LOCUS       sequence-452338-       16609 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  T-DNA vector for SpRY mediated mutagenesis for dicot plants; NG or 
            NA PAM preference; SpRY was driven by ZmUbi1 and the sgRNA was 
            driven by AtU3 promoter; BASTA for plants selection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 16609)
  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 16609)
  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: pYPQAT08
FEATURES             Location/Qualifiers
     source          1..16609
                     /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             2628..2693
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2700..2720
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     RBS             6758..6766
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             6846..6893
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     protein_bind    7568..7588
                     /gene="mutant version of attB"
                     /label=attB5
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     CDS             complement(7949..8254)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGAGL"
     primer_bind     8119..8135
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8145..8164
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        8145..8163
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8211..8231)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(8211..8229)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(8250..8266)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    8274..8290
                     /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(8298..8304,8305..8322,8323..8328))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8343..8364
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_RNA        8406..8481
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(8684..8708)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(8776..8792)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    8800..8816
                     /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(8824..8830,8831..8848,8849..8854))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8869..8890
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        9081..9757
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     primer_bind     9694..9714
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             9802..10353
                     /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    10360..10534
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    10612..10636
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             11061..11855
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      11942..12530
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     12431..12450
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     12684..12701
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    12716..12856
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(12782..12801)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     12942..12964
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     rep_origin      13200..13394
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(13460..14533)
                     /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(14962..15591)
                     /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 ctcgcccgcc gtaataaata gacaccccct ccacaccctc
