LOCUS       sequence-404904-       17611 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For cytosine base editing using miniSdd7 at PPO2 site in soybean 
            (Agrobacterium-mediated transformation).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 17611)
  AUTHORS   Huang J, Lin Q, Fei H, He Z, Xu H, Li Y, Qu K, Han P, Gao Q, Li B, 
            Liu G, Zhang L, Hu J, Zhang R, Zuo E, Luo Y, Ran Y, Qiu JL, Zhao KT,
            Gao C
  TITLE     Discovery of deaminase functions by structure-based protein 
            clustering.
  JOURNAL   Cell. 2023 Jul 20;186(15):3182-3195.e14. doi: 
            10.1016/j.cell.2023.05.041. Epub 2023 Jun 27.
  PUBMED    37379837
REFERENCE   2  (bases 1 to 17611)
  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: 
            "10.1016/j.cell.2023.05.041"; journalName: "Cell"; date: 
            "2023-07-20- 20"; volume: "186"; issue: "15"; pages: "3182-3195.e14"
COMMENT     Sequence Label: pE-35S-miniSdd7-nCas9-PBE-GmPPO2
FEATURES             Location/Qualifiers
     source          1..17611
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..25
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     primer_bind     213..235
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     227..244
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     228..244
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_RNA        699..774
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        1388..1732
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     CDS             1886..1906
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(2432..2455,2456..2458,2459..6532)
                     /codon_start=1
                     /product="nickase mutant of the Cas9 endonuclease from the 
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(D10A)
                     /note="generates RNA-guided single strand nicks in DNA due 
                     to the D10A mutation in the RuvC catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     RBS             6445..6453
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             6533..6580
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6602..6850
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             6863..6883
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6884..7132
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             7145..7165
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7175..7231
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             7253..7948
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDLLSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     7654..7673
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     promoter        8693..9369
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     misc_feature    11332..11356
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             11781..12575
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      12662..13250
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     13151..13170
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     13404..13421
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    13436..13576
