LOCUS       sequence-310773-       16442 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Proof-of-concept of CRISPR/dSpCas9-VPR activation, encoding 
            PelcA-mCherry along with the sgRNA E1 expression cassette, targeting
            PelcA..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 16442)
  AUTHORS   Roux I, Woodcraft C, Hu J, Wolters R, Gilchrist CLM, Chooi YH
  TITLE     CRISPR-mediated activation of biosynthetic gene clusters for 
            bioactive molecule discovery in filamentous fungi.
  JOURNAL   ACS Synth Biol. 2020 Jun 11. doi: 10.1021/acssynbio.0c00197.
  PUBMED    32526136
REFERENCE   2  (bases 1 to 16442)
  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: "ACS Synth 
            Biol. 2020 Jun 11. doi: 10.1021/acssynbio.0c00197."
COMMENT     Sequence Label: 
            pCRI018-pKW20088-PelcA-mCherry-TtrpC-PU3-E1-Cas9Scaffold-8xT
FEATURES             Location/Qualifiers
     source          1..16442
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     2802..2823
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             3982..4692
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKGGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(4127..4145)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4395..4414
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_RNA        5489..5564
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             join(8526..8594,8595..9386)
                     /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     complement(8744..8763)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9557..10145
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10046..10065
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10299..10316
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    10433..10454
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(10469..10474,10475..10492,10493..10499)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10507..10523
                     /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)."
     primer_bind     10512..10534
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     10531..10547
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     10531..10547
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      10564..11906
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(11527..11574)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     primer_bind     11947..11965
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(12003..12025)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     12125..12144
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        12168..12383
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             12384..13187
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     primer_bind     complement(13469..13486)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(13469..13485)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(13478..13500)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(13785..13804)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      join(13826..16442,1..2632)
                     /label=AMA1
                     /note="replication origin from Aspergillus nidulans 
                     (Aleksenko and Clutterbuck, 1996)"
ORIGIN
        1 tgcagatttt gacctagtta acaaggcaca tcatgctatc acttccaaca ctcttgctga
