LOCUS       sequence-438352-       15099 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Single component CRISPR-mediated base-editor for Agrobacterium 
            genetic engineering visually marked with sfGFP for guide insertion 
            and AeBlue for plasmid eviction..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 5328 to 5362)
  AUTHORS   Pennetti VJ, LaFayette PR, Parrott WA
  TITLE     Single component CRISPR-mediated base-editors for Agrobacterium and 
            their use to develop an improved suite of strains
  JOURNAL   Biodes Res. 2025 March, 7:100001. doi: 10.1016/j.bidere.2025.100001
REFERENCE   2  (bases 1 to 15099)
  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: "Biodes Res.
            2025 March, 7:100001. doi: 10.1016/j.bidere.2025.100001"
COMMENT     Sequence Label: pVP143
FEATURES             Location/Qualifiers
     source          1..15099
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1289..1877
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1778..1797
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2031..2048
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2063..2203
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2129..2148)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2289..2311
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     rep_origin      2547..2741
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(2807..3880)
                     /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(4309..4938)
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     promoter        5328..5362
                     /label=﻿BBa_J23100
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 1.0
                     "
                     /citation=[1]
     RBS             5374..5385
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      6107..6153
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             join(6420..6446,6447..6449,6450..8936,8937..8939,
                     8940..10523)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the H840A mutation in the HNH 
                     catalytic domain"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             10731..10796
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             10815..10880
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             10887..11510
                     /codon_start=1
                     /product="cytidine deaminase from the sea lamprey 
                     Petromyzon marinus (Rogozin et al., 2007)"
                     /label=PmCDA1
                     /translation="MTDAEYVRIHEKLDIYTFKKQFFNNKKSVSHRCYVLFELKRRGER
                     RACFWGYAVNKPQSGTERGIHAEIFSIRKVEEYLRDNPGQFTINWYSSWSPCADCAEKI
                     LEWYNQELRGNGHTLKIWACKLYYEKNARNQIGLWNLRDNGVGLNVMVSEHYQCCRKIF
                     IQSSHNQLNENRWLEKTLKRAEKRRSELSIMIQVKILHTTKSPAV"
     CDS             11517..11768
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="MTNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHT
                     AYDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     promoter        11840..11874
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     RBS             12030..12041
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             12048..12761
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      12773..12844
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      12860..12887
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_RNA        12903..12978
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        12992..13437
                     /gene="sacR"
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             join(13438..13524,13525..14859)
                     /codon_start=1
                     /gene="Bacillus subtilis sacB"
