LOCUS       sequence-454166-       17340 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  T-DNA vector for Mb2Cas12a-RVRR based mutagenesis for monocot 
            plants; more relaxed PAM requirements; Mb2Cas12a-RVRR and the crRNA 
            were driven by separate ZmUbi1; BASTA for plants selection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 17340)
  AUTHORS   Cheng Y, Li G, Qi A, Mandlik R, Pan C, Wang D, Ge S, Qi Y
  TITLE     A comprehensive all-in-one CRISPR toolbox for large-scale screens in
            plants.
  JOURNAL   Plant Cell. 2025 Apr 9:koaf081. doi: 10.1093/plcell/koaf081.
  PUBMED    40202185
REFERENCE   2  (bases 1 to 17340)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Plant Cell.
            2025 Apr 9:koaf081. doi: 10.1093/plcell/koaf081."
COMMENT     Sequence Label: pYPQAT46
FEATURES             Location/Qualifiers
     source          1..17340
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    283..307
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     primer_bind     510..526
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        542..2539
                     /label=Ubi promoter
                     /note="maize polyubiquitin gene promoter"
     protein_bind    2582..2606
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2631..2651
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6426..6473
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     terminator      6504..6756
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     protein_bind    6801..6821
                     /gene="mutant version of attB"
                     /label=attB5
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     ncRNA           8817..8853
                     /product="self-cleaving hammerhead ribozyme (Ferre-D'Amare 
                     and Scott, 2010)"
                     /label=hammerhead ribozyme
                     /ncRNA_class="hammerhead_ribozyme"
     CDS             complement(8853..9176)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGAGLIYN
                     SRN"
     primer_bind     9041..9057
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9067..9086
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        9067..9085
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(9133..9153)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(9133..9151)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(9172..9188)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    9196..9212
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(9220..9226,9227..9244,9245..9250))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9265..9286
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     ncRNA           9328..9395
                     /product="hepatitis delta virus self-cleaving ribozyme 
                     (Ferre-D'Amare and Scott, 2010)"
                     /label=HDV ribozyme
                     /ncRNA_class="ribozyme"
     protein_bind    complement(9415..9439)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(9507..9523)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    9531..9547
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(9555..9561,9562..9579,9580..9585))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9600..9621
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        9812..10488
