LOCUS       sequence-449292-       20952 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cas12a and Csy4 with 5' target (I) for Nicotiana leaf assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 20952)
  AUTHORS   Bircheneder M, Schreiber T, Tissier A, Parniske M
  TITLE     A quantitative assay for the efficiency of RNA-guided genome editing
            in plants.
  JOURNAL   Plant J. 2024 Sep;119(5):2564-2577. doi: 10.1111/tpj.16931. Epub 
            2024 Jul 20.
  PUBMED    39032106
REFERENCE   2  (bases 1 to 20952)
  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"; doi: "10.1111/tpj.16931";
            journalName: "Plant J"; date: "2024-09"; volume: "119"; issue: "5"; 
            pages: "2564-2577"
COMMENT     Sequence Label: pMB26
FEATURES             Location/Qualifiers
     source          1..20952
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    18..158
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     73..92
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(173..190)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(344..932)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(424..443)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(1002..1021)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1003..1021)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1026..1042)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1026..1042)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1144..1163)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1155..1964
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1222..1241)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     misc_feature    2220..2244
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        2371..2550
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        2403..2586
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     CDS             4972..4992
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8695..8715
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     ncRNA           10640..10676
                     /product="self-cleaving hammerhead ribozyme (Ferre-D'Amare 
                     and Scott, 2010)"
                     /label=hammerhead ribozyme
                     /ncRNA_class="hammerhead_ribozyme"
     ncRNA           10718..10785
                     /product="hepatitis delta virus self-cleaving ribozyme 
                     (Ferre-D'Amare and Scott, 2010)"
                     /label=HDV ribozyme
                     /ncRNA_class="ribozyme"
     ncRNA           11203..11239
                     /product="self-cleaving hammerhead ribozyme (Ferre-D'Amare 
                     and Scott, 2010)"
                     /label=hammerhead ribozyme
                     /ncRNA_class="hammerhead_ribozyme"
     ncRNA           11281..11348
                     /product="hepatitis delta virus self-cleaving ribozyme 
                     (Ferre-D'Amare and Scott, 2010)"
                     /label=HDV ribozyme
                     /ncRNA_class="ribozyme"
     CDS             12505..12522
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    12533..12552
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
     terminator      14254..14506
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     terminator      16598..16846
                     /label=HSP terminator
                     /note="efficient transcription terminator from the 
                     Arabidopsis thaliana heat shock protein 18.2 gene (Nagaya 
                     et al., 2010)"
     misc_feature    16910..16934
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             18235..18864
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             19293..20366
