LOCUS       sequence-456266-       19247 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The backbone vector for rice genome editing is based on HkCas12a.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 19247)
  AUTHORS   Liu S, He Y, Fan T, Zhu M, Qi C, Ma Y, Yang M, Yang L, Tang X, Zhou 
            J, Zhong Z, An X, Qi Y, Zhang Y
  TITLE     PAM-relaxed and temperature-tolerant CRISPR-Mb3Cas12a single 
            transcript unit systems for efficient singular and multiplexed 
            genome editing in rice, maize, and tomato.
  JOURNAL   Plant Biotechnol J. 2024 Oct 10. doi: 10.1111/pbi.14486.
  PUBMED    39387219
REFERENCE   2  (bases 1 to 19247)
  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 
            Biotechnol J. 2024 Oct 10. doi: 10.1111/pbi.14486."
COMMENT     Sequence Label: pGEL1022
FEATURES             Location/Qualifiers
     source          1..19247
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     17..39
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     31..48
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     32..48
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     ncRNA           complement(404..471)
                     /product="hepatitis delta virus self-cleaving ribozyme 
                     (Ferre-D'Amare and Scott, 2010)"
                     /label=HDV ribozyme
                     /ncRNA_class="ribozyme"
     CDS             798..1103
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     982..1001
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     ncRNA           complement(1131..1167)
                     /product="self-cleaving hammerhead ribozyme (Ferre-D'Amare 
                     and Scott, 2010)"
                     /label=hammerhead ribozyme
                     /ncRNA_class="hammerhead_ribozyme"
     terminator      complement(2731..2979)
                     /label=HSP terminator
                     /note="efficient transcription terminator from the 
                     Arabidopsis thaliana heat shock protein 18.2 gene (Nagaya 
                     et al., 2010)"
     CDS             complement(2998..3045)
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             complement(6997..7017)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      7020..7029
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(9068..9084)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9068..9084)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9081..9103)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9092..9108
                     /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(9116..9122,9123..9140,9141..9146))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9161..9182
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        9373..10049
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     primer_bind     9986..10006
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             10116..11141
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     K"
     CDS             10116..11141
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     K"
     polyA_signal    11182..11356
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    11434..11458
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             11883..12677
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      14102..14690
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     14591..14610
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     14844..14861
