LOCUS       sequence-449332-       20416 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cas12a and Csy4 with 5' and 3' targets (I) for Lotus callus assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 20416)
  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 20416)
  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: pMB45
FEATURES             Location/Qualifiers
     source          1..20416
                     /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             2607..3632
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRSQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     K"
     CDS             5062..5082
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8797..8844
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             12848..12865
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    12876..12895
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
     CDS             join(12935..12937,12938..12940,12941..13651)
                     /codon_start=1
                     /product="enhanced YFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(12980..13001)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(13241..13260)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     13588..13609
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             13664..13684
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      13713..13965
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     terminator      16062..16310
                     /label=HSP terminator
                     /note="efficient transcription terminator from the 
                     Arabidopsis thaliana heat shock protein 18.2 gene (Nagaya 
                     et al., 2010)"
     misc_feature    16374..16398
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             17699..18328
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             18757..19830
                     /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      19896..20090
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(20326..20348)
                     /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 aaaagcctga actcaccgcg acgtctgtcg
     2641 agaagtttct gatcgaaaag ttcgacagcg tgtccgacct gatgcagctc tcggagggcg
     2701 aagaatctcg tgctttcagc ttcgatgtag gagggcgtgg atatgtcctg cgggtaaata
     2761 gctgcgccga tggtttctac aaagatcgtt atgtttatcg gcactttgca tcggccgcgc
     2821 tcccgattcc ggaagtgctt gacattgggg agtttagcga gagcctgacc tattgcatct
     2881 cccgccgttc acagggtgtc acgttgcaag acctgcctga aaccgaactg cccgctgttc
     2941 tacaaccggt cgcggaggct atggatgcga tcgctgcggc cgatcttagc cagacgagcg
     3001 ggttcggccc attcggaccg caaggaatcg gtcaatacac tacatggcgt gatttcatat
     3061 gcgcgattgc tgatccccat gtgtatcact ggcaaactgt gatggacgac accgtcagtg
     3121 cgtccgtcgc gcaggctctc gatgagctga tgctttgggc cgaggactgc cccgaagtcc
     3181 ggcacctcgt gcacgcggat ttcggctcca acaatgtcct gacggacaat ggccgcataa
     3241 cagcggtcat tgactggagc gaggcgatgt tcggggattc ccaatacgag gtcgccaaca
