LOCUS       sequence-430353-       19585 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  FrCas9 genome editing plasmid in rice.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 19585)
  AUTHORS   He Y, Han Y, Ma Y, Liu S, Fan T, Liang Y, Tang X, Zheng X, Wu Y, 
            Zhang T, Qi Y, Zhang Y
  TITLE     Expanding plant genome editing scope and profiles with CRISPR-FrCas9
            systems targeting palindromic TA sites.
  JOURNAL   Plant Biotechnol J. 2024 Sep;22(9):2488-2503. doi: 
            10.1111/pbi.14363. Epub 2024 May 7.
  PUBMED    38713743
REFERENCE   2  (bases 1 to 19585)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1111/pbi.14363";
            journalName: "Plant Biotechnol J"; date: "2024-09"; volume: "22"; 
            issue: "9"; pages: "2488-2503"
COMMENT     Sequence Label: pGEL846
FEATURES             Location/Qualifiers
     source          1..19585
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     164..184
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     promoter        173..219
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     CDS             294..1001
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     707..726
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1023..2048
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     K"
     CDS             1023..2048
                     /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    2089..2263
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    2341..2365
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             2790..3584
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      5009..5597
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5498..5517
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5751..5768
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5783..5923
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5849..5868)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6009..6031
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     rep_origin      6267..6461
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(6527..7600)
                     /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(8029..8658)
                     /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    9959..9983
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     primer_bind     10186..10202
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    10773..10794
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(10809..10814,10815..10832,10833..10839)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10847..10863
                     /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     10871..10887
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             10883..11290
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGARLQGG
                     TRQCIARCMHCLHCGHAATFCNLHKLACVQL"
     primer_bind     10906..10926
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        10908..10926
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(10973..10992)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(10974..10992)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(11002..11018)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      complement(12936..13184)
                     /label=HSP terminator
                     /note="efficient transcription terminator from the 
                     Arabidopsis thaliana heat shock protein 18.2 gene (Nagaya 
                     et al., 2010)"
     CDS             complement(13203..13250)
