LOCUS       sequence-413976-       15653 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cloning in complete guide expression cassettes, with Cas12a (D156R).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15653)
  AUTHORS   Lawrenson T, Clarke M, Kirby R, Forner M, Steuernagel B, Harwood W
  TITLE     An Optimised CRISPR Cas9 and Cas12a Toolkit for Barley and Wheat
  JOURNAL   Research Square, 12 March 2024, PREPRINT
REFERENCE   2  (bases 1 to 15653)
  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: "Research 
            Square, 12 March 2024, PREPRINT"
COMMENT     Sequence Label: EC67842
FEATURES             Location/Qualifiers
     source          1..15653
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    32..56
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             365..994
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             1423..2496
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGASSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     rep_origin      2562..2756
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     promoter        2818..2922
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2923..3717
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      3888..4476
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4377..4396
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    4612..4636
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     polyA_signal    4724..4898
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     intron          5620..5809
                     /label=cat1 intron
                     /note="castor bean catalase intron, modified"
     promoter        complement(6239..6584)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(6276..6296)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     terminator      complement(7600..7852)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(7868..7915)
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     protein_bind    14646..14667
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(14682..14687,14688..14705,14706..14712)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    14720..14736
                     /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)."
     RBS             14754..14765
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             14772..15449
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      15494..15565
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      15581..15608
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
ORIGIN
        1 cacgaagtga tccgtttaaa ctatcagtgt ttgacaggat atattggcgg gtaaacctaa
       61 gagaaaagag cgtttattag aataatcgga tatttaaaag ggcgtgaaaa ggtttatccg
      121 ttcgtccatt tgtatgtgca tgccaaccac agggttcccc tcgggagtca gccgtgcggc
      181 tgcatgaaat cctggccggt ttgtctgatg ccaagctggc ggcctggccg gccagcttgg
      241 ccgctgaaga aaccgagcgc cgccgtctaa aaaggtgatg tgtatttgag taaaacagct
      301 tgcgtcatgc ggtcgctgcg tatatgatgc gatgagtaaa taaacaaata cgcaagggga
      361 acgcatgaag gttatcgctg tacttaacca gaaaggcggg tcaggcaaga cgaccatcgc
      421 aacccatcta gcccgcgccc tgcaactcgc cggggccgat gttctgttag tcgattccga