       61 gcgcgttctg ccttgccttt ctggttctac gaagtatcca gtcccgcctg caacgccgca
      121 gaaacgccaa aaggtggggt tgtcaaccgc cagctagagc tccatcggga ttctttggca
      181 ttccgccgtt cctcggtggt cgcagataac tccctaaatt ccctgagcta actctaagtc
      241 gaccatgccg tttatggtta gcgcctccca aaaaggaatg gccgacttag agttacctct
      301 tgaccgactt tttctttcct cccccttaca tttcgttacc acaacacatt cctatatcca
      361 aactccaggt acataactag tcgaaatctc ttttaaatct agtcaagaac tagatactta
      421 accttcatac tgcttagaga tagcctttga agctcttatt tcgctctttg tctcactctc
      481 accttcctcc tcctcctcca atcctttttg ctcgggacta gtccaagact agtcaacgat
      541 tagtatgccg agcattcgcg ataacgatct ccgaaagtcc ccagagtact gtcactatct
      601 cgaggcagtt aaggacgggg agcttacgct gccggatttc aagatagtaa gccgacccga
      661 ctagctttaa gcctagttac tgactagtcc acaggacgac aatggcgtgc ctgatatcca
      721 tccatatgaa gtctactgcc gagtgaaggg atgcctcaag cgtacagtga gtctactgct
      781 atactagttt ctgactagtt attaactagt tgacaggttc cctctgccaa cagaaacata
      841 ttggtcaagc acttgaagga caagaactcc cacggcatgg agtttacatt gcacaatgga
      901 cctcccacta tgaaggaact gatggaggcc aaaggcaagt cctatctaga ttacttggtg
      961 actagtccct gactagtcaa gtagcatggt atgaaggctt gtttgaaggc actgttctcc
     1021 caaccccgac tcctaccaag aagcgcaagc gagctgcgta agtttctgtg agtctaacta
     1081 gtgtattagc taatatatag cagcaccaag tccaaggacc acaatactaa gggtgtcgag
     1141 aattcgtgag tttcttctcc catttcaact agtccttgac tagtcactaa ctactttgca
     1201 gcaacgaggg tgaagctgga aatgatcagg acaatggcga gggcccgtaa gtacagccat
     1261 tcaatgcaga ctagttgcta actaatctgt gactagttca agtggtccgt acgccgtgca
     1321 tacccctgtg actggtagga atttgagcaa gcctgtcttg ccgcgcgatg agaaaggaaa
     1381 ggcaagttac attcagcccc gtacctagga tcagtgctaa tttataacct ctagccactc
     1441 tttatgcaga tccgccgtga gggtagcaag gcagctaaat cagctggtga gaaaggaacc
     1501 ataccctgca agacctgtcg caacgcaaag ggcaaaggta agctatccaa gctagtttgg
     1561 gactagattc taactagtct cagcaccgtg tggttcaaag ccatattgcg agttttggcg
     1621 ctttttctca tcgattgacg aggcaaaggg agcgagtatg caacctcgta agtcagactc
     1681 agacaaagac caactagtta ttgaccagtc ttctagaagg ctctgttgtg gatcttgagg
     1741 ccctggagag ttcctccaac aatccggaga caagcaagtc ctcgtcggac tagtcactaa
     1801 ctagactcta actagttgca gacatggata atgcaaaaga gacaagcaat gaagaaagtg
     1861 gtaagacatt tctcctttgt ggttctggac tagtctttga ctagtcacag tcttaaacaa
     1921 ggaaaatgag catgaaaatg aggaggaaaa ggctgctgag cccgaggaag tgcagggtga
     1981 tggcagacat ggtaggttaa taccttgtta gttattgcta gtcactgact agtcaataac
     2041 tagtctctga acaccttgca atcactccgt ttgcgcagct gaacagtggt gaggataata
     2101 gtagtaagtt attctagctt cagagttata ggagactaga tactaactag tattagttgc
     2161 aactaacctg gatctcagag actttggcct caatctagaa tctatctagt tgtcaactag
     2221 actgtggtat cattgtcttt tattttccta gtcctggaac tagcttctaa ctagtctccc
     2281 taatatgtgg ctgtcttgtt tttttttttt gtttccctac ccggatatct agtccccttc
     2341 taggttctgt taacctctcg ggctctgatt tagtttaacg caaacctgag attagtttct
     2401 aactagtctc taggttttct atccaccttt aattgtaata ataaatacaa gcaacgttta
     2461 tacgtcaaaa gcatttataa acttttaccc taaagtagct tgcttgtgtg tttagtttat
     2521 aattagtctc ttattaattt gatgtaggta agcccgccac aaatatatat ttttaacaag
     2581 ataccgtgga aaaacttcgt gctatcacaa aacagtatac aaaaaataag ctggctttcc
     2641 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     2701 cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac