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             complement(17388..17408)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      17411..17420
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(19459..19475)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    19483..19499
                     /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(19507..19513,19514..19531,19532..19537))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    19552..19573
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 aaggaaaggc tatcgttcaa gatgcctctg ccgacagtgg tcccaaagat ggacccccac
       61 ccacgaggag catcgtggaa aaagaagacg ttccaaccac gtcttcaaag caagtggatt
      121 gatgtgatat ctccactgac gtaagggatg acgcacaatc ccactatcct tcgcaagacc
      181 cttcctctat ataaggaagt tcatttcatt tggagaggac acgctgaaat caccagtctc
      241 tctctacaaa tctatctctc tcgagctttc gcagatccgg ggggcaatga gatatggtga
      301 gcaagggcga ggaggataac atggccatca tcaaggagtt catgcgcttc aaggtgcaca
      361 tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct
      421 acgagggcac ccagaccgcc aagctgaagg tgaccaaggg tggccccctg cccttcgcct
      481 gggacatcct gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag caccccgccg
      541 acatccccga ctacttgaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga
      601 acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag gacggcgagt
      661 tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc gtaatgcaga
      721 agaagaccat gggctgggag gcctcctccg agcggatgta ccccgaggac ggcgccctga
      781 agggcgagat caagcagagg ctgaagctga aggacggcgg ccactacgac gctgaggtca
      841 agaccaccta caaggccaag aagcccgtgc agctgcccgg cgcctacaac gtcaacatca
      901 agttggacat cacctcccac aacgaggact acaccatcgt ggaacagtac gaacgcgccg
      961 agggccgcca ctccaccggc ggcatggacg agctttacaa gtcgggcgga ggtggctctg
     1021 atatgaaaaa gcctgaactc accgcgacgt ctgtcgagaa gtttctgatc gaaaagttcg
     1081 acagcgtgtc cgacctgatg cagctctcgg agggcgaaga atctcgtgct ttcagcttcg
     1141 atgtaggagg gcgtggatat gtcctgcggg taaatagctg cgccgatggt ttctacaaag
     1201 atcgttatgt ttatcggcac tttgcatcgg ccgcgctccc gattccggaa gtgcttgaca
     1261 ttggggaatt cagcgagagc ctgacctatt gcatctcccg ccgtgcacag ggtgtcacgt
     1321 tgcaagacct gcctgaaacc gaactgcccg ctgttctgca gccggtcgcg gaggccatgg
     1381 atgcgatcgc tgcggccgat cttagccaga cgagcgggtt cggcccattc ggaccgcaag
     1441 gaatcggtca atacactaca tggcgtgatt tcatatgcgc gattgctgat ccccatgtgt
     1501 atcactggca aactgtgatg gacgacaccg tcagtgcgtc cgtcgcgcag gctctcgatg
     1561 agctgatgct ttgggccgag gactgccccg aagtccggca cctcgtgcac gcggatttcg
     1621 gctccaacaa tgtcctgacg gacaatggcc gcataacagc ggtcattgac tggagcgagg
     1681 cgatgttcgg ggattcccaa tacgaggtcg ccaacatctt cttctggagg ccgtggttgg
     1741 cttgtatgga gcagcagacg cgctacttcg agcggaggca tccggagctt gcaggatcgc
     1801 cacgactccg ggcgtatatg ctccgcattg gtcttgacca actctatcag agcttggttg
     1861 acggcaattt cgatgatgca gcttgggcgc agggtcgatg cgacgcaatc gtccgatccg
     1921 gagccgggac tgtcgggcgt acacaaatcg cccgcagaag cgcggccgtc tggaccgatg
     1981 gctgtgtaga agtactcgcc gatagtggaa accgacgccc cagcactcgt ccgagggcaa
     2041 agaaatagag tagatgccga ccgggatctg tcgatcgaca agctcgagtt tctccataat
     2101 aatgtgtgag tagttcccag ataagggaat tagggttcct atagggtttc gctcatgtgt
     2161 tgagcatata agaaaccctt agtatgtatt tgtatttgta aaatacttct atcaataaaa
     2221 tttctaattc ctaaaaccaa aatccagtac taaaatccag atcccccgaa ttaattcggc
     2281 gttaattcag tacattaaaa acgtccgcaa tgtgttatta agttgtctaa gcgtcaattt
     2341 gtttacacca caatatatcc tgccaccagc cagccaacag ctccccgacc ggcagctcgg
