LOCUS       sequence-458115-       22650 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Over expresses Cas9 and GR-GRF-GIF. Must receive guide cassettes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 22650)
  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 22650)
  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: EC67907
FEATURES             Location/Qualifiers
     source          1..22650
                     /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"
     terminator      complement(8037..8289)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(8305..8325)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(14058..14078)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    16202..16223
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(16238..16243,16244..16261,16262..16268)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    16276..16292
                     /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             16310..16321
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             16328..17005
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      17050..17121
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      17137..17164
                     /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 tttttcattc tgcaaaagag aaccagacaa cagggtaagt
     6181 gcctagcagt aaacaaacag aactcatcac aagcaaacag caacatcata ttcataccaa
     6241 caggtcatgt gtgttcatca catcattagt actaagcatg ccatcatcca agtatatcaa
     6301 agtaagggca aagagcatgc atgatcatca ggtgcacaaa agaatcatca aattgtagca
     6361 gtacatatct tcatctatca tgcatatcta tccataacag gacgatgcat gttgaccagg
     6421 taaaagctac aggatcctat aggaacagca gcgtatatat cttcaccaat cttagcatct
     6481 taatcatgtg gcacatgcag tttcaattta agcacatgag ctagttgatt atgaggtacc
     6541 agagaatcat caaactgatg tagcagcata tatcctcatc tatcatgcaa tctaatctaa
     6601 tctaactaaa caggaaaggt gtgctattca gttaaaagct accgcatcat atacaaacgg
     6661 cagcataaga aaaagcataa tcatcttaat catgcaacaa acgcagattc ataataagcc
     6721 caagagctag cttgtgatga tcttattcta ctctgatcta cagcaatcag ataacgacct
     6781 aaccttgcac atggcaacaa aacaatcgat cggacgaatc agttgtttgt tcctagctag
     6841 caccatcgaa ccagataata gatgcacgta cagatcccga aaacgaaccc aaaaacaggg
     6901 cagacctagc tgaacctagg cagcgaccca gcagatcgtg agaacgatct catctacgaa
     6961 cagcctagaa gcaaccccac gattcccgga caaacgacct aaaatcccca caaatcacat
     7021 gagcatgaca ggataaacag cggaaccgat cagatctaca cgaaaacccc acctcccagc
     7081 cacccacgat caggaaacac gcggatctag catgatttcg tcaacgcctc agcctagttc
     7141 ctagccacag accaagcaga accaccaaac cacgccgagc gaggagatgg ggcaagagga
     7201 cgggggacac gatcgccgta ccttgaagcg ggggaaggat cgccgagggt cgcgagaaga
     7261 gcaatttgga tttggaaccc gggggttgtg cgctccgaac gatcagacga tgtgagattg