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    14876..15016
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(14942..14961)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     15102..15124
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     rep_origin      15360..15554
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(15620..16693)
                     /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(17122..17751)
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     misc_feature    19052..19076
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
ORIGIN
        1 gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtgc cacactgagc
       61 tcgatatcgg cgcgccccgg gagctcgcaa aacacaccta gactagattt gttttgctaa
      121 cccaattgat attaattata tatgattaat atttatatgt atatggaatt ggttaataaa
      181 atgcatctgg ttcatcaaag aattataaag acacgtgaca ttcatttagg ataagaaata
      241 tggatgatct ctttctctta ttcagataat tagtaattac acataacaca caactttgat
      301 gcccacatta tagtgattag catgtcacta tgtgtgcatc cttttatttc atacattaat
      361 taacttggcc aatccagaag atggacaagt ctaggactcg agtgtcccat tcgccatgcc
      421 gaagcatgtt gcccagccgg cgccagcgag gaggctggga ccatgccggc ctgagaccaa
      481 gccagataac agtatgcgta tttgcgcgct gatttttgcg gtataagaat atatactgat
      541 atgtataccc gaagtatgtc aaaaagaggt atgctatgaa gcagcgtatt acagtgacag
      601 ttgacagcga cagctatcag ttgctcaagg catatatgat gtcaatatct ccggtctggt
      661 aagcacaacc atgcagaatg aagcccgtcg tctgcgtgcc gaacgctgga aagcggaaaa
      721 tcaggaaggg atggctgagg tcgcccggtt tattgaaatg aacggctctt ttgctgacga
      781 gaacaggggc tggtgaaatg cagtttaagg tttacaccta taaaagagag agccgttatc
      841 gtctgtttgt ggatgtacag agtgatatta ttgacacgcc cgggcgacgg atggtgatcc
      901 ccctggccag tgcacgtctg ctgtcagata aagtctcccg tgaactttac ccggtggtgc
      961 atatcgggga tgaaagctgg cgcatgatga ccaccgatat ggccagtgtg ccggtttccg
     1021 ttatcgggga agaagtggct gatctcagcc accgcgaaaa tgacatcaaa aacgccatta
     1081 acctgatgtt ctggggaata taaggtctcc atctacaata gtagaaatta gacgagctta
     1141 ctcgtttcgt cctcacggac tcatcagtaa tttcctgcag gctgcacaca caattaccga
     1201 agcatggtca gcaactatga acatgaaatt ataacaatac caagcaagta cagcagaaat
     1261 cagtcaatca aattattcca atatgttaga tacacgatct aaaccacaat aacatcagtg
     1321 tacctatcaa caccaacagt gacctaacaa aatagagagt cgcaccgtgc tcgagttcac
     1381 aaaaataatc tgcaggtcta tgaactacta gaattccatc ggcaaactga ttcaataact
     1441 actagcacta tcccgagagc acataaggca ggttaattaa cataggctag aatttacagg
     1501 tgcaacagtt gataagatag gcataagcaa gtaaaatatc ctgaaagcta tgctatgtaa
     1561 tcactaatca acaagacgca gcccttagat taaacatgaa cagagtagaa tctatgcaac
     1621 tagagggact gaatcaaaaa caacctaagc aaaatatacc tgaagctgct acttggatca
     1681 acgagaaaaa cagagactaa atcctactga ttgaacagat cggaacaaat cacaacccta
     1741 gcacagttct acttctagat ccgaaccaat aaatacttgg accacgaaca gcatcgctca
     1801 taatcacaga tccagcatac cacgcgaacc ccatctaacc ctgttcaaat ctaccacaga
     1861 tcgcaaaacg gagactaaca acaacatctc acaatcacga acaggccggg aaatcaaatc
     1921 aaacgcgagg aacggagagg atcagaaaat cgcatacctt gagaaggcga ttcgacgctt
     1981 cgcttcgagg ggagattttt ttttctctgc gattttgggg attcgattga tggaggaaga
     2041 ggcgaggagg gggtggggta tttataggtg gggagggggg gactcgtgag tgggcccagg
     2101 ggtgcggtgg gtggggccca gtgagaatat agacaaataa agagagggag aggcggtttg
     2161 gggagtccgt ccccaccacg caaacgcgag aaggaaagaa aggaacggaa agggggcggg
     2221 tccagacggc gccgcggaga tatcgaggag gtggaaagct gatgccgttt gctcacggcg
     2281 gggttggtgg ccgttaacac gcagcgaagc ggcgttactg ccgtgtacgt tttgtgacct