     2401 cacaaaatca ccactcgata caggcagccc atcagtccgg gacggcgtca gcgggagagc
     2461 cgttgtaagg cggcagactt tgctcatgtt accgatgcta ttcggaagaa cggcaactaa
     2521 gctgccgggt ttgaaacacg gatgatctcg cggagggtag catgttgatt gtaacgatga
     2581 cagagcgttg ctgcctgtga tcaccgcggt ttcaaaatcg gctccgtcga tactatgtta
     2641 tacgccaact ttgaaaacaa ctttgaaaaa gctgttttct ggtatttaag gttttagaat
     2701 gcaaggaaca gtgaattgga gttcgtcttg ttataattag cttcttgggg tatctttaaa
     2761 tactgtagaa aagaggaagg aaataataaa tggctaaaat gagaatatca ccggaattga
     2821 aaaaactgat cgaaaaatac cgctgcgtaa aagatacgga aggaatgtct cctgctaagg
     2881 tatataagct ggtgggagaa aatgaaaacc tatatttaaa aatgacggac agccggtata
     2941 aagggaccac ctatgatgtg gaacgggaaa aggacatgat gctatggctg gaaggaaagc
     3001 tgcctgttcc aaaggtcctg cactttgaac ggcatgatgg ctggagcaat ctgctcatga
     3061 gtgaggccga tggcgtcctt tgctcggaag agtatgaaga tgaacaaagc cctgaaaaga
     3121 ttatcgagct gtatgcggag tgcatcaggc tctttcactc catcgacata tcggattgtc
     3181 cctatacgaa tagcttagac agccgcttag ccgaattgga ttacttactg aataacgatc
     3241 tggccgatgt ggattgcgaa aactgggaag aagacactcc atttaaagat ccgcgcgagc
     3301 tgtatgattt tttaaagacg gaaaagcccg aagaggaact tgtcttttcc cacggcgacc
     3361 tgggagacag caacatcttt gtgaaagatg gcaaagtaag tggctttatt gatcttggga
     3421 gaagcggcag ggcggacaag tggtatgaca ttgccttctg cgtccggtcg atcagggagg
     3481 atatcgggga agaacagtat gtcgagctat tttttgactt actggggatc aagcctgatt
     3541 gggagaaaat aaaatattat attttactgg atgaattgtt ttagtaccta gaatgcatga
     3601 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgct gatgaatccc ctaatgattt
     3661 ttatcaaaat cattaagtta aggtagatac acatcttgtc atatgatcaa atggtttcgc
     3721 caaaaatcaa taatcagaca acaaaatgtg cgaactcgat attttacacg actctcttta
     3781 ccaattctgc cccgaattac acttaaaacg actcaacagc ttaacgttgg cttgccacgc
     3841 cttacttgac tgtaaaactc tcactcttac cgaacttggc cgtaacctgc caaccaaagc
     3901 gagaacaaaa cataacatca aacgaatcga ccgattgtta ggtaatcgtc acctccacaa
     3961 agagcgactc gctgtatacc gttggcatgc tagctttatc tgttcgggca atacgatgcc
     4021 cattgtactt gttgactggt ctgatatccg tgagcaaaaa cggcttatgg tattgcgagc
     4081 ttcagtcgca ctacacggtc gttctgttac tctttatgag aaagcgttcc cgctttcaga
     4141 gcaatgttca aagaaagctc atgaccaatt tctagccgac cttgcgagca ttctaccgag
     4201 taacaccaca ccgctcattg tcagtgatgc tggctttaaa gtgccatggt ataaatccgt
     4261 tgagaagctg ggttggtact ggttaagtcg agtaagagga aaagtacaat atgcagacct
     4321 aggagcggaa aactggaaac ctatcagcaa cttacatgat atgtcatcta gtcactcaaa
     4381 gactttaggc tataagaggc tgactaaaag caatccaatc tcatgccaaa ttctattgta
     4441 taaatctcgc tctaaaggcc gaaaaaatca gcgctcgaca cggactcatt gtcaccaccc
     4501 gtcacctaaa atctactcag cgtcggcaaa ggagccatgg attctagcaa ctaacttacc
     4561 tgttgaaatt cgaacaccca aacaacttgt taatatctat tcgaagcgaa tgcagattga
     4621 agaaaccttc cgagacttga aaagtcctgc ctacggacta ggcctacgcc atagccgaac
     4681 gagcagctca gagcgttttg atatcatgct gctaatcgcc ctgatgcttc aactaacatg
     4741 ttggcttgcg ggcgttcatg ctcagaaaca aggttgggac aagcacttcc aggctaacac
     4801 agtcagaaat cgaaacgtac tctcaacagt tcgcttaggc atggaagttt tgcggcattc
     4861 tggctacaca ataacaaggg aagacttact cgtggctgca accctactag ctcaaaattt
     4921 attcacacat ggttacgctt tggggaaatt atgaggggat ctctcagcac tgagcgtcag
