pAC1750

LOCUS       pAC1750            15408 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  AMA1 plasmid with Aspergillus optimized Cpf1, ble selection marker, 
            and USER cassette (PacI/Nt.BbvCI).
ACCESSION   .
VERSION     .
KEYWORDS    pAC1750
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15408)
  AUTHORS   Vanegas KG, Jarczynska ZD, Strucko T, Mortensen UH
  TITLE     Cpf1 enables fast and efficient genome editing in Aspergilli.
  JOURNAL   Fungal Biol Biotechnol. 2019 May 1;6:6. doi: 
            10.1186/s40694-019-0069-6. eCollection 2019.
  PUBMED    31061713
REFERENCE   2  (bases 1 to 15408)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..15408
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     622..641
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(860..951)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     993..1012
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1018..1037
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     protein_bind    1244..1277
                     /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)"
     rep_origin      1590..6838
                     /label=AMA1
                     /note="replication origin from Aspergillus nidulans 
                     (Aleksenko and Clutterbuck, 1996)"
     CDS             8023..11706
                     /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             11707..11754
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             11761..11841
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     CDS             11845..11865
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             12705..13079
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     protein_bind    complement(13908..13941)
                     /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(14411..14428)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(14411..14427)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(14526..14543)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(14697..15285)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(14777..14796)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(15407..859)
                     /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"
ORIGIN
        1 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
       61 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
      121 gcgctgcgat gataccgcga gaaccacgct caccggctcc ggatttatca gcaataaacc
      181 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
      241 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
      301 ttgttgccat cgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
      361 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
      421 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
      481 tggttatggc agcgctacat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
      541 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
      601 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
