LOCUS       sequence-390456-       11136 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses two gRNAs targeting Arabidopsis NIA1 gene for knockout. 
            Each gRNA is fused to TLS1 mobility signal that confers graft 
            mobility..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11136)
  AUTHORS   Yang L, Machin F, Wang S, Saplaoura E, Kragler F
  TITLE     Heritable transgene-free genome editing in plants by grafting of 
            wild-type shoots to transgenic donor rootstocks.
  JOURNAL   Nat Biotechnol. 2023 Jan 2. doi: 10.1038/s41587-022-01585-8.
  PUBMED    36593415
REFERENCE   2  (bases 1 to 11136)
  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: "Nat 
            Biotechnol. 2023 Jan 2. doi: 10.1038/s41587-022-01585-8."
COMMENT     Sequence Label: gRNA NIA TLS1
FEATURES             Location/Qualifiers
     source          1..11136
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    327..348
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(363..368,369..386,387..393)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    401..417
                     /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)."
     primer_bind     406..428
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     425..441
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     425..441
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    509..533
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_RNA        complement(817..892)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_RNA        complement(1468..1543)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(2022..2046)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2127..2144)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2127..2143)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2136..2158)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     misc_feature    2346..2370
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             3671..4300
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             4729..5802
                     /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      5868..6062
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(6298..6320)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    6406..6546
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     6461..6480
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6561..6578)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6732..7320)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6812..6831)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     mobile_element  7390..8157
                     /mobile_element_type="insertion sequence:IS1"
                     /label=IS1
                     /note="prokaryotic transposable element"
     CDS             complement(8184..8978)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     misc_feature    9403..9427
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     polyA_signal    9505..9679
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(9736..10533)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MGIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR
                     PVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLL
                     SSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEH
                     QGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDI
                     ALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(9847..9866)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     10457..10476