     4981 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     5041 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     5101 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     5161 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     5221 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     5281 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     5341 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     5401 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     5461 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     5521 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     5581 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     5641 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     5701 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     5761 tgagcgagga agcggaagag ggcctgatgc ggtattttct ccttacgcat ctgtgcggta
     5821 tttcacaccg catatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     5881 agtatacact ccgctatcgc tacgtgactg ggtcatggct gcgccccgac acccgccaac
     5941 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     6001 gaccgtttcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     6061 gcagggtgcc ttgatgtggg cgccggcggt cgagtggcga cggcgcggct tgtccgcgcc
     6121 ctggtagatt gcctggccgt aggccagcca tttttgagcg gccagcggcc gcgataggcc
     6181 gacgcgaagc ggcggggcgt agggagcgca gcgaccgaag ggtaggcgct ttttgcagct
     6241 cttgggctgt gcgctggcca gacagttatg cacaggccag gcgggtttta agagttttaa
     6301 taagttttaa agagttttag gcggaaaaat cgcctttttt ctcttttata tcagtcactt
     6361 acatgtgtga ccggttccca atgtacggct ttgggttccc aatgtacggg ttccggttcc
     6421 caatgtacgg ctttgggttc ccaatgtacg tgctatccac aggaaagagg ccttttcgac
     6481 ctttttcccc tgctagggca atttgcccta gcatctgctc cgtacattag gaaccggcgg
     6541 atgcttcgcc ctcgatcagg ttgcggtagc gcatgactag gatcgggcca gcctgccccg
     6601 cctcctcctt caaatcgtac tccggcaggt catttgaccc gatcagcttg cgcacggtga
     6661 aacagaactt cttgaactct ccggcgctgc cactgcgttc gtagatcgtc ttgaacaacc
     6721 atctggcttc tgccttgcct gcggcgcggc gtgccaggcg gtagagaaaa cggccgatgc
     6781 cgggatcgat caaaaagtaa tcggggtgaa ccgtcagcac gtccgggttc ttgccttctg
     6841 tgatctcgcg gtacatccaa tcagctagct cgatctcgat gtactccggc cgcccggttt
     6901 cgctctttac gatcttgtag cggctaatca aggcttcacc ctcggatacc gtcaccaggc
     6961 ggccgttctt ggccttcttc gtacgctgca tggcaacgtg cgtggtgttt aaccgaatgc
     7021 aggtttctac caggtcgtct ttctgctttc cgccatcggc tcgccggcag aacttgagta
     7081 cgtccgcaac gtgtggacgg aacacgcggc cgggcttgtc tcccttccct tcccggtatc
     7141 ggttcatgga ttcggttaga tgggaaaccg ccatcagtac caggtcgtaa tcccacacac
     7201 tggccatgcc ggccggccct gcggaaacct ctacgtgccc gtctggaagc tcgtagcgga
     7261 tcacctcgcc agctcgtcgg tcacgcttcg acagacggaa aacggccacg tccatgatgc
     7321 tgcgactatc gcgggtgccc acgtcataga gcatcggaac gaaaaaatct ggttgctcgt
     7381 cgcccttggg cggcttccta atcgacggcg caccggctgc cggcggttgc cgggattctt
     7441 tgcggattcg atcagcggcc gcttgccacg attcaccggg gcgtgcttct gcctcgatgc
     7501 gttgccgctg ggcggcctgc gcggccttca acttctccac caggtcatca cccagcgccg
     7561 cgccgatttg taccgggccg gatggtttgc gaccgctcac gccgattcct cgggcttggg
     7621 ggttccagtg ccattgcagg gccggcaggc aacccagccg cttacgcctg gccaaccgcc
     7681 cgttcctcca cacatggggc attccacggc gtcggtgcct ggttgttctt gattttccat
     7741 gccgcctcct ttagccgcta aaattcatct actcatttat tcatttgctc atttactctg
     7801 gtagctgcgc gatgtattca gatagcagct cggtaatggt cttgccttgg cgtaccgcgt
     7861 acatcttcag cttggtgtga tcctccgccg gcaactgaaa gttgacccgc ttcatggctg
     7921 gcgtgtctgc caggctggcc aacgttgcag ccttgctgct gcgtgcgctc ggacggccgg
     7981 cacttagcgt gtttgtgctt ttgctcattt tctctttacc tcattaactc aaatgagttt
     8041 tgatttaatt tcagcggcca gcgcctggac ctcgcgggca gcgtcgccct cgggttctga
     8101 ttcaagaacg gttgtgccgg cggcggcagt gcctgggtag ctcacgcgct gcgtgatacg
     8161 ggactcaaga atgggcagct cgtacccggc cagcgcctcg gcaacctcac cgccgatgcg