     3301 tcttcttctg gaggccgtgg ttggcttgta tggagcagca gacgcgctac ttcgagcgga
     3361 ggcatccgga gcttgcagga tcgccacgac tccgggcgta tatgctccgc attggtcttg
     3421 accaactcta tcagagcttg gttgacggca atttcgatga tgcagcttgg gcgcagggtc
     3481 gatgcgacgc aatcgtccga tccggagccg ggactgtcgg gcgtacacaa atcgcccgca
     3541 gaagcgcggc cgtctggacc gatggctgtg tagaagtact cgccgatagt ggaaaccgac
     3601 gccccagcac tcgtccgagg gcaaagaaat agaagggcta agacctaagc tagcctagaa
     3661 tccggccatg ctagagtccg caaaaatcac cagtctctct ctacaaatct atctctctct
     3721 atttttctcc agaataatgt gtgagtagtt cccagataag ggaattaggg ttcttatagg
     3781 gtttcgctca tgtgttgagc atataagaaa cccttagtat gtatttgtat ttgtaaaata
     3841 cttctatcaa taaaatttct aattcctaaa accaaaatcc agtgaccttg agtgacctaa
     3901 gctagcctag tgtcgggcgc ggggagagag gattttgagg aaataattaa ttgaattact
     3961 tgtattattg ataaagtaat ttagataagt tgttagtaca acttattgca actatggata
     4021 gacaaaaatc acttatttta aggggagaag tatataacaa cttattataa aatttctgag
     4081 cagccggcct cctccaatca atcatagaga gtggagccca ttctggaaaa ccaaggaacc
     4141 cccaacttgc actggtgcgg tggcccaatt caaaaaaaac ggggcccagc aaagtaacgc
     4201 cgttgtaacg atttatcaat ccaaactcaa aaggcggcga gatgcgttta actccgtgaa
     4261 attaacaaac cgccaacaac ttgcaatttg caactaccgt ttccaagaag aactcaacca
     4321 cacaacgtat cctatcccaa accacacgca actagtgacg cgtcataagg acacgtgtca
     4381 caatttgact ggttaataat ttcaccgctt ttgctataaa ttacctccaa tccccttagc
     4441 ttcttcacaa ttcagttccc aaccctaaca attcttgttc atatcgcttc tctctacttt
     4501 caaggtatga tccaatttct ctcttctttc tctgtaatcc tttcgttgag ttttgtttcc
     4561 gatcaatcat aggtagtttt cttgtttcga agcatgagat ctaggaattt tttgtgattt
     4621 tccaaaattg agatcggttt gaaattgaat tttacagctt gaatctcaga tcttgtttta
     4681 tcaatgtttt cgatggcttg cgatgtagat ctatgataat tgtggttcag ttttgttagg
     4741 aatcgatttc ggtttagcaa ttgcagatta attagggttt ccaattgaat tcttcagatc
     4801 cgttatggaa ttatgtcaaa taatttattc aaattggaaa ttattgttag atccactctt
     4861 aatctgtttg atccaagctt caattagggt tttcacttgt ttcaatttct tgttatggat
     4921 tctgatttat ctgttgatgt tagatccact cttaatctgt ttgatccaag cgttaattag
     4981 ggttttcact tgtttcaatt tcgtgtgttg gattctgatt tatctgttgt tgatgtgatt
     5041 acagtctgaa caatgctgca gcctaagaag aagagaaagg ttggaggagt cgactcgagt
     5101 gcggccgcca ccatgagcaa gctcgagaag tttaccaact gctacagcct ctctaagacc
     5161 ctcaggttca aggctatccc tgtgggaaag acccaagaga atatcgacaa caagaggctc
     5221 ctcgtcgagg atgagaagag agctgaagat tacaagggcg tgaagaagct cctcgacagg
     5281 tactacctca gcttcatcaa cgatgtgctc cacagcatca agctcaagaa cctcaacaac
     5341 tacatcagcc tcttccgtaa gaaaaccagg accgagaaag agaacaaaga gcttgagaac
     5401 ctcgagatca acctccgtaa agagatcgcc aaggctttca agggaaacga gggatacaag
     5461 agcctcttca agaaggatat tatcgagaca atcctgcctg agttcctgga cgataaggat
     5521 gagatcgctc tcgtgaacag cttcaacgga ttcactactg ccttcaccgg attcttcaga
     5581 aacagggaaa acatgttcag cgaagaggcc aagagcacct ctatcgcttt cagatgcatc
     5641 aacgagaacc tcacgcgtta catcagcaac atggacatct tcgagaaggt ggacgccatc
     5701 ttcgataagc acgaggtgca agaaatcaaa gagaagatcc tcaacagcga ctacgacgtc