     2761 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     2821 tggaacaaca ctcaactcta tctcgggcta ttcttttgat ttataaggga ttttgccgat
     2881 ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     2941 aatggggtga cgatgagccg ctcttgcatc tttgtttgta ttatactgtc tttcttgtta
     3001 cacataatta ttctagaatg ccccaccgtt acatacggga cacagccatt tacatatgca
     3061 tgtggattac gagctaacga gttcattcaa atctcagaac tatcacataa tcatcattcc
     3121 cctatcgtca aagaccgtaa gacaaatccg gttcatgcac tgaacccatt cgggtagtga
     3181 gtcatttact cagcacactc gcgctgacgc tcgtcgaaca ccttcaatgc ctcctcggca
     3241 gccttgacac cactgagaac catggcaccg aaggtagggc ccatgcggtt aaagccatca
     3301 atttcagaca gctccatacc gccgattatc aagcccttag taacctcgcg ggtgttcttg
     3361 acgatggcat cctgccgatg tacagtgctg aggcacgtca tactatgtat tagtagagga
     3421 tctagttgcc aatatcacgg agagcggaca gcttggtgtt gacaagcgcg taagctttac
     3481 catgatgtta gtgatgatct cgatctaccc agatgcatgg cttgagacac gtacccatgg
     3541 tggtcttggg tagctggaat ggggccagcg gccagtgccg gctcaccaat cgccagtgga
     3601 tgaagctcga tcgcgaccag ctagatcgat gctcagtact atgatgagtg gaatgaaagc
     3661 cttacaacca ggttttcggc caaaattggc cgcagttgcc agtacagtgt tgagtaaact
     3721 tcgtcatgga gcgtttcgat ggtggtgcta gcttctcgat catcagtgcc atgatacttt
     3781 cacccgtgcc aaagtgacga ccagctccat ataatatgac ggtatggacg gtgctgtcta
     3841 tccagattca tctctgatca gcgcaatcgt ttcaacatcc ttgtgtgaat attcaccaga
     3901 cagtcgttaa taccctttta gtcacccttc gaagctatta ttgtctcagc cactacgtca
     3961 ctcaacttgc gttatctcaa tatggtgagc aagggcgagg aggataacat ggccatcatc
     4021 aaggagttca tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca cgagttcgag
     4081 atcgagggcg agggcgaggg ccgcccctac gagggcaccc agaccgccaa gctgaaggtg
     4141 accaagggtg gccccctgcc cttcgcctgg gacatcctgt cccctcagtt catgtacggc
     4201 tccaaggcct acgtgaagca ccccgccgac atccccgact acttgaagct gtccttcccc
     4261 gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc
     4321 caggactcct ccctgcagga cggcgagttc atctacaagg tgaagctgcg cggcaccaac
     4381 ttcccctccg acggccccgt aatgcagaag aagaccatgg gctgggaggc ctcctccgag
     4441 cggatgtacc ccgaggacgg cgccctgaag ggcgagatca agcagaggct gaagctgaag
     4501 ggcggcggcc actacgacgc tgaggtcaag accacctaca aggccaagaa gcccgtgcag
     4561 ctgcccggcg cctacaacgt caacatcaag ttggacatca cctcccacaa cgaggactac
     4621 accatcgtgg aacagtacga acgcgccgag ggccgccact ccaccggcgg catggacgag
     4681 ctgtacaagt aatgggcgac gacgacgttc gagggggggc ccggtaccgc cccgtccggt
     4741 cctgcccgtc accgagatcc acttaacgtt actgaaatca tcaaacagct tgacgaatct
     4801 ggatataaga tcgttggtgt cgatgtcagc tccggagttg agacaaatgg tgttcaggat
     4861 ctcgataaga tacgttcatt tgtccaagca gcaaagagtg ccttctagtg atttaatagc
     4921 tccatgtcaa caagaataaa acgcgtttcg ggtttacctc ttccagatac agctcatctg
     4981 caatgcatga cgttaattaa gatcacatag atgctcggtt gacaggacca cctatagcaa
     5041 gtactttgta ggaggttctc ctaacgcttg gcttcaaatc caatcaggat atcagtgagt
     5101 tactgtttgt catcgcatcc tcatgaagtc gctcaacagc cactgagaag caaatatttg
     5161 ggagaccatc cctgatgttg aaattttacg ctggagccca ttcaccggtg agctcgggga
     5221 cagtctggcc agtggagcgg aaaatcttca actaactctg attggtacca caggtagccc
     5281 cacagaacaa accgagcaaa catgaaaatt ttcgcttgag gttagcgcac tcgctagcgc
     5341 ctgcccgcaa atataaaagg ccccgaaatc ccatgcattt tggaaatcaa gcgactctac