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(10596..11136,1..136))
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     primer_bind     complement(10639..10659)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
ORIGIN
        1 agcctttcct ttatcgcaat gatggcattt gtaggtgcca ccttcctttt ctactgtcct
       61 tttgatgaag tgacagatag ctgggcaatg gaatccgagg aggtttcccg atattaccct
      121 ttgttgaaaa gtctcaatag ccctttggtc ttctgagact gtatctttga tattcttgga
      181 gtagacgaga gtgtcgtgct ccaccatgtt ggcaagctgc tctagccaat acgcaaaccg
      241 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
      301 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
      361 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
      421 cacacaggaa acagctatga catgattacg aattcgagct ccaccgcggt ggcggccgct
      481 ctagaactag ttaattaaga attatcgaac cactttgtac aagaaagctg ggtctagata
      541 tctcgagtat tggtttatct catcggaact gcaaaactca actaactgaa aagtacaaaa
      601 ccactgaatc atattatttg agattttttt taggtcaaat tttaggtttc agttacagaa
      661 aacgaagaga aaaccccaga aattgaacgc cgaagaacag aggaagaaga aatcgatctg
      721 gaaaattttg caaaaaaaat ttatcagagc caggtttcga tcctgggacc tgtgggttat
      781 gggcccacca cgcttccgct gcgccactct gataatgcac cgactcggtg ccactttttc
      841 aagttgataa cggactagcc ttattttaac ttgctatttc tagctctaaa acccttcacg
      901 ttgtaaccca tcaatctctt agtcgactct accaatatat aaacagagct tctattttca
      961 actgaacgat gtgggatgtt tttgacatcg tttacagctt aaacgggctc gtttattaaa
     1021 agccctcttc tttcgatcca tcaacattat tggccttaag taaaacaagc ctctttatga
     1081 ttgagaaaac gccgtcgttg gtgcaccaga gagaagaaac aaaacagcta gcgttagata
     1141 aactttaaaa ttgtttgcat cctcatttgt ctgtcaggct gcagtagttt ggattaatat
     1201 tggtttatct catcggaact gcaaaactca actaactgaa aagtacaaaa ccactgaatc
     1261 atattatttg agattttttt taggtcaaat tttaggtttc agttacagaa aacgaagaga
     1321 aaaccccaga aattgaacgc cgaagaacag aggaagaaga aatcgatctg gaaaattttg
     1381 caaaaaaaat tttatcagag ccaggtttcg atcctgggac ctgtgggtta tgggcccacc
     1441 acgcttccgc tgcgccactc tgataatgca ccgactcggt gccacttttt caagttgata
     1501 acggactagc cttattttaa cttgctattt ctagctctaa aactgcagag tcagcagtgt
     1561 tgtcaatcac tacttcgact ctagctgtat ataaactcag cttcgttttc ttatctaagc
     1621 gatgtgggac ttttgaagat tgttttcaac ttaaatgggc ctatataaga aatactattg
     1681 ttctttccca tataaatggg cctgcttctc ttctttcaga ttcccagggg ccttttgaag
     1741 attatcttca tatcttaaga atgaagatgt tttattcaat caaattcttg aaggttcgat
     1801 gcctaatcat tctaatcctg ggacaaacta tgaaacaaga tacaaaaact ccgaatggaa
     1861 agttaaaaag aagaaaacga aagctacggt tcaagaaaat gtaagctgat aaacaaaaaa
     1921 aaactgtatg aacgaagaag aagaaaaaaa gctaagaaga aatgatgtat tgtgcggaag
     1981 gcaagtcggg atccagtcga ctgaattggt tcccatggtg gagcctgctt ttttgtacaa
     2041 acttgtttga tagcttggcg cgcctcgagg gggggcccgg tacccgggga tcctctagag
     2101 tcgacctgca ggcatgcaag cttggcactg gccgtcgttt tacaacgtcg tgactgggaa
     2161 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
     2221 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     2281 tgctagagca gcttgagctt ggatcagatt gtcgtttccc gccttcagtt taaactatca
     2341 gtgtttgaca ggatatattg gcgggtaaac ctaagagaaa agagcgttta ttagaataat
     2401 cggatattta aaagggcgtg aaaaggttta tccgttcgtc catttgtatg tgcatgccaa
     2461 ccacagggtt cccctcggga tcaaagtact ttgatccaac ccctccgctg ctatagtgca
     2521 gtcggcttct gacgttcagt gcagccgtct tctgaaaacg acatgtcgca caagtcctaa