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(13502..13521)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     13662..13684
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     rep_origin      13920..14114
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(14180..15253)
                     /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(15682..16311)
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
ORIGIN
        1 gtttacccgc caatatatcc tgtcaaacac tgatagttta aactgaaggc gggaaacgac
       61 aatctgatcc aagctcaagc tgctctagca ttcgccattc aggctgcgca actgttggga
      121 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc
      181 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc
      241 cagtgccaag cttcgacttg ccttccgcac aatacatcat ttcttcttag ctttttttct
      301 tcttcttcgt tcatacagtt tttttttgtt tatcagctta cattttcttg aaccgtagct
      361 ttcgttttct tctttttaac tttccattcg gagtttttgt atcttgtttc atagtttgtc
      421 ccaggattag aatgattagg catcgaacct tcaagaattt gattgaataa aacatcttca
      481 ttcttaagat atgaagataa tcttcaaaag gcccctggga atctgaaaga agagaagcag
      541 gcccatttat atgggaaaga acaatagtat ttcttatata ggcccattta agttgaaaac
      601 aatcttcaaa agtcccacat cgcttagata agaaaacgaa gctgagttta tatacagcta
      661 gagtcgaagt agtgattgaa gcgctatatt gtgaaaaagt tttagagcta gaaatagcaa
      721 gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg gtgctttttt
      781 ttgcaaaatt ttccagatcg atttcttctt cctctgttct tcggcgttca atttctgggg
      841 ttttctcttc gttttctgta actgaaacct aaaatttgac ctaaaaaaaa tctcaaataa
      901 tatgattcag tggttttgta cttttcagtt agttgagttt tgcagttccg atgagataaa
      961 ccaataagct tgcatgcctg caggtcaaca tggtggagca cgacacactt gtctactcca
     1021 aaaatatcaa agatacagtc tcagaagacc aaagggcaat tgagactttt caacaaaggg
     1081 taatatccgg aaacctcctc ggattccatt gcccagctat ctgtcacttt attgtgaaga
     1141 tagtggaaaa ggaaggtggc tcctacaaat gccatcattg cgataaagga aaggccatcg
     1201 ttgaagatgc ctctgccgac agtggtccca aagatggacc cccacccacg aggagcatcg
     1261 tggaaaaaga agacgttcca accacgtctt caaagcaagt ggattgatgt gataacatgg
     1321 tggagcacga cacacttgtc tactccaaaa atatcaaaga tacagtctca gaagaccaaa
     1381 gggcaattga gacttttcaa caaagggtaa tattcggaaa cctcctcgga ttccattgcc
     1441 cagctatctg tcactttatt gtgaagatag tggaaaagga aggtggctcc tacaaatgcc
     1501 atcattgcga taaaggaaag gccatcgttg aagatgcctc tgccgacagt ggtcccaaag
     1561 atggaccccc acccacgagg agcatcgtgg aaaaagaaga cgttccaacc acgtcttcaa
     1621 agcaagtgga ttgatgtgat atctccactg acgtaaggga tgacgcacaa tcccactatc
     1681 cttcgcaaga cccttcctct atataaggaa gttcatttca tttggagagg acctcgacct
     1741 caacacaaca tatacaaaac aaacgaatct caagcaatca agcattctac ttctattgca
     1801 gcaatttaaa tcatttcttt taaagcaaaa gcaattttct gaaaattttc accatttacg
     1861 aacgatactc gagtaatcta gaatgccaaa gaagaagagg aaggttgagg gaggagggcc
     1921 tggggcggtt cctgaaggag gcgacggccc gccagccgtg ccggcggagg aggtggagag
     1981 gttgagagga gaacttcctc cacctgttgt cccgggcact ggtcaaaaaa cgcatgggcg
     2041 ctggattggt ccagatggtc gcgtccgcgc cattgtttca ggaagggatg aagatgctgc
     2101 actggtgcat gcacagctcg ccgccaaggg gataccagac gagccaacaa gaaattctga
     2161 tgttgagcag aagctggccg cgcacatggt ggcaaatggc atccgccacg tcacgctcgt