     5761 gaggactttt ttgaagggga gttcttcaac ttcgttctca cccaagaggg catcgacgtg
     5821 tacaacgcta ttatcggagg attcgtgacc gagtctgggg agaagattaa gggactcaac
     5881 gagtacatca acctgtacaa ccagaaaacg aagcagaagc tcccgaagtt caagccgctc
     5941 tacaagcagg ttctctctga tcgtgagagc ctctcatttt acggtgaggg ttacacctct
     6001 gacgaggaag tgcttgaggt tttccgtaac accctcaaca agaacagcga gatcttctcg
     6061 tccatcaaga agttggagaa gcttttcaag aacttcgacg agtacagcag cgctgggatc
     6121 ttcgttaaga acggacctgc tatcagcacc atcagcaagg atattttcgg cgagtggaac
     6181 gtgatcaggg acaagtggaa tgctgagtac gatgacatcc acctcaagaa gaaggctgtc
     6241 gtcactgaga agtacgagga tgacaggcgt aagtcgttca agaagatcgg ctctttcagc
     6301 ctcgagcagc ttcaagaata cgctgatgct gatctcagcg tggtcgagaa gctcaaagag
     6361 atcatcatcc agaaggtcga cgagatctac aaggtgtacg ggtcctctga gaagttgttc
     6421 gatgctgatt tcgtcctcga gaagagtctg aagaagaacg acgctgtcgt cgcgatcatg
     6481 aaggatttgc tcgacagcgt gaagtccttc gagaactata tcaaggcctt cttcggagag
     6541 ggcaaagaga ctaataggga cgagtctttc tacggggatt tcgtgctcgc ttacgatatc
     6601 ctcctcaagg tggaccatat ctacgacgcc atcagaaact acgtgaccca gaagccttac
     6661 agcaaggaca agttcaagtt gtactttcag aacccgcagt tcatgggcgg atgggacaaa
     6721 gacaaagaga cagattacag ggccaccatc ctcaggtacg ggtctaagta ctacctggcc
     6781 atcatggaca agaaatacgc caagtgcctc caaaagatcg acaaggatga cgtgaacggg
     6841 aactatgaga agatcaacta caagctcctt ccgggaccga acaagatgct tcctaaggtg
     6901 ttcttcagca agaaatggat ggcctactac aacccgtctg aggacatcca gaaaatctac
     6961 aagaacggga ccttcaagaa aggcgacatg ttcaacctca acgactgcca caagctcatc
     7021 gatttcttca aggacagcat ctcgcgttac ccgaagtggt ctaacgctta cgactttaac
     7081 ttcagcgaga cagaaaagta caaggatatc gccgggttct accgtgaggt tgaggaacag
     7141 ggttacaagg ttagcttcga gagcgcctcc aagaaagagg ttgacaagtt ggtcgaagag
     7201 ggcaagctct acatgttcca gatctataac aaggacttct ccgacaagag ccacggaact
     7261 cctaacctcc atacgatgta cttcaagctg cttttcgacg agaacaacca cgggcagatc
     7321 agactttctg gtggtgctga actcttcatg cgtagggcct cactcaagaa agaagagttg
     7381 gttgttcacc cggccaactc tccaatcgct aacaagaatc ctgacaaccc gaaaaagacc
     7441 accacgctgt cttacgacgt ctacaaggac aaaaggttca gcgaggacca gtacgagctt
     7501 catatcccga tcgctatcaa caagtgcccg aagaacatct tcaagatcaa taccgaggtg
     7561 agggtgctgc tcaagcacga tgataaccct tacgtgatcg gaatcgatcg tggtgagaga
     7621 aacctcctct acatcgttgt ggtggacgga aagggaaaca tcgtcgagca gtacagcctg
     7681 aacgagatta tcaacaattt caacggcatc aggatcaaga ccgactacca ctcactcctc
     7741 gataagaaag aaaaagagcg tttcgaggcc aggcagaact ggacttctat cgaaaacatc
     7801 aaagagttga aggccggcta catctctcag gtggtgcata agatctgcga gctggtggaa
     7861 aagtacgatg ctgtgatcgc tcttgaggac ctcaactctg ggttcaagaa cagtagagtg
     7921 aaggttgaga agcaggtcta ccaaaagttc gagaagatgc tcatcgacaa gctcaactac
     7981 atggtggaca aaaagagcaa cccttgcgct accggtggtg ctcttaaggg ataccagatc
     8041 acgaacaagt tcgagtcctt caagagcatg agcacccaga acggcttcat cttctatatc
     8101 cctgcttggc tcaccagcaa gatcgatcct tctactggtt tcgtgaacct gctcaagacc
     8161 aagtacacct cgatcgccga cagcaagaag ttcatctcgt ctttcgacag gatcatgtac