     8221 cgtgcctttg atcgcccgcg acacgacaaa ggccgcttgt agccttccat ccgtgacctc
     8281 aatgcgctgc ttaaccagct ccaccaggtc ggcggtggcc catatgtcgt aagggcttgg
     8341 ctgcaccgga atcagcacga agtcggctgc cttgatcgcg gacacagcca agtccgccgc
     8401 ctggggcgct ccgtcgatca ctacgaagtc gcgccggccg atggccttca cgtcgcggtc
     8461 aatcgtcggg cggtcgatgc cgacaacggt tagcggttga tcttcccgca cggccgccca
     8521 atcgcgggca ctgccctggg gatcggaatc gactaacaga acatcggccc cggcgagttg
     8581 cagggcgcgg gctagatggg ttgcgatggt cgtcttgcct gacccgcctt tctggttaag
     8641 tacagcgata accttcatgc gttccccttg cgtatttgtt tatttactca tcgcatcata
     8701 tacgcagcga ccgcatgacg caagctgttt tactcaaata cacatcacct ttttagacgg
     8761 cggcgctcgg tttcttcagc ggccaagctg gccggccagg ccgccagctt ggcatcagac
     8821 aaaccggcca ggatttcatg cagccgcacg gttgagccgt gcgcgggcgg ctcgaacacg
     8881 tacccggccg cgatcatctc cgcctcgatc tcttcggtaa tgaaaaacgg ttcgtcctgg
     8941 ccgtcctggt gcggtttcat gcttgttcct cttggcgttc attctcggcg gccgccaggg
     9001 cgtcggcctc ggtcaatgcg tcctcacgga aggcaccgcg ccgcctggcc tcggtgggcg
     9061 tcacttcctc gctgcgctca agtgcgcggt acagggtcga gcgatgcacg ccaagcagtg
     9121 cagccgcctc tttcacggtg cggccttcct ggtcgatcag ctcgcgggcg tgcgcgatct
     9181 gtgccggggt gagggtaggg cgggggccaa acttcacgcc tcgggccttg gcggcctcgc
     9241 gcccgctccg ggtgcggtcg atgattaggg aacgctcgaa ctcggcaatg ccggcgaaca
     9301 cggtcaacac catgcggccg gccggcgtgg tggtgtcggc ccacggctct gccaggctac
     9361 gcaggcccgc gccggcctcc tggatgcgct cggcaatgtc cagtaggtcg cgggtgctgc
     9421 gggccaggcg gtctagcctg gtcactgtca caacgtcgcc agggcgtagg tggtcaagca
     9481 tcctggccag ctccgggcgg tcgcgcctgg tgccggtgat cttctcggaa aacagcttgg
     9541 tgcagccggc cgcgtgcagt tcggcccgtt ggttggtcaa gtcctggtcg tcggtgctga
     9601 cgcgggcata gcccagcagg ccagcggcgg cgctcttgtt catggcgtaa tgtctccggt
     9661 tctagtcgca agtattctac tttatgcgac taaaacacgc gacaagaaaa cgccaggaaa
     9721 agggcagggc ggcagcctgt cgcgtaactt aggacttgtg cgacatgtcg ttttcagaag
     9781 acggctgcac tgaacgtcag aagccgactg cactatagca gcggaggggt tggatcaaag
     9841 tactttgatc ccgaggggaa ccctgtggtt ggcatgcaca tacaaatgga cgaacggata
     9901 aaccttttca cgccctttta aatatccgat tattctaata aacgctcttt tctcttaggt
     9961 ttacccgcca atatatcctg tcaaacactg atagtttaaa ctgaaggcgg gaaacgacaa
    10021 tctgatccaa gctcaagctg ctctagcatt cgccattcag gctgcgcaac tgttgggaag
    10081 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
    10141 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
    10201 gtgccacact gagctcgata tcggcgcgcc ccgggagctc agcggcaaaa cacacctaga
    10261 ctagatttgt tttgctaacc caattgatat taattatata tgattaatat ttatatgtat
    10321 atggaattgg ttaataaaat gcatctggtt catcaaagaa ttataaagac acgtgacatt
    10381 catttaggat aagaaatatg gatgatctct ttctcttatt cagataatta gtaattacac
    10441 ataacacaca actttgatgc ccacattata gtgattagca tgtcactatg tgtgcatcct
    10501 tttatttcat acattaatta acttggccaa tccagaagat ggacaagtct aggctcgagt
    10561 acctgcacca gccgggaatc gaacccgggt ctgtaccgtg gcagggtact attctaccac
    10621 tagaccactg gtgctttgtt gggcacaatg cccgccatga aggtaagttc ggatctctct
    10681 aggagcctga tttgaactca tcttgttaat ttcttaacaa gacactcaaa ctgagaccac
    10741 cggtggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
    10801 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
    10861 caatttcaca caggaaacag ctatgaccat gattacgcca agcgcgcaat taaccctcac
    10921 taaagggaac aaaagctgga ggacctcgag ggggggcccg gtacccaatt cgccctatag
    10981 tgagtcgtat tacgcgcgct cactggccgt cgttttacaa cgtcgtgact gggaaaaccc
    11041 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag
    11101 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggga
    11161 cgcgccctgt agcggcgccc gtctccaggg gggcacccgt cagtgcattg cgcgatgcat
    11221 gcactgcctc cattgcggtc acgccgccac gttttgcaat cttcacaagt tggcctgtgt
    11281 ccaactgtaa ggtctcctgc accagccggg aatcgaaccc gggtctgtac cgtggcaggg
    11341 tactattcta ccactagacc actggtgctt tgttcctgca ggctgcacac acaattaccg
    11401 aagcatggtc agcaactatg aacatgaaat tataacaata ccaagcaagt acagcagaaa
    11461 tcagtcaatc aaattattcc aatatgttag atacacgatc taaaccacaa taacatcagt
    11521 gtacctatca acaccaacag tgacctaaca aaatagagag tcgcaccgtg ctcgagttca
    11581 caaaaataat ctgcaggtct atgaactact agaattccat cggcaaactg attcaataac
    11641 tactagcact atcccgagag cacataaggc aggttaatta acataggcta gaatttacag
    11701 gtgcaacagt tgataagata ggcataagca agtaaaatat cctgaaagct atgctatgta
    11761 atcactaatc aacaagacgc agcccttaga ttaaacatga acagagtaga atctatgcaa
    11821 ctagagggac tgaatcaaaa acaacctaag caaaatatac ctgaagctgc tacttggatc
    11881 aacgagaaaa acagagacta aatcctactg attgaacaga tcggaacaaa tcacaaccct
    11941 agcacagttc tacttctaga tccgaaccaa taaatacttg gaccacgaac agcatcgctc
    12001 ataatcacag atccagcata ccacgcgaac cccatctaac cctgttcaaa tctaccacag
    12061 atcgcaaaac ggagactaac aacaacatct cacaatcacg aacaggccgg gaaatcaaat
    12121 caaacgcgag gaacggagag gatcagaaaa tcgcatacct tgagaaggcg attcgacgct
    12181 tcgcttcgag gggagatttt tttttctctg cgattttggg gattcgattg atggaggaag
    12241 aggcgaggag ggggtggggt atttataggt ggggaggggg ggactcgtga gtgggcccag
    12301 gggtgcggtg ggtggggccc agtgagaata tagacaaata aagagaggga gaggcggttt
    12361 ggggagtccg tccccaccac gcaaacgcga gaaggaaaga aaggaacgga aagggggcgg
    12421 gtccagacgg cgccgcggag atatcgagga ggtggaaagc tgatgccgtt tgctcacggc
    12481 ggggttggtg gccgttaaca cgcagcgaag cggcgttact gccgtgtacg ttttgtgacc
    12541 tttcgaaaga gagccccagc ccaagaaagg ctggccaggt ttttttgtac tgtgtacgat
    12601 tacgacggaa gaaaacatac tcgttgtttt gtattggtag atactgggcc gtagtaaaat
    12661 tagaatcagt tgttaattgg ttattatata taccatctta agctaccaaa attaataaag
    12721 ttgtgatggc tatggtatat tttggtagtg aattatattg atttggtata tattagaatc
    12781 aatccaacca caacacaacc aaaaatacct gaagctcaat attgatggtg cttttcgact
    12841 atctagcaaa tgtggtgaat ggggctttat aattgaagca gcccaggatg cactttgtgc
    12901 agagactgat gtgctccggc gcgccgtccg agctccttat ctttaatcat attccatagt
    12961 ccataccata gcacatacag tagttatatg ctgcagaaga gatccaacaa aacattcaca
    13021 atggattata gaaacatttg tttattcatt ataatgtgat cttacattca tttaatatta
    13081 gaaaaagcca caaattcata acacaacaag ccaagaaaaa aacacaaact taagcacaca
    13141 agctttttat ttgacacacc aaatatttca tcttcatctt catatactcg agtacgcgtt
    13201 cacttctttt tcttagcctg tccggccttt ttggtggcag caggacgctt tatagtcagc
    13261 aggttctttc gagtctcata aattccgcta ggggactgac aaattaggta aaattctgca
    13321 tacttttcag ctcgtaggag cttggaaata ctcatgacgc cttcttgccc tgaacctccg
    13381 ataagcttaa ggtcagcaga agttttgcag tttgtgctgg tcaacttcaa tatctgtaaa
    13441 ataaccttgc attgattctc gatagtttgt ttcacaaatg cctctctgta cttcagcagc
    13501 ttgggtccga tggggttctt gcgtttcgaa aaaatagttg accggtgctt ttcagcgagt
    13561 tcgtcataga gccgaagatt tttcttgcga aaatcgtcct tggcctccgc agacaaagcg
    13621 tccgttgagc gccccccggc tatttttgaa atcatcctga tatgtgcctg ctccttggct
    13681 ggcaagcaga gctcaaccgc attagagaga gacagattgc cacccgtacg tccggtcagg
    13741 taatagtaat agccatcaat caatagaaga gagtacattg ggatgcgagt cagacggaca
    13801 tcaggctgca ccatgccgag cacttgtgca cagtagtgtt cgagttcctc cttggtcttt
    13861 aacttctctt taagatggat cgggacattc tctatgctcc ttattgtttt tttcttctgc