     1441 tttccccaac ctcgtgttgt tcggagcgca cacacacaca accagatctc ccccaaatcc
     1501 acccgtcggc acctccgctt caaggtacgc cgctcgtccy 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 caccatggat tacaaggacc
     2641 acgacgggga ttacaaggac cacgacattg attacaagga tgatgatgac aagatggctc
     2701 cgaagaagaa gaggaaggtt ggcatccacg gggtgccagc tgctgacaag aagtactcga
     2761 tcggcctcga tattgggact aactctgttg gctgggccgt gatcaccgac gagtacaagg
     2821 tgccctcaaa gaagttcaag gtcctgggca acaccgatcg gcattccatc aagaagaatc
     2881 tcattggcgc tctcctgttc gacagcggcg agacggctga gaggacgcgg ctcaagcgca
     2941 ccgcccgcag gcggtacacg cgcaggaaga atcgcatctg ctacctgcag gagattttct
     3001 ccaacgagat ggcgaaggtt gacgattctt tcttccacag gctggaggag tcattcctcg
     3061 tggaggagga taagaagcac gagcggcatc caatcttcgg caacattgtc gacgaggttg
     3121 cctaccacga gaagtaccct acgatctacc atctgcggaa gaagctcgtg gactccacag
     3181 ataaggcgga cctccgcctg atctacctcg ctctggccca catgattaag ttcaggggcc
     3241 atttcctgat cgagggggat ctcaacccgg acaatagcga tgttgacaag ctgttcatcc
     3301 agctcgtgca gacgtacaac cagctcttcg aggagaaccc cattaatgcg tcaggcgtcg
     3361 acgcgaaggc tatcctgtcc gctaggctct cgaagtctcg gcgcctcgag aacctgatcg
     3421 cccagctgcc gggcgagaag aagaacggcc tgttcgggaa tctcattgcg ctcagcctgg
     3481 ggctcacgcc caacttcaag tcgaatttcg atctcgctga ggacgccaag ctgcagctct
     3541 ccaaggacac atacgacgat gacctggata acctcctggc ccagatcggc gatcagtacg
     3601 cggacctgtt cctcgctgcc aagaatctgt cggacgccat cctcctgtct gatattctca
     3661 gggtgaacac cgagattacg aaggctccgc tctcagcctc catgatcaag cgctacgacg
     3721 agcaccatca ggatctgacc ctcctgaagg cgctggtcag gcagcagctc cccgagaagt
     3781 acaaggagat cttcttcgat cagtcgaaga acggctacgc tgggtacatt gacggcgggg
     3841 cctctcagga ggagttctac aagttcatca agccgattct ggagaagatg gacggcacgg
     3901 aggagctgct ggtgaagctc aatcgcgagg acctcctgag gaagcagcgg acattcgata
     3961 acggcagcat cccacaccag attcatctcg gggagctgca cgctatcctg aggaggcagg
     4021 aggacttcta ccctttcctc aaggataacc gcgagaagat cgagaagatt ctgactttca
     4081 ggatcccgta ctacgtcggc ccactcgcta ggggcaactc ccgcttcgct tggatgaccc
     4141 gcaagtcaga ggagacgatc acgccgtgga acttcgagga ggtggtcgac aagggcgcta
     4201 gcgctcagtc gttcatcgag aggatgacga atttcgacaa gaacctgcca aatgagaagg
     4261 tgctccctaa gcactcgctc ctgtacgagt acttcacagt ctacaacgag ctgactaagg
     4321 tgaagtatgt gaccgagggc atgaggaagc cggctttcct gtctggggag cagaagaagg
     4381 ccatcgtgga cctcctgttc aagaccaacc ggaaggtcac ggttaagcag ctcaaggagg
     4441 actacttcaa gaagattgag tgcttcgatt cggtcgagat ctctggcgtt gaggaccgct
     4501 tcaacgcctc cctggggacc taccacgatc tcctgaagat cattaaggat aaggacttcc
     4561 tggacaacga ggagaatgag gatatcctcg aggacattgt gctgacactc actctgttcg
     4621 aggaccggga gatgatcgag gagcgcctga agacttacgc ccatctcttc gatgacaagg
     4681 tcatgaagca gctcaagagg aggaggtaca ccggctgggg gaggctgagc aggaagctca
     4741 tcaacggcat tcgggacaag cagtccggga agacgatcct cgacttcctg aagagcgatg
     4801 gcttcgcgaa ccgcaatttc atgcagctga ttcacgatga cagcctcaca ttcaaggagg
     4861 atatccagaa ggctcaggtg agcggccagg gggactcgct gcacgagcat atcgcgaacc
     4921 tcgctggctc gccagctatc aagaagggga ttctgcagac cgtgaaggtt gtggacgagc
     4981 tggtgaaggt catgggcagg cacaagcctg agaacatcgt cattgagatg gcccgggaga
     5041 atcagaccac gcagaagggc cagaagaact cacgcgagag gatgaagagg atcgaggagg
     5101 gcattaagga gctggggtcc cagatcctca aggagcaccc ggtggagaac acgcagctgc
     5161 agaatgagaa gctctacctg tactacctcc agaatggccg cgatatgtat gtggaccagg
     5221 agctggatat taacaggctc agcgattacg acgtcgatca tatcgttcca cagtcattcc