     2341 ttcgaaagag agccccagcc caagaaaggc tggccaggtt tttttgtact gtgtacgatt
     2401 acgacggaag aaaacatact cgttgttttg tattggtaga tactgggccg tagtaaaatt
     2461 agaatcagtt gttaattggt tattatatat accatcttaa gctaccaaaa ttaataaagt
     2521 tgtgatggct atggtatatt ttggtagtga attatattga tttggtatat attagaatca
     2581 atccaaccac aacacaacca aaaatacctg aagctcaata ttgatggtgc ttttcgacta
     2641 tctagcaaat gtggtgaatg gggctttata attgaagcag cccaggatgc actttgtgca
     2701 gagactgatg tgggcgcgcc gtccgagctc cttatcttta atcatattcc atagtccata
     2761 ccatagcaca tacagtagtt atatgctgca gaagagatcc aacaaaacat tcacaatgga
     2821 ttatagaaac atttgtttat tcattataat gtgatcttac attcatttaa tattagaaaa
     2881 agccacaaat tcataacaca acaagccaag aaaaaaacac aaacttaagc acacaagctt
     2941 tttatttgac acaccaaata tttcatcttc atcttcatat actcgagtac gcgttcactt
     3001 ctttttctta gcctgtccgg cctttttggt ggcagcagga cgcttggaaa tgatgaaatt
     3061 aatccaatcc tcgttcttga tcttgaggtc gaccttggac tcattagagt tcacaatcct
     3121 atccttgagc aggaggccct tccgcgcgat attgaacgcg ccgttggcgt cgccgttatt
     3181 tgggagattg gtcttatcaa tgtcgatctc atttgagttg tagtaattgc cgttatcgtc
     3241 ggctgtcggg ctaatgatgt aatcattctc gaagtcgtag ttgcggagct gcacgctgta
     3301 cttgaaggta tcgaagaggg ccttgagaat cttcctcttg gccttctcct ccaggttgtc
     3361 gatgagagga atgatgtcaa tattctcgta atcgatgttc agctccttca gcttattgat
     3421 gaggaactcg gttggctgca cgtagaagta ctccttcttc ttggagttgt agctaatcct
     3481 ctcgccccgt gttgagatct tccaatcctt gctcttaatg ttattgagga tgaccagatt
     3541 cgccttggcg aagtccttgt agttgaagat gaacttgaag tcctcctcct cattatcgaa
     3601 gtagatgttc ttgaacttct tgatgaactc ctcgtactta gatgaattga tggacttgag
     3661 cctgaacagg tttgtgaagc cggtgacagg gtcaatcaca gaggtgtacg cggctggcag
     3721 gtagaacacg atgccattct ggccctgcag ctcctggtac gcatcgaccg ggtagcaggc
     3781 ctgaatagga ttcaggatgc cggatggctc caggttcttc ctctcgtcgt acttctcctt
     3841 gaaggagagc gcagacagct tgttcatgag ggccagctcg aacttctggt agacctgcct
     3901 ctccaccttg aaccggccgc gcttgaagcc ctgattgagg ttctccataa tcacgatcgc
     3961 ctcgtactta atgatgagcc gggacagctt atggaccacg tgagacaggt agccatcctt
     4021 cagctcctta atcttattga tgttggtcca attcttcctg gcccgatcgc ggtcctcctc
     4081 ccttgtcctc agcttgttct ggtaatcgac cttcacggtg gaattgtcga agcggttctg
     4141 ctccagggta ttcagagagc cctgctccac aatgttgccc tccaggtcga tgacgctgta
     4201 gtacaggaga tggcgctcgc ccctatcgat gccaatgatg tgcttaatct cgccgtcctt
     4261 gatcatgccc tggacaatct gattcacgtt cttattggtc ttaatatccc agttgaatgt
     4321 gatcggcaca tggaagaaga actgtgactc gctgtaccgg cgatccttaa tgatgtcgaa
     4381 cctggcctcc ttgtacttca ggacgtcctt gttgaagagg gcgatcgcct catcggacag
     4441 gtcctcgcgg ccgatcttct tgttgaggta gcggtagcac tcatggtaga tattctccgg
     4501 gatggtgttg ccctccttat cgatcttgtt caggagaata gagccctcct tgtgaatgat
     4561 aggcttcttc tccagcttct tgcggtagaa gatctccgcc tggccattga gcttcagctt
     4621 gcgggtcctg aggttctcct ctgagaacat gctgagccag tacagtgtgt ggagattctt
     4681 cttgcccttg gactctggag agaagtcctt gttgtagatc tggaacagga gcagcttgcc
     4741 gtcctccacg gcattatcga ttgtgttctc gtcaatctta ttgaacagga ccctgtacgc
     4801 gcatgagttc acatcggcgt agaagtcgtt cagattgacg tagagggatg tatccttgag
     4861 ggtagagaag tcgtacatct ccttgtagct ctccttgttc gggcccttgt acttgtacag
     4921 gaactgcttg cagaactcaa tccacttgta gatggtgtag aggtagttct tatttgtgtt
     4981 attatccttg tagaagtcct tcttgtactt cttctccttc ttaatgtcgt agatctcctt
     5041 ggtaatgatc agtggtgaca cgtagccatc gatgagctgg aagtcgctca cgttattctt