     8221 gtgccggaag aggatctttt cgagttcgct ctcgactata agaacttcag caggaccgac
     8281 gccgactaca ttaagaagtg gaagctctac tcctacggga accgtatcag gatcttccga
     8341 aatccgaaga aaaacaacgt gttcgactgg gaagaagtgt gcctcacctc tgcctacaaa
     8401 gaactgttca acaagtacgg catcaactac cagcagggtg atatcagggc tcttttgtgc
     8461 gagcagagcg acaaggcatt ctacagctca ttcatggccc tcatgtctct catgctccag
     8521 atgaggaact ctatcaccgg aaggaccgat gtggacttcc ttatctctcc ggtcaagaac
     8581 tctgacggga tcttctacga cagccgtaac tatgaggctc aagagaacgc tatcctgccg
     8641 aagaatgctg atgcaaacgg ggcttacaac attgcgagaa aggttctctg ggctatcggg
     8701 cagtttaaga aagcggaaga tgagaagctg gacaaggtga agatcgccat ctccaacaaa
     8761 gagtggcttg agtacgctca gacctccgtt aagcacaaga ggcctgctgc tactaagaaa
     8821 gctggtcagg ctaagaagaa gaaatgaaag ggctaagacc taagctagcc tagaatcata
     8881 cagcattccc agaaagagaa acagaagaaa tatacaaact ttcattttga gagcagcacc
     8941 tcgtctattg attgcagata atatgcttct catttgtatt tccttttgat tatttttgtt
     9001 tctatccctt tgtttgagtc aatctcaaat attcggtcat tgttggtatg aaaaatcaag
     9061 cagttcatgt taagagtcaa tttaaaatta atatttttat atagagttgt atgtgaaatg
     9121 atgttgtgat ttggtatata tggataaaga gcttgtcagt tcattttgct ctcatttttt
     9181 tggtatccaa ataagaaaca caaaagggat atgtccctct actatcaaat attagttata
     9241 agtattcatg ttatactatt cgatattttc taccccaatc gttacctatt taaaagtatt
     9301 tacccctcca tctatcaaac ccctggaccc agctttccta ttacatgtgg cttcatctta
     9361 agcccccaaa cctttttctt atttttgatt tttaaaggct catcttaaaa tttattactc
     9421 aaattaatac ctcttaataa cccacctcaa ggacccagta attaaatatc caattagctc
     9481 cagtaattgg ggttcatatt agctccagtc ttaaatttta aaggcgatga tcgtattcct
     9541 ccacttggtt catttatact caaagaatac tcaatgtctt tagtgtttag ataacttttt
     9601 gtaaatcata tagattgttt taacaaaaaa caattcaata gtagattttc acatgaaagt
     9661 tacataaaaa ttctttaaaa ttactttctc aaaaaattgt tccaaacata ttatcccaca
     9721 attaaactca atctgttttt cgaaacctaa atcaaaacca atccaactac cttatataat
     9781 atataatcaa tacattgtaa agaactgcat gttcttttaa attttggggg caaagttatt
     9841 ccgtacgttc acacatgtac taataggagg taataaatga tatgtgaaac aatcgaggtg
     9901 taaacaagct agcattgagc aggcctccca gctttcgtcc gtatcatcgg tttcgacaac
     9961 gttcgtcaag ttcaatgcat cagtttcatt gcccacacac cagaatccta ctaagtttga
    10021 gtattatggc attggaaaag ctgttttctt ctatcatttg ttctgcttgt aatttactgt
    10081 gttctttcag tttttgtttt cggacatcaa aatgcaaatg gatggataag agttaataaa
    10141 tgatatggtc cttttgttca ttctcaaatt attattatct gttgttttta ctttaatggg
    10201 ttgaatttaa gtaagaaagg aactaacagt gtgatattaa ggtgcaatgt tagacatata
    10261 aaacagtctt tcacctctct ttggttatgt cttgaattgg tttgtttctt cacttatctg
    10321 tgtaatcaag tttactatga gtctatgatc aagtaattat gcaatcaagt taagtacagt
    10381 ataggctttg tcattggcgg aatctttgag agggaataaa agaaatactt ccaaaaatcc
    10441 attagaagaa ctataaatct tctgggcatg taaatataac gtaaaatata tgttatgtta
    10501 caaccttatt tcattcacat tttaattttt tttcacacaa tttttattta attttgtttt
    10561 tatctctatt ttattttcat atgataatac atattatctc tttattttac tttatgtgtc