     5401 gtatgtcatc cccctcccta cctttgggaa aacctctcaa ctgcaagtca gaacattttg
     5461 ctaacagcac cagctccata taatatgagt tttagagcta gaaatagcaa gttaaaataa
     5521 ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg gtgctttttt ttcaattcgc
     5581 ggccgcccag aaaattacgg gtatgtcctc cactgtgttg aagtcagaaa aacaatcgat
     5641 aatttgaagc tcattaaaat gccagttccg tcgtcgccgt tgccctagga gctattccag
     5701 tgcttagctt cattcatgga tgtatcgtga ctggccttcg gaaggatgcc aaggtcccct
     5761 atccgcacag ctatgcgacc gttgagcaat gcaagtcaaa tgtgtgcata cctggtcttt
     5821 ggtttggtac accgctatgt gaagattatt aatgctgaag tcatgataca ggccaaagcc
     5881 gagcaattta actgcgcgca gcgggcccac gctaactacc tcgagaatat gcctcaaaca
     5941 atgctcttca ccctcttcgc gggtctgaaa taccctcacc tggcaacagc aattggcgct
     6001 tcatggctgt ttttccgatc tctctacttg tacggctatg tgtactcggg taagccacaa
     6061 ggcaagggca gattgctggg aggtttcttc tggttttctc aaggcgctct gtgggctctg
     6121 agtgtgtttg gtgttgccaa agacatgatc tcttactgag agttattctg tgtctgacga
     6181 aatatgttgt gtatatatat atatgtacgt taaaagttcc gtggagttac cagtgattga
     6241 ccaatgtttt atcttctaca gttctgcctg tctaccccat tctagctgta cctgactaca
     6301 gagtagttta attgtggttg accccacagt cggaggcgga ggaatacagc accgatgtgg
     6361 cctgtctcca tccagattgg cacgcaattt ttacacgcgg aaaagatcga gatagagtac
     6421 gactttaaat ttagtccccg gcggcttcta ttttagaata tttgagattt gattctcaag
     6481 caattgattt ggttgggtca ccctcaattg gataatatac ctcattgctc ggctacttca
     6541 actcatcaat caccgtcata ccccgcatat aaccctccat tcccacgatg tcgtccaagt
     6601 cgcaattgac ttacggtgct cgagccagca agcaccccaa tcctctggca aagagacttt
     6661 ttgagattgc cgaagcaaag aagacaaacg ttaccgtctc tgctgatgtg acgacaaccc
     6721 gagaactcct ggacctcgct gaccgtacgg aagctgttgg atccaataca tatgccgtct
     6781 agcaatggac taatcaactt ttgatgatac aggtctcggt ccctacatcg ccgtcatcaa
     6841 gacacacatc gacatcctca ccgatttcag cgtcgacact atcaatggcc tgaatgtgct
     6901 ggctcaaaag cacaactttt tgatcttcga ggaccgcaaa ttcatcgaca tcggcaatac
     6961 cgtccagaag caataccacg gcggtgctct gaggatctcc gaatgggccc acattatcaa
     7021 ctgcagcgtt ctccctggcg agggcatcgt cgaggctctg gcccagaccg catctgcgca
     7081 agacttcccc tatggtcctg agagaggact gttggtcctg gcagagatga cctccaaagg
     7141 atcgctggct acgggcgagt ataccaaggc atcggttgac tacgctcgca aatacaagaa
     7201 cttcgttatg ggtttcgtgt cgacgcgggc cctgacggaa gtgcagtcgg atgtgtcttc
     7261 agcctcggag gatgaagatt tcgtggtctt cacgacgggt gtgaacctct cttccaaagg
     7321 agataagctt ggacagcaat accagactcc tgcatcggct attggacgcg gtgccgactt
     7381 tatcatcgcc ggtcgaggca tctacgctgc tcccgacccg gttgaagctg cacagcggta
     7441 ccagaaagaa ggctgggaag cttatatggc cagagtatgc ggcaagtcat gatttcctct
     7501 tggagcaaaa gtgtagtgcc agtacgagtg ttgtggagga aggctgcata cattgtgcct
     7561 gtcattaaac gatgagctcg tccgtattgg cccctgtaat gccatgtttt ccgcccccaa
     7621 tcgtcaaggt tttccctttg ttagattcct accagtcatc tagcaagtga ggtaagcttt
     7681 gccagaaacg ccaaggcttt atctatgtag tcgataagca aagtggactg atagcttaat
     7741 atggaaggtc cctcaggaca agtcgacctg tgcagaagag ataacagctt ggcatcacgc
     7801 atcagtgcct cctctcagac agaataagtc gccataagtt atcgaccgaa cctaggtagg
     7861 gtatatttgg tgacgacaat acctcccgac gatacctgga ttagacaccg tatgccgcga
     7921 attaacgcct tccgaccaag tcccttttgg tgatcgacac tttaggatca tctcctgcca
     7981 gtcgcccaga ctgacggtgt tatggtggcg cgctatggct ttcctagccg gtcaagctcc