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     rep_origin      20432..20626
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(20862..20884)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gttggcgggt gtcggggcgc agccatgacc cagtcacgta gcgatagcgg agtgtatact
       61 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa
      121 taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ctcttccgct tcctcgctca
      181 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
      241 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
      301 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
      361 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
      421 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
      481 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
      541 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
      601 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
      661 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
      721 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
      781 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
      841 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
      901 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
      961 ctgacgctca taaaatcatt atttgccatc cagctgatat cccctatagt gagtcgtatt
     1021 acatggtcat agctgtttcc tggcagctct ggcccgtgtc tcaaaatctc tgatgttaca
     1081 ttgcacaaga taaaaatata tcatcatgaa caataaaact gtctgcttac ataaacagta
     1141 atacaagggg tgttatgagc catattcaac gggaaacgtc gaggccgcga ttaaattcca
     1201 acatggatgc tgatttatat gggtataaat gggctcgcga taatgtcggg caatcaggtg
     1261 cgacaatcta tcgcttgtat gggaagcccg atgcgccaga gttgtttctg aaacatggca
     1321 aaggtagcgt tgccaatgat gttacagatg agatggtcag actaaactgg ctgacggaat
     1381 ttatgcctct tccgaccatc aagcatttta tccgtactcc tgatgatgca tggttactca
     1441 ccactgcgat ccccggaaaa acagcattcc aggtattaga agaatatcct gattcaggtg
     1501 aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta
     1561 attgtccttt taacagcgat cgcgtatttc gtcttgctca ggcgcaatca cgaatgaata
     1621 acggtttggt tgatgcgagt gattttgatg acgagcgtaa tggctggcct gttgaacaag
     1681 tctggaaaga aatgcataaa cttttgccat tctcaccgga ttcagtcgtc actcatggtg
     1741 atttctcact tgataacctt atttttgacg aggggaaatt aataggttgt attgatgttg
     1801 gacgagtcgg aatcgcagac cgataccagg atcttgccat cctatggaac tgcctcggtg
     1861 agttttctcc ttcattacag aaacggcttt ttcaaaaata tggtattgat aatcctgata
     1921 tgaataaatt gcagtttcat ttgatgctcg atgagttttt ctaatcagaa ttggttaatt
     1981 ggttgtaaca ctggcagagc attacgctga cttgacggga cggcgcaagc tcatgaccaa
     2041 aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc agacccctgc cggtaacatg
     2101 agcaaagtct gccgccttac aacggctctc ccgctgacgc cgtcccggac tgatgggctg
     2161 cctgtatcga gtggtgattt tgtgccgagc tgccggtcgg ggagctgttg gctggctggt
     2221 ggcaggatat attgtggtgt aaacaaattg acgcttagac ggattacggg tctcagcggt
     2281 acggcgggat catgagcgga gaattaaggg agtcacgtta tgacccccgc cgatgacgcg
     2341 ggacaagccg ttttacgttt ggaactgaca gaaccgcaac gttgaaggag ccactcagcc
     2401 gcgggtttct ggagtttaat gagctaagca catacgtcag aaaccattat tgcgcgttca
     2461 aaagtcgcct aaggtcacta tcagctagca aatatttctt gtcaaaaatg ctccactgac
     2521 gttccataaa ttcccctcgg tatccaatta gagtctcata ttcactctca atccaaataa
     2581 tctgcaccgg atcttctgga agcaccatga cgagtaaagt gtatgatcct gagcaaagga
     2641 agagaatgat cacaggtcct caatggtggg ctaggtgtaa acagatgaat gttcttgact
     2701 ctttcatcaa ctactacgat tctgagaaac atgccgaaaa cgcagtgatt ttcctccatg
     2761 gaaatgctgc ttcaagctat ctctggagac acgttgtccc acatatagaa ccagtagcta
     2821 gatgcattat accggatttg attggtatgg gcaaaagtgg gaaatcaggg aatggatcct
     2881 acaggttatt ggaccactac aagtatctca cagcttggtt tgaactactt aacttgccta
     2941 agaagatcat attcgtggga catgattggg gagcatgtct tgcctttcac tattcctatg