    10621 tctttgctct cacattttat ttcttaacta acatattctc attagcagta gctatcgtaa
    10681 gactaatctc acacaataat taattatcca ttctatttat gaatttttat catttaataa
    10741 ttctttacct gtgcaaattt tgaaggaaaa agtttctgaa tataaaataa caataatact
    10801 aattagatgt caaatcaaat cacccaggct agacaatagt ggtaatgaag aaggctagag
    10861 gtagctctgt gcctctaggg gagtgagaga cgtttttcat ttattggtgt gttgaaactt
    10921 gaaacgttga tgttctccac agctccacgt attctttcgt cctagaacga actagagcaa
    10981 cataatctgt ctcttcctga agcttcattg tttttctttc cttctagcag catcttctta
    11041 acagacgaag aagggactca gatgcttgct tgcatgcttc caaagtttgg agctgaggag
    11101 acttgtgcta tttgagtctg ttttagcatt cgcaagcccc acaccggata aacatacaga
    11161 aatctgagtg tttataacca aatagtcagc aacaaggttc gaatttctac taagtgtaga
    11221 tagggggact atgtgagtcg taatttctac taagtgtaga tatgagtacc ggaatcacca
    11281 gaatttctac taagtgtaga ttttttttgg caaaaatttt cagatttttt cttcatctgt
    11341 agatttctgg gttttttttt ccgtttcgtg aatcataagt gaagttttgg atgcaaatct
    11401 gcgcgaaaaa agttggacct gcaatgagct tatttagata gctaagacaa agtgattggt
    11461 ccgttgtttc agttctgatt gtcagagagt ttgtttcgag tcggcgacac caatgcgttt
    11521 tgttaaccag atttcgggta agaaatgtat cgagagtttg tttccagacg gctacatcat
    11581 tttcttatga aggatgaaat tagatagacc aaagattgaa acacaacatt tctttcacaa
    11641 aaatataata aacttgatag catttaggat cagctgagtg acctaagcta gcctagtgtc
    11701 tgtcgcgggg agagaggatt ttgaggaaat aattaattga attacttgta ttattgataa
    11761 agtaatttag ataagttgtt agtacaactt attgcaacta tggatagaca aaaatcactt
    11821 attttaaggg gagaagtata taacaactta ttataaaatt tctgagcagc cggcctcctc
    11881 caatcaatca tagagagtgg agcccattct ggaaaaccaa ggaaccccca acttgcactg
    11941 gtgcggtggc ccaattcaaa aaaaacgggg cccagcaaag taacgccgtt gtaacgattt
    12001 atcaatccaa actcaaaagg cggcgagatg cgtttaactc cgtgaaatta acaaaccgcc
    12061 aacaacttgc aatttgcaac taccgtttcc aagaagaact caaccacaca acgtatccta
    12121 tcccaaacca cacgcaacta gtgacgcgtc ataaggacac gtgtcacaat ttgactggtt
    12181 aataatttca ccgcttttgc tataaattac ctccaatccc cttagcttct tcacaattca
    12241 gttcccaacc ctaacaattc ttgttcatat cgcttctctc tactttcaag gtatgatcca
    12301 atttctctct tctttctctg taatcctttc gttgagtttt gtttccgatc aatcataggt
    12361 agttttcttg tttcgaagca tgagatctag gaattttttg tgattttcca aaattgagat
    12421 cggtttgaaa ttgaatttta cagcttgaat ctcagatctt gttttatcaa tgttttcgat
    12481 ggcttgcgat gtagatctat gataattgtg gttcagtttt gttaggaatc gatttcggtt
    12541 tagcaattgc agattaatta gggtttccaa ttgaattctt cagatccgtt atggaattat
    12601 gtcaaataat ttattcaaat tggaaattat tgttagatcc actcttaatc tgtttgatcc
    12661 aagcttcaat tagggttttc acttgtttca atttcttgtt atggattctg atttatctgt
    12721 tgatgttaga tccactctta atctgtttga tccaagcgtt aattagggtt ttcacttgtt
    12781 tcaatttcgt gtgttggatt ctgatttatc tgttgttgat gtgattacag tctgaacaat
    12841 gggcagccac caccaccacc accacagcgg aatgagttca ctgccgtata ggcagctaag
    12901 aaaatcaggt ggcaccggcg gcagcggcgg atccatggtg agcaagggcg aggagctgtt
    12961 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
    13021 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg
    13081 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccttcg gctacggcct
    13141 gatgtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
    13201 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac
    13261 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
    13321 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca
    13381 caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg
    13441 ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat
    13501 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agctaccagt ccaagctgag
    13561 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg
    13621 gatcactctc ggcatggacg agctgtacaa gaagggactg cagcctaaga agaagagaaa
    13681 ggttggagga gtcgactcga gtgcgtaaaa tcgatcgttc aaacatttgg caataaagtt
    13741 tcttaagatt gaatcctgtt gccggtcttg cgatgattat catataattt ctgttgaatt
    13801 acgttaagca tgtaataatt aacatgtaat gcatgacgtt atttatgaga tgggttttta
    13861 tgattagagt cccgcaatta tacatttaat acgcgataga aaacaaaata tagcgcgcaa
    13921 actaggataa attatcgcgc gcggtgtcat ctatgttact agatctgagt gacctaagct
    13981 agcctagtgt ctgaggcggc caacaaacta ttcgtcacgt cgacaaaatt tagaacgaac
    14041 ttaattatga tctcaaatac attgatacat atctcatcta gatctaggtt atcattatgt
    14101 aagaaagttt tgacgaatat ggcacgacaa aatggctaga ctcgatgtaa ttggtatctc
    14161 aactcaacat tatacttata ccaaacatta gttagacaaa atttaaacaa ctatttttta
    14221 tgtatgcaag agtcagcata tgtataattg attcagaatc gttttgacga gttcggatgt
    14281 agtagtagcc attatttaat gtacatacta atcgtgaata gtgaatatga tgaaacattg
    14341 tatcttattg tataaatatc cataaacaca tcatgaaaga cactttcttt cacggtctga
    14401 attaattatg atacaattct aatagaaaac gaattaaatt acgttgaatt gtatgaaatc
    14461 taattgaaca agccaaccac gacgacgact aacgttgcct ggattgactc ggtttaagtt
    14521 aaccactaaa aaaacggagc tgtcatgtaa cacgcggatc gagcaggtca cagtcatgaa
    14581 gccatcaaag caaaagaact aatccaaggg ctgagatgat taattagttt aaaaattagt
    14641 taacacgagg gaaaaggctg tctgacagcc aggtcacgtt atctttacct gtggtcgaaa
    14701 tgattcgtgt ctgtcgattt taattatttt tttgaaaggc cgaaaataaa gttgtaagag
    14761 ataaacccgc ctatataaat tcatatattt tcctctccgc tttgaattgt ctcgttgtcc
    14821 tcctcacttt catcagccgt tttgaatctc cggcgacttg acagagaaga acaaggaaga
    14881 agtctaagag agaaagtaag agataatcca ggagattcat tctccgtttt gaatcttcct
    14941 caatctcatc ttcttccgct ctttctttcc aaggtaatag gaactttctg gatctacttt
    15001 atttgctgga tctcgatctt gttttctcaa tttccttgag atctggaatt cgtttaattt
    15061 ggatctgtga acctccacta aatcttttgg ttttactaga atcgatctaa gttgaccgat
    15121 cagttagctc gattatagct accagaattt ggcttgacct tgatggagag atccatgttc
    15181 atgttacctg ggaaatgatt tgtatatgtg aattgaaatc tgaactgttg aagttagatt
    15241 gaatctgaac actgtcaatg ttagattgaa tctgaacact gtttaaggtt agatgaagtt
    15301 tgtgtataga ttcttcgaaa ctttaggatt tgtagtgtcg tacgttgaac agaaagctat
    15361 ttctgattca atcagggttt atttgactgt attgaactct ttttgtgtgt ttgcagctca
    15421 taaaaatctg cccacgactc acatagtccc cctcaaacac catggatcat tatcttgata
    15481 ttagacttag acctgatcct gaatttcctc ctgctcaact tatgtctgtt ctttttggaa
