kim J, Pomraning KR
  TITLE     CRISPR-Cas9/Cas12a systems for efficient genome editing and large 
            genomic fragment deletions in Aspergillus niger.
  JOURNAL   Front Bioeng Biotechnol. 2024 Oct 16;12:1452496. doi: 
            10.3389/fbioe.2024.1452496. eCollection 2024.
  PUBMED    39479294
REFERENCE   2  (bases 1 to 16663)
  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: "Front 
            Bioeng Biotechnol."; date: "2024-10-16"; volume: "12:1452496. doi"; 
            pages: " 10.3389/fbioe.2024.1452496. eCollection 2024"
COMMENT     Sequence Label: pGY53
FEATURES             Location/Qualifiers
     source          1..16663
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      104..5351
                     /label=AMA1
                     /note="replication origin from Aspergillus nidulans 
                     (Aleksenko and Clutterbuck, 1996)"
     CDS             7137..10820
                     /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             10821..10868
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             10875..10955
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     CDS             10959..10979
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             12452..13246
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(12506..12525)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     13116..13135
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     protein_bind    complement(13681..13714)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     primer_bind     complement(14184..14200)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(14299..14316)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(14470..15058)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(14550..14569)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(15180..15971,15972..16040))
                     /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     15803..15822
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(16041..16132)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     16174..16193
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     16199..16218
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     protein_bind    16425..16458
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
ORIGIN
        1 ccaactgacc agaggcaggg tagcggattg acggctccat ctccttcact tacaaggtct
       61 attgaaagcc ctttagcatc accaagcgga gaatagattg ttaagcttat tttttgtata
      121 ctgttttgtg atagcacgaa gtttttccac ggtatcttgt taaaaatata tatttgtggc
      181 gggcttacct acatcaaatt aataagagac taattataaa ctaaacacac aagcaagcta
      241 ctttagggta aaagtttata aatgcttttg acgtataaac gttgcttgta tttattatta
      301 caattaaagg tggatagaaa acctagagac tagttagaaa ctaatctcag gtttgcgtta
      361 aactaaatca gagcccgaga ggttaacaga acctagaagg ggactagata tccgggtagg
      421 gaaacaaaaa aaaaaaacaa gacagccaca tattagggag actagttaga agctagttcc
      481 aggactagga aaataaaaga caatgatacc acagtctagt tgacaactag atagattcta
      541 gattgaggcc aaagtctctg agatccaggt tagttgcaac taatactagt tagtatctag
      601 tctcctataa ctctgaagct agaataactt actactatta tcctcaccac tgttcagctg
      661 cgcaaacgga gtgattgcaa ggtgttcaga gactagttat tgactagtca gtgactagca
      721 ataactaaca aggtattaac ctaccatgtc tgccatcacc ctgcacttcc tcgggctcag
