LOCUS       sequence-454834-       11643 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression plasmid with a ccdB cassette flanked with two inverted 
            BsaI restriction sites. For expressing syn-tasiRNAs in Nicotiana 
            benthamiana..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11643)
  AUTHORS   Cisneros AE, Alarcia A, Llorens-Gamez JJ, Puertes A, Juarez-Molina 
            M, Primc A, Carbonell A
  TITLE     Syn-tasiR-VIGS: virus-based targeted RNAi in plants by synthetic 
            trans-acting small interfering RNAs derived from minimal precursors.
  JOURNAL   Nucleic Acids Res. 2025 Feb 27;53(5):gkaf183. doi: 
            10.1093/nar/gkaf183.
  PUBMED    40105245
REFERENCE   2  (bases 1 to 11643)
  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: "Nucleic 
            Acids Res."; date: "2025-02-27"; volume: "53(5):gkaf183. doi"; 
            pages: " 10.1093/nar/gkaf183"
COMMENT     Sequence Label: pMDC32B-NbmiR482aTS-B/c
FEATURES             Location/Qualifiers
     source          1..11643
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             425..1219
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      1306..1894
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1795..1814
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2048..2065
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2080..2220
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2146..2165)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2306..2328
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     rep_origin      2564..2758
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(2824..3897)
                     /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(4326..4955)
                     /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    6256..6280
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     primer_bind     6468..6490
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6482..6499
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6483..6499
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6925..7269
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     protein_bind    7329..7353
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        join(7434..7439,7440..7457,7458..7464)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             7518..8177
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(7584..7603)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             8519..8824
                     /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     8703..8722
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(8896..8920)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      8984..9236
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(9253..9269)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9253..9269)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9266..9288)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9277..9293
                     /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(9301..9307,9308..9325,9326..9331))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9346..9367
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        9558..10234
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     CDS             10301..11326
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRSQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     K"
     polyA_signal    11367..11541
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    11619..11643
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
ORIGIN