    15541 agcttcatca agctcttgtt gctcaaggag gagatagaat tggagtttct tttcctgatc
    15601 ttgatgaatc tagatctaga cttggagaaa gacttagaat tcatgcttct gctgatgatc
    15661 ttagagctct tcttgctaga ccttggcttg aaggacttag agatcatctt caatttggag
    15721 aacctgctgt tgttcctcat cctactcctt atagacaagt ttctagagtt caagctaagt
    15781 ctaatcctga aagacttaga agaagattga tgaggagaca tgatctttct gaagaagaag
    15841 ctagaaagag aattcctgat actgttgcta gagctcttga tcttcctttt gttactctta
    15901 gatctcaatc tactggacaa cattttagac tttttattag acatggacct cttcaagtta
    15961 ctgctgaaga aggaggattt acttgttatg gactttctaa gggaggattt gttccttggt
    16021 tttgaaaggt ttgatgagta ccggaatcac caggggaatc atatgaagat gaagatgaaa
    16081 tatttggtgt gtcaaataaa aagcttgtgt gcttaagttt gtgttttttt cttggcttgt
    16141 tgtgttatga atttgtggct ttttctaata ttaaatgaat gtaagatctc attataatga
    16201 ataaacaaat gtttctataa tccattgtga atgttttgtt ggatctcttc tgcagcatat
    16261 aactactgta tgtgctatgg tatggactat ggaatatgat taaagataag tgagtgacct
    16321 aagctagcct agtgtctctg tctgtgagac ctctggttta aactatcagt gtttgacagg
    16381 atatattggc gggtaaacct aagagaaaag agcgtttatt agaataatcg gatatttaaa
    16441 agggcgtgaa aaggtttatc cgttcgtcca tttgtatgtg catgccaacc acagggttcc
    16501 cctcgggatc aaagtacttt gatccaaccc ctccgctgct atagtgcagt cggcttctga
    16561 cgttcagtgc agccttcttc tgaaaacgac atgtcgcaca agtcctaagt tacgcgacag
    16621 gctgccgccc tgcccttttc ctggcgtttt cttgtcgcgt gttttagtcg cataaagtag
    16681 aatacttgcg actagaaccg gagacattac gccatgaaca agagcgccgc cgctggcctg
    16741 ctgggctatg cccgcgtcag caccgacgac caggacttga ccaaccaacg ggccgaactg
    16801 cacgcggccg gctgcaccaa gctgttttcc gagaagatca ccggcaccag gcgcgaccgc
    16861 ccggagctgg ccaggatgct tgaccaccta cgccctggcg acgttgtgac agtgaccagg
    16921 ctagaccgcc tggcccgcag cacccgcgac ctactggaca ttgccgagcg catccaggag
    16981 gccggcgcgg gcctgcgtag cctggcagag ccgtgggccg acaccaccac gccggccggc
    17041 cgcatggtgt tgaccgtgtt cgccggcatt gccgagttcg agcgttccct aatcatcgac
    17101 cgcacccgga gcgggcgcga ggccgccaag gcccgaggcg tgaagtttgg cccccgccct
    17161 accctcaccc cggcacagat cgcgcacgcc cgcgagctga tcgaccagga aggccgcacc
    17221 gtgaaagagg cggctgcact gcttggcgtg catcgctcga ccctgtaccg cgcacttgag
    17281 cgcagcgagg aagtgacgcc caccgaggcc aggcggcgcg gtgccttccg tgaggacgca
    17341 ttgaccgagg ccgacgccct ggcggccgcc gagaatgaac gccaagagga acaagcatga
    17401 aaccgcacca ggacggccag gacgaaccgt ttttcattac cgaagagatc gaggcggaga
    17461 tgatcgcggc cgggtacgtg ttcgagccgc ccgcgcacat ctcaaccgtg cggctgcatg
    17521 aaatcctggc cggtttgtct gatgccaagc tggcggcctg gccggccagc ttggccgctg
    17581 aagaaaccga gcgccgccgt ctaaaaaggt gatgtgtatt tgagtaaaac agcttgcgtc
    17641 atgcggtcgc tgcgtatatg atgcgatgag taaataaaca aatacgcaag gggaacgcat
    17701 gaaggttatc gctgtactta accagaaagg cgggtcaggc aagacgacca tcgcaaccca
    17761 tctagcccgc gccctgcaac tcgccggggc cgatgttctg ttagtcgatt ccgatcccca
    17821 gggcagtgcc cgcgattggg cggccgtgcg ggaagatcaa ccgctaaccg ttgtcggcat
    17881 cgaccgcccg acgattgacc gcgacgtgaa ggccatcggc cggcgcgact tcgtagtgat
    17941 cgacggagcg ccccaggcgg cggacttggc tgtgtccgcg atcaaggcag ccgacttcgt