      781 cagccttttc ctcctcattt tcatgctcat tttccttgtt taagactgtg actagtcaaa
      841 gactagtcca gaaccacaaa ggagaaatgt cttaccactt tcttcattgc ttgtctcttt
      901 tgcattatcc atgtctgcaa ctagttagag tctagttagt gactagtccg acgaggactt
      961 gcttgtctcc ggattgttgg aggaactctc cagggcctca agatccacaa cagagccttc
     1021 tagaagactg gtcaataact agttggtctt tgtctgagtc tgacttacga ggttgcatac
     1081 tcgctccctt tgcctcgtca atcgatgaga aaaagcgcca aaactcgcaa tatggctttg
     1141 aaccacacgg tgctgagact agttagaatc tagtcccaaa ctagcttgga tagcttacct
     1201 ttgccctttg cgttgcgaca ggtcttgcag ggtatggttc ctttctcacc agctgattta
     1261 gctgccttgc taccctcacg gcggatctgc ataaagagtg gctagaggtt ataaattagc
     1321 actgatccta ggtacggggc tgaatgtaac ttgcccttcc tttctcatcg cgcggcaaga
     1381 caggcttgct caaattccta ccagtcacag gggtatgcac ggcgtacgga ccacttgaac
     1441 tagtcacaga ttagttagca actagtctgc attgaatggc tgtacttacg ggccctcgcc
     1501 attgtcctga tcatttccag cttcaccctc gttgctgcaa agtagttagt gactagtcaa
     1561 ggactagttg aaatgggaga agaaactcac gaattctcga cacccttagt attgtggtcc
     1621 ttggacttgg tgctgctata tattagctaa tacactagtt agactcacag aaacttacgc
     1681 agctcgcttg cgcttcttgg taggagtcgg ggttgggaga acagtgcctt caaacaagcc
     1741 ttcataccat gctacttgac tagtcaggga ctagtcacca agtaatctag ataggacttg
     1801 cctttggcct ccatcagttc cttcatagtg ggaggtccat tgtgcaatgt aaactccatg
     1861 ccgtgggagt tcttgtcctt caagtgcttg accaatatgt ttctgttggc agagggaacc
     1921 tgtcaactag ttaataacta gtcagaaact agtatagcag tagactcact gtacgcttga
     1981 ggcatccctt cactcggcag tagacttcat atggatggat atcaggcacg ccattgtcgt
     2041 cctgtggact agtcagtaac taggcttaaa gctagtcggg tcggcttact atcttgaaat
     2101 ccggcagcgt aagctccccg tccttaactg cctcgagata gtgacagtac tctggggact
     2161 ttcggagatc gttatcgcga atgctcggca tactaatcgt tgactagtct tggactagtc
     2221 ccgagcaaaa aggattggag gaggaggagg aaggtgagag tgagacaaag agcgaaataa
     2281 gagcttcaaa ggctatctct aagcagtatg aaggttaagt atctagttct tgactagatt
     2341 taaaagagat ttcgactagt tatgtacctg gagtttggat ataggaatgt gttgtggtaa
     2401 cgaaatgtaa gggggaggaa agaaaaagtc ggtcaagagg taactctaag tcggccattc
     2461 ctttttggga ggcgctaacc ataaacggca tggtcgactt agagttagct cagggaattt
     2521 agggagttat ctgcgaccac tggactctca ccgtttccat tggggtacta acatagccat
     2581 caaatgccgg cccgtgttta atctcctcga gatggctggg ctcacatgcg ttctactcaa
     2641 aaactcttat acctaggccc tctcccaatc gtgtatgttt ggaccttcct ctctcacact
     2701 ttggataaca tcaagcatac tgcagatttt gacctagtta acaaggcaca tcatgctatc
     2761 acttccaaca ctcttgctga gcgcgttctg ccttgccttt ctggttctac gaagtatcca
     2821 gtcccgcctg caacgccgca gaaacgccaa aaggtggggt tgtcaaccgc cagctagagc
     2881 tccatcggga ttctttggca ttccgccgtt cctcggtggt cgcagataac tccctaaatt
     2941 ccctgagcta actctaagtc gaccatgccg tttatggtta gcgcctccca aaaaggaatg
     3001 gccgacttag agttacctct tgaccgactt tttctttcct cccccttaca tttcgttacc
     3061 acaacacatt cctatatcca aactccaggt acataactag tcgaaatctc ttttaaatct
     3121 agtcaagaac tagatactta accttcatac tgcttagaga tagcctttga agctcttatt
     3181 tcgctctttg tctcactctc accttcctcc tcctcctcca atcctttttg ctcgggacta
     3241 gtccaagact agtcaacgat tagtatgccg agcattcgcg ataacgatct ccgaaagtcc
     3301 ccagagtact gtcactatct cgaggcagtt aaggacgggg agcttacgct gccggatttc