        1 ccagccagcc aacagctccc cgaccggcag ctcggcacaa aatcaccact cgatacaggc
       61 agcccatcag tccgggacgg cgtcagcggg agagccgttg taaggcggca gactttgctc
      121 atgttaccga tgctattcgg aagaacggca actaagctgc cgggtttgaa acacggatga
      181 tctcgcggag ggtagcatgt tgattgtaac gatgacagag cgttgctgcc tgtgatcacc
      241 gcggtttcaa aatcggctcc gtcgatacta tgttatacgc caactttgaa aacaactttg
      301 aaaaagctgt tttctggtat ttaaggtttt agaatgcaag gaacagtgaa ttggagttcg
      361 tcttgttata attagcttct tggggtatct ttaaatactg tagaaaagag gaaggaaata
      421 ataaatggct aaaatgagaa tatcaccgga attgaaaaaa ctgatcgaaa aataccgctg
      481 cgtaaaagat acggaaggaa tgtctcctgc taaggtatat aagctggtgg gagaaaatga
      541 aaacctatat ttaaaaatga cggacagccg gtataaaggg accacctatg atgtggaacg
      601 ggaaaaggac atgatgctat ggctggaagg aaagctgcct gttccaaagg tcctgcactt
      661 tgaacggcat gatggctgga gcaatctgct catgagtgag gccgatggcg tcctttgctc
      721 ggaagagtat gaagatgaac aaagccctga aaagattatc gagctgtatg cggagtgcat
      781 caggctcttt cactccatcg acatatcgga ttgtccctat acgaatagct tagacagccg
      841 cttagccgaa ttggattact tactgaataa cgatctggcc gatgtggatt gcgaaaactg
      901 ggaagaagac actccattta aagatccgcg cgagctgtat gattttttaa agacggaaaa
      961 gcccgaagag gaacttgtct tttcccacgg cgacctggga gacagcaaca tctttgtgaa
     1021 agatggcaaa gtaagtggct ttattgatct tgggagaagc ggcagggcgg acaagtggta
     1081 tgacattgcc ttctgcgtcc ggtcgatcag ggaggatatc ggggaagaac agtatgtcga
     1141 gctatttttt gacttactgg ggatcaagcc tgattgggag aaaataaaat attatatttt
     1201 actggatgaa ttgttttagt acctagaatg catgaccaaa atcccttaac gtgagttttc
     1261 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     1321 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     1381 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     1441 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     1501 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     1561 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     1621 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     1681 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     1741 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     1801 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     1861 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     1921 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     1981 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     2041 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt attttctcct
     2101 tacgcatctg tgcggtattt cacaccgcat atggtgcact ctcagtacaa tctgctctga
     2161 tgccgcatag ttaagccagt atacactccg ctatcgctac gtgactgggt catggctgcg
     2221 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     2281 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     2341 tcaccgaaac gcgcgaggca gggtgccttg atgtgggcgc cggcggtcga gtggcgacgg
     2401 cgcggcttgt ccgcgccctg gtagattgcc tggccgtagg ccagccattt ttgagcggcc
     2461 agcggccgcg ataggccgac gcgaagcggc ggggcgtagg gagcgcagcg accgaagggt
     2521 aggcgctttt tgcagctctt cggctgtgcg ctggccagac agttatgcac aggccaggcg
     2581 ggttttaaga gttttaataa gttttaaaga gttttaggcg gaaaaatcgc cttttttctc
     2641 ttttatatca gtcacttaca tgtgtgaccg gttcccaatg tacggctttg ggttcccaat
     2701 gtacgggttc cggttcccaa tgtacggctt tgggttccca atgtacgtgc tatccacagg
     2761 aaagagaact tttcgacctt tttcccctgc tagggcaatt tgccctagca tctgctccgt
     2821 acattaggaa ccggcggatg cttcgccctc gatcaggttg cggtagcgca tgactaggat
     2881 cgggccagcc tgccccgcct cctccttcaa atcgtactcc ggcaggtcat ttgacccgat
     2941 cagcttgcgc acggtgaaac agaacttctt gaactctccg gcgctgccac tgcgttcgta