     3001 aacatcagga caaaataaag gccattgtcc atgcagagtc tgttgttgac gtgattgaaa
     3061 gctgggatga atggcctgac attgaggagg atattgcgtt aatcaagagt gaagaaggcg
     3121 agaaaatggt tcttgaaaac aatttctttg tcgaaactat gttgccgtct aaaatcatga
     3181 gaaaactgga accagaagaa ttcgcagcat acttagaacc ctttaaggag aaaggagaag
     3241 tcagacgacc aactctgtct tggcctagag agattccatt ggttaagggt ggcaaaccag
     3301 atgttgtaca aattgtgcga aattataacg cttatctgcg tgcttcagat gatcttccta
     3361 agatgtttat tgagtcagat cccggtttct ttagcaatgc gattgtagag ggtgctaaga
     3421 aatttccaaa taccgaattc gtgaaggtta aaggactaca cttttctcaa gaagatgcac
     3481 ctgatgagat ggggaagtac ataaagtcct ttgtagagcg tgttcttaag aatgagcagt
     3541 aaaagggcta agacctaagc tagcctagaa tccggccatg ctagagtccg caaaaatcac
     3601 cagtctctct ctacaaatct atctctctct atttttctcc agaataatgt gtgagtagtt
     3661 cccagataag ggaattaggg ttcttatagg gtttcgctca tgtgttgagc atataagaaa
     3721 cccttagtat gtatttgtat ttgtaaaata cttctatcaa taaaatttct aattcctaaa
     3781 accaaaatcc agtgaccttg agtgacctaa gctagcctag tgtcgggcgc ggggagagag
     3841 gattttgagg aaataattaa ttgaattact tgtattattg ataaagtaat ttagataagt
     3901 tgttagtaca acttattgca actatggata gacaaaaatc acttatttta aggggagaag
     3961 tatataacaa cttattataa aatttctgag cagccggcct cctccaatca atcatagaga
     4021 gtggagccca ttctggaaaa ccaaggaacc cccaacttgc actggtgcgg tggcccaatt
     4081 caaaaaaaac ggggcccagc aaagtaacgc cgttgtaacg atttatcaat ccaaactcaa
     4141 aaggcggcga gatgcgttta actccgtgaa attaacaaac cgccaacaac ttgcaatttg
     4201 caactaccgt ttccaagaag aactcaacca cacaacgtat cctatcccaa accacacgca
     4261 actagtgacg cgtcataagg acacgtgtca caatttgact ggttaataat ttcaccgctt
     4321 ttgctataaa ttacctccaa tccccttagc ttcttcacaa ttcagttccc aaccctaaca
     4381 attcttgttc atatcgcttc tctctacttt caaggtatga tccaatttct ctcttctttc
     4441 tctgtaatcc tttcgttgag ttttgtttcc gatcaatcat aggtagtttt cttgtttcga
     4501 agcatgagat ctaggaattt tttgtgattt tccaaaattg agatcggttt gaaattgaat
     4561 tttacagctt gaatctcaga tcttgtttta tcaatgtttt cgatggcttg cgatgtagat
     4621 ctatgataat tgtggttcag ttttgttagg aatcgatttc ggtttagcaa ttgcagatta
     4681 attagggttt ccaattgaat tcttcagatc cgttatggaa ttatgtcaaa taatttattc
     4741 aaattggaaa ttattgttag atccactctt aatctgtttg atccaagctt caattagggt
     4801 tttcacttgt ttcaatttct tgttatggat tctgatttat ctgttgatgt tagatccact
     4861 cttaatctgt ttgatccaag cgttaattag ggttttcact tgtttcaatt tcgtgtgttg
     4921 gattctgatt tatctgttgt tgatgtgatt acagtctgaa caatgcttca acctaagaag
     4981 aagagaaagg ttggaggaaa caccatgagc aagctcgaga agtttaccaa ctgctacagc
     5041 ctctctaaga ccctcaggtt caaggctatc cctgtgggaa agacccaaga gaatatcgac
     5101 aacaagaggc tcctcgtcga ggatgagaag agagctgaag attacaaggg cgtgaagaag
     5161 ctcctcgaca ggtactacct cagcttcatc aacgatgtgc tccacagcat caagctcaag
     5221 aacctcaaca actacatcag cctcttccgt aagaaaacca ggaccgagaa agagaacaaa
     5281 gagcttgaga acctcgagat caacctccgt aaagagatcg ccaaggcttt caagggaaac
     5341 gagggataca agagcctctt caagaaggat attatcgaga caatcctgcc tgagttcctg
     5401 gacgataagg atgagatcgc tctcgtgaac agcttcaacg gattcactac tgccttcacc
     5461 ggattcttca gaaacaggga aaacatgttc agcgaagagg ccaagagcac ctctatcgct
     5521 ttcagatgca tcaacgagaa cctcacgcgt tacatcagca acatggacat cttcgagaag
     5581 gtggacgcca tcttcgataa gcacgaggtg caagaaatca aagagaagat cctcaacagc
     5641 gactacgacg tcgaggactt ttttgaaggg gagttcttca acttcgttct cacccaagag
     5701 ggcatcgacg tgtacaacgc tattatcgga ggattcgtga ccgagtctgg ggagaagatt
     5761 aagggactca acgagtacat caacctgtac aaccagaaaa cgaagcagaa gctcccgaag
     5821 ttcaagccgc tctacaagca ggttctctct gatcgtgaga gcctctcatt ttacggtgag
     5881 ggttacacct ctgacgagga agtgcttgag gttttccgta acaccctcaa caagaacagc
     5941 gagatcttct cgtccatcaa gaagttggag aagcttttca agaacttcga cgagtacagc
