LOCUS       sequence-444778-       20903 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cas12a and Csy4 with 5' and 3' targets (I) for Nicotiana leaf assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 20903)
  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 20903)
  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: pMB61
FEATURES             Location/Qualifiers
     source          1..20903
                     /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"
     promoter        4320..4665
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     4608..4628
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             4685..4705
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8432..8452
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             12451..12468
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    12479..12498
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
     terminator      14200..14452
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     terminator      16549..16797
                     /label=HSP terminator
                     /note="efficient transcription terminator from the 
                     Arabidopsis thaliana heat shock protein 18.2 gene (Nagaya 
                     et al., 2010)"
     misc_feature    16861..16885
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             18186..18815
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             19244..20317
                     /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      20383..20577
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(20813..20835)
                     /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 ggagattagc
     3841 cttttcaatt tcagaaagaa tgctaaccca cagatggtta gagaggctta cgcagcacga
     3901 ctcatcaaga cgatctaccc gagcaataat ctccaggaaa tcaaatacct tcccaagaag
     3961 gttaaagatg cagtcaaaag attcaggact aactgcatca agaacacaga gaaagatata
     4021 tttctcaaga tcagaagtac tattccagta tggacgattc aaggcttgct tcacaaacca
     4081 aggcaagtaa tagagattgg agtctctaaa aaggtagttc ccactgaatc aaaggccatg
     4141 gagtcaaaga ttcaaataga ggacctaaca gaactcgccg taaagactgg cgaacagttc
     4201 atacagagtc tcttacgact caatgacaag aagaaaatct tcgtcaacat ggtggagcac
     4261 gacacacttg tctactccaa aaatatcaaa gatacagtct cagaagagca aagggcaatt
     4321 gagacttttc aacaaagggt aatatccgga aacctcctcg gattccattg cccagctatc
     4381 tgtcacttta ttgtgaagat agtggaaaag gaaggtggct cctacaaatg ccatcattgc
     4441 gataaaggaa aggccatcgt tgaagatgcc tctgccgaca gtggtcccaa agatggaccc
     4501 ccacccacga ggagcatcgt ggaaaaagaa cacgttccaa ccacgacttc aaagcaagtg
     4561 gattgatgtg atatctccac tgacgtaagg gatgacgcac aatcccacta tccttcgcaa
     4621 gacccttcct ctatataagg aagttcattt catttggaga gaacacgtct gaacaatgct
     4681 gcagcctaag aagaagagaa aggttggagg agtcgactcg agtgcggccg ccaccatgag
     4741 caagctcgag aagtttacca actgctacag cctctctaag accctcaggt tcaaggctat