      481 tccccagggc agtgcccgcg attgggcggc cgtgcgggaa gatcaaccgc taaccgttgt
      541 cggcatcgac cgcccgacga ttgaccgcga cgtgaaggcc atcggccggc gcgacttcgt
      601 agtgatcgac ggagcgcccc aggcggcgga cttggctgtg tccgcgatca aggcagccga
      661 cttcgtgctg attccggtgc agccaagccc ttacgacata tgggccaccg ccgacctggt
      721 ggagctggtt aagcagcgca ttgaggtcac ggatggaagg ctacaagcgg cctttgtcgt
      781 gtcgcgggcg atcaaaggca cgcgcatcgg cggtgaggtt gccgaggcgc tggccgggta
      841 cgagctgccc attcttgagt cccgtatcac gcagcgcgtg agctacccag gcactgccgc
      901 cgccggcaca accgttcttg aatcagaacc cgagggcgac gctgcccgcg aggtccaggc
      961 gctggccgct gaaattaaat caaaactcat ttgagttaat gaggtaaaga gaaaatgagc
     1021 aaaagcacaa acacgctaag tgccggccgt ccgagcgcac gcagcagcaa ggctgcaacg
     1081 ttggccagcc tggcagacac gccagccatg aagcgggtca actttcagtt gccggcggag
     1141 gatcacacca agctgaagat gtacgcggta cgccaaggca agaccattac cgagctgcta
     1201 tctgaataca tcgcgcagct accagagtaa atgagcaaat gaataaatga gtagatgaat
     1261 tttagcggct aaaggaggcg gcatggaaaa tcaagaacaa ccaggcaccg acgccgtgga
     1321 atgccccatg tgtggaggaa cgggcggttg gccaggcgta agcggctggg ttgtctgccg
     1381 gccctgcaat ggcactggaa cccccaagcc cgaggaatcg gcgtgagcgg tcgcaaacca
     1441 tccggcccgg tacaaatcgg cgcggcgctg ggtgatgacc tggtggagaa gttgaaggcc
     1501 gcgcaggccg cccagcggca acgcatcgag gcagaagcac gccccggtga atcgtggcaa
     1561 gcggccgctg atcgaatccg caaagaatcc cggcaaccgc cggcagccgg tgcgtcgtcg
     1621 attaggaagc cgcccaaggg cgacgagcaa ccagattttt tcgttccgat gctctatgac
     1681 gtgggcaccc gcgatagtcg cagcatcatg gacgtggccg ttttccgtct gtcgaagcgt
     1741 gaccgacgag ctggcgaggt gatccgctac gagcttccag acgggcacgt agaggtttcc
     1801 gcagggccgg ccggcatggc cagtgtgtgg gattacgacc tggtactgat ggcggtttcc
     1861 catctaaccg aatccatgaa ccgataccgg gaagggaagg gagacaagcc cggccgcgtg
     1921 ttccgtccac acgttgcgga cgtactcaag ttctgccggc gagccgatgg cggaaagcag
     1981 aaagacgacc tggtagaaac ctgcattcgg ttaaacacca cgcacgttgc catgcagcgt
     2041 acgaagaagg ccaagaacgg ccgcctggtg acggtatccg agggtgaagc cttgattagc
     2101 cgctacaaga tcgtaaagag cgaaaccggg cggccggagt acatcgagat cgagctagct
     2161 gattggatgt accgcgagat tacagaaggc aagaacccgg acgtgctgac ggttcacccc
     2221 gattactttt tgatcgatcc cggcatcggc cgttttctct accgcctggc acgccgcgcc
     2281 gcaggcaagg cagaagccag atggttgttc aagacgatct acgaacgcag tggcagcgcc
     2341 ggagagttca agaagttctg tttcaccgtg cgcaagctga tcgggtcaaa tgacctgccg
     2401 gagtacgatt tgaaggagga ggcggggcag gctggcccga tcctagtcat gcgctaccgc
     2461 aacctgatcg agggcgaagc atccgccggt tcctaatgta cggagcagat gctagggcaa
     2521 attgccctag caggggaaaa aggtcgaaaa agcttctttc ctgtggatag cacgtacatt
     2581 gggaacccaa agccgtacat tgggaaccgg aacccgtaca ttgggaaccc aaagccgtac
     2641 attgggaacc ggtcacacat gtaagtgact gatataaaag agaaaaaagg cgatttttcc
     2701 gcctaaaact ctttaaaact tattaaaact cttaaaaccc gcctggcctg tgcataactg
     2761 tctggccagc gcacagccga acagctgcaa aaagcgccca catcaaggct ccgagtgcgc
     2821 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     2881 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatggctaa aatgagaata
     2941 tcaccggaat tgaaaaaact gatcgaaaaa taccgctgcg taaaagatac ggaaggaatg