      661 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
      721 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
      781 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
      841 ggaaatgttg aatactcata ttcttccttt ttcaatatta ttgaagcatt tatcagggtt
      901 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggtca
      961 gtgttacaac caattaacca attctgaaca ttatcgcgag cccatttata cctgaatatg
     1021 gctcataaca ccccttgttt gcctggcggc agtagcgcgg tggtcccacc tgaccccatg
     1081 ccgaactcag aagtgaaacg ccgtagcgcc gatggtagtg tggggactcc ccatgcgaga
     1141 gtagggaact gccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg
     1201 cccgggctaa ttatggggtg tcgcccttat tcgactctat agtgaagttc ctattctcta
     1261 gaaagtatag gaacttctga agtggggatt taaatgcggc cgcgctgagg gtttaatcga
     1321 cgaagcagct gacggccagt gccaagctta acgcgtaccg ggcccagtat atgttccgca
     1381 gatgactgga gctctgccat acgtgccctc tcaagcacca tttgttccat ctacagagac
     1441 tagtcaccaa ctagtctatc aagactcaca gggtacattg ctgagaccaa ctgaccagag
     1501 gcagggtagc ggattgacgg ctccatctcc ttcacttaca aggtctattg aaagcccttt
     1561 agcatcacca agcggagaat agattgttaa gcttattttt tgtatactgt tttgtgatag
     1621 cacgaagttt ttccacggta tcttgttaaa aatatatatt tgtggcgggc ttacctacat
     1681 caaattaata agagactaat tataaactaa acacacaagc aagctacttt agggtaaaag
     1741 tttataaatg cttttgacgt ataaacgttg cttgtattta ttattacaat taaaggtgga
     1801 tagaaaacct agagactagt tagaaactaa tctcaggttt gcgttaaact aaatcagagc
     1861 ccgagaggtt aacagaacct agaaggggac tagatatccg ggtagggaaa caaaaaaaaa
     1921 aaacaagaca kccacatatt agggagacta gttagaagct agttccagga ctaggaaaat
     1981 aaaagacaat gataccacag tctagttgac aactagatag attctagatt gaggccaaag
     2041 tctctgagat ccaggttagt tgcaactaat actagttagt atctagtctc ctataactct
     2101 gaagctagaa taacttacta ctattatcct caccactgtt cagctgcgca aacggagtga
     2161 ttgcaaggtg ttcagagact agttattgac tagtcagtga ctagcaataa ctaacaaggt
     2221 attaacctac catgtctgcc atcaccctgc acttcctcgg gctcagcagc cttttcctcc
     2281 tcattttcat gctcattttc cttgtttaag actgtgacta gtcaaagact agtccagaac
     2341 cacaaaggag aaatgtctta ccactttctt cattgcttgt ctcttttgca ttatccatgt
     2401 ctgcaactag ttagagtcta gttagtgact agtccgacga ggacttgctt gtctccggat
     2461 tgttggagga actctccagg gcctcaagat ccacaacaga gccttctaga agactggtca
     2521 ataactagtt ggtctttgtc tgagtctgac ttacgaggtt gcatactcgc tccctttgcc
     2581 tcgtcaatcg atgagaaaaa gcgccaaaac tcgcaatatg gctttgaacc acacggtgct
     2641 gagactagtt agaatctagt cccaaactag cttggatagc ttacctttgy cctttgcgtt
     2701 gcgacaggtc ttgcagggta tggttccttt ctcaccagct gatttagctg ccttgctacc
     2761 ctcacggcgg atctgcataa agagtggcta gaggttataa attagcactg atcctaggta
     2821 cggggctgaa tgtaacttgc cyttcctttc tcatcgcgcg gcaagacagg cttgctcaaa
     2881 ttcctaccag tcacaggggt atgcacggcg tacggaccac ttgaactagt cacagattag
     2941 ttagcaacta gtctgcattg aatggctgta cttacgggcc ctcgccattg tcctgatcat
     3001 ttccagcttc accctcgttg ctgcaaagta gttagtgact agtcaaggac tagttgaaat
     3061 gggagaagaa actcacgaat tctcgacacc cttagtattg tggtccttgg acttggtgct