     2221 catcaaccat agaccctgcc gcggcttcga cgacagctgt gatacacttg tgcccataat
     2281 tctgccggag ggctgcaccc tcacggtgca cgggcaaact gacaagggca tgagggtgcg
     2341 ggtaaggtac accggcggcg cgcggccgtg gtggtccaag tccggcagcg agacgccagg
     2401 cacctccgag agcgctacgc ctgaaaggcc tgacaagaag tactcgatcg gcctcgccat
     2461 tgggactaac tctgttggct gggccgtgat caccgacgag tacaaggtgc cctcaaagaa
     2521 gttcaaggtc ctgggcaaca ccgatcggca ttccatcaag aagaatctca ttggcgctct
     2581 cctgttcgac agcggcgaga cggctgaggc tacgcggctc aagcgcaccg cccgcaggcg
     2641 gtacacgcgc aggaagaatc gcatctgcta cctgcaggag attttctcca acgagatggc
     2701 gaaggttgac gattctttct tccacaggct ggaggagtca ttcctcgtgg aggaggataa
     2761 gaagcacgag cggcatccaa tcttcggcaa cattgtcgac gaggttgcct accacgagaa
     2821 gtaccctacg atctaccatc tgcggaagaa gctcgtggac tccacagata aggcggacct
     2881 ccgcctgatc tacctcgctc tggcccacat gattaagttc aggggccatt tcctgatcga
     2941 gggggatctc aacccggaca atagcgatgt tgacaagctg ttcatccagc tcgtgcagac
     3001 gtacaaccag ctcttcgagg agaaccccat taatgcgtca ggcgtcgacg cgaaggctat
     3061 cctgtccgct aggctctcga agtctcggcg cctcgagaac ctgatcgccc agctgccggg
     3121 cgagaagaag aacggcctgt tcgggaatct cattgcgctc agcctggggc tcacgcccaa
     3181 cttcaagtcg aatttcgatc tcgctgagga cgccaagctg cagctctcca aggacacata
     3241 cgacgatgac ctggataacc tcctggccca gatcggcgat cagtacgcgg acctgttcct
     3301 cgctgccaag aatctgtcgg acgccatcct cctgtctgat attctcaggg tgaacaccga
     3361 gattacgaag gctccgctct cagcctccat gatcaagcgc tacgacgagc accatcagga
     3421 tctgaccctc ctgaaggcgc tggtcaggca gcagctcccc gagaagtaca aggagatttt
     3481 cttcgatcag tcgaagaacg gctacgctgg gtacattgac ggcggggcct ctcaggagga
     3541 gttctacaag ttcatcaagc cgattctgga gaagatggac ggcacggagg agctgctggt
     3601 gaagctcaat cgcgaggacc tcctgaggaa gcagcggaca ttcgataacg gcagcatccc
     3661 acaccagatt catctcgggg agctgcacgc tatcctgagg aggcaggagg acttctaccc
     3721 tttcctcaag gataaccgcg agaagatcga gaagattctg actttcagga tcccgtacta
     3781 cgtcggccca ctcgctaggg gcaactcccg cttcgcttgg atgacccgca agtcagagga
     3841 gacgatcacg ccgtggaact tcgaggaggt ggtcgacaag ggcgctagcg ctcagtcgtt
     3901 catcgagagg atgacgaatt tcgacaagaa cctgccaaat gagaaggtgc tccctaagca
     3961 ctcgctcctg tacgagtact tcacagtcta caacgagctg actaaggtga agtatgtgac
     4021 cgagggcatg aggaagccgg ctttcctgtc tggggagcag aagaaggcca tcgtggacct
     4081 cctgttcaag accaaccgga aggtcacggt taagcagctc aaggaggact acttcaagaa
     4141 gattgagtgc ttcgattcgg tcgagatctc tggcgttgag gaccgcttca acgcctccct
     4201 ggggacctac cacgatctcc tgaagatcat taaggataag gacttcctgg acaacgagga
     4261 gaatgaggat atcctcgagg acattgtgct gacactcact ctgttcgagg accgggagat
     4321 gatcgaggag cgcctgaaga cttacgccca tctcttcgat gacaaggtca tgaagcagct
     4381 caagaggagg aggtacaccg gctgggggag gctgagcagg aagctcatca acggcattcg
     4441 ggacaagcag tccgggaaga cgatcctcga cttcctgaag agcgatggct tcgcgaaccg
     4501 caatttcatg cagctgattc acgatgacag cctcacattc aaggaggata tccagaaggc
     4561 tcaggtgagc ggccaggggg actcgctgca cgagcatatc gcgaacctcg ctggctcgcc
     4621 agctatcaag aaggggattc tgcagaccgt gaaggttgtg gacgagctgg tgaaggtcat
     4681 gggcaggcac aagcctgaga acatcgtcat tgagatggcc cgggagaatc agaccacgca