    13921 tgatgctcaa caagaaagaa gtggcttccc ttcacgttgt cgtagccgcc gtaggtggac
    13981 acctggtttg caaagcgtga gtctgaagtt ttcactggaa gatggacccc aggctttgcc
    14041 ttcttggcgg ctatcaggac ggaatctgcc aacccacctt tcctctcgta cgtccgtttt
    14101 gtcaccatgg gcgaggtctt aaaagctgtc ttcttcactg tttttaacgt cgtgtcattt
    14161 tcggcaaccc acgcggtctg atctccgcgc tggacattga aatcatacat ccttcttaaa
    14221 ctataggagt gcaaattacc agatttcctc atcttcttaa taaaattgat tggatttttg
    14281 gtaaatttgg tgtcataaac gttgccgaca acgatgttaa gataggcgtc tacggcgtgg
    14341 tggtaattgt tcaagtcccg cgactttgga atgtcgaacc gctgccggaa gtccgaggtg
    14401 tttccggcct tcacgtacac aacttctgag tcagggaaac actggttgat gagtgatata
    14461 attgcttttg ttccctgtct agtctccacc agttgtctgt tgacgaactc cgccagctcc
    14521 tcatcggtga acggagtcgt ccgcatgagt cggttgtatt tctcctgaga catgaaacct
    14581 tgcttcagga gcatggccca aaatcctttt tgtctcgatt gaatttgctc aggaatgggg
    14641 tagacgtctt gtttaatatg attattgaag tccttgcttg tgagtaccag attgttttct
    14701 aaggagtcat ctttcttctt gctttgcggg tggatgtggt ctatgtcgta ccgattgccg
    14761 tggagtacat ccatgaaatc aattagcttt ccactataaa tatccctccc catctgccgg
    14821 aaataaagat agagcttctt ggacctcaat tgggcctctt ccaggctctc gagatctcca
    14881 aacaattcat cgttcttcat gcctgcctcc ttgtagagat ctatcaactg tttcttccta
    14941 gacgcagtcc tcttcttctc ctgctccccc cgcgccatct ccatgaaaat cttcttaggt
    15001 gaataaccaa tggtatcgac cacttctctg aggatcaaaa atgtctgcca tatcatccgg
    15061 cgaacaggag cagacagata cattccatcg aggtcgtccg gctctattga ccagatgtcg
    15121 cgcgtcacgt agtcatttaa ctcctgcagc ttcttagcga atgctgagtc gttggagagg
    15181 agctgcatca agtttttttg agtatcacga agcgctctaa ttatggtaac aggatttcct
    15241 gcttcgtcct tgtacccctc catctcgagg aatgccctgc tcaggttccc ccactcggag
    15301 aacttgaagc cagcaatccg tcgaatttgt tgatctgtaa gttccggata gttatcctcg
    15361 agctttctct taagaaactt gcgcgaatcg ccatagacag tggaccattc aataatctgt
    15421 tcagccatca ccttctgtgt atcgttcagt gtcttaacgc caaaaagctc gcagaatttt
    15481 ttgtaactcg acagggacga tttaaagtcc ttgtcaatgc ctgttagaac aaactcctcc
    15541 gttgaagaat tggtgagcac actcctgagg aataactgca gtttttttcc agtgactttt
    15601 ttgccagaat agaagcactg ctcgaagatc atacgcttta ggcctggctg aatttcatct
    15661 gggtacatcc catttatcct gaggttgttg agctcattca ggacttcgaa ttcttgatac
    15721 agcaagctag attttgggag taccttctcg gtcctgaggt aagtacaact gttcaccaaa
    15781 ttcttgatga acagatcggc gctagcggac tcatcaatga tttcttcgaa gttccacggc
    15841 cgtaccggga tttgctcgtg cccgtccctc ctaacagccc aaccgtgccc ttgattgtcc
    15901 gtgttcttga gtgggccgat gtagtacggg attctaaagc tgaacagcgc ctcgattctg
    15961 tcaatggcag tcatatctct ttcatccttc tctgttagaa aggggtagtg cttggacgca
    16021 ttttgcagga tctttcgtag ttcttttagc tgtaactgat tcgggacaaa gccgttctga
    16081 ggcccagttt gcttaaggag gaaagtgccc tctattatct ccttctggat ctccgttttt
    16141 tcactgcact cagggatctt ttcgagcatt ttgttgatac tcttgtagaa gtcctcttgc
    16201 ttggctttgg ctgttctaca acgcttctca ccatcatact cggtgctgcc aatataatgc
    16261 gaatagttgg ccttctccat cttgcggaag aagttgtcgt attcttccgg gatatactcg
    16321 cggaacaatg ttttcagctt aacaagatca ctatgatgct tatcgtacgc cgccacgcgg
    16381 gcctcagcta gatacgagta ataatgaccg gtcaatgtat ctttcatgta gctgttgagg
    16441 aggctccatg aatatagcga gtagatagat tcgatatact caagcgcaag accgtcgatg
    16501 acgccctcta tatcgggaag aatctcctcc agggtctttt cgcggaaaga aaatgaatac
    16561 ttgtactcct ccgtatcttt atcttccatc ataatgagat cgcctaactt ggctttttga
    16621 cctgtgagaa ccttcacgaa ctgggaaacg gcctttttgg actctttgtt gggttgcaat
    16681 atctcaagaa tttgctgttc tttgtcacgc ctcgttattc ggctatccgt caagatatca