     5281 tgaaggatga ctccattgac aacaaggtcc tcaccaggtc ggacaagaac cggggcaagt
     5341 ctgataatgt tccttcagag gaggtcgtta agaagatgaa gaactactgg cgccagctcc
     5401 tgaatgccaa gctgatcacg cagcggaagt tcgataacct cacaaaggct gagaggggcg
     5461 ggctctctga gctggacaag gcgggcttca tcaagaggca gctggtcgag acacggcaga
     5521 tcactaagca cgttgcgcag attctcgact cacggatgaa cactaagtac gatgagaatg
     5581 acaagctgat ccgcgaggtg aaggtcatca ccctgaagtc aaagctcgtc tccgacttca
     5641 ggaaggattt ccagttctac aaggttcggg agatcaacaa ttaccaccat gcccatgacg
     5701 cgtacctgaa cgcggtggtc ggcacagctc tgatcaagaa gtacccaaag ctcgagagcg
     5761 agttcgtgta cggggactac aaggtttacg atgtgaggaa gatgatcgcc aagtcggagc
     5821 aggagattgg caaggctacc gccaagtact tcttctactc taacattatg aatttcttca
     5881 agacagagat cactctggcc aatggcgaga tccggaagcg ccccctcatc gagacgaacg
     5941 gcgagacggg ggagatcgtg tgggacaagg gcagggattt cgcgaccgtc aggaaggttc
     6001 tctccatgcc acaagtgaat atcgtcaaga agacagaggt ccagactggc gggttctcta
     6061 aggagtcaat taggcctaag cggaacagcg acaagctcat cgcccgcaag aaggactggg
     6121 atccgaagaa gtacggcggg ttcctgtggc ccactgtggc ctactcggtc ctggttgtgg
     6181 cgaaggttga gaagggcaag tccaagaagc tcaagagcgt gaaggagctg ctggggatca
     6241 cgattatgga gcgctccagc ttcgagaaga acccgatcga tttcctggag gcgaagggct
     6301 acaaggaggt gaagaaggac ctgatcatta agctccccaa gtactcactc ttcgagctgg
     6361 agaacggcag gaagcggatg ctggcttccg ctaagcagct gcagaagggg aacgagctgg
     6421 ctctgccgtc caagtatgtg aacttcctct acctggcctc ccactacgag aagctcaagg
     6481 gcagccccga ggacaacgag cagaagcagc tgttcgtcga gcagcacaag cattacctcg
     6541 acgagatcat tgagcagatt tccgagttct ccaagcgcgt gatcctggcc gacgcgaatc
     6601 tggataaggt cctctccgcg tacaacaagc accgcgacaa gccaatcagg gagcaggctg
     6661 agaatatcat tcatctcttc accctgacga ggctcggcgc ccctagggct ttcaagtact
     6721 tcgacacaac tatcgatccg aagcagtaca ggagcactaa ggaggtcctg gacgcgaccc
     6781 tcatccacca gtcgattacc ggcctctacg agacgcgcat cgacctgtct cagctcgggg
     6841 gcgacaagcg gccagcggcg acgaagaagg cggggcaggc gaagaagaag aagtgagctc
     6901 agagctttcg ttcgtatcat cggtttcgac aacgttcgtc aagttcaatg catcagtttc
     6961 attgcgcaca caccagaatc ctactgagtt tgagtattat ggcattggga aaactgtttt
     7021 tcttgtacca tttgttgtgc ttgtaattta ctgtgttttt tattcggttt tcgctatcga
     7081 actgtgaaat ggaaatggat ggagaagagt taatgaatga tatggtcctt ttgttcattc
     7141 tcaaattaat attatttgtt ttttctctta tttgttgtgt gttgaatttg aaattataag
     7201 agatatgcaa acattttgtt ttgagtaaaa atgtgtcaaa tcgtggcctc taatgaccga
     7261 agttaatatg aggagtaaaa cacttgtagt tgtaccatta tgcttattca ctaggcaaca
     7321 aatatatttt cagacctaga aaagctgcaa atgttactga atacaagtat gtcctcttgt
     7381 gttttagaca tttatgaact ttcctttatg taattttcca gaatccttgt cagattctaa
     7441 tcattgcttt ataattatag ttatactcat ggatttgtag ttgagtatga aaatattttt
     7501 taatgcattt tatgacttgc caattgttcc ggaactctag ataagcttac cggaaagggc
     7561 gaattcgcaa ctttgtatac aaaagttgcc ccatggcgtt ccctctagat aacgcaggat
     7621 cctatttact ttaaattttt cttatggctc agcctgtgat ggataactga atcaaacaaa
     7681 tggcgtctgg gtttaagaac atctgttttg gctatgttgg acgaaacaag tgaactttta
     7741 ggatcaactt ccgtttatat acggagctta tatcgagcaa taagataagt gggcttttta
     7801 tgtaatttaa tgggctatcg tccatatatt cactaatacc catgcccagt acccatgtat
     7861 gcgtttcata taagctccta atttctccca catcgctcaa atctaaacaa atcttgttgt
     7921 atatataaca ctgagggagc accattggtc agagaccggc gccgctacag ggcgcgtccc
     7981 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