     3001 tctcctgcta aggtatataa gctggtggga gaaaatgaaa acctatattt aaaaatgacg
     3061 gacagccggt ataaagggac cacctatgat gtggaacggg aaaaggacat gatgctatgg
     3121 ctggaaggaa agctgcctgt tccaaaggtc ctgcactttg aacggcatga tggctggagc
     3181 aatctgctca tgagtgaggc cgatggcgtc ctttgctcgg aagagtatga agatgaacaa
     3241 agccctgaaa agattatcga gctgtatgcg gagtgcatca ggctctttca ctccatcgac
     3301 atatcggatt gtccctatac gaatagctta gacagccgct tagccgaatt ggattactta
     3361 ctgaataacg atctggccga tgtggattgc gaaaactggg aagaggacac tccatttaaa
     3421 gatccgcgcg agctgtatga ttttttaaag acggaaaagc ccgaagagga acttgtcttt
     3481 tcccacggcg acctgggaga cagcaacatc tttgtgaaag atggcaaagt aagtggcttt
     3541 attgatcttg ggagaagcgg cagggcggac aagtggtatg acattgcctt ctgcgtccgg
     3601 tcgctcaggg aggatatcgg ggaagaacag tatgtcgagc tattttttga cttactgggg
     3661 atcaagcctg attgggagaa aataaaatat tatattttac tggatgaatt gttttagctg
     3721 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     3781 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     3841 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     3901 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     3961 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     4021 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     4081 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     4141 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     4201 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     4261 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     4321 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     4381 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     4441 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     4501 cggttcctgc tcggatctgt tggaccggac agtagtcatg gttgatgggc tgcctgtatc
     4561 gagtggtgat tttgtgccga gctgccggtc ggggagctgt tggctggctg gtggcaggat
     4621 atattgtggt gtaaacaaat tgacgcttag acaacttaat aacacattgc ggacgttttt
     4681 aatgtactgg ggttgaacac tctgtgccga attcggatcc agcgatctgg attttagtac
     4741 tggattttgg ttttaggaat tagaaatttt attgatagaa gtattttaca aatacaaata
     4801 catactaagg gtttcttata tgctcaacac atgagcgaaa ccctatagga accctaattc
     4861 ccttatctgg gaactactca cacattatta tggagaaact cgagcttgtc gatcgactct
     4921 agctagagaa gcctattcct ttgccctcgg acgagtgctg gggcgtcggt ttccactatc
     4981 ggcgagtact tctacacagc catcggtcca gacggccgcg cttctgcggg cgatttgtgt
     5041 acgcccgaca gtcccggctc cggatcggac gattgcgtcg catcgaccct gcgcccaagc
     5101 tgcatcatcg aaattgccgt caaccaagct ctgatagagt tggtcaagac caatgcggag
     5161 catatacgcc cggagccgcg gcgatcctgc aagctccgga tgcctccgct cgaagtagcg
     5221 cgtctgctgc tccatacaag ccaaccacgg cctccagaag aagatgttgg cgacctcgta
     5281 ttgggaatcc ccgaacatcg cctcgctcca gtcaatgacc gctgttatgc ggccattgtc
     5341 cgtcaggaca ttgttggagc cgaaatccgc gtgcacgagg tgccggactt cggggcagtc
     5401 ctcggcccaa agcatcagct catcgagagc ctgcgcgacg gacgcactga cggtgtcgtc
     5461 catcacagtt tgccagtgat acacatgggg atcagcaatc gcgcatatga aatcacgcca
     5521 tgtagtgtat tgaccgattc cttgcggtcc gaatgggccg aacccgctcg tctggctaag
     5581 atcggccgca gcgatcgcat ccatggcctc cgcgaccggc tgcagttatc atcatcatca