     7321 tgggaagagg cgcggagggc cctgtattta tgggctgcga cggggggagg agaggtgggg
     7381 agggttgggg aaggaatccc ccacccgtgc cgtgacggtt ccgggccgtg tgagaggagc
     7441 ccgctcgtgt ccgccacgca atttccgcga tcggagcgga gctttcgaga ggcggctgga
     7501 tggttggtgg ccgttagatt tgtagacgcc gttaacgcct cgcctccacc gggaagagtt
     7561 ttgagcagcc gcttatgaca atggcttaac gacgttagac ggagcgttag tggcaggcca
     7621 tgatagaata gacgtattgc aatgggatta taattaaata aataagaata taataagata
     7681 tggcaagtcg gcactcatga catggtgtcc gaaatgatag tgctcacttt cttagccgag
     7741 aaagttgacg cgactgattt agaagttaag atattatttc tctcttcttt tctttcctcg
     7801 tcatataagg atgaaataaa ctttagagat tgccggtagt gattttggat ttcggcgatc
     7861 aggcttggtt tgccggtttc ggacggtgtg ccttaggcca cccgcagtgt atcttgtaat
     7921 gttcaaccga taagcaaggg tggggctcaa gcaagtagta aacaactatg tcaaatgtca
     7981 ccatggttat ggtcttgttt agttggcttc ctccgcaaga attcaagctt agcgtcgatc
     8041 tagtaacata gatgacaccg cgcgcgataa tttatcctag tttgcgcgct atattttgtt
     8101 ttctatcgcg tattaaatgt ataattgcgg gactctaatc ataaaaaccc atctcataaa
     8161 taacgtcatg cattacatgt taattattac atgcttaacg taattcaaca gaaattatat
     8221 gataatcatc gcaagaccgg caacaggatt caatcttaag aaactttatt gccaaatgtt
     8281 tgaacgatct gcttgacaag ctcagacttt acgcttcttc tttgggtcag ccctagagtc
     8341 accacccaac tgcgacagat ctatccttgt ctcatagagc cctgtaatag attggtgaat
     8401 aagggtagcg tccaacactt ccttggttga tgtgtagcgc tttctgtcga tagtggtatc
     8461 gaagtactta aaggctgctg gggctcccag attggtcagt gtgaacaggt ggatgatgtt
     8521 ttccgcttgt tcccttatcg gcttatctct gtgcttgttg tacgcggaaa gcacctttaa
     8581 ttgtagaaaa ccagtaaaac atggacaatt aggaggcatc gattatctaa ccaaagactt
     8641 taacaacact aatctgccac taatattcat gagaagtcct taccttatcg aggttagcgt
     8701 ctgcaagaat gacccgcttg gaaaattcgc tgatctgctc tatgatctca tcaagatagt
     8761 gcttgtgctg ctcgacaaac aactgcttct gctcgttgtc ctctggtgac cccttcagct
     8821 tctcatagtg tgaggccaag tacaggaagt tgacgtactt actgggcaat gccagctcgt
     8881 tgcccttctg cagttctccg gcagatgcca gcatgcgttt tctgccattt tccaattcga
     8941 aaagcgaata cttgggcagt ttgataatga ggtctttctt cacctctttg taacctggtt
     9001 tgtataattt tggttcagtt aattaggaaa attacataat ggctccaaaa aaaaattatc
     9061 gaccttttac caaaaaaaat aaaatttatc gaccttgaca ggaaaaagtt acaaatcaaa
     9121 agaaagaata tcgtctggca tccaatattt tacctttagc ctcgaggaag tcgatcggat
     9181 tcttctcgaa cgaggacctt tccattatgg taattccaag cagttccttg acgcttttca
     9241 gtttcttgct cttgcccttc tccaccttgg ccactaccag tacgctgtat gccacggtag
     9301 ggctatcaaa gcctccgtat ttcttcggat cccaatcctt cttcctcgca atcagtttgt
     9361 cggagttgcg cttagggaga atggactcct tggagaaacc accggtttga acctccgttt
     9421 tcttgactat gttgacttgg ggcatgctga gaaccttacg cacagtcgca aaatccctgc