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     primer_bind     10425..10445
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             10533..11084
                     /codon_start=1
                     /gene="Streptomyces hygroscopicus bar"
                     /product="phosphinothricin acetyltransferase"
                     /label=BlpR
                     /note="confers resistance to bialophos or phosphinothricin"
                     /translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
                     WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
                     TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
                     WQLDFSLPVPPRPVLPVTEI"
     polyA_signal    11091..11265
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    11343..11367
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             11792..12586
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      12673..13261
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     13162..13181
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     13415..13432
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    13447..13587
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(13513..13532)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     13673..13695
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     rep_origin      13931..14125
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(14191..15264)
                     /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"
     CDS             complement(15693..16322)
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
ORIGIN
        1 ctactttatg cgactaaaac acgcgacaag aaaacgccag gaaaagggca gggcggcagc
       61 ctgtcgcgta acttaggact tgtgcgacat gtcgttttca gaagacggct gcactgaacg
      121 tcagaagccg actgcactat agcagcggag gggttggatc aaagtacttt gatcccgagg
      181 ggaaccctgt ggttggcatg cacatacaaa tggacgaacg gataaacctt ttcacgccct
      241 tttaaatatc cgattattct aataaacgct cttttctctt aggtttaccc gccaatatat
      301 cctgtcaaac actgatagtt taaactgaag gcgggaaacg acaatctgat ccaagctcaa
      361 gctgctctag cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
      421 ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt
      481 aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgcca agcttgcatg
      541 cctgcagtgc agcgtgaccc ggtcgtgccc ctctctagag ataatgagca ttgcatgtct
      601 aagttataaa aaattaccac atattttttt tgtcacactt gtttgaagtg cagtttatct
      661 atctttatac atatatttaa actttactct acgaataata taatctatag tactacaata
      721 atatcagtgt tttagagaat catataaatg aacagttaga catggtctaa aggacaattg
      781 agtattttga caacaggact ctacagtttt atctttttag tgtgcatgtg ttctcctttt
      841 tttttgcaaa tagcttcacc tatataatac ttcatccatt ttattagtac atccatttag
      901 ggtttagggt taatggtttt tatagactaa tttttttagt acatctattt tattctattt
      961 tagcctctaa attaagaaaa ctaaaactct attttagttt ttttatttaa taatttagat
     1021 ataaaataga ataaaataaa gtgactaaaa attaaacaaa taccctttaa gaaattaaaa
     1081 aaactaagga aacatttttc ttgtttcgag tagataatgc cagcctgtta aacgccgtcg
     1141 acgagtctaa cggacaccaa ccagcgaacc agcagcgtcg cgtcgggcca agcgaagcag
     1201 acggcacggc atctctgtcg ctgcctctgg acccctctcg agagttccgc tccaccgttg
     1261 gacttgctcc gctgtcggca tccagaaatt gcgtggcgga gcggcagacg tgagccggca