     5641 tagacacacg aaataaagta atcagattat cagttaaagc tatgtaatat ttacaccata
     5701 accaatcaat taaaaaatag atcagtttaa agaaagatca aagctcaaaa aaataaaaag
     5761 agaaaagggt cctaaccaag aaaatgaagg agaaaaacta gaaatttacc tgcagaacag
     5821 cgggcagttc ggtttcaggc aggtcttgca acgtgacacc ctgtgcacgg cgggagatgc
     5881 aataggtcag gctctcgctg aattccccaa tgtcaagcac ttccggaatc gggagcgcgg
     5941 ccgatgcaaa gtgccgataa acataacgat ctttgtagaa accatcggcg cagctattta
     6001 cccgcaggac atatccacgc cctcctacat cgaagctgaa agcacgagat tcttcgccct
     6061 ccgagagctg catcaggtcg gagacgctgt cgaacttttc gatcagaaac ttctcgacag
     6121 acgtcgcggt gagttcaggc tttttcattg tatcgataat tgtaaatgta attgtaatgt
     6181 tgtttgttgt ttgttgttgt tggtaattgt tgtaaaaatg agctcttata ctcgagcgtg
     6241 tcctctccaa atgaaatgaa cttccttata tagaggaagg gtcttgcgaa ggatagtggg
     6301 attgtgcgtc atcccttacg tcagtggaga tgtcacatca atccacttgc tttgtagacg
     6361 tggttggaac ctcttctttt tccacgatgc tcctcgtggg tgggggtcca tctttgggac
     6421 cactgtcggc agagagatct tgaatgatag cctttccttt atcgcaatga tggcatttgt
     6481 aggagccacc ttccttttct actgtccttt cgatgaagtg acagatagct gggcaatgga
     6541 atccgaggag gtttcccgaa attatccttt gttgaaaagt ctcaatagcc ctttgatctt
     6601 ctgagactgt atctttgaca tttttggagt agaccagagt gtcgtgctcc accatgttga
     6661 cgaagatttt cttcttgtca ttgagtcgta aaagactctg tatgaactgt tcgccagtct
     6721 tgacggcgag ttctgttaga tcctcgattt gaatcttaga ctccatgcat ggccttagat
     6781 tcagtaggaa ctaccttttt agagactcca atctctatta cttgccttgg tttatgaagc
     6841 aagccttgaa tcgtccatac tggaatagta cttctgatct tgagaaatat gtctttctct
     6901 gtgttcttga tgcaattagt cctgaatctt ttgactgcat ctttaacctt cttgggaagg
     6961 tatttgatct cctggagatt gttactcggg tagatcgtct tgatgagact tgctgcgtag
     7021 gcctctctaa ccatctgtgg gtcagcattc tttctgaaat tgaagaggct aaccttctca
     7081 ttatcagtgg tgaacatagt gtcgtcacct tcaccttcga acttccttcc tagatcgtaa
     7141 agatagagga aatcgtccat tgtaatctcc ggggcaaagg agatctcttt tggggctgga
     7201 tcactgctgg gccttttggt tcctagcgtg agccagtggg ctttttgctt tggtgggctt
     7261 gttagggcct tagcaaagct cttgggcttg agttgagctt ctcctttggg gatgaagttc
     7321 aacctgtctg tttgctgact tgttgtgtac gcgtcagctg ctgctcttgc ctctgtaata
     7381 gtggcaaatt tcttgtgtgc aactccggga acgccgtttg ttgccgcctt tgtacaaccc
     7441 cagtcatcgt atataccggc atgtggaccg ttatacacaa cgtagtagtt gatatgaggg
     7501 tgttgaatac ccgattctgc tctgagagga gcaactgtgc tgttaagctc agatttttgt
     7561 gggattggaa ttcctccgca agaattcaag cttagcgtcg atctagtaac atagatgaca
     7621 ccgcgcgcga taatttatcc tagtttgcgc gctatatttt gttttctatc gcgtattaaa
     7681 tgtataattg cgggactcta atcataaaaa cccatctcat aaataacgtc atgcattaca
     7741 tgttaattat tacatgctta acgtaattca acagaaatta tatgataatc atcgcaagac
     7801 cggcaacagg attcaatctt aagaaacttt attgccaaat gtttgaacga tctgcttgac
     7861 aagctcattt cttcttctta gcctgaccag ctttcttagt agcagcaggc ctcttgtgct
     7921 taacggaggt ctgagcgtac tcaagccact ctttgttgga gatggcgatc ttcaccttgt
     7981 ccagcttctc atcttccgct ttcttaaact gcccgatagc ccagagaacc tataacacgg
     8041 gagaaaccaa aaagcaaact catttcacgc caaatactac agatttctca aaaattaagt