     3361 aagatagtaa gccgacccga ctagctttaa gcctagttac tgactagtcc acaggacgac
     3421 aatggcgtgc ctgatatcca tccatatgaa gtctactgcc gagtgaaggg atgcctcaag
     3481 cgtacagtga gtctactgct atactagttt ctgactagtt attaactagt tgacaggttc
     3541 cctctgccaa cagaaacata ttggtcaagc acttgaagga taagaactcc cacggcatgg
     3601 agtttacatt gcacaatgga cctcccacta tgaaggaact gatggaggcc aaaggcaagt
     3661 cctatctaga ttacttggtg actagtccct gactagtcaa gtagcatggt atgaaggctt
     3721 gtttgaaggc actgttctcc caaccccgac tcctaccaag aagcgcaagc gagctgcgta
     3781 agtttctgtg agtctaacta gtgtattagc taatatatag cagcaccaag tccaaggacc
     3841 acaatactaa gggtgtcgag aattcgtgag tttcttctcc catttcaact agtccttgac
     3901 tagtcactaa ctactttgca gcaacgaggg tgaagctgga aatgatcagg acaatggcga
     3961 gggcccgtaa gtacagccat tcaatgcaga ctagttgcta actaatctgt gactagttca
     4021 agtggtccgt acgccgtgca tacccctgtg actggtagga atttgagcaa gcctgtcttg
     4081 ccgcgcgatg agaaaggaaa ggcaagttac attcagcccc gtacctagga tcagtgctaa
     4141 tttataacct ctagccactc tttatgcaga tccgccgtga gggtagcaag gcagctaaat
     4201 cagctggtga gaaaggaacc ataccctgca agacctgtcg caacgcaaag ggcaaaggta
     4261 agctatccaa gctagtttgg gactagattc taactagtct cagcaccgtg tggttcaaag
     4321 ccatattgcg agttttggcg ctttttctca tcgattgacg aggcaaaggg agcgagtatg
     4381 caacctcgta agtcagactc agacaaagac caactagtta ttgaccagtc ttctagaagg
     4441 ctctgttgtg gatcttgagg ccctggagag ttcctccaac aatccggaga caagcaagtc
     4501 ctcgtcggac tagtcactaa ctagactcta actagttgca gacatggata atgcaaaaga
     4561 gacaagcaat gaagaaagtg gtaagacatt tctcctttgt ggttctggac tagtctttga
     4621 ctagtcacag tcttaaacaa ggaaaatgag catgaaaatg aggaggaaaa ggctgctgag
     4681 cccgaggaag tgcagggtga tggcagacat ggtaggttaa taccttgtta gttattgcta
     4741 gtcactgact agtcaataac tagtctctga acaccttgca atcactccgt ttgcgcagct
     4801 gaacagtggt gaggataata gtagtaagtt attctagctt cagagttata ggagactaga
     4861 tactaactag tattagttgc aactaacctg gatctcagag actttggcct caatctagaa
     4921 tctatctagt tgtcaactag actgtggtat cattgtcttt tattttccta gtcctggaac
     4981 tagcttctaa ctagtctccc taatatgtgg ctgtcttgtt tttttttttg tttccctacc
     5041 cggatatcta gtccccttct aggttctgtt aacctctcgg gctctgattt agtttaacgc
     5101 aaacctgaga ttagtttcta actagtctct aggttttcta tccaccttta attgtaataa
     5161 taaatacaag caacgtttat acgtcaaaag catttataaa cttttaccct aaagtagctt
     5221 gcttgtgtgt ttagtttata attagtctct tattaatttg atgtaggtaa gcccgccaca
     5281 aatatatatt tttaacaaga taccgtggaa aaacttcgtg ctatcacaaa acagtataca
     5341 aaaaataagc ttaacaatct attctccgct tggtgatgct aaagggcttt caatagacct
     5401 tgtaagtgaa ggagatggag ccgtcaatcc gctaccctgc ctctggtcag ttgggatcac
     5461 atagatgctc ggttgacagg accacctata gcaagtactt tgtaggaggt tctcctaacg
     5521 cttggcttca aatccaatca ggatatcagt gagttactgt ttgtcatcgc atcctcatga
     5581 agtcgctcaa cagccactga gaagcaaata tttgggacac catccctgat gttgaaattt
     5641 tacgctggag cccattcacc ggtgagctcg gggacagtct ggccagtgga gcggaaaatc
     5701 ttcaactaac tctgattggt accacaggta gccccacaga acaaaccgag caaacatgaa
     5761 aattttcgct tgaggttagc gcactcgcta gcgcctgccc gcaaatataa aaggccccga
     5821 aatcccatgc attttggaaa tcaagcgact ctacgtatgt catccccctc cctacctttg
     5881 ggaaaacctc tcaactgcaa gtcagaacat tttgctaaca gcaggggctg tggtttagtg