     4741 gaagggccag aagaactcac gcgagaggat gaagaggatc gaggagggca ttaaggagct
     4801 ggggtcccag atcctcaagg agcacccggt ggagaacacg cagctgcaga atgagaagct
     4861 ctacctgtac tacctccaga atggccgcga tatgtatgtg gaccaggagc tggatattaa
     4921 caggctcagc gattacgacg tcgatcatat cgttccacag tcattcctga aggatgactc
     4981 cattgacaac aaggtcctca ccaggtcgga caagaaccgg ggcaagtctg ataatgttcc
     5041 ttcagaggag gtcgttaaga agatgaagaa ctactggcgc cagctcctga atgccaagct
     5101 gatcacgcag cggaagttcg ataacctcac aaaggctgag aggggcgggc tctctgagct
     5161 ggacaaggcg ggcttcatca agaggcagct ggtcgagaca cggcagatca ctaagcacgt
     5221 tgcgcagatt ctcgactcac ggatgaacac taagtacgat gagaatgaca agctgatccg
     5281 cgaggtgaag gtcatcaccc tgaagtcaaa gctcgtctcc gacttcagga aggatttcca
     5341 gttctacaag gttcgggaga tcaacaatta ccaccatgcc catgacgcgt acctgaacgc
     5401 ggtggtcggc acagctctga tcaagaagta cccaaagctc gagagcgagt tcgtgtacgg
     5461 ggactacaag gtttacgatg tgaggaagat gatcgccaag tcggagcagg agattggcaa
     5521 ggctaccgcc aagtacttct tctactctaa cattatgaat ttcttcaaga cagagatcac
     5581 tctggccaat ggcgagatcc ggaagcgccc cctcatcgag acgaacggcg agacggggga
     5641 gatcgtgtgg gacaagggca gggatttcgc gaccgtcagg aaggttctct ccatgccaca
     5701 agtgaatatc gtcaagaaga cagaggtcca gactggcggg ttctctaagg agtcaattct
     5761 gcctaagcgg aacagcgaca agctcatcgc ccgcaagaag gactgggatc cgaagaagta
     5821 cggcgggttc gacagcccca ctgtggccta ctcggtcctg gttgtggcga aggttgagaa
     5881 gggcaagtcc aagaagctca agagcgtgaa ggagctgctg gggatcacga ttatggagcg
     5941 ctccagcttc gagaagaacc cgatcgattt cctggaggcg aagggctaca aggaggtgaa
     6001 gaaggacctg atcattaagc tccccaagta ctcactcttc gagctggaga acggcaggaa
     6061 gcggatgctg gcttccgctg gcgagctgca gaaggggaac gagctggctc tgccgtccaa
     6121 gtatgtgaac ttcctctacc tggcctccca ctacgagaag ctcaagggca gccccgagga
     6181 caacgagcag aagcagctgt tcgtcgagca gcacaagcat tacctcgacg agatcattga
     6241 gcagatttcc gagttctcca agcgcgtgat cctggccgac gcgaatctgg ataaggtcct
     6301 ctccgcgtac aacaagcacc gcgacaagcc aatcagggag caggctgaga atatcattca
     6361 tctcttcacc ctgacgaacc tcggcgcccc tgctgctttc aagtacttcg acacaactat
     6421 cgatcgcaag aggtacacaa gcactaagga ggtcctggac gcgaccctca tccaccagtc
     6481 gattaccggc ctctacgaga cgcgcatcga cctgtctcag ctcgggggcg acaagcggcc
     6541 agcggcgacg aagaaggcgg ggcaggcgaa gaagaagaag acccgcgact ccggcggcag
     6601 cacgaacctc tccgacatca tcgagaagga gacgggcaag cagctcgtga tccaggagag
     6661 catcctcatg ctgccggagg aggtggagga ggtcatcggc aacaagcccg agtccgacat
     6721 cctcgtgcac accgcctacg acgagtccac ggacgagaac gtcatgctcc tgacgagcga
     6781 cgctccagag tacaagccat gggctctcgt gatccaggac agcaacggcg agaacaagat
     6841 caagatgctg tccggcggct ccccaaagaa gaagcggaag gtgaccaacc tgtccgacat
     6901 catcgagaag gagacgggca agcaactcgt gatccaggag agcatcctca tgctgccaga
     6961 ggaggtggag gaggtcatcg gcaacaagcc agagtccgac atcctggtgc acaccgccta
     7021 cgacgagtcc accgacgaga acgtcatgct cctgaccagc gacgccccag agtacaagcc
     7081 atgggccctc gtcatccagg acagcaacgg ggagaacaag atcaagatgc tgtcgggggg
     7141 gagcccaaag aagaagcgga aggtgggatc tggagctact aatttttctc ttcttaagca
     7201 agctggagat gttgaagaaa atcctggacc ctctctgcag ggatctggtt cgatggtgag
     7261 caagggcgag gcagtgatca aggagttcat gcggttcaag gtgcacatgg agggctccat