     8041 gtgcactaga gagcggagaa ggagggggag ttgataggga atgatatcta ggtagagaag
     8101 tactccaacc agagatatgc ggatcaatgc tccggcagtt gttagcatac cccaacctcg
     8161 tttaagtatc ctcttcttcc atccatcttt atcattattt gtctatgtgc atccctagat
     8221 cacgtcaatt atttcgataa atggatacgt gcactcggga atgcgtgatt ggctcgtctc
     8281 gtgttctcag ttcctcattc ccccccacgt tacattgaca acggagtatt agcctatggt
     8341 cctgttcgat aagcttgctc catcgcctcc actacgtcaa gcggagtatc agacatgagc
     8401 tctaccccag atatcgcctg cttatcgctg gttgctttgt catgcaagta cagatcacat
     8461 cggcaattct gataacagat ggtgccattg tcccgtaatc aattgcccat tgaaaaagga
     8521 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     8581 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     8641 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     8701 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     8761 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     8821 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     8881 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     8941 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     9001 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     9061 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     9121 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     9181 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     9241 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     9301 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     9361 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     9421 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     9481 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     9541 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     9601 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     9661 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     9721 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     9781 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     9841 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     9901 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     9961 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
    10021 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
    10081 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
    10141 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
    10201 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
    10261 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
    10321 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
    10381 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
    10441 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
    10501 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca
    10561 ctgaacgaag catctgtgct tcattttgta gaacaaaaat gcaacgcgag agcgctaatt
    10621 tttcaaacaa agaatctgag ctgcattttt acagaacaga aatgcaacgc gaaagcgcta
    10681 ttttaccaac gaagaatctg tgcttcattt ttgtaaaaca aaaatgcaac gcgagagcgc
    10741 taatttttca aacaaagaat ctgagctgca tttttacaga acagaaatgc aacgcgagag
    10801 cgctatttta ccaacaaaga atctatactt cttttttgtt ctacaaaaat gcatcccgag
    10861 agcgctattt ttctaacaaa gcatcttaga ttactttttt tctcctttgt gcgctctata
    10921 atgcagtctc ttgataactt tttgcactgt aggtccgtta aggttagaag aaggctactt
    10981 tggtgtctat tttctcttcc ataaaaaaag cctgactcca cttcccgcgt ttactgatta