     6001 agcgctggga tcttcgttaa gaacggacct gctatcagca ccatcagcaa ggatattttc
     6061 ggcgagtgga acgtgatcag ggacaagtgg aatgctgagt acgatgacat ccacctcaag
     6121 aagaaggctg tcgtcactga gaagtacgag gatgacaggc gtaagtcgtt caagaagatc
     6181 ggctctttca gcctcgagca gcttcaagaa tacgctgatg ctgatctcag cgtggtcgag
     6241 aagctcaaag agatcatcat ccagaaggtc gacgagatct acaaggtgta cgggtcctct
     6301 gagaagttgt tcgatgctga tttcgtcctc gagaagagtc tgaagaagaa cgacgctgtc
     6361 gtcgcgatca tgaaggattt gctcgacagc gtgaagtcct tcgagaacta tatcaaggcc
     6421 ttcttcggag agggcaaaga gactaatagg gacgagtctt tctacgggga tttcgtgctc
     6481 gcttacgata tcctcctcaa ggtggaccat atctacgacg ccatcagaaa ctacgtgacc
     6541 cagaagcctt acagcaagga caagttcaag ttgtactttc agaacccgca gttcatgggc
     6601 ggatgggaca aagacaaaga gacagattac agggccacca tcctcaggta cgggtctaag
     6661 tactacctgg ccatcatgga caagaaatac gccaagtgcc tccaaaagat cgacaaggat
     6721 gacgtgaacg ggaactatga gaagatcaac tacaagctcc ttccgggacc gaacaagatg
     6781 cttcctaagg tgttcttcag caagaaatgg atggcctact acaacccgtc tgaggacatc
     6841 cagaaaatct acaagaacgg gaccttcaag aaaggcgaca tgttcaacct caacgactgc
     6901 cacaagctca tcgatttctt caaggacagc atctcgcgtt acccgaagtg gtctaacgct
     6961 tacgacttta acttcagcga gacagaaaag tacaaggata tcgccgggtt ctaccgtgag
     7021 gttgaggaac agggttacaa ggttagcttc gagagcgcct ccaagaaaga ggttgacaag
     7081 ttggtcgaag agggcaagct ctacatgttc cagatctata acaaggactt ctccgacaag
     7141 agccacggaa ctcctaacct ccatacgatg tacttcaagc tgcttttcga cgagaacaac
     7201 cacgggcaga tcagactttc tggtggtgct gaactcttca tgcgtagggc ctcactcaag
     7261 aaagaagagt tggttgttca cccggccaac tctccaatcg ctaacaagaa tcctgacaac
     7321 ccgaaaaaga ccaccacgct gtcttacgac gtctacaagg acaaaaggtt cagcgaggac
     7381 cagtacgagc ttcatatccc gatcgctatc aacaagtgcc cgaagaacat cttcaagatc
     7441 aataccgagg tgagggtgct gctcaagcac gatgataacc cttacgtgat cggaatcgat
     7501 cgtggtgaga gaaacctcct ctacatcgtt gtggtggacg gaaagggaaa catcgtcgag
     7561 cagtacagcc tgaacgagat tatcaacaat ttcaacggca tcaggatcaa gaccgactac
     7621 cactcactcc tcgataagaa agaaaaagag cgtttcgagg ccaggcagaa ctggacttct
     7681 atcgaaaaca tcaaagagtt gaaggccggc tacatctctc aggtggtgca taagatctgc
     7741 gagctggtgg aaaagtacga tgctgtgatc gctcttgagg acctcaactc tgggttcaag
     7801 aacagtagag tgaaggttga gaagcaggtc taccaaaagt tcgagaagat gctcatcgac
     7861 aagctcaact acatggtgga caaaaagagc aacccttgcg ctaccggtgg tgctcttaag
     7921 ggataccaga tcacgaacaa gttcgagtcc ttcaagagca tgagcaccca gaacggcttc
     7981 atcttctata tccctgcttg gctcaccagc aagatcgatc cttctactgg tttcgtgaac
     8041 ctgctcaaga ccaagtacac ctcgatcgcc gacagcaaga agttcatctc gtctttcgac
     8101 aggatcatgt acgtgccgga agaggatctt ttcgagttcg ctctcgacta taagaacttc
     8161 agcaggaccg acgccgacta cattaagaag tggaagctct actcctacgg gaaccgtatc
     8221 aggatcttcc gaaatccgaa gaaaaacaac gtgttcgact gggaagaagt gtgcctcacc
     8281 tctgcctaca aagaactgtt caacaagtac ggcatcaact accagcaggg tgatatcagg
     8341 gctcttttgt gcgagcagag cgacaaggca ttctacagct cattcatggc cctcatgtct
     8401 ctcatgctcc agatgaggaa ctctatcacc ggaaggaccg atgtggactt ccttatctct
     8461 ccggtcaaga actctgacgg gatcttctac gacagccgta actatgaggc tcaagagaac
     8521 gctatcctgc cgaagaatgc tgatgcaaac ggggcttaca acattgcgag aaaggttctc
     8581 tgggctatcg ggcagtttaa gaaagcggaa gatgagaagc tggacaaggt gaagatcgcc
     8641 atctccaaca aagagtggct tgagtacgct cagacctccg ttaagcacaa gggacctaag
     8701 aagaagagaa aggtttgaaa tcatacagca ttcccagaaa gagaaacaga agaaatatac
     8761 aaactttcat tttgagagca gcacctcgtc tattgattgc agataatatg cttctcattt
     8821 gtatttcctt ttgattattt ttgtttctat ccctttgttt gagtcaatct caaatattcg
     8881 gtcattgttg gtatgaaaaa tcaagcagtt catgttaaga gtcaatttaa aattaatatt
     8941 tttatataga gttgtatgtg aaatgatgtt gtgatttggt atatatggat aaagagcttg