     5941 gtataatatt cccttagcat gggagaggtc cggggttcga ttccccgcag ctccaggaga
     6001 ccatggtctc ctgtcaattt ttacacttga tgttggtgta atcaggcgat cagagtttgc
     6061 tcaaattctc tcttgtttta gattctgaag aatataattt ttacatgact agttcatttt
     6121 acttccaagt agactatttg tagattgaga ttgccgtcaa ctttattcat agaatcacat
     6181 taaattgctc attgacctgc atgtctatgc acacattcac atctatcttc tcagggtatt
     6241 aagacctcag ccgagacagc agaatcaccg cccaagttaa gcctttgtgc tgatcatgct
     6301 ctcgaacggg ccaagttcgg gaaaagcaaa ggagcgttta gtgaggggca atttgactca
     6361 cctcccaggc aacagatgag gggggcaaaa agaaagaaat tttcgtgagt caatatggat
     6421 tccgagcatc attttcttgc ggtctatctt gctacgtatg ttgatcttga cgctgtggat
     6481 caagcaacgc cactcgctcg ctccatcgca ggctggtcgc agacaaatta aaaggcggca
     6541 aactcgtaca gccgcggggt tgtccgctgc aaagtacaga gtgataaaag ccgccatgcg
     6601 accatcaacg cgttgatgcc cagctttttc gatccgagaa tccaccgtag aggcgatagc
     6661 aagtaaagaa aagctaaaca aaaaaaaatt tctgccccta agccatgaaa acgagatggg
     6721 gtggagcaga accaaggaaa gagtcgcgct gggctgccgt tccggaaggt gttgtaaagg
     6781 ctcgacgccc aaggtgggag tctaggagaa gaatttgcat cgggagtggg gcgggttacc
     6841 cctccatatc caatgacaga tatctaccag ccaagggttt gagcccgccc gcttagtcgt
     6901 cgtcctcgct tgcccctcca taaaaggatt tcccctcccc ctcccacaaa attttctttc
     6961 ccttcctctc cttgtccgct tcagtacgta tatcttccct tccctcgctt ctctcctcca
     7021 tccttctttc atccatctcc tgctaacttc tctgctcagc acctctacgc attactagcc
     7081 gtagtatctg agcacttctc ccttttatat tccacaaaac ataacacaga cttcgtatgt
     7141 ccaagctcga gaagttcacc aactgctact ccctcagcaa gaccctccgc ttcaaggcta
     7201 tccccgtcgg caagacccaa gagaatatcg ataacaagcg cctcctcgtc gaggatgaga
     7261 agcgcgccga ggattacaag ggcgtcaaga agctcctcga tcgctactac ctctccttca
     7321 tcaacgatgt cctccactcc atcaagctca agaacctcaa caactacatc tccctgttcc
     7381 gcaagaaaac ccgcaccgag aaagagaaca aagagctgga aaacctcgag atcaacctcc
     7441 gcaaagagat cgccaaggcc ttcaagggca acgagggcta caagtccctg ttcaagaagg
     7501 atatcatcga aaccatcctg cctgagttcc tcgatgataa ggatgagatc gccctcgtca
     7561 actccttcaa cggcttcact accgccttca ccggtttctt cgataaccgc gaaaacatgt
     7621 tctccgagga agccaagtcc acctctatcg ccttccgctg catcaacgag aacttgaccc
     7681 gctacatcag caacatggat atcttcgaga aggtcgatgc catcttcgac aagcacgagg
     7741 tccaagagat caaagaaaag atcctcaact ccgattacga tgtcgaggat ttcttcgagg
     7801 gcgagttctt caacttcgtg ctcacccaag agggcatcga tgtctacaac gccatcatcg
     7861 gcggcttcgt caccgagtcc ggcgagaaga tcaagggcct caacgagtac atcaacctct
     7921 acaatcaaaa gaccaagcag aagctcccca agttcaagcc cctctacaag caggtcctct
     7981 ccgatcgcga gtccctgtcc ttctacggcg agggttacac ctccgatgag gaagtcctcg
     8041 aggtctttcg caacaccctc aacaagaact ccgagatctt ctccagcatc aagaagttgg
     8101 agaagctttt caagaacttc gatgagtact cctccgccgg catcttcgtc aagaacggcc
     8161 ctgctatctc caccatctcc aaggacatct tcggcgagtg gaacgtcatc cgcgataagt
     8221 ggaacgccga gtacgatgac atccacctca agaagaaggc cgtcgttacc gagaagtacg
     8281 aggatgatcg ccgcaagtcc ttcaagaaga tcggctcctt cagcctcgag cagctccaag
     8341 agtacgccga tgccgatctc tccgtcgtcg agaagctgaa agagatcatc atccagaagg
     8401 tggacgagat ctacaaggtc tacggctcct ctgagaagct gttcgatgcc gacttcgtcc
     8461 tcgaaaagtc cctgaagaag aacgacgccg tcgtcgctat catgaaggat ctgctcgatt
     8521 ccgtgaagtc gttcgagaac tacatcaagg ctttcttcgg tgagggcaaa gagactaacc