     3121 gctatatatt agctaataca ctagttagac tcacagaaac ttacgcagct cgcttgcgct
     3181 tcttggtagg agtcggggtt gggagaacag tgccttcaaa caagccttca taccatgcta
     3241 cttgactagt cagggactag tcaccaagta atctagatag gacttgcctt tggcctccat
     3301 cagttccttc atagtgggag gtccattgtg caatgtaaac tccatgccgt gggagttctt
     3361 gtccttcaag tgcttgacca atatgtttct gttggcagag ggaacctgtc aactagttaa
     3421 taactagtca gaaactagta tagcagtaga ctcactgtac gcttgaggca tcccttcact
     3481 cggcagtaga cttcatatgg atggatatca ggcacgccat tgtcgtcctg tggactagtc
     3541 agtaactagg cttaaagcta gtcgggtcgg cttactatct tgaaatccgg cagcgtaagc
     3601 tccccgtcct taactgcctc gagatagtga cagtactctg gggactttcg gagatcgtta
     3661 tcgcgaatgc tcggcatact aatcgttgac tagtcttgga ctagtcccga gcaaaaagga
     3721 ttggaggagr aggaggaagg tgagagtgag acaaagagcg aaataagagc ttcaaaggct
     3781 atctctaagc agtatgaagg ttaagtatct agttcttgac tagatttaaa agagatttcg
     3841 actagttatg tacctggagt ttggatatag gaatgtgttg tggtaacgaa atgtaagggg
     3901 gaggaaagaa aaagtcggtc aataggtaac tctaagtcgg ccattccttt ttgggagtcg
     3961 ctaaccataa acggcatggt cgacttagag ttagctcagg gaatttaggg agttatctgc
     4021 gaccaccgag gaacggcgga atgccaaaga atcccgatgg agctctagct ggcggttgac
     4081 aaccccacct tttggcgttt ctgcggcgtt gcaggcggga ctggatactt cgtagaacca
     4141 gaaaggcaag gcagaacgcg ctcagcaaga gtgttggaag tgatagcatg atgtgccttg
     4201 ttaactaggt caaaatctgc agtatgcttg atgttatcca aagtgtgaga gaggaaggtc
     4261 caaacataca cgattgggag agggcctagg tataagagtt tttgagtaga acgcatgtga
     4321 gcccagccat ctcgaggaga ttaaacacgg gccggcattt gatggctatg ttagtacccc
     4381 aatggaaacg gtgagagtcc agtggtcgca gataactccc taaattccct gagctaactc
     4441 taagtcgacc atgccgttta tggttagcgc ctcccaaaaa ggaatggccg acttagagtt
     4501 acctcttgac cgactttttc tttcctcccc cttacatttc gttaccacaa cacattccta
     4561 tatccaaact ccaggtacat aactagtcga aatctctttt aaatctagtc aagaactaga
     4621 tacttaacct tcatactgct tagagatagc ctttgaagct cttatttcgc tctttgtctc
     4681 actctcacct tcctcctcct cctccaatcc tttttgctcg ggactagtcc aagactagtc
     4741 aacgattagt atgccgagca ttcgcgataa cgatctccga aagtccccag agtactgtca
     4801 ctatctcgag gcagttaagg acggggagct tacgctgccg gatttcaaga tagtaagccg
     4861 acccgactag ctttaagcct agttactgac tagtccacag gacgacaatg gcgtgcctga
     4921 tatccatcca tatgaagtct actgccgagt gaagggatgc ctcaagcgta cagtgagtct
     4981 actgctatac tagtttctga ctagttatta actagttgac aggttccctc tgccaacaga
     5041 aacatattgg tcaagcactt gaaggacaag aactcccacg gcatggagtt tacattgcac
     5101 aatggacctc ccactatgaa ggaactgatg gaggccaaag gcaagtccta tctagattac
     5161 ttggtgacta gtccctgact agtcaagtag catggtatga aggcttgttt gaaggcactg
     5221 ttctcccaac cccgactcct accaagaagc gcaagcgagc tgcgtaagtt tctgtgagtc
     5281 taactagtgt attagctaat atatagcagc accaagtcca aggaccacaa tactaagggt
     5341 gtcgagaatt cgtgagtttc ttctcccatt tcaactagtc cttgactagt cactaactac
     5401 tttgcagcaa cgagggtgaa gctggaaatg atcaggacaa tggcgagggc ccgtaagtac
     5461 agccattcaa tgcagactag ttgctaacta atctgtgact agttcaagtg gtccgtacgc
     5521 cgtgcatacc cctgtgactg gtaggaattt gagcaagcct gtcttgccgc gcgatgagaa