     9001 tcagttcatt ttgctctcat ttttttggta tccaaataag aaacacaaaa gggatatgtc
     9061 cctctactat caaatattag ttataagtat tcatgttata ctattcgata ttttctaccc
     9121 caatcgttac ctatttaaaa gtatttaccc ctccatctat caaacccctg gacccagctt
     9181 tcctattaca tgtggcttca tcttaagccc ccaaaccttt ttcttatttt tgatttttaa
     9241 aggctcatct taaaatttat tactcaaatt aatacctctt aataacccac ctcaaggacc
     9301 cagtaattaa atatccaatt agctccagta attggggttc atattagctc cagtcttaaa
     9361 ttttaaaggc gatgatcgta ttcctccact tggttcattt atactcaaag aatactcaat
     9421 gtctttagtg tttagataac tttttgtaaa tcatatagat tgttttaaca aaaaacaatt
     9481 caatagtaga ttttcacatg aaagttacat aaaaattctt taaaattact ttctcaaaaa
     9541 attgttccaa acatattatc ccacaattaa actcaatctg tttttcgaaa cctaaatcaa
     9601 aaccaatcca actaccttat ataatatata atcaatacat tgtaaagaac tgcatgttct
     9661 tttaaatttt gggggcaaag ttattccgta cgttcacaca tgtactaata ggaggtaata
     9721 aatgatatgt gaaacaatcg aggtgtaaac aagctagcat tgagcaggcc tcccagcttt
     9781 cgtccgtatc atcggtttcg acaacgttcg tcaagttcaa tgcatcagtt tcattgccca
     9841 cacaccagaa tcctactaag tttgagtatt atggcattgg aaaagctgtt ttcttctatc
     9901 atttgttctg cttgtaattt actgtgttct ttcagttttt gttttcggac atcaaaatgc
     9961 aaatggatgg ataagagtta ataaatgata tggtcctttt gttcattctc aaattattat
    10021 tatctgttgt ttttacttta atgggttgaa tttaagtaag aaaggaacta acagtgtgat
    10081 attaaggtgc aatgttagac atataaaaca gtctttcacc tctctttggt tatgtcttga
    10141 attggtttgt ttcttcactt atctgtgtaa tcaagtttac tatgagtcta tgatcaagta
    10201 attatgcaat caagttaagt acagtatagg ctttgtcatt ggcggctttt tttcttcttc
    10261 ttcgttcata cagttttttt ttgtttatca gcttacattt tcttgaaccg tagctttcgt
    10321 tttcttcttt ttaactttcc attcggagtt tttgtatctt gtttcatagt ttgtcccagg
    10381 attagaatga ttaggcatcg aaccttcaag aatttgattg aataaaacat cttcattctt
    10441 aagatatgaa gataatcttc aaaaggcccc tgggaatctg aaagaagaga agcaggccca
    10501 tttatatggg aaagaacaat agtatttctt atataggccc atttaagttg aaaacaatct
    10561 tcaaaagtcc cacatcgctt agataagaaa acgaagctga gtttatatac agctagagtc
    10621 gaagtagtga ttgaaattac tgatgagtcc gtgaggacga aacgagtaag ctcgtctaat
    10681 ttctactaag tgtagatagg gggactatgt gagtcgtggc cggcatggtc ccagcctcct
    10741 cgctggcgcc ggctgggcaa catgcttcgg catggcgaat gggacttttt tgacgctagc
    10801 ctagttcgct ttttttcttc ttcttcgttc atacagtttt tttttgttta tcagcttaca
    10861 ttttcttgaa ccgtagcttt cgttttcttc tttttaactt tccattcgga gtttttgtat
    10921 cttgtttcat agtttgtccc aggattagaa tgattaggca tcgaaccttc aagaatttga
    10981 ttgaataaaa catcttcatt cttaagatat gaagataatc ttcaaaaggc ccctgggaat
    11041 ctgaaagaag agaagcaggc ccatttatat gggaaagaac aatagtattt cttatatagg
    11101 cccatttaag ttgaaaacaa tcttcaaaag tcccacatcg cttagataag aaaacgaagc
    11161 tgagtttata tacagctaga gtcgaagtag tgattgaaat tactgatgag tccgtgagga
    11221 cgaaacgagt aagctcgtct aatttctact aagtgtagat atgagtaccg gaatcaccag
    11281 ggccggcatg gtcccagcct cctcgctggc gccggctggg caacatgctt cggcatggcg
    11341 aatgggactt ttttgtctgt cgcggggaga gaggattttg aggaaataat taattgaatt
    11401 acttgtatta ttgataaagt aatttagata agttgttagt acaacttatt gcaactatgg
    11461 atagacaaaa atcacttatt ttaaggggag aagtatataa caacttatta taaaatttct
    11521 gagcagccgg cctcctccaa tcaatcatag agagtggagc ccattctgga aaaccaagga
    11581 acccccaact tgcactggtg cggtggccca attcaaaaaa aacggggccc agcaaagtaa
    11641 cgccgttgta acgatttatc aatccaaact caaaaggcgg cgagatgcgt ttaactccgt
    11701 gaaattaaca aaccgccaac aacttgcaat ttgcaactac cgtttccaag aagaactcaa
    11761 ccacacaacg tatcctatcc caaaccacac gcaactagtg acgcgtcata aggacacgtg
    11821 tcacaatttg actggttaat aatttcaccg cttttgctat aaattacctc caatcccctt
    11881 agcttcttca caattcagtt cccaacccta acaattcttg ttcatatcgc ttctctctac
    11941 tttcaaggta tgatccaatt tctctcttct ttctctgtaa tcctttcgtt gagttttgtt