                     /product="secreted levansucrase that renders bacterial 
                     growth sensitive to sucrose"
                     /label=SacB
                     /note="negative selection marker"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     primer_bind     complement(14966..14985)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             join(14977..15099,1..693)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             join(14977..15099,1..693)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(15050..15069)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 cgattgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa aggtagcgtt
       61 gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt tatgcctctt
      121 ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac cactgcgatc
      181 cccgggaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga aaatattgtt
      241 gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt
      301 aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa cggtttggtt
      361 gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa
      421 atgcataaac ttttgccatt ctcaccggat tcagtcgtca ctcatggtga tttctcactt
      481 gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg acgagtcgga
      541 atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga gttttctcct
      601 tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat gaataaattg
      661 cagtttcatt tgatgctcga tgagtttttc taacagcggc ggaggaactc tttgatccgg
      721 ttcctgaaca ggatctattt gaggcgctaa atgaaacctt aacgctatgg aactcgccgc
      781 ccgactgggc tggcgatgag cgaaatgtag tgcttacgtt gtcccgcatt tggtacagcg
      841 cagtaaccgg caaaatcgcg ccgaaggatg tcgctgccga ctgggcaatg gagcgcctgc
      901 cggcccagta tcagcccgtc atacttgaag ctagacaggc ttatcttgga caagaagaag
      961 atcgcttggc ctcgcgcgca gatcagttgg aagaatttgt ccactacgtg aaaggcgaga
     1021 tcaccaaggt agtcggcaaa taatgtctag ctagaaattc gttcaagccg acgccgcttc
     1081 gcggcgcggc ttaactcaag cgttagatgc actaagcaca taattgctca cagccaaact
     1141 atcaggtcaa gtctgctttt attattttta agcgtgcata ataagcccta cacaaattgg
     1201 gagatatatc atgcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     1261 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     1321 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     1381 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     1441 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     1501 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     1561 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     1621 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     1681 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     1741 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     1801 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     1861 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     1921 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     1981 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     2041 tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     2101 tttcacaccg catatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     2161 agtatacact ccgctatcgc tacgtgactg ggtcatggct gcgccccgac acccgccaac
     2221 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     2281 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     2341 gcagggtgcc ttgatgtggg cgccggcggt cgagtggcga cggcgcggct tgtccgcgcc
     2401 ctggtagatt gcctggccgt aggccagcca tttttgagcg gccagcggcc gcgataggcc
     2461 gacgcgaagc ggcggggcgt agggagcgca gcgaccgaag ggtaggcgct ttttgcagct
     2521 cttcggctgt gcgctggcca gacagttatg cacaggccag gcgggtttta agagttttaa
     2581 taagttttaa agagttttag gcggaaaaat cgcctttttt ctcttttata tcagtcactt
     2641 acatgtgtga ccggttccca atgtacggct ttgggttccc aatgtacggg ttccggttcc