     5101 gaaatgctcc ttgaccttct ttgtgaatgg gtaattcgga atggacttag agaactccgg
     5161 gatctggttg tagatgagga tctcgtagat cttcttggac ttattcttct tgttgatgag
     5221 cagcttgtta tcaatcagga tgccgaggta gtagtagccg cccttcctca gaatgatgga
     5281 ccgattgtca gaaatctttg actctgacca tccatcggcc agggttggga agtcgaagtt
     5341 caccttgatc ttatccttgt cggagtgctt gcgtgtgagg taattcctgg tcaggttgta
     5401 gagtggaatg atgtagtagt cagaaatgac ctcattgagc agcgggtaga atgtctcatc
     5461 ggtgtccaga tcgtcgttgc cgatgagcag gagcttccac ttatggtact cctccaggtg
     5521 cttctccagg tactccttga tcgcctccag cttgatcggg gagccgtaga taggggcctc
     5581 gaactcgctg cgctcgatgt tattcatctt gaacttctcc aggagcacgg agaacacatc
     5641 cacctcctcg aagccctcga ccagattcac cagctcctca atctcccgga tgttgaggta
     5701 gcgcttggcc ttggatgtct cgtccagctc caggtactcc ttgtacttct tcttctcctc
     5761 cttagagccc tctgtaatct tgacggtgta gcccctccgg atcacatccc actggccgaa
     5821 cagcatctgt gagaacttct gtgtattcct cgcgtcaatg tagacctcct ccttcttgat
     5881 ggacttgagg gcgttgtaga gccgctcgat ctgctcgaac agggaaggag acacctcggt
     5941 tgactcgaag aagctgacat tgaacttgtt gatccgatcc acgaggtcgt tatcgtcctc
     6001 aatctgatcg atcaggaatg atgtgctctc ggagtactcc aggatctgct tgcgcagctt
     6061 ggtgaggacg cccagcttca cattctttgt ctctggcttg cgcctgagct gctgctgggt
     6121 gagattcaga atctcgttga tgccctgcac cttctcgccg ttctcctttg tgaagccgcc
     6181 cctgatcgcc tggtagacat caatgccgtc ctgggtgatc acgtaattga agccgttcgg
     6241 tgtgaagtaa tccttgactg agcggttatc gtccaggatg tcggcctcct tcagctcatt
     6301 ctccaggtta ggaatcttat tctccaggta ctcgatgttc ttattgaaaa tcttgatgtt
     6361 gtcgtagtag aagccgaagt tctcattcac cgcgcggaac ggaatggcgg tgacgatatc
     6421 cttgtctgtg aacacattct tccgtgtctc ccagaagttc ttcaggcgtg tggtgaactt
     6481 agagtagagg ttcaggccct ccaggtcctc ctcagagtat gagtcgttat tgttgacgaa
     6541 atcagggagc acctccagaa tgaactgctt cttgaacagc ttgccgtaga gggccttatt
     6601 gttcttgatc agatcctttg aattcttctc gccgctcttg tccagctcgc ctgagagaat
     6661 gttgaacagg accttcctgt actgcgcctg gatcatctcc aggttcttct ggctttcctt
     6721 cttggaggag tcgagcagct cgtgggcatc gtacagcttc tgccaatcga gattcaggtc
     6781 tgagagcttc tccttgatgt actgccggta gtaatcgtcg gaaatgttct tgaggatagg
     6841 gtagaaatca gagcgctcga agtccttctc cagcttgccc agcttctcga tattctccag
     6901 ggtcttgccc actggaatga gcttgaagcg gattgtcttg ctaatggaca cgatgttgct
     6961 gagcttctcg aaagccgcag gcaccccgtg aataccaacc ttccgcttct tcttaggagc
     7021 catggcggcc tgcaggaagt aacaccaaac aacagggtga gcatcgacaa aagaaacagt
     7081 accaagcaaa taaatagcgt atgaaggcag ggctaaaaaa atccacatat agctgctgca
     7141 tatgccatca tccaagtata tcaagatcaa aataattata aaacatactt gtttattata
     7201 atagataggt actcaaggtt agagcatatg aatagatgct gcatatgcca tcatgtatat
     7261 gcatcagtaa aacccacatc aacatgtata cctatcctag atcgatattt ccatccatct
     7321 taaactcgta actatgaaga tgtatgacac acacatacag ttccaaaatt aataaataca
     7381 ccaggtagtt tgaaacagta ttctactccg atctagaacg aatgaacgac cgcccaacca
     7441 caccacatca tcacaaccaa gcgaacaaaa agcatctctg tatatgcatc agtaaaaccc
     7501 gcatcaacat gtatacctat cctagatcga tatttccatc catcatcttc aattcgtaac
     7561 tatgaatatg tatggcacac acatacagat ccaaaattaa taaatccacc aggtagtttg
     7621 aaacagaatt aattctactc cgatctagaa cgaccgccca accagaccac atcatcacaa
     7681 ccaagacaaa aaaaagcatg aaaagatgac ccgacaaaca agtgcacggc atatattgaa
     7741 ataaaggaaa agggcaaacc aaaccctatg caacgaaaca aaaaaaatca tgaaatcgat
     7801 cccgtctgcg gaacggctag agccatccca ggattcccca aagagaaaca ctggcaagtt
     7861 agcaatcaga acgtgtctga cgtacaggtc gcatccgtgt acgaacgcta gcagcacgga