     9481 ccttatccca gacaatctca cctgcaagaa acagcaaaga gagagttaag atataagtga
     9541 acaagaaagt cttgggatcg acatacatac aatcaattga tcaaattcct aggattaaac
     9601 atatgaaata ggaaagacag acctgtttcg ccatttgtct cgatcagagg cctcttccgg
     9661 atttcaccat tcgccagagt gatctctgtc ttgaagaagt tcatgatatt cgagtaaaag
     9721 aagtatttcg cggtagcctt gccgatctct tgttctgact tcgcgatcat cttcctaacg
     9781 tcgtacacct tgtaatcccc gtacacaaac tcagactcca gtttcggata cttcttgatg
     9841 agagcagtgc caacaacggc gttcaggtag gcgtcatggg catgatggta gttgttgatc
     9901 tcgcgaacct tgtaaaactg gaagtccttc cggaagtcac taaccagttt gctcttgagg
     9961 gtgatcacct gcgtaaccat caaaagtcaa tatggattca tcacaaatta atcaggaata
    10021 gaaagatcgg gtggccttga agttgatcaa ataaaaagtt tcgaacttta gtttgatatg
    10081 gtaaaggaaa gttggaaact ttacctttac ctccctgata agtttgtcat tctcatcata
    10141 cttcgtattc atacgcgaat cgagtatctg ggccacatgt ttcgtaatct gacgagtttc
    10201 aaccaactgg cgcttgatga agccggcttt gtcgagctca gacagtccgc ccctttctgc
    10261 cttcgtgaga ttgtcgaatt tccgttgggt tatgagctta gcattcaaaa gctgcctcca
    10321 atagttcttc atcttcttca caacctcctc gctgggtaca ttgtcggatt ttccgcggtt
    10381 cttgtctgag cgcgtaagga cctataacac gggagaaacc aaaaagcaaa ctcatttcac
    10441 gccaaatact acagatttct caaaaattaa gttataatcc atatatcctg acaagaaatt
    10501 gattaaaaca cagttgcttt accttgttgt caatactatc gtccttcagg aaagattgag
    10561 ggacaatgtg atccacatca tagtcggaga gtctgttaat gtcgagctct tggtcgacgt
    10621 acatatctct cccattctgg aggtaataga ggtacagctt ctcgttctgg agttgagtat
    10681 tttcgactgg gtgctctttc agaatctgac tcccaagctc cttgatgcct tcctctatcc
    10741 gcttcatgcg ctcccgtgag ttcttctgcc ctttctgagt cgtctggttt tccctcgcca
    10801 tctcgatgac gatgttctcc ggcttatggc ggcccatgac ctgtagcaat aattaaaaca
    10861 aaaataccac aaacagttag aactgaggca agaagaatga atgcagagaa aaaaaatctt
    10921 agctattcat ggaaatggaa gtgcgacgaa cacctaaaat gcaaggagca taaccaaatg
    10981 cagaacttac cttcaccagc tcatccacaa ccttcactgt ctgcagtatg cccttcttta
    11041 tagctggcga tccagccagg ttggctatgt gttcgtgaag gctatcgccc tgacctgaca
    11101 cttgggcctt ctggatatct tctttgaagg tcagtgagtc atcgtgaata agctgcatga
    11161 agttccgatt cgcaaaaccg tcggatttga gaaagtccag tatagtcttg ccggattgct
    11221 tgtcgcggat gccgttgatg agcttcctgg agagccttcc ccaacctaaa acatagaaca
    11281 aacatttgtc tcatcacaaa tatgcatcaa ggaacactca gaagttagtc atatacccaa
    11341 attcttgaaa ctagagaaac aaaaaaagag ggatcattgc ggaaaccttt tgacctctta
    11401 cctgtatatc tcctcctctt caactgcttc atgaccttgt catcgaacag atgcgcataa
    11461 gtcttaaggc gctcctcgat catttctctg tcctcaaaca gcgtgagagt cagcacgatg
    11521 tcctcgagga tgtcctcgtt ctcctcgttg tcaaggaagt ccttgtcctt gatgattttc
    11581 aacaggtcat ggtatgtgcc aagagaggca ttgaacctat cctcaacgcc gctgatttcc
    11641 acagaatcaa agcattcgat cttcttaaag tagtcctctt taagctgttt cacagtgacc