    12001 tccgatcaat cataggtagt tttcttgttt cgaagcatga gatctaggaa ttttttgtga
    12061 ttttccaaaa ttgagatcgg tttgaaattg aattttacag cttgaatctc agatcttgtt
    12121 ttatcaatgt tttcgatggc ttgcgatgta gatctatgat aattgtggtt cagttttgtt
    12181 aggaatcgat ttcggtttag caattgcaga ttaattaggg tttccaattg aattcttcag
    12241 atccgttatg gaattatgtc aaataattta ttcaaattgg aaattattgt tagatccact
    12301 cttaatctgt ttgatccaag cttcaattag ggttttcact tgtttcaatt tcttgttatg
    12361 gattctgatt tatctgttga tgttagatcc actcttaatc tgtttgatcc aagcgttaat
    12421 tagggttttc acttgtttca atttcgtgtg ttggattctg atttatctgt tgttgatgtg
    12481 attacagtct gaacaatggg cagccaccac caccaccacc acagcggaat gagttcactg
    12541 ccgtataggc agctaagaaa atcaggtggc accgaagatg caaagaacat taagaaaggg
    12601 cctgcaccat tctatccact tgaggacggt acagccggtg agcaattgca taaggccatg
    12661 aagagatatg cacttgtacc agggacgata gcctttactg atgctcacat tgaggtcaat
    12721 atcacttatg cggaatattt tgaaatgtca gtcagattgg cagaagctat gaaaaggtat
    12781 ggcttaaata ccaatcacag aattgtggtg tgttctgaga atagtttaca gttcttcatg
    12841 ccagtgttgg gtgcactatt tattggtgta gctgttgctc cagctaacga catttacaat
    12901 gagagagaac tactcaactc tatgaatatt tcacagccca cagtcgtatt tgtgagcaag
    12961 aaaggattgc agaagatact taacgttcag aagaaacttc cgattataca gaagatcatt
    13021 attatggaca gcaaaactga ctaccaaggc tttcagtcta tgtatacatt tgtgacatct
    13081 catttacctc caggctttaa tgagtatgac ttcgttccag aatcatttga tcgtgataag
    13141 actatcgctc ttattatgaa tagctctgga tccactggtc tgccgaaggg agtagcattg
    13201 cctcatagga cagcttgtgt taggttttca cacgcaagag atccgatatt tggaaatcag
    13261 atcattcctg atacggctat attgagtgtt gtaccatttc accatggctt tggaatgttt
    13321 accacacttg ggtacttgat ttgcggcttt agagtggtac tgatgtacag gttcgaggag
    13381 gaacttttcc tacgaagtct ccaagattac aaaatccaat ccgctttgtt agttcctaca
    13441 ctgttttcct tcttcgcgaa atctaccctg attgacaaat atgatctctc taatctccac
    13501 gaaatagcaa gtggtggtgc acctctttca aaagaggttg gagaagcagt tgccaaacgt
    13561 tttcatctac caggtatcag acaaggatat ggtcttactg agacaacatc agccatatta
    13621 attacccctg aaggcgatga taagcctggt gctgttggaa aagttgttcc attcttcgaa
    13681 gcaaaagtgg tagacttgga tactggcaag acgcttggag tcaaccaaag aggtgaactg
    13741 tgcgttagag ggcctatgat catgtctggg tacgttaaca atcctgaggc aaccaatgcg
    13801 ttaattgaca aagatggctg gttacatagt ggagatattg cgtactggga cgaagatgag
    13861 cacttcttca tagtggatag gttgaagtct ctcatcaagt acaaaggtta tcaagtagct
    13921 cctgctgaat tagagtcaat ccttcttcaa catcccaaca ttttcgatgc tggagtggcc
    13981 ggattgcccg atgatgatgc cggtgaattg cccgcagctg ttgttgtgct ggaacatggt
    14041 aagactatga cagagaagga aatagtggac tatgtcgcta gccaagttac tactgctaag
    14101 aaactccgtg gaggggttgt ctttgttgat gaagtcccaa aaggattgac tgggaaactt
    14161 gatgctcgaa agataagaga gattctcatt aaagctaaga agggtggaaa atccaaacta
    14221 taaaagggct aagacctaag ctagcctaga atcgatcgtt caaacatttg gcaataaagt
    14281 ttcttaagat tgaatcctgt tgccggtctt gcgatgatta tcatataatt tctgttgaat
    14341 tacgttaagc atgtaataat taacatgtaa tgcatgacgt tatttatgag atgggttttt
    14401 atgattagag tcccgcaatt atacatttaa tacgcgatag aaaacaaaat atagcgcgca
    14461 aactaggata aattatcgcg cgcggtgtca tctatgttac tagatctgag tgacctaagc
    14521 tagcctagtg tctgaggcgg ccaacaaact attcgtcacg tcgacaaaat ttagaacgaa
    14581 cttaattatg atctcaaata cattgataca tatctcatct agatctaggt tatcattatg
    14641 taagaaagtt ttgacgaata tggcacgaca aaatggctag actcgatgta attggtatct
    14701 caactcaaca ttatacttat accaaacatt agttagacaa aatttaaaca actatttttt
    14761 atgtatgcaa gagtcagcat atgtataatt gattcagaat cgttttgacg agttcggatg
    14821 tagtagtagc cattatttaa tgtacatact aatcgtgaat agtgaatatg atgaaacatt
    14881 gtatcttatt gtataaatat ccataaacac atcatgaaag acactttctt tcacggtctg
    14941 aattaattat gatacaattc taatagaaaa cgaattaaat tacgttgaat tgtatgaaat