     5581 aggaarggca agttacattc agccccgtac ctaggatcag tgctaattta taacctctag
     5641 ccactcttta tgcagatccg ccgtgagggt agcaaggcag ctaaatcagc tggtgagaaa
     5701 ggaaccatac cctgcaagac ctgtcgcaac gcaaaggrca aaggtaagct atccaagcta
     5761 gtttgggact agattctaac tagtctcagc accgtgtggt tcaaagccat attgcgagtt
     5821 ttggcgcttt ttctcatcga ttgacgaggc aaagggagcg agtatgcaac ctcgtaagtc
     5881 agactcagac aaagaccaac tagttattga ccagtcttct agaaggctct gttgtggatc
     5941 ttgaggccct ggagagttcc tccaacaatc cggagacaag caagtcctcg tcggactagt
     6001 cactaactag actctaacta gttgcagaca tggataatgc aaaagagaca agcaatgaag
     6061 aaagtggtaa gacatttctc ctttgtggtt ctggactagt ctttgactag tcacagtctt
     6121 aaacaaggaa aatgagcatg aaaatgagga ggaaaaggct gctgagcccg aggaagtgca
     6181 gggtgatggc agacatggta ggttaatacc ttgttagtta ttgctagtca ctgactagtc
     6241 aataactagt ctctgaacac cttgcaatca ctccgtttgc gcagctgaac agtggtgagg
     6301 ataatagtag taagttattc tagcttcaga gttataggag actagatact aactagtatt
     6361 agttgcaact aacctggatc tcagagactt tggcctcaat ctagaatcta tctagttgtc
     6421 aactagactg tggtatcatt gtcttttatt ttcctagtcc tggaactagc ttctaactag
     6481 tctccctaat atgtggmtgt cttgtttttt ttttttgttt ccctacccgg atatctagtc
     6541 cccttctagg ttctgttaac ctctcgggct ctgatttagt ttaacgcaaa cctgagatta
     6601 gtttctaact agtctctagg ttttctatcc acctttaatt gtaataataa atacaagcaa
     6661 cgtttatacg tcaaaagcat ttataaactt ttaccctaaa gtagcttgct tgtgtattta
     6721 gtttataatt agtctcttat taatttgatg taggtaagcc cgccacaaat atatattttt
     6781 aacaagatac cgtggaaaaa cttcgtgcta tcacaaaaca gtatacaaaa aataagctta
     6841 acaatctatt ctccgcttgg tgatgctaaa gggctttcaa tagaccttgt aagtgaagga
     6901 gatggagccg tcaatccgct accctgcctc tggtcagttg gtctcagcaa tgtaccctgt
     6961 gagtcttgat agactagttg gtgactagtc tctgtagatg gaacaaatgg tgcttgagag
     7021 ggcacgtatg gcagagctcc agtcatctgc ggaacatata ctgggcccgg gaagatctca
     7081 tggtcatagc tgtttccgct gagggtttaa tgctgagggt ttaattaaga cctcagccga
     7141 gacagcagaa tcaccgccca agttaagcct ttgtgctgat catgctctcg aacgggccaa
     7201 gttcgggaaa agcaaaggag cgtttagtga ggggcaattt gactcacctc ccaggcaaca
     7261 gatgaggggg gcaaaaagaa agaaattttc gtgagtcaat atggattccg agcatcattt
     7321 tcttgcggtc tatcttgcta cgtatgttga tcttgacgct gtggatcaag caacgccact
     7381 cgctcgctcc atcgcaggct ggtcgcagac aaattaaaag gcggcaaact cgtacagccg
     7441 cggggttgtc cgctgcaaag tacagagtga taaaagccgc catgcgacca tcaacgcgtt
     7501 gatgcccagc tttttcgatc cgagaatcca ccgtagaggc gatagcaagt aaagaaaagc
     7561 taaacaaaaa aaaatttctg cccctaagcc atgaaaacga gatggggtgg agcagaacca
     7621 aggaaagagt cgcgctgggc tgccgttccg gaaggtgttg taaaggctcg acgcccaagg
     7681 tgggagtcta ggagaagaat ttgcatcggg agtggggcgg gttacccctc catatccaat
     7741 gacagatatc taccagccaa gggtttgagc ccgcccgctt agtcgtcgtc ctcgcttgcc
     7801 cctccataaa aggatttccc ctccccctcc cacaaaattt tctttccctt cctctccttg
     7861 tccgcttcag tacgtatatc ttcccttccc tcgcttctct cctccatcct tctttcatcc
     7921 atctcctgct aacttctctg ctcagcacct ctacgcatta ctagccgtag tatctgagca