     8041 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
     8101 tttcccagtc acgacgttgt aaaacgacgg ccagtgagcg cgcgtaatac gactcactat
     8161 agggcgaatt gggtaccggg ccccccctcg aggtcctcca gcttttgttc cctttagtga
     8221 gggttaattg cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     8281 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     8341 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccaccggtgg
     8401 tctctgtttt agagctagaa atagcaagtt aaaataaggc tagtccgtta tcaacttgaa
     8461 aaagtggcac cgagtcggtg cttttttttc cctttccttt tttctttttt ttgccataaa
     8521 cttaaatttg tatatcgatc attgtagata ttgaaaacct agaacaaacc aacatccatg
     8581 tgaatgtctt tcatgactga tttagagata attcttgaat tttggaacta gaatctataa
     8641 tgagcctaaa ttaaaacatt gtgaactagt aagggcgaat tcgacccagc tttcttgtac
     8701 aaagtggttc gataattctt aattaactag ttctagagcg gccgccaccg cggtggagct
     8761 cgaattcgta atcatgtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     8821 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     8881 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     8941 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg gctagagcag
     9001 cttgccaaca tggtggagca cgacactctc gtctactcca agaatatcaa agatacagtc
     9061 tcagaagacc aaagggctat tgagactttt caacaaaggg taatatcggg aaacctcctc
     9121 ggattccatt gcccagctat ctgtcacttc atcaaaagga cagtagaaaa ggaaggtggc
     9181 acctacaaat gccatcattg cgataaagga aaggctatcg ttcaagatgc ctctgccgac
     9241 agtggtccca aagatggacc cccacccacg aggagcatcg tggaaaaaga agacgttcca
     9301 accacgtctt caaagcaagt ggattgatgt gaacatggtg gagcacgaca ctctcgtcta
     9361 ctccaagaat atcaaagata cagtctcaga agaccaaagg gctattgaga cttttcaaca
     9421 aagggtaata tcgggaaacc tcctcggatt ccattgccca gctatctgtc acttcatcaa
     9481 aaggacagta gaaaaggaag gtggcaccta caaatgccat cattgcgata aaggaaaggc
     9541 tatcgttcaa gatgcctctg ccgacagtgg tcccaaagat ggacccccac ccacgaggag
     9601 catcgtggaa aaagaagacg ttccaaccac gtcttcaaag caagtggatt gatgtgatat
     9661 ctccactgac gtaagggatg acgcacaatc ccactatcct tcgcaagacc cttcctctat
     9721 ataaggaagt tcatttcatt tggagaggac acgctgaaat caccagtctc tctctacaaa
     9781 tctatctctc tcgagtctac catgagccca gaacgacgcc cggccgacat ccgccgtgcc
     9841 accgaggcgg acatgccggc ggtctgcacc atcgtcaacc actacatcga gacaagcacg
     9901 gtcaacttcc gtaccgagcc gcaggaaccg caggagtgga cggacgacct cgtccgtctg
     9961 cgggagcgct atccctggct cgtcgccgag gtggacggcg aggtcgccgg catcgcctac
    10021 gcgggcccct ggaaggcacg caacgcctac gactggacgg ccgagtcgac cgtgtacgtc
    10081 tccccccgcc accagcggac gggactgggc tccacgctct acacccacct gctgaagtcc
    10141 ctggaggcac agggcttcaa gagcgtggtc gctgtcatcg ggctgcccaa cgacccgagc
    10201 gtgcgcatgc acgaggcgct cggatatgcc ccccgcggca tgctgcgggc ggccggcttc
    10261 aagcacggga actggcatga cgtgggtttc tggcagctgg acttcagcct gccggtaccg
    10321 ccccgtccgg tcctgcccgt caccgagatt tgactcgagt ttctccataa taatgtgtga
    10381 gtagttccca gataagggaa ttagggttcc tatagggttt cgctcatgtg ttgagcatat
    10441 aagaaaccct tagtatgtat ttgtatttgt aaaatacttc tatcaataaa atttctaatt
    10501 cctaaaacca aaatccagta ctaaaatcca gatcccccga attaattcgg cgttaattca
    10561 gtacattaaa aacgtccgca atgtgttatt aagttgtcta agcgtcaatt tgtttacacc
    10621 acaatatatc ctgccaccag ccagccaaca gctccccgac cggcagctcg gcacaaaatc
    10681 accactcgat acaggcagcc catcagtccg ggacggcgtc agcgggagag ccgttgtaag
    10741 gcggcagact ttgctcatgt taccgatgct attcggaaga acggcaacta agctgccggg
    10801 tttgaaacac ggatgatctc gcggagggta gcatgttgat tgtaacgatg acagagcgtt
    10861 gctgcctgtg atcaccgcgg tttcaaaatc ggctccgtcg atactatgtt atacgccaac