     8101 tataatccat atatcctgac aagaaattga ttaaaacaca gttgctttac ctttctcgca
     8161 atgttgtaag ccccgtttgc atcagcattc ttcggcagga tagcgttctc ttgagcctca
     8221 tagttacggc tgtcgtagaa gatcccgtca gagttcttga ccggagagat aaggaagtcc
     8281 acatcggtcc ttccggtgat agagttcctc atctggagca tgagagacat gagggccatg
     8341 aatgagctgt agaatgcctt gtcgctctgc tcgcacaaaa gagccctgat atcaccctgc
     8401 tggtagttga tgccgtactt gttgaacagt tctttgtagg cagaggtgag gcacacttct
     8461 tcccagtcga acacgttgtt tttcttcgga tttcggaaga tcctgatacg gttcccgtag
     8521 gagtagagct tccacttctt aatgtagtcg gcgtcggtcc tgctgaagtt cttatagtcg
     8581 agagcgaact cgaaaagatc ctcttccggc acgtacatga tcctgtcgaa agacgagatg
     8641 aacttcttgc tgtcggcgat cgaggtgtac ttggtcttga gcaggttcac gaaaccagta
     8701 gaaggatcga tcttgctggt gagccaagca gggatataga agatgaagcc gttctgggtg
     8761 ctcatgctct tgaaggactc gaacttgttc gtgatctggt atcccttaag agcaccaccg
     8821 gtagcgcaag ggttgctctt tttgtccacc atgtagttga gcttgtcgat gagcatcttc
     8881 tcgaactttt ggtagacctg cttctcaacc tgtagcaata attaaaacaa aaataccaca
     8941 aacagttaga actgaggcaa gaagaatgaa tgcagagaaa aaaaatctta gctattcatg
     9001 gaaatggaag tgcgacgaac acctaaaatg caaggagcat aaccaaatgc agaacttacc
     9061 ttcactctac tgttcttgaa cccagagttg aggtcctcaa gagcgatcac agcatcgtac
     9121 ttttccacca gctcgcagat cttatgcacc acctgagaga tgtagccggc cttcaactct
     9181 ttgatgtttt cgatagaagt ccagttctgc ctggcctcga aacgctcttt ttctttctta
     9241 tcgaggagtg agtggtagtc ggtcttgatc ctgatgccgt tgaaattgtt gataatctcg
     9301 ttcaggctgt actgctcgac gatgtttccc tttccgtcca ccacaacgat gtagaggagg
     9361 tttctctcac cacgatcgat tccgatcacg taagggttat catcgtgctt gagcagcacc
     9421 ctcaccttta attgtagaaa accagtaaaa catggacaat taggaggcat cgattatcta
     9481 accaaagact ttaacaacac taatctgcca ctaatattca tgagaagtcc ttacctcggt
     9541 attgatcttg aagatgttct tcgggcactt gttgatagcg atcgggatat gaagctcgta
     9601 ctggtcctcg ctgaaccttt tgtccttgta gacgtcgtaa gacagcgtgg tggtcttttt
     9661 cgggttgtca ggattcttgt tagcgattgg agagttggcc gggtgaacaa ccaactcttc
     9721 tttcttgagt gaggccctac gcatgaagag ttcagcacca ccagaaagtc tgatctgccc
     9781 gtggttgttc tcgtcgaaaa gcagcttgaa gtacatcgta tggaggttag gagttccgtg
     9841 gctcttgtcg gagaagtcct tgttatagat ctggaacatg tagagcttgc cctcttcgac
     9901 caacttgtca acctgttata acagtaaaaa ttactttaga aacaaaacaa aagtgtggtc
     9961 ttttaggaat gattattgat cagttcatag tttaaaatct tgtgttttta tcgtaaaagt
    10021 gtggaccagg atttacctct ttcttggagg cgctctcgaa gctaactttg taaccctgtt
    10081 cctcaacctc acggtagaac ccggcgatat ccttgtactt ttctgtctcg ctgaagttaa
    10141 agtcgtaagc gttagaccac ttcgggtaac gcgagatgct gtccttgaag aaatcgatga
    10201 gcttgtggca gtcgttgagg ttgaacatgt cgcctttctt gaaggtcccg ttcttgtaga
    10261 ttttctggat gtcctcagac gggttgtagt aggccatcca tttcttgctg aagaacacct
    10321 taggaagcat cttgttcggt cccggaagga gcttgtagtt gatcttctca tagttcccgt
    10381 tcacgtcatc cttgtcgatc ttttggaggc acttggcgta tttcttgtcc atgatggcca
    10441 ggtagtactt agacccgtac ctaatgaatt aatagaaaaa tcaggcaaat tagaaaagaa
    10501 ataattctca aaagagaaca actgaggtca gaaaatagca ttcatcaact gaaactaggt
    10561 atttcttcat atgatactaa cctgaggatg gtggccctgt aatctgtctc tttgtctttg