     4801 ccctgtggga aagacccaag agaatatcga caacaagagg ctcctcgtcg aggatgagaa
     4861 gagagctgaa gattacaagg gcgtgaagaa gctcctcgac aggtactacc tcagcttcat
     4921 caacgatgtg ctccacagca tcaagctcaa gaacctcaac aactacatca gcctcttccg
     4981 taagaaaacc aggaccgaga aagagaacaa agagcttgag aacctcgaga tcaacctccg
     5041 taaagagatc gccaaggctt tcaagggaaa cgagggatac aagagcctct tcaagaagga
     5101 tattatcgag acaatcctgc ctgagttcct ggacgataag gatgagatcg ctctcgtgaa
     5161 cagcttcaac ggattcacta ctgccttcac cggattcttc agaaacaggg aaaacatgtt
     5221 cagcgaagag gccaagagca cctctatcgc tttcagatgc atcaacgaga acctcacgcg
     5281 ttacatcagc aacatggaca tcttcgagaa ggtggacgcc atcttcgata agcacgaggt
     5341 gcaagaaatc aaagagaaga tcctcaacag cgactacgac gtcgaggact tttttgaagg
     5401 ggagttcttc aacttcgttc tcacccaaga gggcatcgac gtgtacaacg ctattatcgg
     5461 aggattcgtg accgagtctg gggagaagat taagggactc aacgagtaca tcaacctgta
     5521 caaccagaaa acgaagcaga agctcccgaa gttcaagccg ctctacaagc aggttctctc
     5581 tgatcgtgag agcctctcat tttacggtga gggttacacc tctgacgagg aagtgcttga
     5641 ggttttccgt aacaccctca acaagaacag cgagatcttc tcgtccatca agaagttgga
     5701 gaagcttttc aagaacttcg acgagtacag cagcgctggg atcttcgtta agaacggacc
     5761 tgctatcagc accatcagca aggatatttt cggcgagtgg aacgtgatca gggacaagtg
     5821 gaatgctgag tacgatgaca tccacctcaa gaagaaggct gtcgtcactg agaagtacga
     5881 ggatgacagg cgtaagtcgt tcaagaagat cggctctttc agcctcgagc agcttcaaga
     5941 atacgctgat gctgatctca gcgtggtcga gaagctcaaa gagatcatca tccagaaggt
     6001 cgacgagatc tacaaggtgt acgggtcctc tgagaagttg ttcgatgctg atttcgtcct
     6061 cgagaagagt ctgaagaaga acgacgctgt cgtcgcgatc atgaaggatt tgctcgacag
     6121 cgtgaagtcc ttcgagaact atatcaaggc cttcttcgga gagggcaaag agactaatag
     6181 ggacgagtct ttctacgggg atttcgtgct cgcttacgat atcctcctca aggtggacca
     6241 tatctacgac gccatcagaa actacgtgac ccagaagcct tacagcaagg acaagttcaa
     6301 gttgtacttt cagaacccgc agttcatggg cggatgggac aaagacaaag agacagatta
     6361 cagggccacc atcctcaggt acgggtctaa gtactacctg gccatcatgg acaagaaata
     6421 cgccaagtgc ctccaaaaga tcgacaagga tgacgtgaac gggaactatg agaagatcaa
     6481 ctacaagctc cttccgggac cgaacaagat gcttcctaag gtgttcttca gcaagaaatg
     6541 gatggcctac tacaacccgt ctgaggacat ccagaaaatc tacaagaacg ggaccttcaa
     6601 gaaaggcgac atgttcaacc tcaacgactg ccacaagctc atcgatttct tcaaggacag
     6661 catctcgcgt tacccgaagt ggtctaacgc ttacgacttt aacttcagcg agacagaaaa
     6721 gtacaaggat atcgccgggt tctaccgtga ggttgaggaa cagggttaca aggttagctt
     6781 cgagagcgcc tccaagaaag aggttgacaa gttggtcgaa gagggcaagc tctacatgtt
     6841 ccagatctat aacaaggact tctccgacaa gagccacgga actcctaacc tccatacgat
     6901 gtacttcaag ctgcttttcg acgagaacaa ccacgggcag atcagacttt ctggtggtgc
     6961 tgaactcttc atgcgtaggg cctcactcaa gaaagaagag ttggttgttc acccggccaa
     7021 ctctccaatc gctaacaaga atcctgacaa cccgaaaaag accaccacgc tgtcttacga