     7981 cttctccctt ttatattcca caaaacataa cacagacttc gtatgtccaa gctcgagaag
     8041 ttcaccaact gctactccct cagcaagacc ctccgcttca aggctatccc cgtcggcaag
     8101 acccaagaga atatcgataa caagcgcctc ctcgtcgagg atgagaagcg cgccgaggat
     8161 tacaagggcg tcaagaagct cctcgatcgc tactacctct ccttcatcaa cgatgtcctc
     8221 cactccatca agctcaagaa cctcaacaac tacatctccc tgttccgcaa gaaaacccgc
     8281 accgagaaag agaacaaaga gctggaaaac ctcgagatca acctccgcaa agagatcgcc
     8341 aaggccttca agggcaacga gggctacaag tccctgttca agaaggatat catcgaaacc
     8401 atcctgcctg agttcctcga tgataaggat gagatcgccc tcgtcaactc cttcaacggc
     8461 ttcactaccg ccttcaccgg tttcttcgat aaccgcgaaa acatgttctc cgaggaagcc
     8521 aagtccacct ctatcgcctt ccgctgcatc aacgagaact tgacccgcta catcagcaac
     8581 atggatatct tcgagaaggt cgatgccatc ttcgacaagc acgaggtcca agagatcaaa
     8641 gaaaagatcc tcaactccga ttacgatgtc gaggatttct tcgagggcga gttcttcaac
     8701 ttcgtgctca cccaagaggg catcgatgtc tacaacgcca tcatcggcgg cttcgtcacc
     8761 gagtccggcg agaagatcaa gggcctcaac gagtacatca acctctacaa tcaaaagacc
     8821 aagcagaagc tccccaagtt caagcccctc tacaagcagg tcctctccga tcgcgagtcc
     8881 ctgtccttct acggcgaggg ttacacctcc gatgaggaag tcctcgaggt ctttcgcaac
     8941 accctcaaca agaactccga gatcttctcc agcatcaaga agttggagaa gcttttcaag
     9001 aacttcgatg agtactcctc cgccggcatc ttcgtcaaga acggccctgc tatctccacc
     9061 atctccaagg acatcttcgg cgagtggaac gtcatccgcg ataagtggaa cgccgagtac
     9121 gatgacatcc acctcaagaa gaaggccgtc gttaccgaga agtacgagga tgatcgccgc
     9181 aagtccttca agaagatcgg ctccttcagc ctcgagcagc tccaagagta cgccgatgcc
     9241 gatctctccg tcgtcgagaa gctgaaagag atcatcatcc agaaggtgga cgagatctac
     9301 aaggtctacg gctcctctga gaagctgttc gatgccgact tcgtcctcga aaagtccctg
     9361 aagaagaacg acgccgtcgt cgctatcatg aaggatctgc tcgattccgt gaagtcgttc
     9421 gagaactaca tcaaggcttt cttcggtgag ggcaaagaga ctaaccgcga cgagagcttc
     9481 tacggtgatt tcgtcctggc ctacgatatc ctcctcaagg tcgatcacat ctacgatgct
     9541 atccgcaact acgtcaccca gaagccttac tccaaggata agttcaagtt gtacttccag
     9601 aatcctcagt tcatgggcgg ctgggacaaa gacaaagaaa ccgattaccg cgccaccatc
     9661 ctccgctacg gcagcaagta ctacctcgcc atcatggata agaaatacgc caagtgcctc
     9721 caaaagatcg ataaggacga tgtcaacggc aactacgaaa agatcaacta caagctgctc
     9781 cccggtccta acaagatgct gcccaaggtg ttcttcagca agaaatggat ggcctactac
     9841 aacccctccg aggatatcca gaagatctat aagaacggca ccttcaaaaa gggcgatatg
     9901 ttcaacctca acgattgcca caagctcatc gatttcttca aggattccat ctcgcgctac
     9961 cccaagtggt ccaacgccta cgatttcaac ttctccgaga ctgagaagta taaggatatc
    10021 gctggcttct accgtgaggt cgaggaacag ggttacaagg tgtccttcga gtccgcctcc
    10081 aagaaagagg tcgataagct cgtcgaagag ggcaagctct acatgttcca aatctacaac
    10141 aaggatttct ccgataagtc ccacggcacc cctaacctcc acaccatgta cttcaagctg
    10201 ctgttcgacg agaacaacca cggccagatc cgcctctccg gcggtgccga gctgttcatg
    10261 cgccgtgcta gcctgaagaa agaagagttg gtcgtccatc ctgctaactc cccgatcgcc
    10321 aacaagaatc ctgataaccc caagaaaact accaccttgt cctacgacgt ctacaaggac
    10381 aagcgcttca gcgaggatca gtacgagctg cacatcccta tcgccatcaa caagtgccct