    10621 tcccatccgc ccatgaactg cgggttctga aagtacaact tgaacttgtc cttgctgtaa
    10681 ggcttctggg tcacgtagtt tctgatggcg tcgtagatat ggtccacctt gaggaggata
    10741 tcgtaagcga gcacgaaatc cccgtagaaa gactcgtccc tattagtctc tttgccctct
    10801 ccgaagaagg ccttgatata gttctcgaag gacttcacgc tgtcgagcaa atccttcatg
    10861 atcgcgacga cagcgtcgtt cttcttcaga ctcttctcga ggacgaaatc agcatcgaac
    10921 aacttctcag aggacccgta cacctaataa tatcatcaga ggaaaactca attcttatat
    10981 cttacagaaa tgaacaacaa aatacatgta gcagagagga aaacaatcat aagtaacaac
    11041 ttaccttgta gatctcgtcg accttctgga tgatgatctc tttgagcttc tcgaccacgc
    11101 tgagatcagc atcagcgtat tcttgaagct gctcgaggct gaaagagccg atcttcttga
    11161 acgacttacg cctgtcatcc tcgtacttct cagtgacgac agccttcttc ttgaggtgga
    11221 tgtcatcgta ctcagcattc cacttgtccc tgatcacgtt ccactcgccg aaaatatcct
    11281 tgctgatggt gctgatagca ggtccgttct taacgaagat cccagcgctg ctgtactcgt
    11341 cgaagttctt gaaaagcttc tccaacttct tgatggacga gaagatctcg ctgttcttgt
    11401 tgagggtgtt acggaaaacc tcaagcactt cctcgtcaga ggtgtaaccc tcaccgtaaa
    11461 atgagaggct ctcacgatca gagagaacct gcaagaaaca gcaaagagag agttaagata
    11521 taagtgaaca agaaagtctt gggatcgaca tacatacaat caattgatca aattcctagg
    11581 attaaacata tgaaatagga aagacagacc tgcttgtaga gcggcttgaa cttcgggagc
    11641 ttctgcttcg ttttctggtt gtacaggttg atgtactcgt tgagtccctt aatcttctcc
    11701 ccagactcgg tcacgaatcc tccgataata gcgttgtaca cgtcgatgcc ctcttgggtg
    11761 agaacgaagt tgaagaactc cccttcaaaa aagtcctcga cgtcgtagtc gctgttgagg
    11821 atcttctctt tgatttcttg cacctcgtgc ttatcgaaga tggcgtccac ctgagattgt
    11881 ttgcaaagaa aaatctcaaa gcatgtacaa tctataaact tatgatggaa aaagaaaaga
    11941 aaggtaacta aggaatgtta ccttctcgaa gatgtccatg ttgctgatgt aacgcgtgag
    12001 gttctcgttg atgcatctga aagcgataga ggtgctcttg gcctcttcgc tgaacatgtt
    12061 ttccctgttt ctgaagaatc cggtgaaggc agtagtgaat ccgttgaagc tgttcacgag
    12121 agcgatctca tccttatcgt ccaggaactc aggcaggatt gtctcgataa tatccttctt
    12181 gaagaggctc ttgtatccct cgtttccctt gaaagccttg gcgatctctt tacggaggtt
    12241 gatctcgagg ttctcaagct ctttgttctc tttctcggtc ctggttttct tacggaagag
    12301 gctgatgtag ttgttgaggt tcttgagctt gatgctgtgg agcacatcgt tgatgaagct
    12361 gaggtagtac ctgtcgagga gcttcttcac gcccttgtaa tcttcagctc tcttctcatc
    12421 ctcgacgagg agcctcttgt tgtcgatatt ctcttgggtc tttcccacag ggatagcctt
    12481 gaacctgagg gtcttagaga ggctgtagca gttggtaaac ttctcgagct tgctcattcc
    12541 tgcagaagta acaccaaaca acagggtgag catcgacaaa agaaacagta ccaagcaaat
    12601 aaatagcgta tgaaggcagg gctaaaaaaa tccacatata gctgctgcat atgccatcat
    12661 ccaagtatat caagatcaaa ataattataa aacatacttg tttattataa tagataggta
    12721 ctcaaggtta gagcatatga atagatgctg catatgccat catgtatatg catcagtaaa
    12781 acccacatca acatgtatac ctatcctaga tcgatatttc catccatctt aaactcgtaa
    12841 ctatgaagat gtatgacaca cacatacagt tccaaaatta ataaatacac caggtagttt
    12901 gaaacagtat tctactccga tctagaacga atgaacgacc gcccaaccac accacatcat
    12961 cacaaccaag cgaacaaaaa gcatctctgt atatgcatca gtaaaacccg catcaacatg
    13021 tatacctatc ctagatcgat atttccatcc atcatcttca attcgtaact atgaatatgt
    13081 atggcacaca catacagatc caaaattaat aaatccacca ggtagtttga aacagaattc