     2581 gttacgcgac aggctgccgc cctgcccttt tcctggcgtt ttcttgtcgc gtgttttagt
     2641 cgcataaagt agaatacttg cgactagaac cggagacatt acgccatgaa caagagcgcc
     2701 gccgctggcc tgctgggcta tgcccgcgtc agcaccgacg accaggactt gaccaaccaa
     2761 cgggccgaac tgcacgcggc cggctgcacc aagctgtttt ccgagaagat caccggcacc
     2821 aggcgcgacc gcccggagct ggccaggatg cttgaccacc tacgccctgg cgacgttgtg
     2881 acagtgacca ggctagaccg cctggcccgc agcacccgcg acctactgga cattgccgag
     2941 cgcatccagg aggccggcgc gggcctgcgt agcctggcag agccgtgggc cgacaccacc
     3001 acgccggccg gccgcatggt gttgaccgtg ttcgccggca ttgccgagtt cgagcgttcc
     3061 ctaatcatcg accgcacccg gagcgggcgc gaggccgcca aggcccgagg cgtgaagttt
     3121 ggcccccgcc ctaccctcac cccggcacag atcgcgcacg cccgcgagct gatcgaccag
     3181 gaaggccgca ccgtgaaaga ggcggctgca ctgcttggcg tgcatcgctc gaccctgtac
     3241 cgcgcacttg agcgcagcga ggaagtgacg cccaccgagg ccaggcggcg cggtgccttc
     3301 cgtgaggacg cattgaccga ggccgacgcc ctggcggccg ccgagaatga acgccaagag
     3361 gaacaagcat gaaaccgcac caggacggcc aggacgaacc gtttttcatt accgaagaga
     3421 tcgaggcgga gatgatcgcg gccgggtacg tgttcgagcc gcccgcgcac gtctcaaccg
     3481 tgcggctgca tgaaatcctg gccggtttgt ctgatgccaa gctggcggcc tggccggcca
     3541 gcttggccgc tgaagaaacc gagcgccgcc gtctaaaaag gtgatgtgta tttgagtaaa
     3601 acagcttgcg tcatgcggtc gctgcgtata tgatgcgatg agtaaataaa caaatacgca
     3661 aggggaacgc atgaaggtta tcgctgtact taaccagaaa ggcgggtcag gcaagacgac
     3721 catcgcaacc catctagccc gcgccctgca actcgccggg gccgatgttc tgttagtcga
     3781 ttccgatccc cagggcagtg cccgcgattg ggcggccgtg cgggaagatc aaccgctaac
     3841 cgttgtcggc atcgaccgcc cgacgattga ccgcgacgtg aaggccatcg gccggcgcga
     3901 cttcgtagtg atcgacggag cgccccaggc ggcggacttg gctgtgtccg cgatcaaggc
     3961 agccgacttc gtgctgattc cggtgcagcc aagcccttac gacatatggg ccaccgccga
     4021 cctggtggag ctggttaagc agcgcattga ggtcacggat ggaaggctac aagcggcctt
     4081 tgtcgtgtcg cgggcgatca aaggcacgcg catcggcggt gaggttgccg aggcgctggc
     4141 cgggtacgag ctgcccattc ttgagtcccg tatcacgcag cgcgtgagct acccaggcac
     4201 tgccgccgcc ggcacaaccg ttcttgaatc agaacccgag ggcgacgctg cccgcgaggt
     4261 ccaggcgctg gccgctgaaa ttaaatcaaa actcatttga gttaatgagg taaagagaaa
     4321 atgagcaaaa gcacaaacac gctaagtgcc ggccgtccga gcgcacgcag cagcaaggct
     4381 gcaacgttgg ccagcctggc agacacgcca gccatgaagc gggtcaactt tcagttgccg
     4441 gcggaggatc acaccaagct gaagatgtac gcggtacgcc aaggcaagac cattaccgag
     4501 ctgctatctg aatacatcgc gcagctacca gagtaaatga gcaaatgaat aaatgagtag
     4561 atgaatttta gcggctaaag gaggcggcat ggaaaatcaa gaacaaccag gcaccgacgc
     4621 cgtggaatgc cccatgtgtg gaggaacggg cggttggcca ggcgtaagcg gctgggttgc
     4681 ctgccggccc tgcaatggca ctggaacccc caagcccgag gaatcggcgt gagcggtcgc
     4741 aaaccatccg gcccggtaca aatcggcgcg gcgctgggtg atgacctggt ggagaagttg
     4801 aaggccgcgc aggccgccca gcggcaacgc atcgaggcag aagcacgccc cggtgaatcg
     4861 tggcaagcgg ccgctgatcg aatccgcaaa gaatcccggc aaccgccggc agccggtgcg
     4921 ccgtcgatta ggaagccgcc caagggcgac gagcaaccag attttttcgt tccgatgctc
     4981 tatgacgtgg gcacccgcga tagtcgcagc atcatggacg tggccgtttt ccgtctgtcg
     5041 aagcgtgacc gacgagctgg cgaggtgatc cgctacgagc ttccagacgg gcacgtagag
     5101 gtttccgcag ggccggccgg catggccagt gtgtgggatt acgacctggt actgatggcg
     5161 gtttcccatc taaccgaatc catgaaccga taccgggaag ggaagggaga caagcccggc
     5221 cgcgtgttcc gtccacacgt tgcggacgta ctcaagttct gccggcgagc cgatggcgga
     5281 aagcagaaag acgacctggt agaaacctgc attcggttaa acaccacgca cgttgccatg
     5341 cagcgtacga agaaggccaa gaacggccgc ctggtgacgg tatccgaggg tgaagccttg