     7921 tctaacacaa acacggatct aacacaaaca tgaacagaag tagaactacc gggccctaac
     7981 catggaccgg aacgccgatc tagagaaggt agagaggggg gggggggagg acgagcggcg
     8041 taccttgaag cggaggtgcc gacgggtgga tttgggggag atctggttgt gtgtgtgtgc
     8101 gctccgaaca acacgaggtt ggggaaagag ggtgtggagg gggtgtctat ttattacggc
     8161 gggcgaggaa gggaaagcga aggagcggtg ggaaaggaat cccccgtagc tgccggtgcc
     8221 gtgagaggag gaggaggccg cctgccgtgc cggctcacgt ctgccgctcc gccacgcaat
     8281 ttctggatgc cgacagcgga gcaagtccaa cggtggagcg gaactctcga gaggggtcca
     8341 gaggcagcga cagagatgcc gtgccgtctg cttcgcttgg cccgacgcga cgctgctggt
     8401 tcgctggttg gtgtccgtta gactcgtcga cggcgtttaa caggctggca ttatctactc
     8461 gaaacaagaa aaatgtttcc ttagtttttt taatttctta aagggtattt gtttaatttt
     8521 tagtcacttt attttattct attttatatc taaattatta aataaaaaaa ctaaaataga
     8581 gttttagttt tcttaattta gaggctaaaa tagaataaaa tagatgtact aaaaaaatta
     8641 gtctataaaa accattaacc ctaaacccta aatggatgta ctaataaaat ggatgaagta
     8701 ttatataggt gaagctattt gcaaaaaaaa aggagaacac atgcacacta aaaagataaa
     8761 actgtagagt cctgttgtca aaatactcaa ttgtccttta gaccatgtct aactgttcat
     8821 ttatatgatt ctctaaaaca ctgatattat tgtagtacta tagattatat tattcgtaga
     8881 gtaaagttta aatatatgta taaagataga taaactgcac ttcaaacaag tgtgacaaaa
     8941 aaaatatgtg gtaatttttt ataacttaga catgcaatgc tcattatctc tagagagggg
     9001 cacgaccggg tcacgctgca ctgcaggcgc gccgatcccc gggtaccgag ctcgaattcg
     9061 taatcatgtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     9121 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     9181 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     9241 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tggctagagc agcttgccaa
     9301 catggtggag cacgacactc tcgtctactc caagaatatc aaagatacag tctcagaaga
     9361 ccaaagggct attgagactt ttcaacaaag ggtaatatcg ggaaacctcc tcggattcca
     9421 ttgcccagct atctgtcact tcatcaaaag gacagtagaa aaggaaggtg gcacctacaa
     9481 atgccatcat tgcgataaag gaaaggctat cgttcaagat gcctctgccg acagtggtcc
     9541 caaagatgga cccccaccca cgaggagcat cgtggaaaaa gaagacgttc caaccacgtc
     9601 ttcaaagcaa gtggattgat gtgaacatgg tggagcacga cactctcgtc tactccaaga
     9661 atatcaaaga tacagtctca gaagaccaaa gggctattga gacttttcaa caaagggtaa
     9721 tatcgggaaa cctcctcgga ttccattgcc cagctatctg tcacttcatc aaaaggacag
     9781 tagaaaagga aggtggcacc tacaaatgcc atcattgcga taaaggaaag gctatcgttc
     9841 aagatgcctc tgccgacagt ggtcccaaag atggaccccc acccacgagg agcatcgtgg
     9901 aaaaagaaga cgttccaacc acgtcttcaa agcaagtgga ttgatgtgat atctccactg
     9961 acgtaaggga tgacgcacaa tcccactatc cttcgcaaga cccttcctct atataaggaa
    10021 gttcatttca tttggagagg acacgctgaa atcaccagtc tctctctaca aatctatctc
    10081 tctcgagctt tcgcagatcc ggggggcaat gagatatgaa aaagcctgaa ctcaccgcga
    10141 cgtctgtcga gaagtttctg atcgaaaagt tcgacagcgt gtccgacctg atgcagctct
    10201 cggagggcga agaatctcgt gctttcagct tcgatgtagg agggcgtgga tatgtcctgc
    10261 gggtaaatag ctgcgccgat ggtttctaca aagatcgtta tgtttatcgg cactttgcat
    10321 cggccgcgct cccgattccg gaagtgcttg acattgggga attcagcgag agcctgacct
    10381 attgcatctc ccgccgtgca cagggtgtca cgttgcaaga cctgcctgaa accgaactgc
    10441 ccgctgttct gcagccggtc gcggaggcca tggatgcgat cgctgcggcc gatcttagcc
    10501 agacgagcgg gttcggccca ttcggaccgc aaggaatcgg tcaatacact acatggcgtg
    10561 atttcatatg cgcgattgct gatccccatg tgtatcactg gcaaactgtg atggacgaca
    10621 ccgtcagtgc gtccgtcgcg caggctctcg atgagctgat gctttgggcc gaggactgcc
    10681 ccgaagtccg gcacctcgtg cacgcggatt tcggctccaa caatgtcctg acggacaatg