    15001 ctaattgaac aagccaacca cgacgacgac taacgttgcc tggattgact cggtttaagt
    15061 taaccactaa aaaaacggag ctgtcatgta acacgcggat cgagcaggtc acagtcatga
    15121 agccatcaaa gcaaaagaac taatccaagg gctgagatga ttaattagtt taaaaattag
    15181 ttaacacgag ggaaaaggct gtctgacagc caggtcacgt tatctttacc tgtggtcgaa
    15241 atgattcgtg tctgtcgatt ttaattattt ttttgaaagg ccgaaaataa agttgtaaga
    15301 gataaacccg cctatataaa ttcatatatt ttcctctccg ctttgaattg tctcgttgtc
    15361 ctcctcactt tcatcagccg ttttgaatct ccggcgactt gacagagaag aacaaggaag
    15421 aagtctaaga gagaaagtaa gagataatcc aggagattca ttctccgttt tgaatcttcc
    15481 tcaatctcat cttcttccgc tctttctttc caaggtaata ggaactttct ggatctactt
    15541 tatttgctgg atctcgatct tgttttctca atttccttga gatctggaat tcgtttaatt
    15601 tggatctgtg aacctccact aaatcttttg gttttactag aatcgatcta agttgaccga
    15661 tcagttagct cgattatagc taccagaatt tggcttgacc ttgatggaga gatccatgtt
    15721 catgttacct gggaaatgat ttgtatatgt gaattgaaat ctgaactgtt gaagttagat
    15781 tgaatctgaa cactgtcaat gttagattga atctgaacac tgtttaaggt tagatgaagt
    15841 ttgtgtatag attcttcgaa actttaggat ttgtagtgtc gtacgttgaa cagaaagcta
    15901 tttctgattc aatcagggtt tatttgactg tattgaactc tttttgtgtg tttgcagctc
    15961 ataaaaatct gcccacgact cacatagtcc ccctcaaaca ccatggatca ttatcttgat
    16021 attagactta gacctgatcc tgaatttcct cctgctcaac ttatgtctgt tctttttgga
    16081 aagcttcatc aagctcttgt tgctcaagga ggagatagaa ttggagtttc ttttcctgat
    16141 cttgatgaat ctagatctag acttggagaa agacttagaa ttcatgcttc tgctgatgat
    16201 cttagagctc ttcttgctag accttggctt gaaggactta gagatcatct tcaatttgga
    16261 gaacctgctg ttgttcctca tcctactcct tatagacaag tttctagagt tcaagctaag
    16321 tctaatcctg aaagacttag aagaagattg atgaggagac atgatctttc tgaagaagaa
    16381 gctagaaaga gaattcctga tactgttgct agagctcttg atcttccttt tgttactctt
    16441 agatctcaat ctactggaca acattttaga ctttttatta gacatggacc tcttcaagtt
    16501 actgctgaag aaggaggatt tacttgttat ggactttcta agggaggatt tgttccttgg
    16561 ttttgaaagg gctaagacct aagctagcct agaatcatat gaagatgaag atgaaatatt
    16621 tggtgtgtca aataaaaagc ttgtgtgctt aagtttgtgt ttttttcttg gcttgttgtg
    16681 ttatgaattt gtggcttttt ctaatattaa atgaatgtaa gatctcatta taatgaataa
    16741 acaaatgttt ctataatcca ttgtgaatgt tttgttggat ctcttctgca gcatataact
    16801 actgtatgtg ctatggtatg gactatggaa tatgattaaa gataagtgag tgacctaagc
    16861 tagcctagtg tctctgtctg tgagacctct ggtttaaact atcagtgttt gacaggatat
    16921 attggcgggt aaacctaaga gaaaagagcg tttattagaa taatcggata tttaaaaggg
    16981 cgtgaaaagg tttatccgtt cgtccatttg tatgtgcatg ccaaccacag ggttcccctc
    17041 gggatcaaag tactttgatc caacccctcc gctgctatag tgcagtcggc ttctgacgtt
    17101 cagtgcagcc ttcttctgaa aacgacatgt cgcacaagtc ctaagttacg cgacaggctg
    17161 ccgccctgcc cttttcctgg cgttttcttg tcgcgtgttt tagtcgcata aagtagaata
    17221 cttgcgacta gaaccggaga cattacgcca tgaacaagag cgccgccgct ggcctgctgg
    17281 gctatgcccg cgtcagcacc gacgaccagg acttgaccaa ccaacgggcc gaactgcacg
    17341 cggccggctg caccaagctg ttttccgaga agatcaccgg caccaggcgc gaccgcccgg
    17401 agctggccag gatgcttgac cacctacgcc ctggcgacgt tgtgacagtg accaggctag
    17461 accgcctggc ccgcagcacc cgcgacctac tggacattgc cgagcgcatc caggaggccg
    17521 gcgcgggcct gcgtagcctg gcagagccgt gggccgacac caccacgccg gccggccgca
    17581 tggtgttgac cgtgttcgcc ggcattgccg agttcgagcg ttccctaatc atcgaccgca
    17641 cccggagcgg gcgcgaggcc gccaaggccc gaggcgtgaa gtttggcccc cgccctaccc
    17701 tcaccccggc acagatcgcg cacgcccgcg agctgatcga ccaggaaggc cgcaccgtga
    17761 aagaggcggc tgcactgctt ggcgtgcatc gctcgaccct gtaccgcgca cttgagcgca
    17821 gcgaggaagt gacgcccacc gaggccaggc ggcgcggtgc cttccgtgag gacgcattga
    17881 ccgaggccga cgccctggcg gccgccgaga atgaacgcca agaggaacaa gcatgaaacc