     3001 gatcgtcttg aacaaccatc tggcttctgc cttgcctgcg gcgcggcgtg ccaggcggta
     3061 gagaaaacgg ccgatgccgg gatcgatcaa aaagtaatcg gggtgaaccg tcagcacgtc
     3121 cgggttcttg ccttctgtga tctcgcggta catccaatca gctagctcga tctcgatgta
     3181 ctccggccgc ccggtttcgc tctttacgat cttgtagcgg ctaatcaagg cttcaccctc
     3241 ggataccgtc accaggcggc cgttcttggc cttcttcgta cgctgcatgg caacgtgcgt
     3301 ggtgtttaac cgaatgcagg tttctaccag gtcgtctttc tgctttccgc catcggctcg
     3361 ccggcagaac ttgagtacgt ccgcaacgtg tggacggaac acgcggccgg gcttgtctcc
     3421 cttcccttcc cggtatcggt tcatggattc ggttagatgg gaaaccgcca tcagtaccag
     3481 gtcgtaatcc cacacactgg ccatgccggc cggccctgcg gaaacctcta cgtgcccgtc
     3541 tggaagctcg tagcggatca cctcgccagc tcgtcggtca cgcttcgaca gacggaaaac
     3601 ggccacgtcc atgatgctgc gactatcgcg ggtgcccacg tcatagagca tcggaacgaa
     3661 aaaatctggt tgctcgtcgc ccttgggcgg cttcctaatc gacggcgcac cggctgccgg
     3721 cggttgccgg gattctttgc ggattcgatc agcggccgct tgccacgatt caccggggcg
     3781 tgcttctgcc tcgatgcgtt gccgctgggc ggcctgcgcg gccttcaact tctccaccag
     3841 gtcatcaccc agcgccgcgc cgatttgtac cgggccggat ggtttgcgac cgctcacgcc
     3901 gattcctcgg gcttgggggt tccagtgcca ttgcagggcc ggcaggcaac ccagccgctt
     3961 acgcctggcc aaccgcccgt tcctccacac atggggcatt ccacggcgtc ggtgcctggt
     4021 tgttcttgat tttccatgcc gcctccttta gccgctaaaa ttcatctact catttattca
     4081 tttgctcatt tactctggta gctgcgcgat gtattcagat agcagctcgg taatggtctt
     4141 gccttggcgt accgcgtaca tcttcagctt ggtgtgatcc tccgccggca actgaaagtt
     4201 gacccgcttc atggctggcg tgtctgccag gctggccaac gttgcagcct tgctgctgcg
     4261 tgcgctcgga cggccggcac ttagcgtgtt tgtgcttttg ctcattttct ctttacctca
     4321 ttaactcaaa tgagttttga tttaatttca gcggccagcg cctggacctc gcgggcagcg
     4381 tcgccctcgg gttctgattc aagaacggtt gtgccggcgg cggcagtgcc tgggtagctc
     4441 acgcgctgcg tgatacggga ctcaagaatg ggcagctcgt acccggccag cgcctcggca
     4501 acctcaccgc cgatgcgcgt gcctttgatc gcccgcgaca cgacaaaggc cgcttgtagc
     4561 cttccatccg tgacctcaat gcgctgctta accagctcca ccaggtcggc ggtggcccat
     4621 atgtcgtaag ggcttggctg caccggaatc agcacgaagt cggctgcctt gatcgcggac
     4681 acagccaagt ccgccgcctg gggcgctccg tcgatcacta cgaagtcgcg ccggccgatg
     4741 gccttcacgt cgcggtcaat cgtcgggcgg tcgatgccga caacggttag cggttgatct
     4801 tcccgcacgg ccgcccaatc gcgggcactg ccctggggat cggaatcgac taacagaaca
     4861 tcggccccgg cgagttgcag ggcgcgggct agatgggttg cgatggtcgt cttgcctgac
     4921 ccgcctttct ggttaagtac agcgataacc ttcatgcgtt ccccttgcgt atttgtttat
     4981 ttactcatcg catcatatac gcagcgaccg catgacgcaa gctgttttac tcaaatacac
     5041 atcacctttt tagacggcgg cgctcggttt cttcagcggc caagctggcc ggccaggccg
     5101 ccagcttggc atcagacaaa ccggccagga tttcatgcag ccgcacggtt gagacgtgcg
     5161 cgggcggctc gaacacgtac ccggccgcga tcatctccgc ctcgatctct tcggtaatga
     5221 aaaacggttc gtcctggccg tcctggtgcg gtttcatgct tgttcctctt ggcgttcatt
     5281 ctcggcggcc gccagggcgt cggcctcggt caatgcgtcc tcacggaagg caccgcgccg
     5341 cctggcctcg gtgggcgtca cttcctcgct gcgctcaagt gcgcggtaca gggtcgagcg
     5401 atgcacgcca agcagtgcag ccgcctcttt cacggtgcgg ccttcctggt cgatcagctc
     5461 gcgggcgtgc gcgatctgtg ccggggtgag ggtagggcgg gggccaaact tcacgcctcg
     5521 ggccttggcg gcctcgcgcc cgctccgggt gcggtcgatg attagggaac gctcgaactc
     5581 ggcaatgccg gcgaacacgg tcaacaccat gcggccggcc ggcgtggtgg tgtcggccca
     5641 cggctctgcc aggctacgca ggcccgcgcc ggcctcctgg atgcgctcgg caatgtccag
     5701 taggtcgcgg gtgctgcggg ccaggcggtc tagcctggtc actgtcacaa cgtcgccagg
     5761 gcgtaggtgg tcaagcatcc tggccagctc cgggcggtcg cgcctggtgc cggtgatctt
     5821 ctcggaaaac agcttggtgc agccggccgc gtgcagttcg gcccgttggt tggtcaagtc