    11701 ttccgattgg ttttgaacaa gagatcgaca atggccttct tctgctcacc tgttataaca
    11761 gtaaaaatta ctttagaaac aaaacaaaag tgtggtcttt taggaatgat tattgatcag
    11821 ttcatagttt aaaatcttgt gtttttatcg taaaagtgtg gaccaggatt tacctgacag
    11881 gaaggccggt ttgcgcatcc cctccgtcac atatttcacc ttcgtcagct cgttgtagac
    11941 cgtgaagtat tcgtacagga gactatgctt gggcaggact ttctcatttg ggagattctt
    12001 atcgaagttg gtcatccttt cgatgaagga ctgtgcactc gcacccttgt ccacgacctc
    12061 ttcgaagttc caaggcgtga tagtctcctc ggatttgcgt gtcatccacg caaaccgcga
    12121 gttcccgcgt gcgagtggac ctaatgaatt aatagaaaaa tcaggcaaat tagaaaagaa
    12181 ataattctca aaagagaaca actgaggtca gaaaatagca ttcatcaact gaaactaggt
    12241 atttcttcat atgatactaa cctacatagt agggaatgcg gaacgtgagg atcttttcaa
    12301 tcttctctct gttgtctttc aggaacggat agaagtcctc ttgcctcctc aagatagcat
    12361 gcagctcacc gaggtggatc tggtgcggta tgctgccatt gtcgaaggta cgctgcttcc
    12421 gaagcaaatc ttcgcggttc aatttaacga gcagctcctc tgtaccgtcc atcttttcca
    12481 gaataggctt gatgaatttg tagaattcct cctgacttgc acctaataat atcatcagag
    12541 gaaaactcaa ttcttatatc ttacagaaat gaacaacaaa atacatgtag cagagaggaa
    12601 aacaatcata agtaacaact tacctccatc aatgtaccct gcgtagccat tcttcgattg
    12661 gtcaaagaaa atctccttgt acttctcggg gagttgctgg cgaaccaaag ccttcagaag
    12721 ggtcagatcc tggtgatgct cgtcgtacct cttgatcatg gaggcgctaa gtggcgcttt
    12781 ggttatctcc gtgttgaccc gcagaatgtc actgagcaag atagcgtctg acagattctt
    12841 ggcagccaag aacaggtcgg catactgatc acctacaaaa gtttaaaaga ctcaatttta
    12901 actctacagc ttcaaataaa tttccgcaaa gaaattgata ggtttagaca acaaaagaag
    12961 agttccaatt tcaagagcac ctatttgggc gagcagattg tccagatcat catcgtatgt
    13021 atcctttgac agctgcagct tcgcgtcctc ggcgaggtca aagttgcttt tgaagttagg
    13081 ggtgaggccc agagacagtg caatcagatt cccgaacaag ccgttcttct tctcgcctgg
    13141 caattgagcg atcagattct caagccgtct ggacttcgaa agtctggcag acagtattgc
    13201 cttggcgtcg acacctgaga ttgtttgcaa agaaaaatct caaagcatgt acaatctata
    13261 aacttatgat ggaaaaagaa aagaaaggta actaaggaat gttacctgat gcattgatgg
    13321 ggttctcctc gaacagctga ttgtaggttt ggacgagctg aatgaacagt ttatccacat
    13381 cggagttatc cgggttgagg tccccttcga tcaagaaatg gccacgaaac tttatcatgt
    13441 gcgcgagagc caggtatatg agccgaaggt ctgccttatc ggttgagtcc accagcttct
    13501 tgcgaaggtg gtatatagtc ggatactttt catgataggc tacttcgtca acaatattcc
    13561 caaagatggg gtgacgctcg tgtttcttat cttcctctac caggaaggat tcctcgaggc
    13621 gatggaagaa cgagtcgtca acctgcaaaa tcaagtttat tgatcatcaa gtgagtttac
    13681 atgtagttaa atgctttcag taatatccca atgaagcagg atatgctttc agtacaaaat
    13741 acataagcat actatatcat aaaaatcctt accttcgcca tctcgttgct aaagatctct
    13801 tggagataac agattctgtt cttcctgcga gtgtaccttc tcctcgcagt acgtttaagc