     1321 cggcaggcgg cctcctcctc ctctcacggc accggcagct acgggggatt cctttcccac
     1381 cgctccttcg ctttcccttc cycscccgcc gtaataaata gacaccccct ccacaccctc
     1441 tttccccaac cycgtgttgt tcggagcgca cacacacaca accagatctc ccccaaatcc
     1501 mcccstcggc mccyccgctt caaggtacgc cgctcgtccc cccccccccc cccctctcta
     1561 ccttctctag atcggcgttc cggtccatgg ttagggcccg gtagttctac ttctgttcat
     1621 gtttgtgtta gatccgtgtt tgtgttagat ccgtgctgct agcgttcgta cacggatgcg
     1681 acctgtacgt cagacacgtt ctgattgcta acttgccagt gtttctcttt ggggaatcct
     1741 gggatggctc tagccgttcc gcagacggga tcgatttcat gatttttttt gtttcgttgc
     1801 atagggtttg gtttgccctt ttcctttatt tcaatatatg ccgtgcactt gtttgtcggg
     1861 tcatcttttc atgctttttt ttgtcttggt tgtgatgatg tggtctggtt gggcggtcgt
     1921 tctagatcgg agtagaatta attctgtttc aaactacctg gtggatttat taattttgga
     1981 tctgtatgtg tgtgccatac atattcatag ttacgaattg aagatgatgg atggaaatat
     2041 cgatctagga taggtataca tgttgatgcg ggttttactg atgcatatac agagatgctt
     2101 tttgttcgct tggttgtgat gatgtggtgt ggttgggcgg tcgttcattc gttctagatc
     2161 ggagtagaat actgtttcaa actacctggt gtatttatta attttggaac tgtatgtgtg
     2221 tgtcatacat cttcatagtt acgagtttaa gatggatgga aatatcgatc taggataggt
     2281 atacatgttg atgtgggttt tactgatgca tatacatgat ggcatatgca gcatctattc
     2341 atatgctcta accttgagta cctatctatt ataataaaca agtatgtttt ataattattt
     2401 tgatcttgat atacttggat gatggcatat gcagcagcta tatgtggatt tttttagccc
     2461 tgccttcata cgctatttat ttgcttggta ctgtttcttt tgtcgatgct caccctgttg
     2521 tttggtgtta cttctgcagg tcgactctag aggatcccct cgaggcgcgc caagctatca
     2581 aacaagtttg tacaaaaaag caggctccga attcgccctt caccatggct cctaagaaga
     2641 agcggaaggt tggtattcac ggggtgcctg cggctctgtt tcaagatttt acacatctgt
     2701 acccgctgag taaaacagtg cggttcgagc tgaaacccat aggaaggacc ctcgagcaca
     2761 tccacgcgaa gaattttctg agccaggatg aaactatggc tgatatgtat caaaaagtta
     2821 aggtcatttt ggacgactat catcgcgatt ttattgccga catgatggga gaggtgaaac
     2881 tcacgaagct tgctgaattt tacgacgtct atctgaagtt caggaaaaat cctaaggacg
     2941 atgggctgca aaaacagctt aaagaccttc aagctgtcct tcggaaggaa tcggtgaagc
     3001 ctatagggtc aggtgggaag tacaaaacag gctacgatag actctttggg gcaaaactct
     3061 tcaaagatgg aaaagagttg ggtgacctcg caaaattcgt tatagcccaa gaaggtgagt
     3121 cttctccgaa gctggctcat cttgctcatt ttgagaagtt cagcacgtat tttactggat
     3181 ttcacgataa tcggaagaat atgtactcgg atgaagacaa gcatactgca atagcgtaca
     3241 ggctcatcca tgagaatttg ccgagattca tcgacaatct gcaaatcttg acaacaatca
     3301 aacaaaagca tagcgccctc tatgatcaga taatcaacga gctcacggcc tccgggctcg
     3361 acgtctcctt ggcttctcat cttgacgggt atcacaagct ccttacacaa gaggggatca
     3421 cggcatacaa caggatcata ggagaggtga atggatatac aaataagcat aaccagatat
     3481 gccacaagag cgagcgcata gcgaaactta gacccttgca caagcaaatc ctttctgacg
     3541 gaatgggagt gtcattcctt ccgtctaagt tcgcggatga tagtgagatg tgccaagcgg
     3601 tcaacgaatt ttatcgccat tatactgacg tgttcgcaaa ggtgcaaagt ctctttgacg
     3661 gatttgatga tcaccagaaa gacgggatct atgttgaaca caaaaacctt aatgaactga
     3721 gcaaacaggc gttcggcgac tttgctttgc tggggagggt ccttgatgga tactacgtgg
     3781 acgttgtcaa tccggagttc aatgagcggt tcgcaaaggc caagactgac aatgcgaaag
     3841 ccaagcttac aaaagaaaag gacaaattca ttaaaggagt ccactcactg gcttccctcg
     3901 aacaagcaat agaacaccat acagctagac acgacgatga gagtgttcaa gccggaaaac