     7321 gaacggccac gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca
     7381 gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc ttctcctggg acctcctgtc
     7441 ccctcagttc atgtacggct ccagggcctt caccaagcac cccgccgaca tccccgacta
     7501 ctataagcag tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg
     7561 cggcgccgtg accgtgaccc aggacacctc cctggaggac ggcaccctga tctacaaggt
     7621 gaagctccgc ggcaccaact tccctcctga cggccccgta atgcagaaga agacaatggg
     7681 ctgggaagcg tccaccgagc ggttgtaccc cgaggacggc gtgctgaagg gcgacattaa
     7741 gatggccctg cgcctgaagg acggcggccg ctacctggcg gacttcaaga ccacctacaa
     7801 ggccaagaag cccgtgcaga tgcccggcgc ctacaacgtc gaccgcaagt tggacatcac
     7861 ctcccacaac gaggactaca ccgtggtgga acagtacgaa cgctccgagg gccgccactc
     7921 caccggcggc atggacgagc tgtacaagta agagctcaga gctttcgttc gtatcatcgg
     7981 tttcgacaac gttcgtcaag ttcaatgcat cagtttcatt gcgcacacac cagaatccta
     8041 ctgagtttga gtattatggc attgggaaaa ctgtttttct tgtaccattt gttgtgcttg
     8101 taatttactg tgttttttat tcggttttcg ctatcgaact gtgaaatgga aatggatgga
     8161 gaagagttaa tgaatgatat ggtccttttg ttcattctca aattaatatt atttgttttt
     8221 tctcttattt gttgtgtgtt gaatttgaaa ttataagaga tatgcaaaca ttttgttttg
     8281 agtaaaaatg tgtcaaatcg tggcctctaa tgaccgaagt taatatgagg agtaaaacac
     8341 ttgtagttgt accattatgc ttattcacta ggcaacaaat atattttcag acctagaaaa
     8401 gctgcaaatg ttactgaata caagtatgtc ctcttgtgtt ttagacattt atgaactttc
     8461 ctttatgtaa ttttccagaa tccttgtcag attctaatca ttgctttata attatagtta
     8521 tactcatgga tttgtagttg agtatgaaaa tattttttaa tgcattttat gacttgccaa
     8581 ttgattgaca acgaattcgt tggctagagc agcttgccaa catggtggag cacgacactc
     8641 tcgtctactc caagaatatc aaagatacag tctcagaaga ccaaagggct attgagactt
     8701 ttcaacaaag ggtaatatcg ggaaacctcc tcggattcca ttgcccagct atctgtcact
     8761 tcatcaaaag gacagtagaa aaggaaggtg gcacctacaa atgccatcat tgcgataaag
     8821 gaaaggctat cgttcaagat gcctctgccg acagtggtcc caaagatgga cccccaccca
     8881 cgaggagcat cgtggaaaaa gaagacgttc caaccacgtc ttcaaagcaa gtggattgat
     8941 gtgaacatgg tggagcacga cactctcgtc tactccaaga atatcaaaga tacagtctca
     9001 gaagaccaaa gggctattga gacttttcaa caaagggtaa tatcgggaaa cctcctcgga
     9061 ttccattgcc cagctatctg tcacttcatc aaaaggacag tagaaaagga aggtggcacc
     9121 tacaaatgcc atcattgcga taaaggaaag gctatcgttc aagatgcctc tgccgacagt
     9181 ggtcccaaag atggaccccc acccacgagg agcatcgtgg aaaaagaaga cgttccaacc
     9241 acgtcttcaa agcaagtgga ttgatgtgat atctccactg acgtaaggga tgacgcacaa
     9301 tcccactatc cttcgcaaga cccttcctct atataaggaa gttcatttca tttggagagg
     9361 acacgctgaa atcaccagtc tctctctaca aatctatctc tctcgagtca acacaacata
     9421 tacaaaacaa acgaatctca agcaatcaag cattctactt ctattgcagc aatttaaatc
     9481 atttctttta aagcaaaagc aattttctga aaattttcac catttacgaa cgatagccat
     9541 ggctcaagtt agcagaatct gcaatggtgt gcagaaccca tctcttatct ccaatctctc
     9601 taaatccagt caaaggaaat ctcccttatc ggtttctctg aagactcagc agcatccacg
     9661 agcttatcca atttcttcat cttggggatt gaagaagagt gggatgactt taattggctc
     9721 tgagcttcgt cctcttaagg tcatgtcttc tgtttccacg gcggagaagg gatccgacgc
     9781 tcttccagct accttcgacg ttatcgtgca tccagctaga gaactcagag gtgaacttag