     5401 attagccgct acaagatcgt aaagagcgaa accgggcggc cggagtacat cgagatcgag
     5461 ctagctgatt ggatgtaccg cgagatcaca gaaggcaaga acccggacgt gctgacggtt
     5521 caccccgatt actttttgat cgatcccggc atcggccgtt ttctctaccg cctggcacgc
     5581 cgcgccgcag gcaaggcaga agccagatgg ttgttcaaga cgatctacga acgcagtggc
     5641 agcgccggag agttcaagaa gttctgtttc accgtgcgca agctgatcgg gtcaaatgac
     5701 ctgccggagt acgatttgaa ggaggaggcg gggcaggctg gcccgatcct agtcatgcgc
     5761 taccgcaacc tgatcgaggg cgaagcatcc gccggttcct aatgtacgga gcagatgcta
     5821 gggcaaattg ccctagcagg ggaaaaaggt cgaaaaggtc tctttcctgt ggatagcacg
     5881 tacattggga acccaaagcc gtacattggg aaccggaacc cgtacattgg gaacccaaag
     5941 ccgtacattg ggaaccggtc acacatgtaa gtgactgata taaaagagaa aaaaggcgat
     6001 ttttccgcct aaaactcttt aaaacttatt aaaactctta aaacccgcct ggcctgtgca
     6061 taactgtctg gccagcgcac agccgaagag ctgcaaaaag cgcctaccct tcggtcgctg
     6121 cgctccctac gccccgccgc ttcgcgtcgg cctatcgcgg ccgctggccg ctcaaaaatg
     6181 gctggcctac ggccaggcaa tctaccaggg cgcggacaag ccgcgccgtc gccactcgac
     6241 cgccggcgcc cacatcaagg caccctgcct cgcgcgtttc ggtgatgacg gtgaaaacct
     6301 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     6361 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca
     6421 gtcacgtagc gatagcggag tgtatactgg cttaactatg cggcatcaga gcagattgta
     6481 ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc
     6541 atcaggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     6601 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     6661 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     6721 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     6781 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     6841 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     6901 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     6961 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     7021 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     7081 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     7141 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
     7201 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     7261 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     7321 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     7381 gggattttgg gtgatgctgc caacttactg atttagtgta tgatggtgtt tttgaggtgc
     7441 tccagtggct tctgtttcta tcagctgtcc ctcctgttca gctactgacg gggtggtgcg
     7501 taacggcaaa agcaccgccg gacatcagcg ctatctctgc tctcactgcc gtaaaacatg
     7561 gcaactgcag ttcacttaca ccgcttctca acccggtacg caccagaaaa tcattgatat
     7621 ggccatgaat ggcgttggat gccgggcaac agcccgcatt atgggcgttg gcctcaacac
     7681 gattttacgt cacttaaaaa actcaggccg cagtcggtaa cctcgcgcat acagccgggc
     7741 agtgacgtca tcgtctgcgc ggaaatggac gaacagtggg gctatgtcgg ggctaaatcg
     7801 cgccagcgct ggctgtttta cgcgtatgac agtctccgga agacggttgt tgcgcacgta
     7861 ttcggtgaac gcactatggc gacgctgggg cgtcttatga gcctgctgtc accctttgac
     7921 gtggtgatat ggatgacgga tggctggccg ctgtatgaat cccgcctgaa gggaaagctg
     7981 cacgtaatca gcaagcgata tacgcagcga attgagcggc ataacctgaa tctgaggcag
     8041 cacctggcac ggctgggacg gaagtcgctg tcgttctcaa aatcggtgga gctgcatgac
     8101 aaagtcatcg ggcattatct gaacataaaa cactatcaat aagttggagt cattaccggg
     8161 attttggtca tgcattctag gtactaaaac aattcatcca gtaaaatata atattttatt
     8221 ttctcccaat caggcttgat ccccagtaag tcaaaaaata gctcgacata ctgttcttcc
     8281 ccgatatcct ccctgatcga ccggacgcag aaggcaatgt cataccactt gtccgccctg