    13861 cttgtcgcct ccgcggtttc tccagagtca aagagcagag ctccgatcaa gttcttctta
    13921 atgctgtgcc tatctgtgtt gcccagcact ttaaatttct tcgaaggtac cttgtattca
    13981 tcggtgataa cggcccaccc aactgagttc gtaccaatat caagtccaat gctgtacttc
    14041 ttgtccattt tccagctgac cttccgcttc ttcttcggtg ggctagaagc cattcctgca
    14101 gaagtaacac caaacaacag ggtgagcatc gacaaaagaa acagtaccaa gcaaataaat
    14161 agcgtatgaa ggcagggcta aaaaaatcca catatagctg ctgcatatgc catcatccaa
    14221 gtatatcaag atcaaaataa ttataaaaca tacttgttta ttataataga taggtactca
    14281 aggttagagc atatgaatag atgctgcata tgccatcatg tatatgcatc agtaaaaccc
    14341 acatcaacat gtatacctat cctagatcga tatttccatc catcttaaac tcgtaactat
    14401 gaagatgtat gacacacaca tacagttcca aaattaataa atacaccagg tagtttgaaa
    14461 cagtattcta ctccgatcta gaacgaatga acgaccgccc aaccacacca catcatcaca
    14521 accaagcgaa caaaaagcat ctctgtatat gcatcagtaa aacccgcatc aacatgtata
    14581 cctatcctag atcgatattt ccatccatca tcttcaattc gtaactatga atatgtatgg
    14641 cacacacata cagatccaaa attaataaat ccaccaggta gtttgaaaca gaattctact
    14701 ccgatctaga acgaccgccc aaccagacca catcatcaca accaagacaa aaaaaagcat
    14761 gaaaagatga cccgacaaac aagtgcacgg catatattga aataaaggaa aagggcaaac
    14821 caaaccctat gcaacgaaac aaaaaaaatc atgaaatcga tcccgtctgc ggaacggcta
    14881 gagccatccc aggattcccc aaagagaaac actggcaagt tagcaatcag aacgtgtctg
    14941 acgtacaggt cgcatccgtg tacgaacgct agcagcacgg atctaacaca aacacggatc
    15001 taacacaaac atgaacagaa gtagaactac cgggccctaa ccatggaccg gaacgccgat
    15061 ctagagaagg tagagagggg ggggggggga ggacgagcgg cgtaccttga agcggaggtg
    15121 ccgacgggtg gatttggggg agatctggtt gtgtgtgtgt gcgctccgaa caacacgagg
    15181 ttggggaaag agggtgtgga gggggtgtct atttattacg gcgggcgagg aagggaaagc
    15241 gaaggagcgg tgggaaagga atcccccgta gctgccgtgc cgtgagagga ggaggaggcc
    15301 gcctgccgtg ccggctcacg tctgccgctc cgccacgcaa tttctggatg ccgacagcgg
    15361 agcaagtcca acggtggagc ggaactctcg agaggggtcc agaggcagcg acagagatgc
    15421 cgtgccgtct gcttcgcttg gcccgacgcg acgctgctgg ttcgctggtt ggtgtccgtt
    15481 agactcgtcg acggcgttta acaggctggc attatctact cgaaacaaga aaaatgtttc
    15541 cttagttttt ttaatttctt aaagggtatt tgtttaattt ttagtcactt tattttattc
    15601 tattttatat ctaaattatt aaataaaaaa actaaaatag agttttagtt ttcttaattt
    15661 agaggctaaa atagaataaa atagatgtac taaaaaaatt agtctataaa aaccattaac
    15721 cctaaaccct aaatggatgt actaataaaa tggatgaagt attatatagg tgaagctatt
    15781 tgcaaaaaaa aaggagaaca catgcacact aaaaagataa aactgtagag tcctgttgtc
    15841 aaaatactca attgtccttt agaccatgtc taactgttca tttatatgat tctctaaaac
    15901 actgatatta ttgtagtact atagattata ttattcgtag agtaaagttt aaatatatgt
    15961 ataaagatag ataaactgca cttcaaacaa gtgtgacaaa aaaaatatgt ggtaattttt
    16021 tataacttag acatgcaatg ctcattatct ctagagaggg gcacgaccgg gtcacgctgc