     8581 gcgacgagag cttctacggt gatttcgtcc tggcctacga tatcctcctc aaggtcgatc
     8641 acatctacga tgctatccgc aactacgtca cccagaagcc ttactccaag gataagttca
     8701 agttgtactt ccagaatcct cagttcatgg gcggctggga caaagacaaa gaaaccgatt
     8761 accgcgccac catcctccgc tacggcagca agtactacct cgccatcatg gataagaaat
     8821 acgccaagtg cctccaaaag atcgataagg acgatgtcaa cggcaactac gaaaagatca
     8881 actacaagct gctccccggt cctaacaaga tgctgcccaa ggtgttcttc agcaagaaat
     8941 ggatggccta ctacaacccc tccgaggata tccagaagat ctataagaac ggcaccttca
     9001 aaaagggcga tatgttcaac ctcaacgatt gccacaagct catcgatttc ttcaaggatt
     9061 ccatctcgcg ctaccccaag tggtccaacg cctacgattt caacttctcc gagactgaga
     9121 agtataagga tatcgctggc ttctaccgtg aggtcgagga acagggttac aaggtgtcct
     9181 tcgagtccgc ctccaagaaa gaggtcgata agctcgtcga agagggcaag ctctacatgt
     9241 tccaaatcta caacaaggat ttctccgata agtcccacgg cacccctaac ctccacacca
     9301 tgtacttcaa gctgctgttc gacgagaaca accacggcca gatccgcctc tccggcggtg
     9361 ccgagctgtt catgcgccgt gctagcctga agaaagaaga gttggtcgtc catcctgcta
     9421 actccccgat cgccaacaag aatcctgata accccaagaa aactaccacc ttgtcctacg
     9481 acgtctacaa ggacaagcgc ttcagcgagg atcagtacga gctgcacatc cctatcgcca
     9541 tcaacaagtg ccctaagaac atcttcaaga tcaataccga ggtccgcgtc ctcctgaagc
     9601 acgatgacaa cccctacgtc atcggtatcg atcgcggcga gcgcaacctc ctctacatcg
     9661 tcgtcgtcga tggcaagggt aacatcgtcg agcagtactc cctgaacgaa atcatcaaca
     9721 acttcaacgg tatccgcatc aagaccgact accacagcct cctcgataag aaagaaaaag
     9781 agcgcttcga ggcccgccag aactggacct ccatcgagaa catcaaagag ttgaaggccg
     9841 gctacatctc gcaggtcgtc cacaagatct gcgagctggt cgagaagtat gatgccgtga
     9901 tcgctctcga ggatctgaac tccggcttca agaacagccg cgtcaaggtt gagaagcagg
     9961 tctaccaaaa gttcgagaag atgctcatcg acaagctcaa ctacatggtc gacaagaagt
    10021 cgaacccctg cgccaccggc ggagccctca agggctacca gatcactaac aagttcgagt
    10081 cgttcaagtc catgagcacc cagaacggtt tcatcttcta catccccgcc tggctcacct
    10141 ccaagatcga tccctcgacc ggcttcgtga acctgctcaa gaccaagtac acctccattg
    10201 ccgattccaa gaagttcatc tccagcttcg atcgcatcat gtacgtcccc gaggaagatc
    10261 tcttcgagtt cgccctggat tataagaact tcagccgcac cgacgccgat tacattaaga
    10321 agtggaagct ctactcctac ggcaaccgca tccgcatctt ccgcaatcct aagaagaaca
    10381 acgttttcga ttgggaagaa gtctgcctca cctccgccta caaagaactg ttcaacaagt
    10441 acggcatcaa ctaccagcag ggcgacatcc gcgctctgct ctgcgagcag tccgataagg
    10501 ccttctactc ctccttcatg gccctcatgt ccctgatgct ccagatgcgc aactccatca
    10561 ccggccggac cgatgtcgat ttcctcatct cccctgtcaa gaactcggat ggtatcttct
    10621 acgattcccg gaactacgag gctcaagaga acgccatctt gcccaagaac gccgacgcca
    10681 acggcgccta caatatcgcc cgcaaggtcc tctgggccat cggccagttc aagaaagctg
    10741 aggacgagaa gctcgataag gtcaagatcg ccatttccaa caaagagtgg ctcgagtacg
    10801 cccagacctc cgtcaagcac aagcgccctg ccgccaccaa gaaggctggc caggcaaaga
    10861 agaagaaagg ctcgtacccc tacgatgtcc ccgattacgc ttacccttac gacgtgcctg
    10921 actacgccta tccgtacgac gtcccggatt atgcccctcc caagaagaag cgcaaggtct
    10981 gactgagcgg acattcgatt tatgccgtta tgacttcctt aaaaaagcct ttacgaatga
    11041 aagaaatgga attagacttg ttatgtagtt gattctacaa tggattatga ttcctgaact
    11101 tcaaatccgc tgttcattat taatctcagc tcttcccgta aagccaatgt tgaaactatt