     3961 ttggccagta cttcaaacac ggtttggcgg gggttgacaa cccgattcag aaaattcaca
     4021 ataaccattc gacgattaaa gggtttctgg aaagggaaag gcctgctggg gaacgggcgc
     4081 tcccgaagat caagtcagga aaaaacccag aaatgacaca gctcaggcag ctgaaggaac
     4141 ttttggacaa cgcattgaat gtggcgcact tcgctaagct gctgacaact aaaacaacct
     4201 tggacaacca ggatggaaat ttttacgggg agtttggggt gctttacgac gagctggcta
     4261 aaattccaac tctctacaat aaggttagag attatctctc tcaaaagccc ttttctaccg
     4321 aaaagtataa gctcaacttc ggcaatccga cccttctccg cgggtgggac ctgaacgtgg
     4381 agaaagatcg ctttggggtt atacttcaga aggatggatg ctattacttg gcgcttcttg
     4441 ataaggctca taaaaaagtt ttcgacaacg cccctaacac tggtaagaac gtctaccaaa
     4501 agatggtcta caaactgttg cccggcccca acaaaatgct tcctagggtg tttttcgcaa
     4561 aatcgaatct cgactattat aatccatctg ccgagctcct tgacaaatat gctaagggga
     4621 cccataaaaa gggtgataat ttcaacctga aggactgcca cgcgcttatc gactttttca
     4681 aagccgggat aaataagcat ccggagtggc aacattttgg ttttaaattt tcgccaacgt
     4741 cgtcctatcg cgacctttcc gatttctata gggaagttga acctcagggg taccaggtca
     4801 aatttgttga cattaatgcg gactacattg atgaattggt ggagcaaggg aagctctacc
     4861 tctttcaaat atataacaaa gatttctcgc caaaagcgca tggtaaaccg aatcttcata
     4921 ccttgtactt taaagcactt ttttcagaag ataacttggc ggacccgatc tacaagctga
     4981 atggggaagc tcagatcttc tacaggaaag cttcgttgga catgaacgag actaccatac
     5041 atcgcgcggg agaggtgctt gagaacaaaa atcccgacaa cccgaaaaag cggcaattcg
     5101 tttacgacat catcaaagac aaacggtaca cgcaggacaa atttatgctc cacgtcccca
     5161 ttaccatgaa ttttggagtc caaggcatga ccattaagga attcaacaaa aaggtcaacc
     5221 aaagtattca gcaatacgat gaagtcaatg tcataggcat agatcgggga gaaaggcatc
     5281 tgttgtatct taccgtgatt aactctaagg gtgaaatact ggagcaacgg tcacttaacg
     5341 atataaccac ggcgtccgcg aacggtacac aagtgaccac tccctaccac aaaatattgg
     5401 ataaaaggga gatagaacgc ttgaatgccc gcgttggctg gggtgagatt gagacgatca
     5461 aagagcttaa atcgggatat ttgtctcacg tcgttcatca aattaaccaa ctcatgctta
     5521 agtacaatgc aatcgttgtg ctcgaggacc tgaactttgg tttcaaaaga gggaggttca
     5581 aggtggaaaa acaaatttac cagaactttg aaaacgcgct tatcaagaaa ttgaatcacc
     5641 ttgttttgaa agataaggca gatgacgaaa tcgggtcgta taaaaatgca ctccagttga
     5701 caaataattt cacggatttg aagtcgatcg gcaagcaaac agggttcctc ttttatgtgc
     5761 cagcgtggaa tacatcaaaa attgatccgg agacgggatt tgtcgacttg ctgaagccta
     5821 ggtatgagaa cattgcccaa tctcaggcct ttttcggcaa attcgataaa atatgctaca
     5881 acacagacaa aggttatttt gaatttcaca ttgattacgc caaatttaca gataaggcga
     5941 aaaacagcag acagaaatgg gctatctgtt ctcatgggga caaacgctat gtctacgata
     6001 agacggctaa tcaaaataaa ggcgccgcaa aaggtattaa tgtgaatgat gagctgaaaa
     6061 gcttgtttgc ccgctaccat atcaatgata aacaaccaaa cttggtgatg gacatatgcc
     6121 agaacaatga caaagaattc cacaagtcac tcatgtgcct gcttaaaacc cttttggcgc
     6181 tgcggtatag caatgcatct agcgatgaag actttatttt gagtcccgtg gccaacgacg
     6241 agggcgtgtt ttttaattca gccttggcgg acgatacgca gccccagaat gcggacgcaa
     6301 acggcgcgta ccacattgca ctgaagggac tgtggcttct gaacgagctg aaaaatagcg
     6361 acgacctgaa taaagtcaag ttggccattg acaatcaaac ctggttgaat ttcgctcaaa
     6421 atagaaagcg tcctgctgcc accaaaaagg ccggacaggc taagaaaaag aagtgagacg
     6481 actagtggcg gccgccgacg tccgatcgtt caaacatttg gcaataaagt ttcttaagat
     6541 tgaatcctgt tgccggtctt gcgatgatta tcatataatt tctgttgaat tacgttaagc