     5881 ctggtcgtcg gtgctgacgc gggcatagcc cagcaggcca gcggcggcgc tcttgttcat
     5941 ggcgtaatgt ctccggttct agtcgcaagt attctacttt atgcgactaa aacacgcgac
     6001 aagaaaacgc caggaaaagg gcagggcggc agcctgtcgc gtaacttagg acttgtgcga
     6061 catgtcgttt tcagaagacg gctgcactga acgtcagaag ccgactgcac tatagcagcg
     6121 gaggggttgg atcaaagtac tttgatcccg aggggaaccc tgtggttggc atgcacatac
     6181 aaatggacga acggataaac cttttcacgc ccttttaaat atccgattat tctaataaac
     6241 gctcttttct cttaggttta cccgccaata tatcctgtca aacactgata gtttaaactg
     6301 aaggcgggaa acgacaatct gatccaagct caagctgctc tagcattcgc cattcaggct
     6361 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
     6421 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
     6481 ttgtaaaacg acggccagtg ccaagcttgc atgcctgcag gtcaacatgg tggagcacga
     6541 cacacttgtc tactccaaaa atatcaaaga tacagtctca gaagaccaaa gggcaattga
     6601 gacttttcaa caaagggtaa tatccggaaa cctcctcgga ttccattgcc cagctatctg
     6661 tcactttatt gtgaagatag tggaaaagga aggtggctcc tacaaatgcc atcattgcga
     6721 taaaggaaag gccatcgttg aagatgcctc tgccgacagt ggtcccaaag atggaccccc
     6781 acccacgagg agcatcgtgg aaaaagaaga cgttccaacc acgtcttcaa agcaagtgga
     6841 ttgatgtgat aacatggtgg agcacgacac acttgtctac tccaaaaata tcaaagatac
     6901 agtctcagaa gaccaaaggg caattgagac ttttcaacaa agggtaatat ccggaaacct
     6961 cctcggattc cattgcccag ctatctgtca ctttattgtg aagatagtgg aaaaggaagg
     7021 tggctcctac aaatgccatc attgcgataa aggaaaggcc atcgttgaag atgcctctgc
     7081 cgacagtggt cccaaagatg gacccccacc cacgaggagc atcgtggaaa aagaagacgt
     7141 tccaaccacg tcttcaaagc aagtggattg atgtgatatc tccactgacg taagggatga
     7201 cgcacaatcc cactatcctt cgcaagaccc ttcctctata taaggaagtt catttcattt
     7261 ggagaggacc tcgactctag aggatccccg ggtaccgggc cccccctcga ggcgcgccaa
     7321 gctatcaaac aagtttgtac aaaaaagcag gctccgcggc cgcccccttc acctgtagtg
     7381 gtatgggggg agtcgggaat agaccattta agagaccatt aggcacccca ggctttacac
     7441 tttatgcttc cggctcgtat aatgtgtgga ttttgagtta ggagccgtcg agattttcag
     7501 gagctaagga agctaaaatg gagaaaaaaa tcactggata taccaccgtt gatatatccc
     7561 aatggcatcg taaagaacat tttgaggcat ttcagtcagt tgctcaatgt acctataacc
     7621 agaccgttca gctggatatt acggcctttt taaagaccgt aaagaaaaat aagcacaagt
     7681 tttatccggc ctttattcac attcttgccc gcctgatgaa tgctcatccg gagttccgta
     7741 tggcaatgaa agacggtgag ctggtgatat gggatagtgt tcacccttgt tacaccgttt
     7801 tccatgagca aactgaaacg ttttcatcgc tctggagtga ataccacgac gatttccggc
     7861 agtttctaca catatattcg caagatgtgg cgtgttacgg tgaaaacctg gcctatttcc
     7921 ctaaagggtt tattgagaat atgtttttcg tctcagccaa tccctgggtg agtttcacca
     7981 gttttgattt aaacgtggcc aatatggaca acttcttcgc ccccgttttc accatgggca
     8041 aatattatac gcaaggcgac aaggtgctga tgccgctggc gattcaggtt catcatgccg
     8101 tttgtgatgg cttccatgtc ggcagaatgc ttaatgaatt acaacagtac tgcgatgagt
     8161 ggcagggcgg ggcgtaaacg cgtggagccg gcttactaaa agccagataa cagtatgcgt
     8221 atttgcgcgc tgatttttgc ggtataagaa tatatactga tatgtatacc cgaagtatgt
     8281 caaaaagagg tatgctatga agcagcgtat tacagtgaca gttgacagcg acagctatca
     8341 gttgctcaag gcatatatga tgtcaatatc tccggtctgg taagcacaac catgcagaat
     8401 gaagcccgtc gtctgcgtgc cgaacgctgg aaagcggaaa atcaggaagg gatggctgag
     8461 gtcgcccggt ttattgaaat gaacggctct tttgctgacg agaacagggg ctggtgaaat
     8521 gcagtttaag gtttacacct ataaaagaga gagccgttat cgtctgtttg tggatgtaca
     8581 gagtgatatt attgacacgc ccggccgacg gatggtgatc cccctggcca gtgcacgtct
     8641 gctgtcagat aaagtctccc gtgaacttta cccggtggtg catatcgggg atgaaagctg
     8701 gcgcatgatg accaccgata tggccagtgt gccggtttcc gttatcgggg aagaagtggc