    11161 cgtaaatgta cctcgttttg cgtgtacctt gcttatcacg tgatattaca tgacctggac
    11221 agagttctgc gcgaaagtca taacgtaaat cccgggcggt aggtgcgtcc cgggcggaag
    11281 gtagttttct cgtccacccc aacgcgttta tcaacctcaa ctttcaacaa ccatcatgcc
    11341 accaaaagcg cgtaaaacaa agcgagattt gattgagcaa gagggcagga tccaatgcgc
    11401 gattcaagac attaaaaatg gaaaatttca aaaaattgcg cccgcagcgc gtgcatacaa
    11461 aattcatccc aatacaagtg taatgcgaac gactccagaa gtgactaagc aaccgtaaac
    11521 cggaagcaac aatccggctc tcatggcaaa gatggcatgt ccggattgcg gaggaaagat
    11581 ttggcgtgtg cctgacggtt ttgttccaca aggaccctgt aaacttggtg atggtaatga
    11641 ggtgctctgg ccttgattga gcaaaagtca gctcgcgggg actacgtgag ttgatggtgg
    11701 aggatcatgt actaaataga gcatacctag gggatggtga acggacgtcc agggttccgt
    11761 agaattgtaa caactctact cgacttgctg cccactgggg tatgtaaata catgcctttt
    11821 caaagttgcc gagtgtacta aatggagtgt atgtttggcc aatgtcattt gggaggggcc
    11881 gggaaccctt tgggacaccc gcggatgggc tggatctcgg tcactggaga catagcctag
    11941 tggtctagta gtctagcagc aggacagctt catggttaac cccacatgtg atcctccgtc
    12001 cggtttccgt agtatataat ggacctgaag cttctccctc tctcgatgcg gttgaaacat
    12061 aaatttgctc ataacgtatg tccatctaac cctttcattg tttcttgacc actgtctcct
    12121 ttctggaaaa aaaaagactc tggtaagtat gggcctagga tcgagctact gatcccccaa
    12181 tccttcctaa tccctctgcc acttgccccg acctgttcac cgtgaccgta ccaccaccac
    12241 cccgccccct cctcaactaa acccctccct tcccctcccc cctgatgttt tctcttttct
    12301 ttaatctgac tgtcacctcc ctctctactt cttttccctc aattttcttc tcctcttctt
    12361 actctctgtc atctgtttcg atacccatac taatctcgcg tgctttttct tcaccttcgt
    12421 gtatagattt gtcacaatcc caaatttcac catgattgaa caagatggat tgcacgcagg
    12481 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
    12541 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
    12601 gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct
    12661 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
    12721 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
    12781 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
    12841 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
    12901 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
    12961 gttcgccagg ctcaaggcgc gcatgcccga cggcgatgat ctcgtcgtga cccatggcga
    13021 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
    13081 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
    13141 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
    13201 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
    13261 ttcgaaatga ccgaccaagc gacgatccac ttaacgttac tgaaatcatc aaacagcttg
    13321 acgaatctgg atataagatc gttggtgtcg atgtcagctc cggagttgag acaaatggtg
    13381 ttcaggatct cgataagata cgttcatttg tccaagcagc aaagagtgcc ttctagtgat
    13441 ttaatagctc catgtcaaca agaataaaac gcgtttcggg tttacctctt ccagatacag
    13501 ctcatctgca atgcattaat gcattggacc tcgcaaccct agtacgccct tcaggctccg
    13561 gcgaagcaga agaatagctt agcagagtcc atgcggattt cggctccaaa gtagatgccg
    13621 accgatcccc actaccgcat taagacctca gcgcggccgc aaatttaaat aaaatgaagt
    13681 gaagttccta tactttctag agaataggaa cttctatagt gagtcgaata agggcgacac
    13741 aaaatttatt ctaaatgcat aataaatact gataacatct tatagtttgt attatatttt
    13801 gtattatcgt tgacatgtat aattttgata tcaaaaactg attttccctt tattattttc