     6601 atgtaataat taacatgtaa tgcatgacgt tatttatgag atgggttttt atgattagag
     6661 tcccgcaatt atacatttaa tacgcgatag aaaacaaaat atagcgcgca aactaggata
     6721 aattatcgcg cgcggtgtca tctatgttac tagatcggga attgatcccc cctcgacagc
     6781 ttccggaaag ggcgaattcg caactttgta tacaaaagtt gccccatggc gttccctcta
     6841 gagagataat gagcattgca tgtctaagtt ataaaaaatt accacatatt ttttttgtca
     6901 cacttgtttg aagtgcagtt tatctatctt tatacatata tttaaacttt actctacgaa
     6961 taatataatc tatagtacta caataatatc agtgttttag agaatcatat aaatgaacag
     7021 ttagacatgg tctaaaggac aattgagtat tttgacaaca ggactctaca gttttatctt
     7081 tttagtgtgc atgtgttctc cttttttttt gcaaatagct tcacctatat aatacttcat
     7141 ccattttatt agtacatcca tttagggttt agggttaatg gtttttatag actaattttt
     7201 ttagtacatc tattttattc tattttagcc tctaaattaa gaaaactaaa actctatttt
     7261 agttttttta tttaataatt tagatataaa atagaataaa ataaagtgac taaaaattaa
     7321 acaaataccc tttaagaaat taaaaaaact aaggaaacat ttttcttgtt tcgagtagat
     7381 aatgccagcc tgttaaacgc cgtcgacgag tctaacggac accaaccagc gaaccagcag
     7441 cgtcgcgtcg ggccaagcga agcagacggc acggcatctc tgtcgctgcc tctggacccc
     7501 tctcgagagt tccgctccac cgttggactt gctccgctgt cggcatccag aaattgcgtg
     7561 gcggagcggc agacgtgagc cggcacggca ggcggcctcc tcctcctctc acggcaccgg
     7621 cagctacggg ggattccttt cccmccgctc cttcgctttc ccttccycgc ccgccgtaat
     7681 aaatagacac cccctccaca ccctctttcc ccaacctcgt gttgttcgga gcgcacacac
     7741 acacaaccag atctccccca aatccacccg tcggcacctc cgcttcaagg tacgccgctc
     7801 gtcctccccc cccccccccy ctctaccttc tctagatcgg cgttccggtc catggttagg
     7861 gcccggtagt tctacttctg ttcatgtttg tgttagatcc gtgtttgtgt tagatccgtg
     7921 ctgctagcgt tcgtacacgg atgcgacctg tacgtcagac acgttctgat tgctaacttg
     7981 ccagtgtttc tctttgggga atcctgggat ggctctagcc gttccgcaga cgggatcgat
     8041 ttcatgattt tttttgtttc gttgcatagg gtttggtttg cccttttcct ttatttcaat
     8101 atatgccgtg cacttgtttg tcgggtcatc ttttcatgct tttttttgtc ttggttgtga
     8161 tgatgtggtc tggttgggcg gtcgttctag atcggagtag aattaattct gtttcaaact
     8221 acctggtgga tttattaatt ttggatctgt atgtgtgtgc catacatatt catagttacg
     8281 aattgaagat gatggatgga aatatcgatc taggataggt atacatgttg atgcgggttt
     8341 tactgatgca tatacagaga tgctttttgt tcgcttggtt gtgatgatgt ggtgtggttg
     8401 ggcggtcgtt cattcgttct agatcggagt agaatactgt ttcaaactac ctggtgtatt
     8461 tattaatttt ggaactgtat gtgtgtgtca tacatcttca tagttacgag tttaagatgg
     8521 atggaaatat cgatctagga taggtataca tgttgatgtg ggttttactg atgcatatac
     8581 atgatggcat atgcagcatc tattcatatg ctctaacctt gagtacctat ctattataat
     8641 aaacaagtat gttttataat tattttgatc ttgatatact tggatgatgg catatgcagc
     8701 agctatatgt ggattttttt agccctgcct tcatacgcta tttatttgct tggtactgtt
     8761 tcttttgtcg atgctcaccc tgttgtttgg tgttacttct gcagggatcc aaattactga
     8821 tgagtccgtg aggacgaaac gagtaagctc gtctaatttc tactgttgta gatgagaccg
     8881 gcgccgctac agggcgcgtc ccattcgcca ttcaggctgc gcaactgttg ggaagggcga
     8941 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga
     9001 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgag
     9061 cgcgcgtaat acgactcact atagggcgaa ttgggtaccg ggccccccct cgaggtcctc
     9121 cagcttttgt tccctttagt gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct
     9181 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     9241 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     9301 actgcccgct ttccaccggt ggtctctggc cggcatggtc ccagcctcct cgctggcgcc
     9361 ggctgggcaa catgcttcgg catggcgaat gggacggtac ccggccggaa ttcgacccag
     9421 ctttcttgta caaagtggtt cgataattct taattaacta gttctagagc ggccgccacc
     9481 gcggtggagc tcgaattcgt aatcatgtca tagctgtttc ctgtgtgaaa ttgttatccg
     9541 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     9601 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     9661 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     9721 ggctagagca gcttgccaac atggtggagc acgacactct cgtctactcc aagaatatca
     9781 aagatacagt ctcagaagac caaagggcta ttgagacttt tcaacaaagg gtaatatcgg
     9841 gaaacctcct cggattccat tgcccagcta tctgtcactt catcaaaagg acagtagaaa
     9901 aggaaggtgg cacctacaaa tgccatcatt gcgataaagg aaaggctatc gttcaagatg
     9961 cctctgccga cagtggtccc aaagatggac ccccacccac gaggagcatc gtggaaaaag
    10021 aagacgttcc aaccacgtct tcaaagcaag tggattgatg tgaacatggt ggagcacgac
    10081 actctcgtct actccaagaa tatcaaagat acagtctcag aagaccaaag ggctattgag
    10141 acttttcaac aaagggtaat atcgggaaac ctcctcggat tccattgccc agctatctgt
    10201 cacttcatca aaaggacagt agaaaaggaa ggtggcacct acaaatgcca tcattgcgat
    10261 aaaggaaagg ctatcgttca agatgcctct gccgacagtg gtcccaaaga tggaccccca
    10321 cccacgagga gcatcgtgga aaaagaagac gttccaacca cgtcttcaaa gcaagtggat
    10381 tgatgtgata tctccactga cgtaagggat gacgcacaat cccactatcc ttcgcaagac
    10441 ccttcctcta tataaggaag ttcatttcat ttggagagga cacgctgaaa tcaccagtct
    10501 ctctctacaa atctatctct ctcgagtcta ccatgagccc agaacgacgc ccggccgaca
    10561 tccgccgtgc caccgaggcg gacatgccgg cggtctgcac catcgtcaac cactacatcg
    10621 agacaagcac ggtcaacttc cgtaccgagc cgcaggaacc gcaggagtgg acggacgacc
    10681 tcgtccgtct gcgggagcgc tatccctggc tcgtcgccga ggtggacggc gaggtcgccg
    10741 gcatcgccta cgcgggcccc tggaaggcac gcaacgccta cgactggacg gccgagtcga
    10801 ccgtgtacgt ctccccccgc caccagcgga cgggactggg ctccacgctc tacacccacc
    10861 tgctgaagtc cctggaggca cagggcttca agagcgtggt cgctgtcatc gggctgccca
    10921 acgacccgag cgtgcgcatg cacgaggcgc tcggatatgc cccccgcggc atgctgcggg
    10981 cggccggctt caagcacggg aactggcatg acgtgggttt ctggcagctg gacttcagcc
    11041 tgccggtacc gccccgtccg gtcctgcccg tcaccgagat ttgactcgag tttctccata
    11101 ataatgtgtg agtagttccc agataaggga attagggttc ctatagggtt tcgctcatgt
    11161 gttgagcata taagaaaccc ttagtatgta tttgtatttg taaaatactt ctatcaataa
    11221 aatttctaat tcctaaaacc aaaatccagt actaaaatcc agatcccccg aattaattcg
    11281 gcgttaattc agtacattaa aaacgtccgc aatgtgttat taagttgtct aagcgtcaat
    11341 ttgtttacac cacaatatat cctgccacca gccagccaac agctccccga ccggcagctc
    11401 ggcacaaaat caccactcga tacaggcagc ccatcagtcc gggacggcgt cagcgggaga
    11461 gccgttgtaa ggcggcagac tttgctcatg ttaccgatgc tattcggaag aacggcaact
    11521 aagctgccgg gtttgaaaca cggatgatct cgcggagggt agcatgttga ttgtaacgat
    11581 gacagagcgt tgctgcctgt gatcaccgcg gtttcaaaat cggctccgtc gatactatgt
    11641 tatacgccaa ctttgaaaac aactttgaaa aagctgtttt ctggtattta aggttttaga
    11701 atgcaaggaa cagtgaattg gagttcgtct tgttataatt agcttcttgg ggtatcttta
    11761 aatactgtag aaaagaggaa ggaaataata aatggctaaa atgagaatat caccggaatt
    11821 gaaaaaactg atcgaaaaat accgctgcgt aaaagatacg gaaggaatgt ctcctgctaa
    11881 ggtatataag ctggtgggag aaaatgaaaa cctatattta aaaatgacgg acagccggta