    13141 tactccgatc tagaacgacc gcccaaccag accacatcat cacaaccaag acaaaaaaaa
    13201 gcatgaaaag atgacccgac aaacaagtgc acggcatata ttgaaataaa ggaaaagggc
    13261 aaaccaaacc ctatgcaacg aaacaaaaaa aatcatgaaa tcgatcccgt ctgcggaacg
    13321 gctagagcca tcccaggatt ccccaaagag aaacactggc aagttagcaa tcagaacgtg
    13381 tctgacgtac aggtcgcatc cgtgtacgaa cgctagcagc acggatctaa cacaaacacg
    13441 gatctaacac aaacatgaac agaagtagaa ctaccgggcc ctaaccatgg accggaacgc
    13501 cgatctagag aaggtagaga gggggggggg gggaggacga gcggcgtacc ttgaagcgga
    13561 ggtgccgacg ggtggatttg ggggagatct ggttgtgtgt gtgtgcgctc cgaacaacac
    13621 gaggttgggg aaagagggtg tggagggggt gtctatttat tacggcgggc gaggaaggga
    13681 aagcgaagga gcggtgggaa aggaatcccc cgtagctgcc gtgccgtgag aggaggagga
    13741 ggccgcctgc cgtgccggct cacgtctgcc gctccgccac gcaatttctg gatgccgaca
    13801 gcggagcaag tccaacggtg gagcggaact ctcgagaggg gtccagaggc agcgacagag
    13861 atgccgtgcc gtctgcttcg cttggcccga cgcgacgctg ctggttcgct ggttggtgtc
    13921 cgttagactc gtcgacggcg tttaacaggc tggcattatc tactcgaaac aagaaaaatg
    13981 tttccttagt ttttttaatt tcttaaaggg tatttgttta atttttagtc actttatttt
    14041 attctatttt atatctaaat tattaaataa aaaaactaaa atagagtttt agttttctta
    14101 atttagaggc taaaatagaa taaaatagat gtactaaaaa aattagtcta taaaaaccat
    14161 taaccctaaa ccctaaatgg atgtactaat aaaatggatg aagtattata taggtgaagc
    14221 tatttgcaaa aaaaaaggag aacacatgca cactaaaaag ataaaactgt agagtcctgt
    14281 tgtcaaaata ctcaattgtc ctttagacca tgtctaactg ttcatttata tgattctcta
    14341 aaacactgat attattgtag tactatagat tatattattc gtagagtaaa gtttaaatat
    14401 atgtataaag atagataaac tgcacttcaa acaagtgtga caaaaaaaat atgtggtaat
    14461 tttttataac ttagacatgc aatgctcatt atctctagag aggggcacga ccgggtcacg
    14521 ctgcacctcc actatgcctg agacccaata cgcaaaccgc ctctccccgc gcgttggccg
    14581 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
    14641 gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg
    14701 gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacatacta gagaaagagg
    14761 agaaatacta gatggcttcc tccgaggacg ttatcaaaga gttcatgcgt ttcaaagttc
    14821 gtatggaagg ttccgttaac ggtcacgagt tcgaaatcga aggtgaaggt gaaggtcgtc
    14881 cgtacgaagg tacccagacc gctaaactga aagttaccaa aggtggtccg ctgccgttcg
    14941 cttgggacat cctgtccccg cagttccagt acggttccaa agcttacgtt aaacacccgg
    15001 ctgacatccc ggactacctg aaactgtcct tcccggaagg tttcaaatgg gaacgtgtta
    15061 tgaacttcga ggacggtggt gttgttaccg ttacccagga ctcctccctg caagacggtg
    15121 agttcatcta caaagttaaa ctgcgtggta ccaacttccc gtccgacggt ccggttatgc
    15181 agaaaaaaac catgggttgg gaagcttcca ccgaacgtat gtacccggag gacggtgctc
    15241 tgaaaggtga aatcaaaatg cgtctgaaac tgaaagacgg tggtcactac gacgctgaag
    15301 ttaaaaccac ctacatggct aaaaaaccgg ttcagctgcc gggtgcttac aaaaccgaca
    15361 tcaaactgga catcacctcc cacaacgagg actacaccat cgttgaacag tacgaacgtg
    15421 ctgaaggtcg tcactccacc ggtgcttaat aacgctgata gtgctagtgt agatcgctac
    15481 tagagccagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat
    15541 ctgttgtttg tcggtgaacg ctctctacta gagtcacact ggctcacctt cgggtgggcc
    15601 tttctgcgtt tataggtctc agggattacg aggatgcaca tgtgaccgag gga
//