    10441 aagaacatct tcaagatcaa taccgaggtc cgcgtcctcc tgaagcacga tgacaacccc
    10501 tacgtcatcg gtatcgatcg cggcgagcgc aacctcctct acatcgtcgt cgtcgatggc
    10561 aagggtaaca tcgtcgagca gtactccctg aacgaaatca tcaacaactt caacggtatc
    10621 cgcatcaaga ccgactacca cagcctcctc gataagaaag aaaaagagcg cttcgaggcc
    10681 cgccagaact ggacctccat cgagaacatc aaagagttga aggccggcta catctcgcag
    10741 gtcgtccaca agatctgcga gctggtcgag aagtatgatg ccgtgatcgc tctcgaggat
    10801 ctgaactccg gcttcaagaa cagccgcgtc aaggttgaga agcaggtcta ccaaaagttc
    10861 gagaagatgc tcatcgacaa gctcaactac atggtcgaca agaagtcgaa cccctgcgcc
    10921 accggcggag ccctcaaggg ctaccagatc actaacaagt tcgagtcgtt caagtccatg
    10981 agcacccaga acggtttcat cttctacatc cccgcctggc tcacctccaa gatcgatccc
    11041 tcgaccggct tcgtgaacct gctcaagacc aagtacacct ccattgccga ttccaagaag
    11101 ttcatctcca gcttcgatcg catcatgtac gtccccgagg aagatctctt cgagttcgcc
    11161 ctggattata agaacttcag ccgcaccgac gccgattaca ttaagaagtg gaagctctac
    11221 tcctacggca accgcatccg catcttccgc aatcctaaga agaacaacgt tttcgattgg
    11281 gaagaagtct gcctcacctc cgcctacaaa gaactgttca acaagtacgg catcaactac
    11341 cagcagggcg acatccgcgc tctgctctgc gagcagtccg ataaggcctt ctactcctcc
    11401 ttcatggccc tcatgtccct gatgctccag atgcgcaact ccatcaccgg ccggaccgat
    11461 gtcgatttcc tcatctcccc tgtcaagaac tcggatggta tcttctacga ttcccggaac
    11521 tacgaggctc aagagaacgc catcttgccc aagaacgccg acgccaacgg cgcctacaat
    11581 atcgcccgca aggtcctctg ggccatcggc cagttcaaga aagctgagga cgagaagctc
    11641 gataaggtca agatcgccat ttccaacaaa gagtggctcg agtacgccca gacctccgtc
    11701 aagcacaagc gccctgccgc caccaagaag gctggccagg caaagaagaa gaaaggctcg
    11761 tacccctacg atgtccccga ttacgcttac ccttacgacg tgcctgacta cgcctatccg
    11821 tacgacgtcc cggattatgc ccctcccaag aagaagcgca aggtctgaaa gtctactgcg
    11881 gacattcgat ttatgccgtt atgacttcct taaaaaagcc tttacgaatg aaagaaatgg
    11941 aattagactt gttatgtagt tgattctaca atggattatg attcctgaac ttcaaatccg
    12001 ctgttcatta ttaatctcag ctcttcccgt aaagccaatg ttgaaactat tcgtaaatgt
    12061 acctcgtttt gcgtgtacct tgcttatcac gtgatattac atgacctgga cagagttctg
    12121 cgcgaaagtc ataacgtaaa tcccgggcgg taggtgcgtc ccgggcggaa ggtagttttc
    12181 tcgtccaccc caacgcgttt atcaacctca actttcaaca accatcatgc caccaaaagc
    12241 gcgtaaaaca aagcgagatt tgattgagca agagggcagg atccaatgcg cgattcaaga
    12301 cattaaaaat ggaaaatttc aaaaaattgc gcccgcagcg cgtgcataca aaattcatcc
    12361 caatacatta agacctcagc gctagtggag gtcaacacat caatgcctat tttggtttag
    12421 tcgtccaggc ggtgagcaca aaatttgtgt cgtttgacaa gatggttcat ttaggcaact
    12481 ggtcagatca gccccacttg tagcagtagc ggcggcgctc gaagtgtgac tcttattagc
    12541 agacaggaac gaggacatta ttatcatctg ctgcttggtg cacgataact tggtgcgttt
    12601 gtcaagcaag gtaagtggac gacccggtca taccttctta agttcgccct tcctcccttt
    12661 atttcagatt caatctgact tacctattct acccaagcat ccaaatggcc aagttgacca
    12721 gtgccgttcc ggtgctcacc gcgcgcgacg tcgccggagc ggtcgagttc tggaccgacc
    12781 ggctcgggtt ctcccgggac ttcgtggagg acgacttcgc cggtgtggtc cgggacgacg