    18001 gctgattccg gtgcagccaa gcccttacga catatgggcc accgccgacc tggtggagct
    18061 ggttaagcag cgcattgagg tcacggatgg aaggctacaa gcggcctttg tcgtgtcgcg
    18121 ggcgatcaaa ggcacgcgca tcggcggtga ggttgccgag gcgctggccg ggtacgagct
    18181 gcccattctt gagtcccgta tcacgcagcg cgtgagctac ccaggcactg ccgccgccgg
    18241 cacaaccgtt cttgaatcag aacccgaggg cgacgctgcc cgcgaggtcc aggcgctggc
    18301 cgctgaaatt aaatcaaaac tcatttgagt taatgaggta aagagaaaat gagcaaaagc
    18361 acaaacacgc taagtgccgg ccgtccgagc gcacgcagca gcaaggctgc aacgttggcc
    18421 agcctggcag acacgccagc catgaagcgg gtcaactttc agttgccggc ggaggatcac
    18481 accaagctga agatgtacgc ggtacgccaa ggcaagacca ttaccgagct gctatctgaa
    18541 tacatcgcgc agctaccaga gtaaatgagc aaatgaataa atgagtagat gaattttagc
    18601 ggctaaagga ggcggcatgg aaaatcaaga acaaccaggc accgacgccg tggaatgccc
    18661 catgtgtgga ggaacgggcg gttggccagg cgtaagcggc tgggttgcct gccggccctg
    18721 caatggcact ggaaccccca agcccgagga atcggcgtga gcggtcgcaa accatccggc
    18781 ccggtacaaa tcggcgcggc gctgggtgat gacctggtgg agaagttgaa ggccgcgcag
    18841 gccgcccagc ggcaacgcat cgaggcagaa gcacgccccg gtgaatcgtg gcaagcggcc
    18901 gctgatcgaa tccgcaaaga atcccggcaa ccgccggcag ccggtgcgcc gtcgattagg
    18961 aagccgccca agggcgacga gcaaccagat tttttcgttc cgatgctcta tgacgtgggc
    19021 acccgcgata gtcgcagcat catggacgtg gccgttttcc gtctgtcgaa gcgtgaccga
    19081 cgagctggcg aggtgatccg ctacgagctt ccagacgggc acgtagaggt ttccgcaggg
    19141 ccggccggca tggccagtgt gtgggattac gacctggtac tgatggcggt ttcccatcta
    19201 accgaatcca tgaaccgata ccgggaaggg aagggagaca agcccggccg cgtgttccgt
    19261 ccacacgttg cggacgtact caagttctgc cggcgagccg atggcggaaa gcagaaagac
    19321 gacctggtag aaacctgcat tcggttaaac accacgcacg ttgccatgca gcgtacgaag
    19381 aaggccaaga acggccgcct ggtgacggta tccgagggtg aagccttgat tagccgctac
    19441 aagatcgtaa agagcgaaac cgggcggccg gagtacatcg agatcgagct agctgattgg
    19501 atgtaccgcg agatcacaga aggcaagaac ccggacgtgc tgacggttca ccccgattac
    19561 tttttgatcg atcccggcat cggccgtttt ctctaccgcc tggcacgccg cgccgcaggc
    19621 aaggcagaag ccagatggtt gttcaagacg atctacgaac gcagtggcag cgccggagag
    19681 ttcaagaagt tctgtttcac cgtgcgcaag ctgatcgggt caaatgacct gccggagtac
    19741 gatttgaagg aggaggcggg gcaggctggc ccgatcctag tcatgcgcta ccgcaacctg
    19801 atcgagggcg aagcatccgc cggttcctaa tgtacggagc agatgctagg gcaaattgcc
    19861 ctagcagggg aaaaaggtcg aaaaggactc tttcctgtgg atagcacgta cattgggaac
    19921 ccaaagccgt acattgggaa ccggaacccg tacattggga acccaaagcc gtacattggg
    19981 aaccggtcac acatgtaagt gactgatata aaagagaaaa aaggcgattt ttccgcctaa
    20041 aactctttaa aacttattaa aactcttaaa acccgcctgg cctgtgcata actgtctggc
    20101 cagcgcacag ccgaagagct gcaaaaagcg cctacccttc ggtcgctgcg ctccctacgc
    20161 cccgccgctt cgcgtcggcc tatcgcggcc gctggccgct caaaaatggc tggcctacgg
    20221 ccaggcaatc taccagggcg cggacaagcc gcgccgtcgc cactcgaccg ccggcgccca
    20281 catcaaggca ccctgcctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
    20341 gctcccggag acagtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
    20401 gggcgcgtca gcgggt
//