    10741 gccgcataac agcggtcatt gactggagcg aggcgatgtt cggggattcc caatacgagg
    10801 tcgccaacat cttcttctgg aggccgtggt tggcttgtat ggagcagcag acgcgctact
    10861 tcgagcggag gcatccggag cttgcaggat cgccacgact ccgggcgtat atgctccgca
    10921 ttggtcttga ccaactctat cagagcttgg ttgacggcaa tttcgatgat gcagcttggg
    10981 cgcagggtcg atgcgacgca atcgtccgat ccggagccgg gactgtcggg cgtacacaaa
    11041 tcgcccgcag aagcgcggcc gtctggaccg atggctgtgt agaagtactc gccgatagtg
    11101 gaaaccgacg ccccagcact cgtccgaggg caaagaaata gagtagatgc cgaccgggat
    11161 ctgtcgatcg acaagctcga gtttctccat aataatgtgt gagtagttcc cagataaggg
    11221 aattagggtt cctatagggt ttcgctcatg tgttgagcat ataagaaacc cttagtatgt
    11281 atttgtattt gtaaaatact tctatcaata aaatttctaa ttcctaaaac caaaatccag
    11341 tactaaaatc cagatccccc gaattaattc ggcgttaatt cagtacatta aaaacgtccg
    11401 caatgtgtta ttaagttgtc taagcgtcaa tttgtttaca ccacaatata tcctgccacc
    11461 agccagccaa cagctccccg accggcagct cggcacaaaa tcaccactcg atacaggcag
    11521 cccatcagtc cgggacggcg tcagcgggag agccgttgta aggcggcaga ctttgctcat
    11581 gttaccgatg ctattcggaa gaacggcaac taagctgccg ggtttgaaac acggatgatc
    11641 tcgcggaggg tagcatgttg attgtaacga tgacagagcg ttgctgcctg tgatcaccgc
    11701 ggtttcaaaa tcggctccgt cgatactatg ttatacgcca actttgaaaa caactttgaa
    11761 aaagctgttt tctggtattt aaggttttag aatgcaagga acagtgaatt ggagttcgtc
    11821 ttgttataat tagcttcttg gggtatcttt aaatactgta gaaaagagga aggaaataat
    11881 aaatggctaa aatgagaata tcaccggaat tgaaaaaact gatcgaaaaa taccgctgcg
    11941 taaaagatac ggaaggaatg tctcctgcta aggtatataa gctggtggga gaaaatgaaa
    12001 acctatattt aaaaatgacg gacagccggt ataaagggac cacctatgat gtggaacggg
    12061 aaaaggacat gatgctatgg ctggaaggaa agctgcctgt tccaaaggtc ctgcactttg
    12121 aacggcatga tggctggagc aatctgctca tgagtgaggc cgatggcgtc ctttgctcgg
    12181 aagagtatga agatgaacaa agccctgaaa agattatcga gctgtatgcg gagtgcatca
    12241 ggctctttca ctccatcgac atatcggatt gtccctatac gaatagctta gacagccgct
    12301 tagccgaatt ggattactta ctgaataacg atctggccga tgtggattgc gaaaactggg
    12361 aagaagacac tccatttaaa gatccgcgcg agctgtatga ttttttaaag acggaaaagc
    12421 ccgaagagga acttgtcttt tcccacggcg acctgggaga cagcaacatc tttgtgaaag
    12481 atggcaaagt aagtggcttt attgatcttg ggagaagcgg cagggcggac aagtggtatg
    12541 acattgcctt ctgcgtccgg tcgatcaggg aggatatcgg ggaagaacag tatgtcgagc
    12601 tattttttga cttactgggg atcaagcctg attgggagaa aataaaatat tatattttac
    12661 tggatgaatt gttttagtac ctagaatgca tgaccaaaat cccttaacgt gagttttcgt
    12721 tccactgagc gctgatgaat cccctaatga tttttatcaa aatcattaag ttaaggtaga
    12781 tacacatctt gtcatatgat caaatggttt cgccaaaaat caataatcag acaacaaaat
    12841 gtgcgaactc gatattttac acgactctct ttaccaattc tgccccgaat tacacttaaa
    12901 acgactcaac agcttaacgt tggcttgcca cgccttactt gactgtaaaa ctctcactct
    12961 taccgaactt ggccgtaacc tgccaaccaa agcgagaaca aaacataaca tcaaacgaat
    13021 cgaccgattg ttaggtaatc gtcacctcca caaagagcga ctcgctgtat accgttggca
    13081 tgctagcttt atctgttcgg gcaatacgat gcccattgta cttgttgact ggtctgatat
    13141 ccgtgagcaa aaacggctta tggtattgcg agcttcagtc gcactacacg gtcgttctgt
    13201 tactctttat gagaaagcgt tcccgctttc agagcaatgt tcaaagaaag ctcatgacca
    13261 atttctagcc gaccttgcga gcattctacc gagtaacacc acaccgctca ttgtcagtga
    13321 tgctggcttt aaagtgccat ggtataaatc cgttgagaag ctgggttggt actggttaag
    13381 tcgagtaaga ggaaaagtac aatatgcaga cctaggagcg gaaaactgga aacctatcag
    13441 caacttacat gatatgtcat ctagtcactc aaagacttta ggctataaga ggctgactaa
    13501 aagcaatcca atctcatgcc aaattctatt gtataaatct cgctctaaag gccgaaaaaa