    11941 taaagggacc acctatgatg tggaacggga aaaggacatg atgctatggc tggaaggaaa
    12001 gctgcctgtt ccaaaggtcc tgcactttga acggcatgat ggctggagca atctgctcat
    12061 gagtgaggcc gatggcgtcc tttgctcgga agagtatgaa gatgaacaaa gccctgaaaa
    12121 gattatcgag ctgtatgcgg agtgcatcag gctctttcac tccatcgaca tatcggattg
    12181 tccctatacg aatagcttag acagccgctt agccgaattg gattacttac tgaataacga
    12241 tctggccgat gtggattgcg aaaactggga agaagacact ccatttaaag atccgcgcga
    12301 gctgtatgat tttttaaaga cggaaaagcc cgaagaggaa cttgtctttt cccacggcga
    12361 cctgggagac agcaacatct ttgtgaaaga tggcaaagta agtggcttta ttgatcttgg
    12421 gagaagcggc agggcggaca agtggtatga cattgccttc tgcgtccggt cgatcaggga
    12481 ggatatcggg gaagaacagt atgtcgagct attttttgac ttactgggga tcaagcctga
    12541 ttgggagaaa ataaaatatt atattttact ggatgaattg ttttagtacc tagaatgcat
    12601 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
    12661 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
    12721 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
    12781 ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt
    12841 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
    12901 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
    12961 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
    13021 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
    13081 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
    13141 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
    13201 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
    13261 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
    13321 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
    13381 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
    13441 agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatatg
    13501 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtata cactccgcta
    13561 tcgctacgtg actgggtcat ggctgcgccc cgacacccgc caacacccgc tgacgcgccc
    13621 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
    13681 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgaggcaggg tgccttgatg
    13741 tgggcgccgg cggtcgagtg gcgacggcgc ggcttgtccg cgccctggta gattgcctgg
    13801 ccgtaggcca gccatttttg agcggccagc ggccgcgata ggccgacgcg aagcggcggg
    13861 gcgtagggag cgcagcgacc gaagggtagg cgctttttgc agctcttcgg ctgtgcgctg
    13921 gccagacagt tatgcacagg ccaggcgggt tttaagagtt ttaataagtt ttaaagagtt
    13981 ttaggcggaa aaatcgcctt ttttctcttt tatatcagtc acttacatgt gtgaccggtt
    14041 cccaatgtac ggctttgggt tcccaatgta cgggttccgg ttcccaatgt acggctttgg
    14101 gttcccaatg tacgtgctat ccacaggaaa cagacctttt cgaccttttt cccctgctag
    14161 ggcaatttgc cctagcatct gctccgtaca ttaggaaccg gcggatgctt cgccctcgat
    14221 caggttgcgg tagcgcatga ctaggatcgg gccagcctgc cccgcctcct ccttcaaatc
    14281 gtactccggc aggtcatttg acccgatcag cttgcgcacg gtgaaacaga acttcttgaa
    14341 ctctccggcg ctgccactgc gttcgtagat cgtcttgaac aaccatctgg cttctgcctt
    14401 gcctgcggcg cggcgtgcca ggcggtagag aaaacggccg atgccgggat cgatcaaaaa
    14461 gtaatcgggg tgaaccgtca gcacgtccgg gttcttgcct tctgtgatct cgcggtacat
    14521 ccaatcagct agctcgatct cgatgtactc cggccgcccg gtttcgctct ttacgatctt
    14581 gtagcggcta atcaaggctt caccctcgga taccgtcacc aggcggccgt tcttggcctt
    14641 cttcgtacgc tgcatggcaa cgtgcgtggt gtttaaccga atgcaggttt ctaccaggtc