     7081 cgtctacaag gacaaaaggt tcagcgagga ccagtacgag cttcatatcc cgatcgctat
     7141 caacaagtgc ccgaagaaca tcttcaagat caataccgag gtgagggtgc tgctcaagca
     7201 cgatgataac ccttacgtga tcggaatcga tcgtggtgag agaaacctcc tctacatcgt
     7261 tgtggtggac ggaaagggaa acatcgtcga gcagtacagc ctgaacgaga ttatcaacaa
     7321 tttcaacggc atcaggatca agaccgacta ccactcactc ctcgataaga aagaaaaaga
     7381 gcgtttcgag gccaggcaga actggacttc tatcgaaaac atcaaagagt tgaaggccgg
     7441 ctacatctct caggtggtgc ataagatctg cgagctggtg gaaaagtacg atgctgtgat
     7501 cgctcttgag gacctcaact ctgggttcaa gaacagtaga gtgaaggttg agaagcaggt
     7561 ctaccaaaag ttcgagaaga tgctcatcga caagctcaac tacatggtgg acaaaaagag
     7621 caacccttgc gctaccggtg gtgctcttaa gggataccag atcacgaaca agttcgagtc
     7681 cttcaagagc atgagcaccc agaacggctt catcttctat atccctgctt ggctcaccag
     7741 caagatcgat ccttctactg gtttcgtgaa cctgctcaag accaagtaca cctcgatcgc
     7801 cgacagcaag aagttcatct cgtctttcga caggatcatg tacgtgccgg aagaggatct
     7861 tttcgagttc gctctcgact ataagaactt cagcaggacc gacgccgact acattaagaa
     7921 gtggaagctc tactcctacg ggaaccgtat caggatcttc cgaaatccga agaaaaacaa
     7981 cgtgttcgac tgggaagaag tgtgcctcac ctctgcctac aaagaactgt tcaacaagta
     8041 cggcatcaac taccagcagg gtgatatcag ggctcttttg tgcgagcaga gcgacaaggc
     8101 attctacagc tcattcatgg ccctcatgtc tctcatgctc cagatgagga actctatcac
     8161 cggaaggacc gatgtggact tccttatctc tccggtcaag aactctgacg ggatcttcta
     8221 cgacagccgt aactatgagg ctcaagagaa cgctatcctg ccgaagaatg ctgatgcaaa
     8281 cggggcttac aacattgcga gaaaggttct ctgggctatc gggcagttta agaaagcgga
     8341 agatgagaag ctggacaagg tgaagatcgc catctccaac aaagagtggc ttgagtacgc
     8401 tcagacctcc gttaagcaca agggactgca gcctaagaag aagagaaagg ttggaggagt
     8461 cgactcgagt gcgtaaaatc atacagcatt cccagaaaga gaaacagaag aaatatacaa
     8521 actttcattt tgagagcagc acctcgtcta ttgattgcag ataatatgct tctcatttgt
     8581 atttcctttt gattattttt gtttctatcc ctttgtttga gtcaatctca aatattcggt
     8641 cattgttggt atgaaaaatc aagcagttca tgttaagagt caatttaaaa ttaatatttt
     8701 tatatagagt tgtatgtgaa atgatgttgt gatttggtat atatggataa agagcttgtc
     8761 agttcatttt gctctcattt ttttggtatc caaataagaa acacaaaagg gatatgtccc
     8821 tctactatca aatattagtt ataagtattc atgttatact attcgatatt ttctacccca
     8881 atcgttacct atttaaaagt atttacccct ccatctatca aacccctgga cccagctttc
     8941 ctattacatg tggcttcatc ttaagccccc aaaccttttt cttatttttg atttttaaag
     9001 gctcatctta aaatttatta ctcaaattaa tacctcttaa taacccacct caaggaccca
     9061 gtaattaaat atccaattag ctccagtaat tggggttcat attagctcca gtcttaaatt
     9121 ttaaaggcga tgatcgtatt cctccacttg gttcatttat actcaaagaa tactcaatgt
     9181 ctttagtgtt tagataactt tttgtaaatc atatagattg ttttaacaaa aaacaattca
     9241 atagtagatt ttcacatgaa agttacataa aaattcttta aaattacttt ctcaaaaaat
     9301 tgttccaaac atattatccc acaattaaac tcaatctgtt tttcgaaacc taaatcaaaa
     9361 ccaatccaac taccttatat aatatataat caatacattg taaagaactg catgttcttt