    10921 tttgaaaaca actttgaaaa agctgttttc tggtatttaa ggttttagaa tgcaaggaac
    10981 agtgaattgg agttcgtctt gttataatta gcttcttggg gtatctttaa atactgtaga
    11041 aaagaggaag gaaataataa atggctaaaa tgagaatatc accggaattg aaaaaactga
    11101 tcgaaaaata ccgctgcgta aaagatacgg aaggaatgtc tcctgctaag gtatataagc
    11161 tggtgggaga aaatgaaaac ctatatttaa aaatgacgga cagccggtat aaagggacca
    11221 cctatgatgt ggaacgggaa aaggacatga tgctatggct ggaaggaaag ctgcctgttc
    11281 caaaggtcct gcactttgaa cggcatgatg gctggagcaa tctgctcatg agtgaggccg
    11341 atggcgtcct ttgctcggaa gagtatgaag atgaacaaag ccctgaaaag attatcgagc
    11401 tgtatgcgga gtgcatcagg ctctttcact ccatcgacat atcggattgt ccctatacga
    11461 atagcttaga cagccgctta gccgaattgg attacttact gaataacgat ctggccgatg
    11521 tggattgcga aaactgggaa gaagacactc catttaaaga tccgcgcgag ctgtatgatt
    11581 ttttaaagac ggaaaagccc gaagaggaac ttgtcttttc ccacggcgac ctgggagaca
    11641 gcaacatctt tgtgaaagat ggcaaagtaa gtggctttat tgatcttggg agaagcggca
    11701 gggcggacaa gtggtatgac attgccttct gcgtccggtc gatcagggag gatatcgggg
    11761 aagaacagta tgtcgagcta ttttttgact tactggggat caagcctgat tgggagaaaa
    11821 taaaatatta tattttactg gatgaattgt tttagtacct agaatgcatg accaaaatcc
    11881 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
    11941 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
    12001 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
    12061 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
    12121 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
    12181 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
    12241 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
    12301 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
    12361 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
    12421 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
    12481 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
    12541 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
    12601 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
    12661 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga
    12721 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatatgg tgcactctca
    12781 gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat cgctacgtga
    12841 ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
    12901 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
    12961 gaggttttca ccgtcatcac cgaaacgcgc gaggcagggt gccttgatgt gggcgccggc
    13021 ggtcgagtgg cgacggcgcg gcttgtccgc gccctggtag attgcctggc cgtaggccag
    13081 ccatttttga gcggccagcg gccgcgatag gccgacgcga agcggcgggg cgtagggagc
    13141 gcagcgaccg aagggtaggc gctttttgca gctcttcggc tgtgcgctgg ccagacagtt
    13201 atgcacaggc caggcgggtt ttaagagttt taataagttt taaagagttt taggcggaaa
    13261 aatcgccttt tttctctttt atatcagtca cttacatgtg tgaccggttc ccaatgtacg
    13321 gctttgggtt cccaatgtac gggttccggt tcccaatgta cggctttggg ttcccaatgt
    13381 acgtgctatc cacaggaaac agaccttttc gacctttttc ccctgctagg gcaatttgcc
    13441 ctagcatctg ctccgtacat taggaaccgg cggatgcttc gccctcgatc aggttgcggt
    13501 agcgcatgac taggatcggg ccagcctgcc ccgcctcctc cttcaaatcg tactccggca
    13561 ggtcatttga cccgatcagc ttgcgcacgg tgaaacagaa cttcttgaac tctccggcgc
    13621 tgccactgcg ttcgtagatc gtcttgaaca accatctggc ttctgccttg cctgcggcgc
    13681 ggcgtgccag gcggtagaga aaacggccga tgccgggatc gatcaaaaag taatcggggt
    13741 gaaccgtcag cacgtccggg ttcttgcctt ctgtgatctc gcggtacatc caatcagcta