     8761 tgatctcagc caccgcgaaa atgacatcaa aaacgccatt aacctgatgt tctggggaat
     8821 ataaatgtca ggctccctta tacacagcca gtctgcacct cgacggtctc atcggcatta
     8881 agggtgggcg cgccgaccca gctttcttgt acaaagtggt tcgataattc ttaattaact
     8941 agttctagag cggccgccac cgcggtggag ctcgaatttc cccgatcgtt caaacatttg
     9001 gcaataaagt ttcttaagat tgaatcctgt tgccggtctt gcgatgatta tcatataatt
     9061 tctgttgaat tacgttaagc atgtaataat taacatgtaa tgcatgacgt tatttatgag
     9121 atgggttttt atgattagag tcccgcaatt atacatttaa tacgcgatag aaaacaaaat
     9181 atagcgcgca aactaggata aattatcgcg cgcggtgtca tctatgttac tagatcggga
     9241 attcgtaatc atgtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     9301 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     9361 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     9421 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattggct agagcagctt
     9481 gccaacatgg tggagcacga cactctcgtc tactccaaga atatcaaaga tacagtctca
     9541 gaagaccaaa gggctattga gacttttcaa caaagggtaa tatcgggaaa cctcctcgga
     9601 ttccattgcc cagctatctg tcacttcatc aaaaggacag tagaaaagga aggtggcacc
     9661 tacaaatgcc atcattgcga taaaggaaag gctatcgttc aagatgcctc tgccgacagt
     9721 ggtcccaaag atggaccccc acccacgagg agcatcgtgg aaaaagaaga cgttccaacc
     9781 acgtcttcaa agcaagtgga ttgatgtgaa catggtggag cacgacactc tcgtctactc
     9841 caagaatatc aaagatacag tctcagaaga ccaaagggct attgagactt ttcaacaaag
     9901 ggtaatatcg ggaaacctcc tcggattcca ttgcccagct atctgtcact tcatcaaaag
     9961 gacagtagaa aaggaaggtg gcacctacaa atgccatcat tgcgataaag gaaaggctat
    10021 cgttcaagat gcctctgccg acagtggtcc caaagatgga cccccaccca cgaggagcat
    10081 cgtggaaaaa gaagacgttc caaccacgtc ttcaaagcaa gtggattgat gtgatatctc
    10141 cactgacgta agggatgacg cacaatccca ctatccttcg caagaccctt cctctatata
    10201 aggaagttca tttcatttgg agaggacacg ctgaaatcac cagtctctct ctacaaatct
    10261 atctctctcg agctttcgca gatccggggg gcaatgagat atgaaaaagc ctgaactcac
    10321 cgcgacgtct gtcgagaagt ttctgatcga aaagttcgac agcgtctccg acctgatgca
    10381 gctctcggag ggcgaagaat ctcgtgcttt cagcttcgat gtaggagggc gtggatatgt
    10441 cctgcgggta aatagctgcg ccgatggttt ctacaaagat cgttatgttt atcggcactt
    10501 tgcatcggcc gcgctcccga ttccggaagt gcttgacatt ggggagttta gcgagagcct
    10561 gacctattgc atctcccgcc gttcacaggg tgtcacgttg caagacctgc ctgaaaccga
    10621 actgcccgct gttctacaac cggtcgcgga ggctatggat gcgatcgctg cggccgatct
    10681 tagccagacg agcgggttcg gcccattcgg accgcaagga atcggtcaat acactacatg
    10741 gcgtgatttc atatgcgcga ttgctgatcc ccatgtgtat cactggcaaa ctgtgatgga
    10801 cgacaccgtc agtgcgtccg tcgcgcaggc tctcgatgag ctgatgcttt gggccgagga
    10861 ctgccccgaa gtccggcacc tcgtgcacgc ggatttcggc tccaacaatg tcctgacgga
    10921 caatggccgc ataacagcgg tcattgactg gagcgaggcg atgttcgggg attcccaata
    10981 cgaggtcgcc aacatcttct tctggaggcc gtggttggct tgtatggagc agcagacgcg
    11041 ctacttcgag cggaggcatc cggagcttgc aggatcgcca cgactccggg cgtatatgct
    11101 ccgcattggt cttgaccaac tctatcagag cttggttgac ggcaatttcg atgatgcagc
    11161 ttgggcgcag ggtcgatgcg acgcaatcgt ccgatccgga gccgggactg tcgggcgtac
    11221 acaaatcgcc cgcagaagcg cggccgtctg gaccgatggc tgtgtagaag tactcgccga
    11281 tagtggaaac cgacgcccca gcactcgtcc gagggcaaag aaatagagta gatgccgacc
    11341 gggatctgtc gatcgacaag ctcgagtttc tccataataa tgtgtgagta gttcccagat
    11401 aagggaatta gggttcctat agggtttcgc tcatgtgttg agcatataag aaacccttag
    11461 tatgtatttg tatttgtaaa atacttctat caataaaatt tctaattcct aaaaccaaaa
    11521 tccagtacta aaatccagat cccccgaatt aattcggcgt taattcagta cattaaaaac
    11581 gtccgcaatg tgttattaag ttgtctaagc gtcaatttgt ttacaccaca atatatcctg
    11641 cca
//