     9421 taaattttgg gggcaaagtt attccgtacg ttcacacatg tactaatagg aggtaataaa
     9481 tgatatgtga aacaatcgag gtgtaaacaa gctagcattg agcaggcctc ccagctttcg
     9541 tccgtatcat cggtttcgac aacgttcgtc aagttcaatg catcagtttc attgcccaca
     9601 caccagaatc ctactaagtt tgagtattat ggcattggaa aagctgtttt cttctatcat
     9661 ttgttctgct tgtaatttac tgtgttcttt cagtttttgt tttcggacat caaaatgcaa
     9721 atggatggat aagagttaat aaatgatatg gtccttttgt tcattctcaa attattatta
     9781 tctgttgttt ttactttaat gggttgaatt taagtaagaa aggaactaac agtgtgatat
     9841 taaggtgcaa tgttagacat ataaaacagt ctttcacctc tctttggtta tgtcttgaat
     9901 tggtttgttt cttcacttat ctgtgtaatc aagtttacta tgagtctatg atcaagtaat
     9961 tatgcaatca agttaagtac agtataggct ttgtcattgg cggaatcttt gagagggaat
    10021 aaaagaaata cttccaaaaa tccattagaa gaactataaa tcttctgggc atgtaaatat
    10081 aacgtaaaat atatgttatg ttacaacctt atttcattca cattttaatt ttttttcaca
    10141 caatttttat ttaattttgt ttttatctct attttatttt catatgataa tacatattat
    10201 ctctttattt tactttatgt gtctctttgc tctcacattt tatttcttaa ctaacatatt
    10261 ctcattagca gtagctatcg taagactaat ctcacacaat aattaattat ccattctatt
    10321 tatgaatttt tatcatttaa taattcttta cctgtgcaaa ttttgaagga aaaagtttct
    10381 gaatataaaa taacaataat actaattaga tgtcaaatca aatcacccag gctagacaat
    10441 agtggtaatg aagaaggcta gaggtagctc tgtgcctcta ggggagtgag agacgttttt
    10501 catttattgg tgtgttgaaa cttgaaacgt tgatgttctc cacagctcca cgtattcttt
    10561 cgtcctagaa cgaactagag caacataatc tgtctcttcc tgaagcttca ttgtttttct
    10621 ttccttctag cagcatcttc ttaacagacg aagaagggac tcagatgctt gcttgcatgc
    10681 ttccaaagtt tggagctgag gagacttgtg ctatttgagt ctgttttagc attcgcaagc
    10741 cccacaccgg ataaacatac agaaatctga gtgtttataa ccaaatagtc agcaacaagg
    10801 ttcgaatttc tactaagtgt agataggggg actatgtgag tcgtaatttc tactaagtgt
    10861 agatatgagt accggaatca ccagaatttc tactaagtgt agattttttt tggcaaaaat
    10921 tttcagattt tttcttcatc tgtagatttc tgggtttttt tttccgtttc gtgaatcata
    10981 agtgaagttt tggatgcaaa tctgcgcgaa aaaagttgga cctgcaatga gcttatttag
    11041 atagctaaga caaagtgatt ggtccgttgt ttcagttctg attgtcagag agtttgtttc
    11101 gagtcggcga caccaatgcg ttttgttaac cagatttcgg gtaagaaatg tatcgagagt
    11161 ttgtttccag acggctacat cattttctta tgaaggatga aattagatag accaaagatt
    11221 gaaacacaac atttctttca caaaaatata ataaacttga tagcatttag gatcagctga
    11281 gtgacctaag ctagcctagt gtctgtcgcg gggagagagg attttgagga aataattaat
    11341 tgaattactt gtattattga taaagtaatt tagataagtt gttagtacaa cttattgcaa
    11401 ctatggatag acaaaaatca cttattttaa ggggagaagt atataacaac ttattataaa
    11461 atttctgagc agccggcctc ctccaatcaa tcatagagag tggagcccat tctggaaaac
    11521 caaggaaccc ccaacttgca ctggtgcggt ggcccaattc aaaaaaaacg gggcccagca
    11581 aagtaacgcc gttgtaacga tttatcaatc caaactcaaa aggcggcgag atgcgtttaa
    11641 ctccgtgaaa ttaacaaacc gccaacaact tgcaatttgc aactaccgtt tccaagaaga