    16081 acctccacta tgcctgagac ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
    16141 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
    16201 ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc
    16261 gtatgttgtg tggaattgtg agcggataac aatttcacac atactagaga aagaggagaa
    16321 atactagatg gcttcctccg aggacgttat caaagagttc atgcgtttca aagttcgtat
    16381 ggaaggttcc gttaacggtc acgagttcga aatcgaaggt gaaggtgaag gtcgtccgta
    16441 cgaaggtacc cagaccgcta aactgaaagt taccaaaggt ggtccgctgc cgttcgcttg
    16501 ggacatcctg tccccgcagt tccagtacgg ttccaaagct tacgttaaac acccggctga
    16561 catcccggac tacctgaaac tgtccttccc ggaaggtttc aaatgggaac gtgttatgaa
    16621 cttcgaggac ggtggtgttg ttaccgttac ccaggactcc tccctgcaag acggtgagtt
    16681 catctacaaa gttaaactgc gtggtaccaa cttcccgtcc gacggtccgg ttatgcagaa
    16741 aaaaaccatg ggttgggaag cttccaccga acgtatgtac ccggaggacg gtgctctgaa
    16801 aggtgaaatc aaaatgcgtc tgaaactgaa agacggtggt cactacgacg ctgaagttaa
    16861 aaccacctac atggctaaaa aaccggttca gctgccgggt gcttacaaaa ccgacatcaa
    16921 actggacatc acctcccaca acgaggacta caccatcgtt gaacagtacg aacgtgctga
    16981 aggtcgtcac tccaccggtg cttaataacg ctgatagtgc tagtgtagat cgctactaga
    17041 gccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt
    17101 tgtttgtcgg tgaacgctct ctactagagt cacactggct caccttcggg tgggcctttc
    17161 tgcgtttata ggtctcaggg attacgaatt cccatgggga ggtgcagcgt gacccggtcg
    17221 tgcccctctc tagagataat gagcattgca tgtctaagtt ataaaaaatt accacatatt
    17281 ttttttgtca cacttgtttg aagtgcagtt tatctatctt tatacatata tttaaacttt
    17341 actctacgaa taatataatc tatagtacta caataatatc agtgttttag agaatcatat
    17401 aaatgaacag ttagacatgg tctaaaggac aattgagtat tttgacaaca ggactctaca
    17461 gttttatctt tttagtgtgc atgtgttctc cttttttttt gcaaatagct tcacctatat
    17521 aatacttcat ccattttatt agtacatcca tttagggttt agggttaatg gtttttatag
    17581 actaattttt ttagtacatc tattttattc tattttagcc tctaaattaa gaaaactaaa
    17641 actctatttt agttttttta tttaataatt tagatataaa atagaataaa ataaagtgac
    17701 taaaaattaa acaaataccc tttaagaaat taaaaaaact aaggaaacat ttttcttgtt
    17761 tcgagtagat aatgccagcc tgttaaacgc cgtcgacgag tctaacggac accaaccagc
    17821 gaaccagcag cgtcgcgtcg ggccaagcga agcagacggc acggcatctc tgtcgctgcc
    17881 tctggacccc tctcgagagt tccgctccac cgttggactt gctccgctgt cggcatccag
    17941 aaattgcgtg gcggagcggc agacgtgagc cggcacggca ggcggcctcc tcctcctctc
    18001 acggcacggc agctacgggg gattcctttc ccaccgctcc ttcgctttcc cttcctcgcc
    18061 cgccgtaata aatagacacc ccctccacac cctctttccc caacctcgtg ttgttcggag
    18121 cgcacacaca cacaaccaga tctcccccaa atccacccgt cggcacctcc gcttcaaggt
    18181 acgccgctcg tcctcccccc cccccccccc cccccyyctc tagatcggcg ttccggtcca
    18241 tggttagggc ccggtagttc tacttctgtt catgtttgtg ttagatccgt gtttgtgtta