    16741 gttaagactt tttggtttga gacgagcggg atgctgattt gtatctcgtt catgcagtct
    16801 tcgagaaact gtatcagttc ttcgggacct gtattgtccg tatccacgtc ccccttgtcg
    16861 aggaaatggc ctggatgctg gaacaacgag tggattccca ggaatacaag gcgaatatca
    16921 aacctctcat tggattcgat taaggctttt ctcagatggt agatggtcgg gtactggttg
    16981 tagtattcta catctgtgta cccggaatca ttgaacaggg tgtacttctc tggcaccctc
    17041 ttgtcgctaa taacaaaccg agaatcctcc attcttttta aaaacccaga atcgatcttt
    17101 gtgatttcat tctgaaattc ctcactcaat agtcgattcc tgacctgctg cctctgtttc
    17161 cttcgacgag cgacccggaa cccccggcgt tccgccgccg ttttcgcttc ttcgaactgg
    17221 cgaacgcccc acaaatcctt gccttttctg cggattagat tgtactcgga gtcactcacc
    17281 gcccagccag ctgaggatgt gcccatgtcc agaccaatgt agtaatccgc gttcttattg
    17341 agcttttccg actccttggt gcaagccgca ggcaccccgt gaataccaac cttccgcttc
    17401 ttcttaggag ccatggcggc ctgcaggaag taacaccaaa caacagggtg agcatcgaca
    17461 aaagaaacag taccaagcaa ataaatagcg tatgaaggca gggctaaaaa aatccacata
    17521 tagctgctgc atatgccatc atccaagtat atcaagatca aaataattat aaaacatact
    17581 tgtttattat aatagatagg tactcaaggt tagagcatat gaatagatgc tgcatatgcc
    17641 atcatgtata tgcatcagta aaacccacat caacatgtat acctatccta gatcgatatt
    17701 tccatccatc ttaaactcgt aactatgaag atgtatgaca cacacataca gttccaaaat
    17761 taataaatac accaggtagt ttgaaacagt attctactcc gatctagaac gaatgaacga
    17821 ccgcccaacc acaccacatc atcacaacca agcgaacaaa aagcatctct gtatatgcat
    17881 cagtaaaacc cgcatcaaca tgtataccta tcctagatcg atatttccat ccatcatctt
    17941 caattcgtaa ctatgaatat gtatggcaca cacatacaga tccaaaatta ataaatccac
    18001 caggtagttt gaaacagaat taattctact ccgatctaga acgaccgccc aaccagacca
    18061 catcatcaca accaagacaa aaaaaagcat gaaaagatga cccgacaaac aagtgcacgg
    18121 catatattga aataaaggaa aagggcaaac caaaccctat gcaacgaaac aaaaaaaatc
    18181 atgaaatcga tcccgtctgc ggaacggcta gagccatccc aggattcccc aaagagaaac
    18241 actggcaagt tagcaatcag aacgtgtctg acgtacaggt cgcatccgtg tacgaacgct
    18301 agcagcacgg atctaacaca aacacggatc taacacaaac atgaacagaa gtagaactac
    18361 cgggccctaa ccatggaccg gaacgccgat ctagagaagg tagagagggg ggggggggag
    18421 gacgagcggc gtaccttgaa gcggaggtgc cgacgggtgg atttggggga gatctggttg
    18481 tgtgtgtgtg cgctccgaac aacacgaggt tggggaaaga gggtgtggag ggggtgtcta
    18541 tttattacgg cgggcgagga agggaaagcg aaggagcggt gggaaaggaa tcccccgtag
    18601 ctgccggtgc cgtgagagga ggaggaggcc gcctgccgtg ccggctcacg tctgccgctc
    18661 cgccacgcaa tttctggatg ccgacagcgg agcaagtcca acggtggagc ggaactctcg
    18721 agaggggtcc agaggcagcg acagagatgc cgtgccgtct gcttcgcttg gcccgacgcg
    18781 acgctgctgg ttcgctggtt ggtgtccgtt agactcgtcg acggcgttta acaggctggc
    18841 attatctact cgaaacaaga aaaatgtttc cttagttttt ttaatttctt aaagggtatt
    18901 tgtttaattt ttagtcactt tattttattc tattttatat ctaaattatt aaataaaaaa
    18961 actaaaatag agttttagtt ttcttaattt agaggctaaa atagaataaa atagatgtac
    19021 taaaaaaatt agtctataaa aaccattaac cctaaaccct aaatggatgt actaataaaa
    19081 tggatgaagt attatatagg tgaagctatt tgcaaaaaaa aaggagaaca catgcacact
    19141 aaaaagataa aactgtagag tcctgttgtc aaaatactca attgtccttt agaccatgtc
    19201 taactgttca tttatatgat tctctaaaac actgatatta ttgtagtact atagattata
    19261 ttattcgtag agtaaagttt aaatatatgt ataaagatag ataaactgca cttcaaacaa
    19321 gtgtgacaaa aaaaatatgt ggtaattttt tataacttag acatgcaatg ctcattatct
    19381 ctagagaggg gcacgaccgg gtcacgctgc actgcaggcg cgccgatccc cgggtaccga
    19441 gctcgaattc gtaatcatgt catagctgtt tcctgtgtga aattgttatc cgctcacaat
    19501 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
    19561 ctaactcaca ttaattgcgt tgcgc
//