    13561 tcagcgctcg acacggactc attgtcacca cccgtcacct aaaatctact cagcgtcggc
    13621 aaaggagcca tggattctag caactaactt acctgttgaa attcgaacac ccaaacaact
    13681 tgttaatatc tattcgaagc gaatgcagat tgaagaaacc ttccgagact tgaaaagtcc
    13741 tgcctacgga ctaggcctac gccatagccg aacgagcagc tcagagcgtt ttgatatcat
    13801 gctgctaatc gccctgatgc ttcaactaac atgttggctt gcgggcgttc atgctcagaa
    13861 acaaggttgg gacaagcact tccaggctaa cacagtcaga aatcgaaacg tactctcaac
    13921 agttcgctta ggcatggaag ttttgcggca ttctggctac acaataacaa gggaagactt
    13981 actcgtggct gcaaccctac tagctcaaaa tttattcaca catggttacg ctttggggaa
    14041 attatgaggg gatctctcag cactgagcgt cagaccccgt agaaaagatc aaaggatctt
    14101 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
    14161 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
    14221 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
    14281 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
    14341 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
    14401 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
    14461 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
    14521 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
    14581 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
    14641 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
    14701 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
    14761 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
    14821 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagggcctga
    14881 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatatgg tgcactctca
    14941 gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat cgctacgtga
    15001 ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
    15061 tctgctcccg gcatccgctt acagacaagc tgtgaccgtt tccgggagct gcatgtgtca
    15121 gaggttttca ccgtcatcac cgaaacgcgc gaggcagggt gccttgatgt gggcgccggc
    15181 ggtcgagtgg cgacggcgcg gcttgtccgc gccctggtag attgcctggc cgtaggccag
    15241 ccatttttga gcggccagcg gccgcgatag gccgacgcga agcggcgggg cgtagggagc
    15301 gcagcgaccg aagggtaggc gctttttgca gctcttgggc tgtgcgctgg ccagacagtt
    15361 atgcacaggc caggcgggtt ttaagagttt taataagttt taaagagttt taggcggaaa
    15421 aatcgccttt tttctctttt atatcagtca cttacatgtg tgaccggttc ccaatgtacg
    15481 gctttgggtt cccaatgtac gggttccggt tcccaatgta cggctttggg ttcccaatgt
    15541 acgtgctatc cacaggaaag aggccttttc gacctttttc ccctgctagg gcaatttgcc
    15601 ctagcatctg ctccgtacat taggaaccgg cggatgcttc gccctcgatc aggttgcggt
    15661 agcgcatgac taggatcggg ccagcctgcc ccgcctcctc cttcaaatcg tactccggca
    15721 ggtcatttga cccgatcagc ttgcgcacgg tgaaacagaa cttcttgaac tctccggcgc
    15781 tgccactgcg ttcgtagatc gtcttgaaca accatctggc ttctgccttg cctgcggcgc
    15841 ggcgtgccag gcggtagaga aaacggccga tgccgggatc gatcaaaaag taatcggggt
    15901 gaaccgtcag cacgtccggg ttcttgcctt ctgtgatctc gcggtacatc caatcagcta
    15961 gctcgatctc gatgtactcc ggccgcccgg tttcgctctt tacgatcttg tagcggctaa
    16021 tcaaggcttc accctcggat accgtcacca ggcggccgtt cttggccttc ttcgtacgct
    16081 gcatggcaac gtgcgtggtg tttaaccgaa tgcaggtttc taccaggtcg tctttctgct
    16141 ttccgccatc ggctcgccgg cagaacttga gtacgtccgc aacgtgtgga cggaacacgc
    16201 ggccgggctt gtctcccttc ccttcccggt atcggttcat ggattcggtt agatgggaaa
    16261 ccgccatcag taccaggtcg taatcccaca cactggccat gccggccggc cctgcggaaa
    16321 cctctacgtg cccgtctgga agctcgtagc ggatcacctc gccagctcgt cggtcacgct
    16381 tcgacagacg gaaaacggcc acgtccatga tgctgcgact atcgcgggtg cccacgtcat