    11701 actcaaccac acaacgtatc ctatcccaaa ccacacgcaa ctagtgacgc gtcataagga
    11761 cacgtgtcac aatttgactg gttaataatt tcaccgcttt tgctataaat tacctccaat
    11821 ccccttagct tcttcacaat tcagttccca accctaacaa ttcttgttca tatcgcttct
    11881 ctctactttc aaggtatgat ccaatttctc tcttctttct ctgtaatcct ttcgttgagt
    11941 tttgtttccg atcaatcata ggtagttttc ttgtttcgaa gcatgagatc taggaatttt
    12001 ttgtgatttt ccaaaattga gatcggtttg aaattgaatt ttacagcttg aatctcagat
    12061 cttgttttat caatgttttc gatggcttgc gatgtagatc tatgataatt gtggttcagt
    12121 tttgttagga atcgatttcg gtttagcaat tgcagattaa ttagggtttc caattgaatt
    12181 cttcagatcc gttatggaat tatgtcaaat aatttattca aattggaaat tattgttaga
    12241 tccactctta atctgtttga tccaagcttc aattagggtt ttcacttgtt tcaatttctt
    12301 gttatggatt ctgatttatc tgttgatgtt agatccactc ttaatctgtt tgatccaagc
    12361 gttaattagg gttttcactt gtttcaattt cgtgtgttgg attctgattt atctgttgtt
    12421 gatgtgatta cagtctgaac aatgggcagc caccaccacc accaccacag cggaatgagt
    12481 tcactgccgt ataggcagct aagaaaatca ggtggcaccg aagatgcaaa gaacattaag
    12541 aaagggcctg caccattcta tccacttgag gacggtacag ccggtgagca attgcataag
    12601 gccatgaaga gatatgcact tgtaccaggg acgatagcct ttactgatgc tcacattgag
    12661 gtcaatatca cttatgcgga atattttgaa atgtcagtca gattggcaga agctatgaaa
    12721 aggtatggct taaataccaa tcacagaatt gtggtgtgtt ctgagaatag tttacagttc
    12781 ttcatgccag tgttgggtgc actatttatt ggtgtagctg ttgctccagc taacgacatt
    12841 tacaatgaga gagaactact caactctatg aatatttcac agcccacagt cgtatttgtg
    12901 agcaagaaag gattgcagaa gatacttaac gttcagaaga aacttccgat tatacagaag
    12961 atcattatta tggacagcaa aactgactac caaggctttc agtctatgta tacatttgtg
    13021 acatctcatt tacctccagg ctttaatgag tatgacttcg ttccagaatc atttgatcgt
    13081 gataagacta tcgctcttat tatgaatagc tctggatcca ctggtctgcc gaagggagta
    13141 gcattgcctc ataggacagc ttgtgttagg ttttcacacg caagagatcc gatatttgga
    13201 aatcagatca ttcctgatac ggctatattg agtgttgtac catttcacca tggctttgga
    13261 atgtttacca cacttgggta cttgatttgc ggctttagag tggtactgat gtacaggttc
    13321 gaggaggaac ttttcctacg aagtctccaa gattacaaaa tccaatccgc tttgttagtt
    13381 cctacactgt tttccttctt cgcgaaatct accctgattg acaaatatga tctctctaat
    13441 ctccacgaaa tagcaagtgg tggtgcacct ctttcaaaag aggttggaga agcagttgcc
    13501 aaacgttttc atctaccagg tatcagacaa ggatatggtc ttactgagac aacatcagcc
    13561 atattaatta cccctgaagg cgatgataag cctggtgctg ttggaaaagt tgttccattc
    13621 ttcgaagcaa aagtggtaga cttggatact ggcaagacgc ttggagtcaa ccaaagaggt
    13681 gaactgtgcg ttagagggcc tatgatcatg tctgggtacg ttaacaatcc tgaggcaacc
    13741 aatgcgttaa ttgacaaaga tggctggtta catagtggag atattgcgta ctgggacgaa
    13801 gatgagcact tcttcatagt ggataggttg aagtctctca tcaagtacaa aggttatcaa
    13861 gtagctcctg ctgaattaga gtcaatcctt cttcaacatc ccaacatttt cgatgctgga
    13921 gtggccggat tgcccgatga tgatgccggt gaattgcccg cagctgttgt tgtgctggaa