    18301 gatccgtgct gctagcgttc gtacacggat gcgacctgta cgtcagacac gttctgattg
    18361 ctaacttgcc agtgtttctc tttggggaat cctgggatgg ctctagccgt tccgcagacg
    18421 ggatcgattt catgattttt tttgtttcgt tgcatagggt ttggtttgcc cttttccttt
    18481 atttcaatat atgccgtgca cttgtttgtc gggtcatctt ttcatgcttt tttttgtctt
    18541 ggttgtgatg atgtggtctg gttgggcggt cgttctagat cggagtagaa ttctgtttca
    18601 aactacctgg tggatttatt aattttggat ctgtatgtgt gtgccataca tattcatagt
    18661 tacgaattga agatgatgga tggaaatatc gatctaggat aggtatacat gttgatgcgg
    18721 gttttactga tgcatataca gagatgcttt ttgttcgctt ggttgtgatg atgtggtgtg
    18781 gttgggcggt cgttcattcg ttctagatcg gagtagaata ctgtttcaaa ctacctggtg
    18841 tatttattaa ttttggaact gtatgtgtgt gtcatacatc ttcatagtta cgagtttaag
    18901 atggatggaa atatcgatct aggataggta tacatgttga tgtgggtttt actgatgcat
    18961 atacatgatg gcatatgcag catctattca tatgctctaa ccttgagtac ctatctatta
    19021 taataaacaa gtatgtttta taattatttt gatcttgata tacttggatg atggcatatg
    19081 cagcagctat atgtggattt ttttagccct gccttcatac gctatttatt tgcttggtac
    19141 tgtttctttt gtcgatgctc accctgttgt ttggtgttac ttctgcagga atggctagtg
    19201 aagctcgaaa aacaaagaaa aaaatcaaag ggattcagca agccactgca ggagtctcac
    19261 aagacacttc ggaaaatcct aacaaaacaa tagttcctgc agcattacca cagctcaccc
    19321 ctaccttggt gtcactgctg gaggtgattg aacccgaggt gttgtatgca ggatatgata
    19381 gctctgttcc agattcagca tggagaatta tgaccacact caacatgtta ggtgggcgtc
    19441 aagtgattgc agcagtgaaa tgggcaaagg cgataccagg cttcagaaac ttacacctgg
    19501 atgaccaaat gaccctgcta cagtactcat ggatgtttct catggcattt gccctgggtt
    19561 ggagatcata cagacaatca agtggaaacc tgctctgctt tgctcctgat ctgattatta
    19621 atgagcagag aatgtctcta ccctgcatgt atgaccaatg taaacacatg ctgtttgtct
    19681 cctctgaatt acaaagattg caggtatcct atgaagagta tctctgtatg aaaaccttac
    19741 tgcttctctc ctcagttcct aaggaaggtc tgaagagcca agagttattt gatgagattc
    19801 gaatgactta tatcaaagag ctaggaaaag ccatcgtcaa aagggaaggg aactccagtc
    19861 agaactggca acggttttac caactgacaa agcttctgga ctccatgcat gaggtggttg
    19921 agaatctcct tacctactgc ttccagacat ttttggataa gaccatgagt attgaattcc
    19981 cagagatgtt agctgaaatc atcactaatc agataccaaa atattcaaat ggaaatatca
    20041 aaaagcttct gtttcatcaa aaatacgacc ccatggcgat gccgtatgcc tctctttccc
    20101 cggcaggcga ccgccgctcc tccccggccg ccaccgccac cgcctccctc ctccccttct
    20161 gccgctcctc ccccttctcc gccggcggca atggcggcat gggggaggag gcgcggatgg
    20221 acgggaggtg gatggcgagg ccggtgccct tcacggcggc gcagtacgag gagctggagc
    20281 accaggcgct catatacaag tacctggtgg ccggcgtgtc cgtcccgccg gatctcgtgc
    20341 tccccatccg ccgcggcatc gagtccctcg ccgcccgctt ctaccacaac cccctcgcca
    20401 tcgggtacgg atcgtacctg ggcaagaagg tggatccgga gccgggccgg tgccggcgca