     9841 agcacagcca tccaagaact acaccactag atacctcctc gccgctgctc tcgctgaggg
     9901 tgaaaccaga gttgttggtg tggctacctc tgaggatgcc gaagctatgc tcagatgcct
     9961 cagagattgg ggtgctggtg ttgagcttgt tggtgatgac gccgtgatca gaggtttcgg
    10021 tgctagacca caggctggtg ttacccttaa cccaggtaac gctgctgcgg tggccagact
    10081 ccttatgggt gttgctgctc tcacctctgg tacaactttc gttaccgatt accctgattc
    10141 ccttggtaaa agacctcagg gtgaccttct tgaagccctc gaaagacttg gtgcttgggt
    10201 gtcctccaac gatggtagac tccctatctc cgtttccggt ccagttagag gtggtacagt
    10261 ggaggtttcc gccgaaagat cctcccagta cgcttccgcc cttatgttcc tcggtcctct
    10321 tcttcctgac ggactcgaac ttagactcac cggtgatatc aagtcccacg ctcctcttag
    10381 acagacactt gacaccctct ctgatttcgg tgttagagct actgcctccg atgaccttag
    10441 aagaatctcc atccctggtg gtcagaagta cagaccaggt agagtgctcg ttcctggtga
    10501 ttaccctggt tccgctgcta tccttaccgc cgctgctctt ctcccaggtg aggttagact
    10561 ttctaacctt agagaacacg acctccaggg tgagaaggaa gctgtgaacg ttcttagaga
    10621 gatgggtgct gatatcgtta gagaaggtga tacccttacc gtgagaggtg gtagacctct
    10681 ccacgctgtt actagagatg gtgattcctt caccgacgcc gtgcaagctc ttaccgctgc
    10741 tgctgccttc gctgagggtg ataccacctg ggaaaacgtt gctactctta gactcaagga
    10801 atgcgataga atctctgaca ccagagctga gcttgaaaga cttggtctta gagcaagaga
    10861 aaccgccgat tctctctccg ttactggttc tgctcacctt gctggtggta tcaccgctga
    10921 tggtcacggt gaccacagaa tgatcatgct tctcaccctt cttgggctca gagcagatgc
    10981 tccacttaga atcaccggtg cacaccacat cagaaagtcc taccctcagt tcttcgctca
    11041 ccttgaagct cttggtgcta gattcgaata cgctgaggct accgccataa taatgtgtga
    11101 gtagttccca gataagggaa ttagggttcc tatagggttt cgctcatgtg ttgagcatat
    11161 aagaaaccct tagtatgtat ttgtatttgt aaaatacttc tatcaataaa atttctaatt
    11221 cctaaaacca aaatccagta ctaaaatcca gatcccccga attaattcgg cgttaattca
    11281 gtacattaaa aacgtccgca atgtgttatt aagttgtcta agcgtcaatt tgtttacacc
    11341 acaatatatc ctgccaccag ccagccaaca gctccccgac cggcagctcg gcacaaaatc
    11401 accactcgat acaggcagcc catcagtccg ggacggcgtc agcgggagag ccgttgtaag
    11461 gcggcagact ttgctcatgt taccgatgct attcggaaga acggcaacta agctgccggg
    11521 tttgaaacac ggatgatctc gcggagggta gcatgttgat tgtaacgatg acagagcgtt
    11581 gctgcctgtg atcaccgcgg tttcaaaatc ggctccgtcg atactatgtt atacgccaac
    11641 tttgaaaaca actttgaaaa agctgttttc tggtatttaa ggttttagaa tgcaaggaac
    11701 agtgaattgg agttcgtctt gttataatta gcttcttggg gtatctttaa atactgtaga
    11761 aaagaggaag gaaataataa atggctaaaa tgagaatatc accggaattg aaaaaactga
    11821 tcgaaaaata ccgctgcgta aaagatacgg aaggaatgtc tcctgctaag gtatataagc
    11881 tggtgggaga aaatgaaaac ctatatttaa aaatgacgga cagccggtat aaagggacca
    11941 cctatgatgt ggaacgggaa aaggacatga tgctatggct ggaaggaaag ctgcctgttc
    12001 caaaggtcct gcactttgaa cggcatgatg gctggagcaa tctgctcatg agtgaggccg
    12061 atggcgtcct ttgctcggaa gagtatgaag atgaacaaag ccctgaaaag attatcgagc
    12121 tgtatgcgga gtgcatcagg ctctttcact ccatcgacat atcggattgt ccctatacga
    12181 atagcttaga cagccgctta gccgaattgg attacttact gaataacgat ctggccgatg
    12241 tggattgcga aaactgggaa gaagacactc catttaaaga tccgcgcgag ctgtatgatt
    12301 ttttaaagac ggaaaagccc gaagaggaac ttgtcttttc ccacggcgac ctgggagaca