    12841 tgaccctgtt catcagcgcg gtccaggacc aggtggtgcc ggacaacacc ctggcctggg
    12901 tgtgggtgcg cggcctggac gagctgtacg ccgagtggtc ggaggtcgtg tccacgaact
    12961 tccgggacgc ctccgggccg gccatgaccg agatcggcga gcagccgtgg gggcgggagt
    13021 tcgccctgcg cgacccggcc ggcaactgcg tgcacttcgt ggccgaggag caggactgac
    13081 cgacgccgac caacaccgcc ggtccgacgc ggcccgacgg gtccgaggag cttgagatcc
    13141 acttaacgtt actgaaatca tcaaacagct tgacgaatct ggatataaga tcgttggtgt
    13201 cgatgtcagc tccggagttg agacaaatgg tgttcaggat ctcgataaga tacgttcatt
    13261 tgtccaagca gcaaagagtg ccttctagtg atttaatagc tccatgtcaa caagaataaa
    13321 acgcgtttcg ggtttacctc ttccagatac agctcatctg caatgcatta atgcattgga
    13381 cctcgcaacc ctagtacgcc cttcaggctc cggcgaagca gaagaatagc ttagcagagt
    13441 ctattttcat tttcgggaga cgagatcaag cagatcaacg gtcgtcaaga gacctacgag
    13501 actgaggaat ccgctcttgg ctccacgcga ctatatattt gtctctaatt gtactttgac
    13561 atgctcctct tctttactct gatagcttga ctatgaaaat tccgtcacca gcccctgggt
    13621 tcgcaaagat aattgcactg tttcttcctt gaactctcaa gcctacagga cacacattca
    13681 tcgtaggtat aaacctcgaa aatcattcct actaagatgg gtatacaata gtaaccatgc
    13741 atggttgcct agtgaatgct ccgtaacacc caatacgccg gccgaaactt ttttacaact
    13801 ctcctatgag tcgtttaccc agaatgcaca ggtacacttg tttagaggta aatccccact
    13861 accgcattaa gacctcagcg cggccgcaaa tttaaataaa atgaagtgaa gttcctatac
    13921 tttctagaga ataggaactt ctatagtgag tcgaataagg gcgacacaaa atttattcta
    13981 aatgcataat aaatactgat aacatcttat agtttgtatt atattttgta ttatcgttga
    14041 catgtataat tttgatatca aaaactgatt ttccctttat tattttcgag atttattttc
    14101 ttaattctct ttaacaaact agaaatattg tatatacaaa aaatcataaa taatagatga
    14161 atagtttaat tataggtgtt catcaatcga aaaagcaacg tatcttattt aaagtgcgtt
    14221 gcttttttct catttataag gttaaataat tctcatatat caagcaaagt gacaggcgcc
    14281 cttaaatatt ctgacaaatg ctctttccct aaactccccc cataaaaaaa cccgccgaag
    14341 cgggttttta cgttatttgc ggattaacga ttactcgtta tcagaaccgc ccagggggcc
    14401 cgagcttaag actggccgtc gttttacaac acagaaagag tttgtagaaa cgcaaaaagg
    14461 ccatccgtca ggggccttct gcttagtttg atgcctggca gttccctact ctcgccttcc
    14521 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
    14581 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
    14641 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
    14701 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
    14761 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
    14821 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
    14881 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
    14941 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
    15001 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
    15061 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtgggctaac
    15121 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
    15181 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
    15241 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
    15301 ttttctacgg ggtctgacgc tcagtggaac gacgcgcgcg taactcacgt taagggattt
    15361 tggtcatgag cttgcgccgt cccgtcaagt cagcgtaatg ctctgctt
//