    17941 gcaccaggac ggccaggacg aaccgttttt cattaccgaa gagatcgagg cggagatgat
    18001 cgcggccggg tacgtgttcg agccgcccgc gcacatctca accgtgcggc tgcatgaaat
    18061 cctggccggt ttgtctgatg ccaagctggc ggcctggccg gccagcttgg ccgctgaaga
    18121 aaccgagcgc cgccgtctaa aaaggtgatg tgtatttgag taaaacagct tgcgtcatgc
    18181 ggtcgctgcg tatatgatgc gatgagtaaa taaacaaata cgcaagggga acgcatgaag
    18241 gttatcgctg tacttaacca gaaaggcggg tcaggcaaga cgaccatcgc aacccatcta
    18301 gcccgcgccc tgcaactcgc cggggccgat gttctgttag tcgattccga tccccagggc
    18361 agtgcccgcg attgggcggc cgtgcgggaa gatcaaccgc taaccgttgt cggcatcgac
    18421 cgcccgacga ttgaccgcga cgtgaaggcc atcggccggc gcgacttcgt agtgatcgac
    18481 ggagcgcccc aggcggcgga cttggctgtg tccgcgatca aggcagccga cttcgtgctg
    18541 attccggtgc agccaagccc ttacgacata tgggccaccg ccgacctggt ggagctggtt
    18601 aagcagcgca ttgaggtcac ggatggaagg ctacaagcgg cctttgtcgt gtcgcgggcg
    18661 atcaaaggca cgcgcatcgg cggtgaggtt gccgaggcgc tggccgggta cgagctgccc
    18721 attcttgagt cccgtatcac gcagcgcgtg agctacccag gcactgccgc cgccggcaca
    18781 accgttcttg aatcagaacc cgagggcgac gctgcccgcg aggtccaggc gctggccgct
    18841 gaaattaaat caaaactcat ttgagttaat gaggtaaaga gaaaatgagc aaaagcacaa
    18901 acacgctaag tgccggccgt ccgagcgcac gcagcagcaa ggctgcaacg ttggccagcc
    18961 tggcagacac gccagccatg aagcgggtca actttcagtt gccggcggag gatcacacca
    19021 agctgaagat gtacgcggta cgccaaggca agaccattac cgagctgcta tctgaataca
    19081 tcgcgcagct accagagtaa atgagcaaat gaataaatga gtagatgaat tttagcggct
    19141 aaaggaggcg gcatggaaaa tcaagaacaa ccaggcaccg acgccgtgga atgccccatg
    19201 tgtggaggaa cgggcggttg gccaggcgta agcggctggg ttgcctgccg gccctgcaat
    19261 ggcactggaa cccccaagcc cgaggaatcg gcgtgagcgg tcgcaaacca tccggcccgg
    19321 tacaaatcgg cgcggcgctg ggtgatgacc tggtggagaa gttgaaggcc gcgcaggccg
    19381 cccagcggca acgcatcgag gcagaagcac gccccggtga atcgtggcaa gcggccgctg
    19441 atcgaatccg caaagaatcc cggcaaccgc cggcagccgg tgcgccgtcg attaggaagc
    19501 cgcccaaggg cgacgagcaa ccagattttt tcgttccgat gctctatgac gtgggcaccc
    19561 gcgatagtcg cagcatcatg gacgtggccg ttttccgtct gtcgaagcgt gaccgacgag
    19621 ctggcgaggt gatccgctac gagcttccag acgggcacgt agaggtttcc gcagggccgg
    19681 ccggcatggc cagtgtgtgg gattacgacc tggtactgat ggcggtttcc catctaaccg
    19741 aatccatgaa ccgataccgg gaagggaagg gagacaagcc cggccgcgtg ttccgtccac
    19801 acgttgcgga cgtactcaag ttctgccggc gagccgatgg cggaaagcag aaagacgacc
    19861 tggtagaaac ctgcattcgg ttaaacacca cgcacgttgc catgcagcgt acgaagaagg
    19921 ccaagaacgg ccgcctggtg acggtatccg agggtgaagc cttgattagc cgctacaaga
    19981 tcgtaaagag cgaaaccggg cggccggagt acatcgagat cgagctagct gattggatgt
    20041 accgcgagat cacagaaggc aagaacccgg acgtgctgac ggttcacccc gattactttt
    20101 tgatcgatcc cggcatcggc cgttttctct accgcctggc acgccgcgcc gcaggcaagg
    20161 cagaagccag atggttgttc aagacgatct acgaacgcag tggcagcgcc ggagagttca
    20221 agaagttctg tttcaccgtg cgcaagctga tcgggtcaaa tgacctgccg gagtacgatt
    20281 tgaaggagga ggcggggcag gctggcccga tcctagtcat gcgctaccgc aacctgatcg
    20341 agggcgaagc atccgccggt tcctaatgta cggagcagat gctagggcaa attgccctag
    20401 caggggaaaa aggtcgaaaa ggactctttc ctgtggatag cacgtacatt gggaacccaa
    20461 agccgtacat tgggaaccgg aacccgtaca ttgggaaccc aaagccgtac attgggaacc
    20521 ggtcacacat gtaagtgact gatataaaag agaaaaaagg cgatttttcc gcctaaaact
    20581 ctttaaaact tattaaaact cttaaaaccc gcctggcctg tgcataactg tctggccagc
    20641 gcacagccga agagctgcaa aaagcgccta cccttcggtc gctgcgctcc ctacgccccg
    20701 ccgcttcgcg tcggcctatc gcggccgctg gccgctcaaa aatggctggc ctacggccag
    20761 gcaatctacc agggcgcgga caagccgcgc cgtcgccact cgaccgccgg cgcccacatc
    20821 aaggcaccct gcctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
    20881 ccggagacag tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
    20941 gcgtcagcgg gt
//