    12361 gcaacatctt tgtgaaagat ggcaaagtaa gtggctttat tgatcttggg agaagcggca
    12421 gggcggacaa gtggtatgac attgccttct gcgtccggtc gatcagggag gatatcgggg
    12481 aagaacagta tgtcgagcta ttttttgact tactggggat caagcctgat tgggagaaaa
    12541 taaaatatta tattttactg gatgaattgt tttagtacct agaatgcatg accaaaatcc
    12601 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
    12661 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
    12721 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
    12781 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
    12841 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
    12901 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
    12961 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
    13021 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
    13081 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
    13141 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
    13201 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
    13261 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
    13321 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
    13381 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga
    13441 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatatgg tgcactctca
    13501 gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat tgctacgtga
    13561 ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
    13621 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
    13681 gaggttttca ccgtcattac cgaaacgcgc gaggcagggt gccttgatgt gggcgccggc
    13741 ggtcgagtgg cgacggcgcg gcttgtccgc gccctggtag attgcctggc cgtaggccag
    13801 ccatttttga gcggccagcg gccgcgatag gccgacgcga agcggcgggg cgtagggagc
    13861 gcagcgaccg aagggtaggc gctttttgca gctcttcggc tgtgcgctgg ccagacagtt
    13921 atgcacaggc caggcgggtt ttaagagttt taataagttt taaagagttt taggcggaaa
    13981 aatcgccttt tttctctttt atatcagtca cttacatgtg tgaccggttc ccaatgtacg
    14041 gctttgggtt cccaatgtac gggttccggt tcccaatgta cggctttggg ttcccaatgt
    14101 acgtgctatc cacaggaaac agaccttttc gacctttttc ccctgctagg gcaatttgcc
    14161 ctagcatctg ctccgtacat taggaaccgg cggatgcttc gccctcgatc aggttgcggt
    14221 agcgcatgac taggatcggg ccagcctgcc ccgcctcctc cttcaaatcg tactccggca
    14281 ggtcatttga cccgatcagc ttgcgcacgg tgaaacagaa cttcttgaac tctccggcgc
    14341 tgccactgcg ttcgtagatc gtcttgaaca accatctggc ttctgccttg cctgcggcgc
    14401 ggcgtgccag gcggtagaga aaacggccga tgccgggatc gatcaaaaag taatcggggt
    14461 gaaccgtcag cacgtccggg ttcttgcctt ctgtgatctc gcggtacatc caatcagcta
    14521 gctcgatctc gatgtactcc ggccgcccgg tttcgctctt tacgatcttg tagcggctaa
    14581 tcaaggcttc accctcggat accgtcacca ggcggccgtt cttggccttc ttcgtacgct
    14641 gcatggcaac gtgcgtggtg tttaaccgaa tgcaggtttc taccaggtcg tctttctgct
    14701 ttccgccatc ggctcgccgg cagaacttga gtacgtccgc aacgtgtgga cggaacacgc
    14761 ggccgggctt gtctcccttc ccttcccggt atcggttcat ggattcggtt agatgggaaa
    14821 ccgccatcag taccaggtcg taatcccaca cactggccat gccggccggc cctgcggaaa
    14881 cctctacgtg cccgtctgga agctcgtagc ggatcacctc gccagctcgt cggtcacgct
    14941 tcgacagacg gaaaacggcc acgtccatga tgctgcgact atcgcgggtg cccacgtcat
    15001 agagcatcgg aacgaaaaaa tctggttgct cgtcgccctt gggcggcttc ctaatcgacg