    11041 ctagcgaagc tgcgggtgca ttttttcaag ataaaggcat ccccgattat attctatacc
    11101 gatgtggatt gcgcatactt tgtgaacaga aagtgatagc gttgatgatt cttcattggt
    11161 cagaaaatta tgaacggttt cttctatttt gtctctatat actacgtata ggaaatgttt
    11221 acattttcgt attgttttcg attcactcta tgaatagttc ttactacaat ttttttgtct
    11281 aaagagtaat actagagata aacataaaaa atgtagaggt cgagtttaga tgcaagttca
    11341 aggagcgaaa ggtggatggg taggttatat agggatatag cacagagata tatagcaaag
    11401 agatactttt gagcaatgtt tgtggaagcg gtattcgcaa tattttagta gctcgttaca
    11461 gtccggtgcg tttttggttt tttgaaagtg cgtcttcaga gcgcttttgg ttttcaaaag
    11521 cgctctgaag ttcctatact ttctagagaa taggaacttc ggaataggaa cttcaaagcg
    11581 tttccgaaaa cgagcgcttc cgaaaatgca acgcgagctg cgcacataca gctcactgtt
    11641 cacgtcgcac ctatatctgc gtgttgcctg tatatatata tacatgagaa gaacggcata
    11701 gtgcgtgttt atgcttaaat gcgtacttat atgcgtctat ttatgtagga tgaaaggtag
    11761 tctagtacct cctgtgatat tatcccattc catgcggggt atcgtatgct tccttcagca
    11821 ctacccttta gctgttctat atgctgccac tcctcaattg gattagtctc atccttcaat
    11881 gctatcattt cctttgatat tggatcatac taagaaacca ttattatcat gacattaacc
    11941 tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa
    12001 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg
    12061 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac
    12121 tatgcggcat cagagcagat tgtactgaga gtgcaccata ccaccttttc aattcatcat
    12181 ttttttttta ttcttttttt tgatttcggt ttccttgaaa tttttttgat tcggtaatct
    12241 ccgaacagaa ggaagaacga aggaaggagc acagacttag attggtatat atacgcatat
    12301 gtagtgttga agaaacatga aattgcccag tattcttaac ccaactgcac agaacaaaaa
    12361 cctgcaggaa acgaagataa atcatgtcga aagctacata taaggaacgt gctgctactc
    12421 atcctagtcc tgttgctgcc aagctattta atatcatgca cgaaaagcaa acaaacttgt
    12481 gtgcttcatt ggatgttcgt accaccaagg aattactgga gttagttgaa gcattaggtc
    12541 ccaaaatttg tttactaaaa acacatgtgg atatcttgac tgatttttcc atggagggca
    12601 cagttaagcc gctaaaggca ttatccgcca agtacaattt tttactcttc gaagacagaa
    12661 aatttgctga cattggtaat acagtcaaat tgcagtactc tgcgggtgta tacagaatag
    12721 cagaatgggc agacattacg aatgcacacg gtgtggtggg cccaggtatt gttagcggtt
    12781 tgaagcaggc ggcagaagaa gtaacaaagg aacctagagg ccttttgatg ttagcagaat
    12841 tgtcatgcaa gggctcccta tctactggag aatatactaa gggtactgtt gacattgcga
    12901 agagcgacaa agattttgtt atcggcttta ttgctcaaag agacatgggt ggaagagatg
    12961 aaggttacga ttggttgatt atgacacccg gtgtgggttt agatgacaag ggagacgcat
    13021 tgggtcaaca gtatagaacc gtggatgatg tggtctctac aggatctgac attattattg
    13081 ttggaagagg actatttgca aagggaaggg atgctaaggt agagggtgaa cgttacagaa
    13141 aagcaggctg ggaagcatat ttgagaagat gcggccagca aaactaaaaa actgtattat
    13201 aagtaaatgc atgtatacta aactcacaaa ttagagcttc aatttaatta tatcagttat
    13261 taccctatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggaaa
    13321 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
    13381 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
    13441 ggttgagtgg ggtaccgagc tcgaattcac tggccgtcgt tttacaacgt cgtgactggg
    13501 aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc
    13561 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
    13621 aatggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac
    13681 gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
    13741 tacgcgcagc gtgaccgcta cacttgccag cgccttagcg cccgctcctt tcgctttctt