    13801 gctcgatctc gatgtactcc ggccgcccgg tttcgctctt tacgatcttg tagcggctaa
    13861 tcaaggcttc accctcggat accgtcacca ggcggccgtt cttggccttc ttcgtacgct
    13921 gcatggcaac gtgcgtggtg tttaaccgaa tgcaggtttc taccaggtcg tctttctgct
    13981 ttccgccatc ggctcgccgg cagaacttga gtacgtccgc aacgtgtgga cggaacacgc
    14041 ggccgggctt gtctcccttc ccttcccggt atcggttcat ggattcggtt agatgggaaa
    14101 ccgccatcag taccaggtcg taatcccaca cactggccat gccggccggc cctgcggaaa
    14161 cctctacgtg cccgtctgga agctcgtagc ggatcacctc gccagctcgt cggtcacgct
    14221 tcgacagacg gaaaacggcc acgtccatga tgctgcgact atcgcgggtg cccacgtcat
    14281 agagcatcgg aacgaaaaaa tctggttgct cgtcgccctt gggcggcttc ctaatcgacg
    14341 gcgcaccggc tgccggcggt tgccgggatt ctttgcggat tcgatcagcg gccgcttgcc
    14401 acgattcacc ggggcgtgct tctgcctcga tgcgttgccg ctgggcggcc tgcgcggcct
    14461 tcaacttctc caccaggtca tcacccagcg ccgcgccgat ttgtaccggg ccggatggtt
    14521 tgcgaccgct cacgccgatt cctcgggctt gggggttcca gtgccattgc agggccggca
    14581 ggcaacccag ccgcttacgc ctggccaacc gcccgttcct ccacacatgg ggcattccac
    14641 ggcgtcggtg cctggttgtt cttgattttc catgccgcct cctttagccg ctaaaattca
    14701 tctactcatt tattcatttg ctcatttact ctggtagctg cgcgatgtat tcagatagca
    14761 gctcggtaat ggtcttgcct tggcgtaccg cgtacatctt cagcttggtg tgatcctccg
    14821 ccggcaactg aaagttgacc cgcttcatgg ctggcgtgtc tgccaggctg gccaacgttg
    14881 cagccttgct gctgcgtgcg ctcggacggc cggcacttag cgtgtttgtg cttttgctca
    14941 ttttctcttt acctcattaa ctcaaatgag ttttgattta atttcagcgg ccagcgcctg
    15001 gacctcgcgg gcagcgtcgc cctcgggttc tgattcaaga acggttgtgc cggcggcggc
    15061 agtgcctggg tagctcacgc gctgcgtgat acgggactca agaatgggca gctcgtaccc
    15121 ggccagcgcc tcggcaacct caccgccgat gcgcgtgcct ttgatcgccc gcgacacgac
    15181 aaaggccgct tgtagccttc catccgtgac ctcaatgcgc tgcttaacca gctccaccag
    15241 gtcggcggtg gcccatatgt cgtaagggct tggctgcacc ggaatcagca cgaagtcggc
    15301 tgccttgatc gcggacacag ccaagtccgc cgcctggggc gctccgtcga tcactacgaa
    15361 gtcgcgccgg ccgatggcct tcacgtcgcg gtcaatcgtc gggcggtcga tgccgacaac
    15421 ggttagcggt tgatcttccc gcacggccgc ccaatcgcgg gcactgccct ggggatcgga
    15481 atcgactaac agaacatcgg ccccggcgag ttgcagggcg cgggctagat gggttgcgat
    15541 ggtcgtcttg cctgacccgc ctttctggtt aagtacagcg ataaccttca tgcgttcccc
    15601 ttgcgtattt gtttatttac tcatcgcatc atatacgcag cgaccgcatg acgcaagctg
    15661 ttttactcaa atacacatca cctttttaga cggcggcgct cggtttcttc agcggccaag
    15721 ctggccggcc aggccgccag cttggcatca gacaaaccgg ccaggatttc atgcagccgc
    15781 acggttgaga cgtgcgcggg cggctcgaac acgtacccgg ccgcgatcat ctccgcctcg
    15841 atctcttcgg taatgaaaaa cggttcgtcc tggccgtcct ggtgcggttt catgcttgtt
    15901 cctcttggcg ttcattctcg gcggccgcca gggcgtcggc ctcggtcaat gcgtcctcac
    15961 ggaaggcacc gcgccgcctg gcctcggtgg gcgtcacttc ctcgctgcgc tcaagtgcgc
    16021 ggtacagggt cgagcgatgc acgccaagca gtgcagccgc ctctttcacg gtgcggcctt
    16081 cctggtcgat cagctcgcgg gcgtgcgcga tctgtgccgg ggtgagggta gggcgggggc
    16141 caaacttcac gcctcgggcc ttggcggcct cgcgcccgct ccgggtgcgg tcgatgatta
    16201 gggaacgctc gaactcggca atgccggcga acacggtcaa caccatgcgg ccggccggcg
    16261 tggtggtgtc ggcccacggc tctgccaggc tacgcaggcc cgcgccggcc tcctggatgc
    16321 gctcggcaat gtccagtagg tcgcgggtgc tgcgggccag gcggtctagc ctggtcactg
    16381 tcacaacgtc gccagggcgt aggtggtcaa gcatcctggc cagctccggg cggtcgcgcc
    16441 tggtgccggt gatcttctcg gaaaacagct tggtgcagcc ggccgcgtgc agttcggccc
    16501 gttggttggt caagtcctgg tcgtcggtgc tgacgcgggc atagcccagc aggccagcgg
    16561 cggcgctctt gttcatggcg taatgtctcc ggttctagtc gcaagtatt
//