    14701 gtctttctgc tttccgccat cggctcgccg gcagaacttg agtacgtccg caacgtgtgg
    14761 acggaacacg cggccgggct tgtctccctt cccttcccgg tatcggttca tggattcggt
    14821 tagatgggaa accgccatca gtaccaggtc gtaatcccac acactggcca tgccggccgg
    14881 ccctgcggaa acctctacgt gcccgtctgg aagctcgtag cggatcacct cgccagctcg
    14941 tcggtcacgc ttcgacagac ggaaaacggc cacgtccatg atgctgcgac tatcgcgggt
    15001 gcccacgtca tagagcatcg gaacgaaaaa atctggttgc tcgtcgccct tgggcggctt
    15061 cctaatcgac ggcgcaccgg ctgccggcgg ttgccgggat tctttgcgga ttcgatcagc
    15121 ggccgcttgc cacgattcac cggggcgtgc ttctgcctcg atgcgttgcc gctgggcggc
    15181 ctgcgcggcc ttcaacttct ccaccaggtc atcacccagc gccgcgccga tttgtaccgg
    15241 gccggatggt ttgcgaccgc tcacgccgat tcctcgggct tgggggttcc agtgccattg
    15301 cagggccggc aggcaaccca gccgcttacg cctggccaac cgcccgttcc tccacacatg
    15361 gggcattcca cggcgtcggt gcctggttgt tcttgatttt ccatgccgcc tcctttagcc
    15421 gctaaaattc atctactcat ttattcattt gctcatttac tctggtagct gcgcgatgta
    15481 ttcagatagc agctcggtaa tggtcttgcc ttggcgtacc gcgtacatct tcagcttggt
    15541 gtgatcctcc gccggcaact gaaagttgac ccgcttcatg gctggcgtgt ctgccaggct
    15601 ggccaacgtt gcagccttgc tgctgcgtgc gctcggacgg ccggcactta gcgtgtttgt
    15661 gcttttgctc attttctctt tacctcatta actcaaatga gttttgattt aatttcagcg
    15721 gccagcgcct ggacctcgcg ggcagcgtcg ccctcgggtt ctgattcaag aacggttgtg
    15781 ccggcggcgg cagtgcctgg gtagctcacg cgctgcgtga tacgggactc aagaatgggc
    15841 agctcgtacc cggccagcgc ctcggcaacc tcaccgccga tgcgcgtgcc tttgatcgcc
    15901 cgcgacacga caaaggccgc ttgtagcctt ccatccgtga cctcaatgcg ctgcttaacc
    15961 agctccacca ggtcggcggt ggcccatatg tcgtaagggc ttggctgcac cggaatcagc
    16021 acgaagtcgg ctgccttgat cgcggacaca gccaagtccg ccgcctgggg cgctccgtcg
    16081 atcactacga agtcgcgccg gccgatggcc ttcacgtcgc ggtcaatcgt cgggcggtcg
    16141 atgccgacaa cggttagcgg ttgatcttcc cgcacggccg cccaatcgcg ggcactgccc
    16201 tggggatcgg aatcgactaa cagaacatcg gccccggcga gttgcagggc gcgggctaga
    16261 tgggttgcga tggtcgtctt gcctgacccg cctttctggt taagtacagc gataaccttc
    16321 atgcgttccc cttgcgtatt tgtttattta ctcatcgcat catatacgca gcgaccgcat
    16381 gacgcaagct gttttactca aatacacatc acctttttag acggcggcgc tcggtttctt
    16441 cagcggccaa gctggccggc caggccgcca gcttggcatc agacaaaccg gccaggattt
    16501 catgcagccg cacggttgag acgtgcgcgg gcggctcgaa cacgtacccg gccgcgatca
    16561 tctccgcctc gatctcttcg gtaatgaaaa acggttcgtc ctggccgtcc tggtgcggtt
    16621 tcatgcttgt tcctcttggc gttcattctc ggcggccgcc agggcgtcgg cctcggtcaa
    16681 tgcgtcctca cggaaggcac cgcgccgcct ggcctcggtg ggcgtcactt cctcgctgcg
    16741 ctcaagtgcg cggtacaggg tcgagcgatg cacgccaagc agtgcagccg cctctttcac
    16801 ggtgcggcct tcctggtcga tcagctcgcg ggcgtgcgcg atctgtgccg gggtgagggt
    16861 agggcggggg ccaaacttca cgcctcgggc cttggcggcc tcgcgcccgc tccgggtgcg
    16921 gtcgatgatt agggaacgct cgaactcggc aatgccggcg aacacggtca acaccatgcg
    16981 gccggccggc gtggtggtgt cggcccacgg ctctgccagg ctacgcaggc ccgcgccggc
    17041 ctcctggatg cgctcggcaa tgtccagtag gtcgcgggtg ctgcgggcca ggcggtctag
    17101 cctggtcact gtcacaacgt cgccagggcg taggtggtca agcatcctgg ccagctccgg
    17161 gcggtcgcgc ctggtgccgg tgatcttctc ggaaaacagc ttggtgcagc cggccgcgtg
    17221 cagttcggcc cgttggttgg tcaagtcctg gtcgtcggtg ctgacgcggg catagcccag
    17281 caggccagcg gcggcgctct tgttcatggc gtaatgtctc cggttctagt cgcaagtatt
//