    13981 catggtaaga ctatgacaga gaaggaaata gtggactatg tcgctagcca agttactact
    14041 gctaagaaac tccgtggagg ggttgtcttt gttgatgaag tcccaaaagg attgactggg
    14101 aaacttgatg ctcgaaagat aagagagatt ctcattaaag ctaagaaggg tggaaaatcc
    14161 aaactataaa agggctaaga cctaagctag cctagaatcg atcgttcaaa catttggcaa
    14221 taaagtttct taagattgaa tcctgttgcc ggtcttgcga tgattatcat ataatttctg
    14281 ttgaattacg ttaagcatgt aataattaac atgtaatgca tgacgttatt tatgagatgg
    14341 gtttttatga ttagagtccc gcaattatac atttaatacg cgatagaaaa caaaatatag
    14401 cgcgcaaact aggataaatt atcgcgcgcg gtgtcatcta tgttactaga tctgagtgac
    14461 ctaagctagc ctagtgtctg aggcggccaa caaactattc gtcacgtcga caaaatttag
    14521 aacgaactta attatgatct caaatacatt gatacatatc tcatctagat ctaggttatc
    14581 attatgtaag aaagttttga cgaatatggc acgacaaaat ggctagactc gatgtaattg
    14641 gtatctcaac tcaacattat acttatacca aacattagtt agacaaaatt taaacaacta
    14701 ttttttatgt atgcaagagt cagcatatgt ataattgatt cagaatcgtt ttgacgagtt
    14761 cggatgtagt agtagccatt atttaatgta catactaatc gtgaatagtg aatatgatga
    14821 aacattgtat cttattgtat aaatatccat aaacacatca tgaaagacac tttctttcac
    14881 ggtctgaatt aattatgata caattctaat agaaaacgaa ttaaattacg ttgaattgta
    14941 tgaaatctaa ttgaacaagc caaccacgac gacgactaac gttgcctgga ttgactcggt
    15001 ttaagttaac cactaaaaaa acggagctgt catgtaacac gcggatcgag caggtcacag
    15061 tcatgaagcc atcaaagcaa aagaactaat ccaagggctg agatgattaa ttagtttaaa
    15121 aattagttaa cacgagggaa aaggctgtct gacagccagg tcacgttatc tttacctgtg
    15181 gtcgaaatga ttcgtgtctg tcgattttaa ttattttttt gaaaggccga aaataaagtt
    15241 gtaagagata aacccgccta tataaattca tatattttcc tctccgcttt gaattgtctc
    15301 gttgtcctcc tcactttcat cagccgtttt gaatctccgg cgacttgaca gagaagaaca
    15361 aggaagaagt ctaagagaga aagtaagaga taatccagga gattcattct ccgttttgaa
    15421 tcttcctcaa tctcatcttc ttccgctctt tctttccaag gtaataggaa ctttctggat
    15481 ctactttatt tgctggatct cgatcttgtt ttctcaattt ccttgagatc tggaattcgt
    15541 ttaatttgga tctgtgaacc tccactaaat cttttggttt tactagaatc gatctaagtt
    15601 gaccgatcag ttagctcgat tatagctacc agaatttggc ttgaccttga tggagagatc
    15661 catgttcatg ttacctggga aatgatttgt atatgtgaat tgaaatctga actgttgaag
    15721 ttagattgaa tctgaacact gtcaatgtta gattgaatct gaacactgtt taaggttaga
    15781 tgaagtttgt gtatagattc ttcgaaactt taggatttgt agtgtcgtac gttgaacaga
    15841 aagctatttc tgattcaatc agggtttatt tgactgtatt gaactctttt tgtgtgtttg
    15901 cagctcataa aaatctgccc acgactcaca tagtccccct caaacaccat ggatcattat
    15961 cttgatatta gacttagacc tgatcctgaa tttcctcctg ctcaacttat gtctgttctt
    16021 tttggaaagc ttcatcaagc tcttgttgct caaggaggag atagaattgg agtttctttt
    16081 cctgatcttg atgaatctag atctagactt ggagaaagac ttagaattca tgcttctgct
    16141 gatgatctta gagctcttct tgctagacct tggcttgaag gacttagaga tcatcttcaa
    16201 tttggagaac ctgctgttgt tcctcatcct actccttata gacaagtttc tagagttcaa
    16261 gctaagtcta atcctgaaag acttagaaga agattgatga ggagacatga tctttctgaa