     2701 caatgtacgg ctttgggttc ccaatgtacg tgctatccac aggaaagaga gcttttcgac
     2761 ctttttcccc tgctagggca atttgcccta gcatctgctc cgtacattag gaaccggcgg
     2821 atgcttcgcc ctcgatcagg ttgcggtagc gcatgactag gatcgggcca gcctgccccg
     2881 cctcctcctt caaatcgtac tccggcaggt catttgaccc gatcagcttg cgcacggtga
     2941 aacagaactt cttgaactct ccggcgctgc cactgcgttc gtagatcgtc ttgaacaacc
     3001 atctggcttc tgccttgcct gcggcgcggc gtgccaggcg gtagagaaaa cggccgatgc
     3061 cgggatcgat caaaaagtaa tcggggtgaa ccgtcagcac gtccgggttc ttgccttctg
     3121 tgatctcgcg gtacatccaa tcagctagct cgatctcgat gtactccggc cgcccggttt
     3181 cgctctttac gatcttgtag cggctaatca aggcttcacc ctcggatacc gtcaccaggc
     3241 ggccgttctt ggccttcttc gtacgctgca tggcaacgtg cgtggtgttt aaccgaatgc
     3301 aggtttctac caggtcgtct ttctgctttc cgccatcggc tcgccggcag aacttgagta
     3361 cgtccgcaac gtgtggacgg aacacgcggc cgggcttgtc tcccttccct tcccggtatc
     3421 ggttcatgga ttcggttaga tgggaaaccg ccatcagtac caggtcgtaa tcccacacac
     3481 tggccatgcc ggccggccct gcggaaacct ctacgtgccc gtctggaagc tcgtagcgga
     3541 tcacctcgcc agctcgtcgg tcacgcttcg acagacggaa aacggccacg tccatgatgc
     3601 tgcgactatc gcgggtgccc acgtcataga gcatcggaac gaaaaaatct ggttgctcgt
     3661 cgcccttggg cggcttccta atcgacggcg caccggctgc cggcggttgc cgggattctt
     3721 tgcggattcg atcagcggcc gcttgccacg attcaccggg gcgtgcttct gcctcgatgc
     3781 gttgccgctg ggcggcctgc gcggccttca acttctccac caggtcatca cccagcgccg
     3841 cgccgatttg taccgggccg gatggtttgc gaccgctcac gccgattcct cgggcttggg
     3901 ggttccagtg ccattgcagg gccggcagac aacccagccg cttacgcctg gccaaccgcc
     3961 cgttcctcca cacatggggc attccacggc gtcggtgcct ggttgttctt gattttccat
     4021 gccgcctcct ttagccgcta aaattcatct actcatttat tcatttgctc atttactctg
     4081 gtagctgcgc gatgtattca gatagcagct cggtaatggt cttgccttgg cgtaccgcgt
     4141 acatcttcag cttggtgtga tcctccgccg gcaactgaaa gttgacccgc ttcatggctg
     4201 gcgtgtctgc caggctggcc aacgttgcag ccttgctgct gcgtgcgctc ggacggccgg
     4261 cacttagcgt gtttgtgctt ttgctcattt tctctttacc tcattaactc aaatgagttt
     4321 tgatttaatt tcagcggcca gcgcctggac ctcgcgggca gcgtcgccct cgggttctga
     4381 ttcaagaacg gttgtgccgg cggcggcagt gcctgggtag ctcacgcgct gcgtgatacg
     4441 ggactcaaga atgggcagct cgtacccggc cagcgcctcg gcaacctcac cgccgatgcg
     4501 cgtgcctttg atcgcccgcg acacgacaaa ggccgcttgt agccttccat ccgtgacctc
     4561 aatgcgctgc ttaaccagct ccaccaggtc ggcggtggcc catatgtcgt aagggcttgg
     4621 ctgcaccgga atcagcacga agtcggctgc cttgatcgcg gacacagcca agtccgccgc
     4681 ctggggcgct ccgtcgatca ctacgaagtc gcgccggccg atggccttca cgtcgcggtc
     4741 aatcgtcggg cggtcgatgc cgacaacggt tagcggttga tcttcccgca cggccgccca
     4801 atcgcgggca ctgccctggg gatcggaatc gactaacaga acatcggccc cggcgagttg
     4861 cagggcgcgg gctagatggg ttgcgatggt cgtcttgcct gacccgcctt tctggttaag
     4921 tacagcgata accttcatgc gttccccttg cgtatttgtt tatttactca tcgcatcata
     4981 tacgcagcga ccgcatgacg caagctgttt tactcaaata cacatcacct ttttagacgg
     5041 cggcgctcgg tttcttcagc ggccaagctg gccggccagg ccgccagctt ggcatcagac
     5101 aaaccggcca ggatttcatg cagccgcacg gttgagacgt gcgcgggcgg ctcgaacacg
     5161 tacccggccg cgatcatctc cgcctcgatc tcttcggtaa tgaaaaacgg ttcgtcctgg
     5221 ccgtcctggt gcggtttcat gcttgttcct cttggcgttc attctcggcg gccgccaggg
     5281 cgtcggcctc ggtcaatgcg tcctcacgga aggcaccgcg ccgcctgttg acggctagct
     5341 cagtcctagg tacagtgcta gctactagag attaaagagg agaaagagtc ccatggcttc
     5401 actggttaaa aaagacatgt gcatcaaaat gacgatggaa ggaacagtaa acggtcacca
     5461 tttcaagtgt gtaggagaag gcgaaggcaa accatttgaa gggacccagg tggaaaagat
     5521 acgcatcact gaaggtgggc ccttaccatt tgcgtatgat attttggccc cttgttgcat
     5581 gtatggcagt aaaaccttca ttaagcatgt gtcgggtatt ccggattact ttaaggagtc
     5641 ttttcctgag ggctttacct gggaaagaac acaaatcttc gaggatggcg gctatctcac
     5701 catacaccag gacacgagcc ttcagggtaa taattttatt ttcaaagtta atgtcatcgg