    16441 agagcatcgg aacgaaaaaa tctggttgct cgtcgccctt gggcggcttc ctaatcgacg
    16501 gcgcaccggc tgccggcggt tgccgggatt ctttgcggat tcgatcagcg gccgcttgcc
    16561 acgattcacc ggggcgtgct tctgcctcga tgcgttgccg ctgggcggcc tgcgcggcct
    16621 tcaacttctc caccaggtca tcacccagcg ccgcgccgat ttgtaccggg ccggatggtt
    16681 tgcgaccgct cacgccgatt cctcgggctt gggggttcca gtgccattgc agggccggca
    16741 ggcaacccag ccgcttacgc ctggccaacc gcccgttcct ccacacatgg ggcattccac
    16801 ggcgtcggtg cctggttgtt cttgattttc catgccgcct cctttagccg ctaaaattca
    16861 tctactcatt tattcatttg ctcatttact ctggtagctg cgcgatgtat tcagatagca
    16921 gctcggtaat ggtcttgcct tggcgtaccg cgtacatctt cagcttggtg tgatcctccg
    16981 ccggcaactg aaagttgacc cgcttcatgg ctggcgtgtc tgccaggctg gccaacgttg
    17041 cagccttgct gctgcgtgcg ctcggacggc cggcacttag cgtgtttgtg cttttgctca
    17101 ttttctcttt acctcattaa ctcaaatgag ttttgattta atttcagcgg ccagcgcctg
    17161 gacctcgcgg gcagcgtcgc cctcgggttc tgattcaaga acggttgtgc cggcggcggc
    17221 agtgcctggg tagctcacgc gctgcgtgat acgggactca agaatgggca gctcgtaccc
    17281 ggccagcgcc tcggcaacct caccgccgat gcgcgtgcct ttgatcgccc gcgacacgac
    17341 aaaggccgct tgtagccttc catccgtgac ctcaatgcgc tgcttaacca gctccaccag
    17401 gtcggcggtg gcccatatgt cgtaagggct tggctgcacc ggaatcagca cgaagtcggc
    17461 tgccttgatc gcggacacag ccaagtccgc cgcctggggc gctccgtcga tcactacgaa
    17521 gtcgcgccgg ccgatggcct tcacgtcgcg gtcaatcgtc gggcggtcga tgccgacaac
    17581 ggttagcggt tgatcttccc gcacggccgc ccaatcgcgg gcactgccct ggggatcgga
    17641 atcgactaac agaacatcgg ccccggcgag ttgcagggcg cgggctagat gggttgcgat
    17701 ggtcgtcttg cctgacccgc ctttctggtt aagtacagcg ataaccttca tgcgttcccc
    17761 ttgcgtattt gtttatttac tcatcgcatc atatacgcag cgaccgcatg acgcaagctg
    17821 ttttactcaa atacacatca cctttttaga cggcggcgct cggtttcttc agcggccaag
    17881 ctggccggcc aggccgccag cttggcatca gacaaaccgg ccaggatttc atgcagccgc
    17941 acggttgagc cgtgcgcggg cggctcgaac acgtacccgg ccgcgatcat ctccgcctcg
    18001 atctcttcgg taatgaaaaa cggttcgtcc tggccgtcct ggtgcggttt catgcttgtt
    18061 cctcttggcg ttcattctcg gcggccgcca gggcgtcggc ctcggtcaat gcgtcctcac
    18121 ggaaggcacc gcgccgcctg gcctcggtgg gcgtcacttc ctcgctgcgc tcaagtgcgc
    18181 ggtacagggt cgagcgatgc acgccaagca gtgcagccgc ctctttcacg gtgcggcctt
    18241 cctggtcgat cagctcgcgg gcgtgcgcga tctgtgccgg ggtgagggta gggcgggggc
    18301 caaacttcac gcctcgggcc ttggcggcct cgcgcccgct ccgggtgcgg tcgatgatta
    18361 gggaacgctc gaactcggca atgccggcga acacggtcaa caccatgcgg ccggccggcg
    18421 tggtggtgtc ggcccacggc tctgccaggc tacgcaggcc cgcgccggcc tcctggatgc
    18481 gctcggcaat gtccagtagg tcgcgggtgc tgcgggccag gcggtctagc ctggtcactg
    18541 tcacaacgtc gccagggcgt aggtggtcaa gcatcctggc cagctccggg cggtcgcgcc
    18601 tggtgccggt gatcttctcg gaaaacagct tggtgcagcc ggccgcgtgc agttcggccc
    18661 gttggttggt caagtcctgg tcgtcggtgc tgacgcgggc atagcccagc aggccagcgg
    18721 cggcgctctt gttcatggcg taatgtctcc ggttctagtc gcaagtattc tactttatgc
    18781 gactaaaaca cgcgacaaga aaacgccagg aaaagggcag ggcggcagcc tgtcgcgtaa
    18841 cttaggactt gtgcgacatg tcgttttcag aagacggctg cactgaacgt cagaagccga
    18901 ctgcactata gcagcggagg ggttggatca aagtactttg atcccgaggg gaaccctgtg
    18961 gttggcatgc acatacaaat ggacgaacgg ataaaccttt tcacgccctt ttaaatatcc
    19021 gattattcta ataaacgctc ttttctctta ggtttacccg ccaatatatc ctgtcaaaca
    19081 ctgatagttt aaactgaagg cgggaaacga caatctgatc caagctcaag ctgctctagc
    19141 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
    19201 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgg
//