    16321 gaagaagcta gaaagagaat tcctgatact gttgctagag ctcttgatct tccttttgtt
    16381 actcttagat ctcaatctac tggacaacat tttagacttt ttattagaca tggacctctt
    16441 caagttactg ctgaagaagg aggatttact tgttatggac tttctaaggg aggatttgtt
    16501 ccttggtttt gaaaggtttg atgagtaccg gaatcaccag gggaatcata tgaagatgaa
    16561 gatgaaatat ttggtgtgtc aaataaaaag cttgtgtgct taagtttgtg tttttttctt
    16621 ggcttgttgt gttatgaatt tgtggctttt tctaatatta aatgaatgta agatctcatt
    16681 ataatgaata aacaaatgtt tctataatcc attgtgaatg ttttgttgga tctcttctgc
    16741 agcatataac tactgtatgt gctatggtat ggactatgga atatgattaa agataagtga
    16801 gtgacctaag ctagcctagt gtctctgtct gtgagacctc tggtttaaac tatcagtgtt
    16861 tgacaggata tattggcggg taaacctaag agaaaagagc gtttattaga ataatcggat
    16921 atttaaaagg gcgtgaaaag gtttatccgt tcgtccattt gtatgtgcat gccaaccaca
    16981 gggttcccct cgggatcaaa gtactttgat ccaacccctc cgctgctata gtgcagtcgg
    17041 cttctgacgt tcagtgcagc cttcttctga aaacgacatg tcgcacaagt cctaagttac
    17101 gcgacaggct gccgccctgc ccttttcctg gcgttttctt gtcgcgtgtt ttagtcgcat
    17161 aaagtagaat acttgcgact agaaccggag acattacgcc atgaacaaga gcgccgccgc
    17221 tggcctgctg ggctatgccc gcgtcagcac cgacgaccag gacttgacca accaacgggc
    17281 cgaactgcac gcggccggct gcaccaagct gttttccgag aagatcaccg gcaccaggcg
    17341 cgaccgcccg gagctggcca ggatgcttga ccacctacgc cctggcgacg ttgtgacagt
    17401 gaccaggcta gaccgcctgg cccgcagcac ccgcgaccta ctggacattg ccgagcgcat
    17461 ccaggaggcc ggcgcgggcc tgcgtagcct ggcagagccg tgggccgaca ccaccacgcc
    17521 ggccggccgc atggtgttga ccgtgttcgc cggcattgcc gagttcgagc gttccctaat
    17581 catcgaccgc acccggagcg ggcgcgaggc cgccaaggcc cgaggcgtga agtttggccc
    17641 ccgccctacc ctcaccccgg cacagatcgc gcacgcccgc gagctgatcg accaggaagg
    17701 ccgcaccgtg aaagaggcgg ctgcactgct tggcgtgcat cgctcgaccc tgtaccgcgc
    17761 acttgagcgc agcgaggaag tgacgcccac cgaggccagg cggcgcggtg ccttccgtga
    17821 ggacgcattg accgaggccg acgccctggc ggccgccgag aatgaacgcc aagaggaaca
    17881 agcatgaaac cgcaccagga cggccaggac gaaccgtttt tcattaccga agagatcgag
    17941 gcggagatga tcgcggccgg gtacgtgttc gagccgcccg cgcacatctc aaccgtgcgg
    18001 ctgcatgaaa tcctggccgg tttgtctgat gccaagctgg cggcctggcc ggccagcttg
    18061 gccgctgaag aaaccgagcg ccgccgtcta aaaaggtgat gtgtatttga gtaaaacagc
    18121 ttgcgtcatg cggtcgctgc gtatatgatg cgatgagtaa ataaacaaat acgcaagggg
    18181 aacgcatgaa ggttatcgct gtacttaacc agaaaggcgg gtcaggcaag acgaccatcg
    18241 caacccatct agcccgcgcc ctgcaactcg ccggggccga tgttctgtta gtcgattccg
    18301 atccccaggg cagtgcccgc gattgggcgg ccgtgcggga agatcaaccg ctaaccgttg
    18361 tcggcatcga ccgcccgacg attgaccgcg acgtgaaggc catcggccgg cgcgacttcg
    18421 tagtgatcga cggagcgccc caggcggcgg acttggctgt gtccgcgatc aaggcagccg
    18481 acttcgtgct gattccggtg cagccaagcc cttacgacat atgggccacc gccgacctgg
    18541 tggagctggt taagcagcgc attgaggtca cggatggaag gctacaagcg gcctttgtcg