    13861 gagatttatt ttcttaattc tctttaacaa actagaaata ttgtatatac aaaaaatcat
    13921 aaataataga tgaatagttt aattataggt gttcatcaat cgaaaaagca acgtatctta
    13981 tttaaagtgc gttgcttttt tctcatttat aaggttaaat aattctcata tatcaagcaa
    14041 agtgacaggc gcccttaaat attctgacaa atgctctttc cctaaactcc ccccataaaa
    14101 aaacccgccg aagcgggttt ttacgttatt tgcggattaa cgattactcg ttatcagaac
    14161 cgcccagggg gcccgagctt aagactggcc gtcgttttac aacacagaaa gagtttgtag
    14221 aaacgcaaaa aggccatccg tcaggggcct tctgcttagt ttgatgcctg gcagttccct
    14281 actctcgcct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
    14341 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
    14401 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
    14461 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
    14521 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
    14581 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
    14641 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
    14701 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
    14761 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
    14821 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
    14881 gtggtgggct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
    14941 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
    15001 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
    15061 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgacgcgc gcgtaactca
    15121 cgttaaggga ttttggtcat gagcttgcgc cgtcccgtca agtcagcgta atgctctgct
    15181 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
    15241 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
    15301 agcgctgcga tgataccgcg agaaccacgc tcaccggctc cggatttatc agcaataaac
    15361 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
    15421 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
    15481 gttgttgcca tcgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
    15541 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
    15601 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
    15661 atggttatgg cagcgctaca taattctctt actgtcatgc catccgtaag atgcttttct
    15721 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
    15781 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
    15841 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
    15901 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
    15961 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
    16021 cggaaatgtt gaatactcat attcttcctt tttcaatatt attgaagcat ttatcagggt
    16081 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtc
    16141 agtgttacaa ccaattaacc aattctgaac attatcgcga gcccatttat acctgaatat
    16201 ggctcataac accccttgtt tgcctggcgg cagtagcgcg gtggtcccac ctgaccccat
    16261 gccgaactca gaagtgaaac gccgtagcgc cgatggtagt gtggggactc cccatgcgag
    16321 agtagggaac tgccaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc
    16381 gcccgggcta attatggggt gtcgccctta ttcgactcta tagtgaagtt cctattctct
    16441 agaaagtata ggaacttctg aagtggggat ttaaatgcgg ccgcgctgag ggtttaatcg
    16501 acgaagcagc tgacggccag tgccaagctt aacgcgtacc gggcccagta tatgttccgc
    16561 agatgactgg agctctgcca tacgtgccct ctcaagcacc atttgttcca tctacagaga
    16621 ctagtcacca actagtctat caagactcac agggtacatt gct
//