    20461 cggacggcaa gaagtggcgg tgcgccaagg aggccgcctc cgactccaag tattgcgagc
    20521 gccacatgca ccgcggccgc aaccgttcaa gaaagcctgt ggaaacgcag ctcgtctcgc
    20581 actcccagcc gccggccgcc tccgtcgtgc cgcccctcgc caccggcttc cacaaccact
    20641 ccctctaccc cgccatcggc ggcaccaacg gtggtggagg cggggggaac aacggcatgt
    20701 ccatgcccgg cacgttctcc tccgcgctgg ggccgcctca gcagcacatg ggcaacaatg
    20761 ccgcctctcc ctacgcggct ctcggcggcg ccggaacatg caaagatttc aggtataccg
    20821 catatggaat aagatctttg gcagacgagc agagtcagct catgacagaa gccatgaaca
    20881 cctccgtgga gaacccatgg cgcctgccgc catcttctca aacgactaca ttcccgctct
    20941 caagctactc tcctcagctt ggagcaacga gtgacctggg tcagaacaac agcagcaaca
    21001 acaacagcgg cgtcaaggcc gagggacagc agcagcagca gccgctctcc ttcccggggt
    21061 gcggcgactt cggcagcggc gactccgcga agcaggagaa ccagacgctg cggccgttct
    21121 tcgacgagtg gcccaagacg agggactcgt ggtcggacct gaccgacgac aactcgaacg
    21181 tcgcctcctt ctcggccacc cagctgtcga tctcgatacc catgacgtcc tccgacttct
    21241 ccgccgccag ctcccagtcg cccaacggca tgctgttcgc cggcgagatg tacgcggccg
    21301 ctgccatgca gcagcaacac ctgatgcaga tgaaccagag catgatgggg ggctacgctt
    21361 cctctaccac tgccaccact gatctcattc agcagtacct ggatgagaac aagcagctga
    21421 tcctggccat cctcgacaac cagaacaacg gcaaggtgga ggagtgcgca cggaaccaag
    21481 ctaagctcca gcagaacctc atgtacctcg ccgccatcgc cgacagccag cctccgcaga
    21541 cggcatcgct gtctcagtac ccgtccaacc tgatgatgca gtccgggccg cggtacatgc
    21601 agcagcagtc ggcgcagatg atgtcgccgc agtcgctgat ggcggcgcgg tcgtcgatga
    21661 tgtacgcgca gcaggccatg tcgccgctcc agcagcagca gcagcagcag cagcaccagg
    21721 cggccgcgca cggccagctg gggatgtcct ccggcgcgac caccgggttc aacctcctgc
    21781 acggtgaggc cagcatgggc ggcggcggcg gcgccactgg caacagcatg atgaacgcca
    21841 gcgtcttttc ggactatggc cgcggcggca gcggcgccaa ggaggggtcg acctcgctgt
    21901 cggccgacgc tcgcggcgcc aactctggcg cgcacagcgg cgacggggag tacctcaagg
    21961 gcaccgagga ggaaggaagc taggcttgct ttcgttcgta tcatcggttt cgacaacgtt
    22021 cgtcaagttc aatgcatcag tttcattgcg cacacaccag aatcctactg agtttgagta
    22081 ttatggcatt gggaaaactg tttttcttgt accatttgtt gtgcttgtaa tttactgtgt
    22141 tttttattcg gttttcgcta tcgaactgtg aaatggaaat ggatggagaa gagttaatga
    22201 atgatatggt ccttttgttc attctcaaat taatattatt tgttttttct cttatttgtt
    22261 gtgtgttgaa tttgaaatta taagagatat gcaaacattt tgttttgagt aaaaatgtgt
    22321 caaatcgtgg cctctaatga ccgaagttaa tatgaggagt aaaacacttg tagttgtacc
    22381 attatgctta ttcactaggc aacaaatata ttttcagacc tagaaaagct gcaaatgtta
    22441 ctgaatacaa gtatgtcctc ttgtgtttta gacatttatg aactttcctt tatgtaattt
    22501 tccagaatcc ttgtcagatt ctaatcattg ctttataatt atagttatac tcatggattt
    22561 gtagttgagt atgaaaatat tttttaatgc attttatgac ttgccaattg attgacaacg
    22621 ctcagagagg atgcacatgt gaccgaggga
//