    18601 tgtcgcgggc gatcaaaggc acgcgcatcg gcggtgaggt tgccgaggcg ctggccgggt
    18661 acgagctgcc cattcttgag tcccgtatca cgcagcgcgt gagctaccca ggcactgccg
    18721 ccgccggcac aaccgttctt gaatcagaac ccgagggcga cgctgcccgc gaggtccagg
    18781 cgctggccgc tgaaattaaa tcaaaactca tttgagttaa tgaggtaaag agaaaatgag
    18841 caaaagcaca aacacgctaa gtgccggccg tccgagcgca cgcagcagca aggctgcaac
    18901 gttggccagc ctggcagaca cgccagccat gaagcgggtc aactttcagt tgccggcgga
    18961 ggatcacacc aagctgaaga tgtacgcggt acgccaaggc aagaccatta ccgagctgct
    19021 atctgaatac atcgcgcagc taccagagta aatgagcaaa tgaataaatg agtagatgaa
    19081 ttttagcggc taaaggaggc ggcatggaaa atcaagaaca accaggcacc gacgccgtgg
    19141 aatgccccat gtgtggagga acgggcggtt ggccaggcgt aagcggctgg gttgcctgcc
    19201 ggccctgcaa tggcactgga acccccaagc ccgaggaatc ggcgtgagcg gtcgcaaacc
    19261 atccggcccg gtacaaatcg gcgcggcgct gggtgatgac ctggtggaga agttgaaggc
    19321 cgcgcaggcc gcccagcggc aacgcatcga ggcagaagca cgccccggtg aatcgtggca
    19381 agcggccgct gatcgaatcc gcaaagaatc ccggcaaccg ccggcagccg gtgcgccgtc
    19441 gattaggaag ccgcccaagg gcgacgagca accagatttt ttcgttccga tgctctatga
    19501 cgtgggcacc cgcgatagtc gcagcatcat ggacgtggcc gttttccgtc tgtcgaagcg
    19561 tgaccgacga gctggcgagg tgatccgcta cgagcttcca gacgggcacg tagaggtttc
    19621 cgcagggccg gccggcatgg ccagtgtgtg ggattacgac ctggtactga tggcggtttc
    19681 ccatctaacc gaatccatga accgataccg ggaagggaag ggagacaagc ccggccgcgt
    19741 gttccgtcca cacgttgcgg acgtactcaa gttctgccgg cgagccgatg gcggaaagca
    19801 gaaagacgac ctggtagaaa cctgcattcg gttaaacacc acgcacgttg ccatgcagcg
    19861 tacgaagaag gccaagaacg gccgcctggt gacggtatcc gagggtgaag ccttgattag
    19921 ccgctacaag atcgtaaaga gcgaaaccgg gcggccggag tacatcgaga tcgagctagc
    19981 tgattggatg taccgcgaga tcacagaagg caagaacccg gacgtgctga cggttcaccc
    20041 cgattacttt ttgatcgatc ccggcatcgg ccgttttctc taccgcctgg cacgccgcgc
    20101 cgcaggcaag gcagaagcca gatggttgtt caagacgatc tacgaacgca gtggcagcgc
    20161 cggagagttc aagaagttct gtttcaccgt gcgcaagctg atcgggtcaa atgacctgcc
    20221 ggagtacgat ttgaaggagg aggcggggca ggctggcccg atcctagtca tgcgctaccg
    20281 caacctgatc gagggcgaag catccgccgg ttcctaatgt acggagcaga tgctagggca
    20341 aattgcccta gcaggggaaa aaggtcgaaa aggactcttt cctgtggata gcacgtacat
    20401 tgggaaccca aagccgtaca ttgggaaccg gaacccgtac attgggaacc caaagccgta
    20461 cattgggaac cggtcacaca tgtaagtgac tgatataaaa gagaaaaaag gcgatttttc
    20521 cgcctaaaac tctttaaaac ttattaaaac tcttaaaacc cgcctggcct gtgcataact
    20581 gtctggccag cgcacagccg aagagctgca aaaagcgcct acccttcggt cgctgcgctc
    20641 cctacgcccc gccgcttcgc gtcggcctat cgcggccgct ggccgctcaa aaatggctgg
    20701 cctacggcca ggcaatctac cagggcgcgg acaagccgcg ccgtcgccac tcgaccgccg
    20761 gcgcccacat caaggcaccc tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac
    20821 acatgcagct cccggagaca gtcacagctt gtctgtaagc ggatgccggg agcagacaag
    20881 cccgtcaggg cgcgtcagcg ggt
//