    13801 cccttccttt ctcgccacgt tcgccagctt attttttgta tactgttttg tgatagcacg
    13861 aagtttttcc acggtatctt gttaaaaata tatatttgtg gcgggcttac ctacatcaaa
    13921 ttaataagag actaattata aactaaacac acaagcaagc tactttaggg taaaagttta
    13981 taaatgcttt tgacgtataa acgttgcttg tatttattat tacaattaaa ggtggataga
    14041 aaacctagag actagttaga aactaatctc aggtttgcgt taaactaaat cagagcccga
    14101 gaggttaaca gaacctagaa ggggactaga tatccgggta gggaaacaaa aaaaaaaaac
    14161 aagacagcca catattaggg agactagtta gaagctagtt ccaggactag gaaaataaaa
    14221 gacaatgata ccacagtcta gttgacaact agatagattc tagattgagg ccaaagtctc
    14281 tgagatccag gttagttgca actaatacta gttagtatct agtctcctat aactctgaag
    14341 ctagaataac ttactactat tatcctcacc actgttcagc tgcgcaaacg gagtgattgc
    14401 aaggtgttca gagactagtt attgactagt cagtgactag caataactaa caaggtatta
    14461 acctaccatg tctgccatca ccctgcactt cctcgggctc agcagccttt tcctcctcat
    14521 tttcatgctc attttccttg tttaagactg tgactagtca aagactagtc cagaaccaca
    14581 aaggagaaat gtcttaccac tttcttcatt gcttgtctct tttgcattat ccatgtctgc
    14641 aactagttag agtctagtta gtgactagtc cgacgaggac ttgcttgtct ccggattgtt
    14701 ggaggaactc tccagggcct caagatccac aacagagcct tctagaagac tggtcaataa
    14761 ctagttggtc tttgtctgag tctgacttac gaggttgcat actcgctccc tttgcctcgt
    14821 caatcgatga gaaaaagcgc caaaactcgc aatatggctt tgaaccacac ggtgctgaga
    14881 ctagttagaa tctagtccca aactagcttg gatagcttac ctttgccctt tgcgttgcga
    14941 caggtcttgc agggtatggt tcctttctca ccagctgatt tagctgcctt gctaccctca
    15001 cggcggatct gcataaagag tggctagagg ttataaatta gcactgatcc taggtacggg
    15061 gctgaatgta acttgccttt cctttctcat cgcgcggcaa gacaggcttg ctcaaattcc
    15121 taccagtcac aggggtatgc acggcgtacg gaccacttga actagtcaca gattagttag
    15181 caactagtct gcattgaatg gctgtactta cgggccctcg ccattgtcct gatcatttcc
    15241 agcttcaccc tcgttgctgc aaagtagtta gtgactagtc aaggactagt tgaaatggga
    15301 gaagaaactc acgaattctc gacaccctta gtattgtggt ccttggactt ggtgctgcta
    15361 tatattagct aatacactag ttagactcac agaaacttac gcagctcgct tgcgcttctt
    15421 ggtaggagtc ggggttggga gaacagtgcc ttcaaacaag ccttcatacc atgctacttg
    15481 actagtcagg gactagtcac caagtaatct agataggact tgcctttggc ctccatcagt
    15541 tccttcatag tgggaggtcc attgtgcaat gtaaactcca tgccgtggga gttcttgtcc
    15601 ttcaagtgct tgaccaatat gtttctgttg gcagagggaa cctgtcaact agttaataac
    15661 tagtcagaaa ctagtatagc agtagactca ctgtacgctt gaggcatccc ttcactcggc
    15721 agtagacttc atatggatgg atatcaggca cgccattgtc gtcctgtgga ctagtcagta
    15781 actaggctta aagctagtcg ggtcggctta ctatcttgaa atccggcagc gtaagctccc
    15841 cgtccttaac tgcctcgaga tagtgacagt actctgggga ctttcggaga tcgttatcgc
    15901 gaatgctcgg catactaatc gttgactagt cttggactag tcccgagcaa aaaggattgg
    15961 aggaggagga ggaaggtgag agtgagacaa agagcgaaat aagagcttca aaggctatct
    16021 ctaagcagta tgaaggttaa gtatctagtt cttgactaga tttaaaagag atttcgacta
    16081 gttatgtacc tggagtttgg atataggaat gtgttgtggt aacgaaatgt aagggggagg
    16141 aaagaaaaag tcggtcaaga ggtaactcta agtcggccat tcctttttgg gaggcgctaa
    16201 ccataaacgg catggtcgac ttagagttag ctcagggaat ttagggagtt atctgcgacc
    16261 actggactct caggctttcc attggggtac taacatagcc atcaaatgcc ggcccgtgtt
    16321 taatctcctc gagatggctg ggctcacatg cgttctactc aaaaactctt atacctaggc
    16381 cctctcccaa tcgtgtatgt ttggaccttc ctctctcaca ctttggataa catcaagcat
    16441 ac
//