     8341 ccgcttctcc caagatcaat aaagccactt actttgccat ctttcacaaa gatgttgctg
     8401 tctcccaggt cgccgtggga aaagacaagt tcctcttcgg gcttttccgt ctttaaaaaa
     8461 tcatacagct cgcgcggatc tttaaatgga gtgtcttctt cccagttttc gcaatccaca
     8521 tcggccagat cgttattcag taagtaatcc aattcggcta agcggctgtc taagctattc
     8581 gtatagggac aatccgatat gtcgatggag tgaaagagcc tgatgcactc cgcatacagc
     8641 tcgataatct tttcagggct ttgttcatct tcatactctt ccgagcaaag gacgccatcg
     8701 gcctcactca tgagcagatt gctccagcca tcatgccgtt caaagtgcag gacctttgga
     8761 acaggcagct ttccttccag ccatagcatc atgtcctttt cccgttccac atcataggtg
     8821 gtccctttat accggctgtc cgtcattttt aaatataggt tttcattttc tcccaccagc
     8881 ttatatacct tagcaggaga cattccttcc gtatctttta cgcagcggta tttttcgatc
     8941 agttttttca attccggtga tattctcatt ttagccattt attatttcct tcctcttttc
     9001 tacagtattt aaagataccc caagaagcta attataacaa gacgaactcc aattcactgt
     9061 tccttgcatt ctaaaacctt aaataccaga aaacagcttt ttcaaagttg ttttcaaagt
     9121 tggcgtataa catagtatcg acggagccga ttttgaaacc gcggtgatca caggcagcaa
     9181 cgctctgtca tcgttacaat caacatgcta ccctccgcga gatcatccgt gtttcaaacc
     9241 cggcagctta gttgccgttc ttccgaatag catcggtaac atgagcaaag tctgccgcct
     9301 tacaacggct ctcccgctga cgccgtcccg gactgatggg ctgcctgtat cgagtggtga
     9361 ttttgtgccg agctgccggt cggggagctg ttggctggct ggtggcagga tatattgtgg
     9421 tgtaaacaaa ttgacgctta gacaacttaa taacacattg cggacgtttt taatgtactg
     9481 aattaacgcc gaattaattc gggggatctg gattttagta ctggattttg gttttaggaa
     9541 ttagaaattt tattgataga agtattttac aaatacaaat acatactaag ggtttcttat
     9601 atgctcaaca catgagcgaa accctatagg aaccctaatt cccttatctg ggaactactc
     9661 acacattatt atggagaaac tcgagcttgt cgatcgactc tagctagagg atcgatccga
     9721 accccagagt cccgctcaga agaactcgtc aagaaggcga tagaaggcga tgcgctgcga
     9781 atcgggagcg gcgataccgt aaagcacgag gaagcggtca gcccattcgc cgccaagctc
     9841 ttcagcaata tcacgggtag ccaacgctat gtcctgatag cggtccgcca cacccagccg
     9901 gccacagtcg atgaatccag aaaagcggcc attttccacc atgatattcg gcaagcaggc
     9961 atcgccatgt gtcacgacga gatcctcgcc gtcgggcatg cgcgccttga gcctggcgaa
    10021 cagttcggct ggcgcgagcc cctgatgctc ttcgtccaga tcatcctgat cgacaagacc
    10081 ggcttccatc cgagtacgtg ctcgctcgat gcgatgtttc gcttggtggt cgaatgggca
    10141 ggtagccgga tcaagcgtat gcagccgccg cattgcatca gccatgatgg atactttctc
    10201 ggcaggagca aggtgagatg acaggagatc ctgccccggc acttcgccca atagcagcca
    10261 gtcccttccc gcttcagtga caacgtcgag cacagctgcg caaggaacgc ccgtcgtggc
    10321 cagccacgat agccgcgctg cctcgtcctg gagttcattc agggcaccgg acaggtcggt
    10381 cttgacaaaa agaaccgggc gcccctgcgc tgacagccga aacacggcgg catcagagca
    10441 gccgattgtc tgttgtgccc agtcatagcc gaatagcctc tccacccaag cggccggaga
    10501 acctgcgtgc aatccatctt gttcaatccc catggtcgat cgacagatct gcgaaagctc
    10561 gagagagata gatttgtaga gagagactgg tgatttcagc gtgtcctctc caaatgaaat
    10621 gaacttcctt atatagagga agggtcttgc gaaggatagt gggattgtgc gtcatccctt
    10681 acgtcagtgg agatatcaca tcaatccact tgctttgaag acgtggttgg aacgtcttct
    10741 ttttccacga tgctcctcgt gggtgggggt ccatctttgg gaccactgtc ggcagaggca
    10801 tcttgaacga tagcctttcc tttatcgcaa tgatggcatt tgtaggtgcc accttccttt
    10861 tctactgtcc ttttgatgaa gtgacagata gctgggcaat ggaatccgag gaggtttccc
    10921 gatattaccc tttgttgaaa agtctcaata gccctttggt cttctgagac tgtatctttg
    10981 atattcttgg agtagacgag agtgtcgtgc tccaccatgt tcacatcaat ccacttgctt
    11041 tgaagacgtg gttggaacgt cttctttttc cacgatgctc ctcgtgggtg ggggtccatc
    11101 tttgggacca ctgtcggcag aggcatcttg aacgat
//