    15061 gcgcaccggc tgccggcggt tgccgggatt ctttgcggat tcgatcagcg gccgcttgcc
    15121 acgattcacc ggggcgtgct tctgcctcga tgcgttgccg ctgggcggcc tgcgcggcct
    15181 tcaacttctc caccaggtca tcacccagcg ccgcgccgat ttgtaccggg ccggatggtt
    15241 tgcgaccgct cacgccgatt cctcgggctt gggggttcca gtgccattgc agggccggca
    15301 ggcaacccag ccgcttacgc ctggccaacc gcccgttcct ccacacatgg ggcattccac
    15361 ggcgtcggtg cctggttgtt cttgattttc catgccgcct cctttagccg ctaaaattca
    15421 tctactcatt tattcatttg ctcatttact ctggtagctg cgcgatgtat tcagatagca
    15481 gctcggtaat ggtcttgcct tggcgtaccg cgtacatctt cagcttggtg tgatcctccg
    15541 ccggcaactg aaagttgacc cgcttcatgg ctggcgtgtc tgccaggctg gccaacgttg
    15601 cagccttgct gctgcgtgcg ctcggacggc cggcacttag cgtgtttgtg cttttgctca
    15661 ttttctcttt acttcattaa ctcaaatgag ttttgattta atttcagcgg ccagcgcctg
    15721 gacctcgcgg gcagcgtcgc cctcgggttc tgattcaaga acggttgtgc cggcggcggc
    15781 agtgcctggg tagctcacgc gttgcgtgat acgggactca agaatgggca gctcgtaccc
    15841 ggccagcgcc tcggcaacct caccgccgat gcgcgtgcct ttgatcgccc gcgacacgac
    15901 aaaggccgct tgtagccttc catccgtgac ctcaatgcgc tgcttaacca gctccaccag
    15961 gtcggcggtg gcccatatgt cgtaagggct tggctgcacc ggaatcagca cgaagtcggc
    16021 tgccttgatc gcggacacag ccaagtccgc cgcctggggc gctccgtcga tcactacgaa
    16081 gtcgcgccgg ccgatggcct tcacgtcgcg gtcaatcgtc gggcggtcga tgccgacaac
    16141 ggttagcggt tgatcttccc gcacggccgc ccaatcgcgg gcactgccct ggggatcgga
    16201 atcgactaac agaacatcgg ccccggcgag ttgcagggcg cgggctagat gggttgcgat
    16261 ggtcgtcttg cctgacccgc ctttctggtt aagtacagcg ataaccttca tgcgttcccc
    16321 ttgcgtattt gtttatttac tcatcgcatc atatacgcag cgaccgcatg acgcaagctg
    16381 ttttactcaa atacacatca cctttttaga cggcggcgct cggtttcttc agcggccaag
    16441 ctggccggcc aggccgccag cttggcatca gacaaaccgg ccaggatttc atgcagccgc
    16501 acggttgaga cgtgcgcggg cggctcgaac acgtacccgg ccgcgatcat ctccgcctcg
    16561 atctcttcgg taatgaaaaa cggttcgtcc tggccgtcct ggtgcggttt catgcttgtt
    16621 cctcttggcg ttcattctcg gcggccgcca gggcgtcggc ctcggtcaat gcgtcctcac
    16681 ggaaggcacc gcgccgcctg gcctcggtgg gcgtcacttc ctcgctgcgc tcaagtgcgc
    16741 ggtacagggt cgagcgatgc acgccaagca gtgcagccgc ctctttcacg gtgcggcctt
    16801 cctggtcgat cagctcgcgg gcgtgcgcga tctgtgccgg ggtgagggta gggcgggggc
    16861 caaacttcac gcctcgggcc ttggcggcct cgcgcccgct ccgggtgcgg tcgatgatta
    16921 gggaacgctc gaactcggca atgccggcga acacggtcaa caccatgcgg ccggccggcg
    16981 tggtggtgtc ggcccacggc tctgccaggc tacgcaggcc cgcgccggcc tcctggatgc
    17041 gctcggcaat gtccagtagg tcgcgggtgc tgcgggccag gcggtctagc ctggtcactg
    17101 tcacaacgtc gccagggcgt aggtggtcaa gcatcctggc cagctccggg cggtcgcgcc
    17161 tggtgccggt gatcttctcg gaaaacagct tggtgcagcc ggccgcgtgc agttcggccc
    17221 gttggttggt caagtcctgg tcgtcggtgc tgacgcgggc atagcccagc aggccagcgg
    17281 cggcgctctt gttcatggcg taatgtctcc ggttctagtc gcaagtattc tactttatgc
    17341 gactaaaaca cgcgacaaga aaacgccagg aaaagggcag ggcggcagcc tgtcgcgtaa
    17401 cttaggactt gtgcgacatg tcgttttcag aagacggctg cactgaacgt cagaagccga
    17461 ctgcactata gcagcggagg ggttggatca aagtactttg atcccgaggg gaaccctgtg
    17521 gttggcatgc acatacaaat ggacgaacgg ataaaccttt tcacgccctt ttaaatatcc
    17581 gattattcta ataaacgctc ttttctctta g
//