     5761 tgccaacttc cctgcaaacg gtcccgtgat gcagaaaaaa acagctggat gggaaccgtg
     5821 cgttgagatg ctttatccgc gggacggcgt cctgtgtggt cagagcctga tggccctgaa
     5881 atgcactgat ggcaatcatc tgacgtccca cctgcgcact acctatcgtt ctcgcaagcc
     5941 atccaatgca gttaacatgc cggaatttca ttttggggat catcgcattg agattttgaa
     6001 agctgaacaa ggtaaatttt atgaacaata cgagtcagcg gtggcccgtt actgtgaggc
     6061 ggcaccgagt aaattagggc atcactaata atagagctca ggacatcaaa taaaacgaaa
     6121 ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttacgagcat tttcgttctg
     6181 agcctgacct gaccaactgc ggcctgaata ggtcgatccg gttgcttagt catggatgcg
     6241 cggttctcgg tccatgttgc gttccaagac gccgggcgag gtttctcgct tcaattgaaa
     6301 tcataaagaa gcaattgaaa attttcgagt aaccgaccct cccgataatc ttcaacataa
     6361 aacaacgcac ttcttccaac gggagaggcg gtgttagttg cgagctaagg agaggctcca
     6421 tggataagaa atactcaata ggcttagcta tcggcacaaa tagcgtcgga tgggcggtga
     6481 tcactgatga atataaggtt ccgtctaaaa agttcaaggt tctgggaaat acagaccgcc
     6541 acagtatcaa aaaaaatctt ataggggctc ttttatttga cagtggagag acagcggaag
     6601 cgactcgtct caaacggaca gctcgtagaa ggtatacacg tcggaagaat cgtatttgtt
     6661 atctacagga gattttttca aatgagatgg cgaaagtaga tgatagtttc tttcatcgac
     6721 ttgaagagtc ttttttggtg gaagaagaca agaagcatga acgtcatcct atttttggaa
     6781 atatagtaga tgaagttgct tatcatgaga aatatccaac tatctatcat ctgcgaaaaa
     6841 aattggtaga ttctactgat aaagcggatt tgcgcttaat ctatttggcc ttagcgcata
     6901 tgattaagtt tcgtggtcat tttttgattg agggagattt aaatcctgat aatagtgatg
     6961 tggacaaact atttatccag ttggtacaaa cctacaatca attatttgaa gaaaacccta
     7021 ttaacgcaag tggagtagat gctaaagcga ttctttctgc acgattgagt aaatcaagac
     7081 gattagaaaa tctcattgct cagctccccg gtgagaagaa aaatggctta tttgggaatc
     7141 tcattgcttt gtcattgggt ttgaccccta attttaaatc aaattttgat ttggcagaag
     7201 atgctaaatt acagctttca aaagatactt acgatgatga tttagataat ttattggcgc
     7261 aaattggaga tcaatatgct gatttgtttt tggcagctaa gaatttatca gatgctattt
     7321 tactttcaga tatcctaaga gtaaatactg aaataactaa ggctccccta tcagcttcaa
     7381 tgattaaacg ctacgatgaa catcatcaag acttgactct tttaaaagct ttagttcgac
     7441 aacaacttcc agaaaagtat aaagaaatct tttttgatca atcaaaaaac ggatatgcag
     7501 gttatattga tgggggagct agccaagaag aattttataa atttatcaaa ccaattttag
     7561 aaaaaatgga tggtactgag gaattattgg tgaaactaaa tcgtgaagat ttgctgcgca
     7621 agcaacggac ctttgacaac ggctctattc cccatcaaat tcacttgggt gagctgcatg
     7681 ctattttgag aagacaagaa gacttttatc catttttaaa agacaatcgt gagaagattg
     7741 aaaaaatctt gacttttcga attccttatt atgttggtcc attggcgcgt ggcaatagtc
     7801 gttttgcatg gatgactcgg aagtctgaag aaacaattac cccatggaat tttgaagaag
     7861 ttgtcgataa aggtgcttca gctcaatcat ttattgaacg catgacaaac tttgataaaa
     7921 atcttccaaa tgaaaaagta ctaccaaaac atagtttgct ttatgagtat tttacggttt
     7981 ataacgaatt gacaaaggtc aaatatgtta ctgaaggaat gcgaaaacca gcatttcttt
     8041 caggtgaaca gaagaaagcc attgttgatt tactcttcaa aacaaatcga aaagtaaccg
     8101 ttaagcaatt aaaagaagat tatttcaaaa aaatagaatg ttttgatagt gttgaaattt
     8161 caggagttga agatagattt aatgcttcat taggtaccta ccatgatttg ctaaaaatta
     8221 ttaaagataa agattttttg gataatgaag aaaatgaaga tatcttagag gatattgttt
     8281 taacattgac cttatttgaa gatagggaga tgattgagga aagacttaaa acatatgctc
     8341 acctctttga tgataaggtg atgaaacagc ttaaacgtcg ccgttatact ggttggggac
     8401 gtttgtctcg aaaattgatt aatggtatta gggataagca atctggcaaa acaatattag
     8461 attttttgaa atcagatggt tttgccaatc gcaattttat gcagctgatc catgatgata
     8521 gtttgacatt taaagaagac attcaaaaag cacaagtgtc tggacaaggc gatagtttac
     8581 atgaacatat tgcaaattta gctggtagcc ctgctattaa aaaaggtatt ttacagactg
     8641 taaaagttgt tgatgaattg gtcaaagtaa tggggcggca taagccagaa aatatcgtta
     8701 ttgaaatggc acgtgaaaat cagacaactc aaaagggcca gaaaaattcg cgagagcgta
     8761 tgaaacgaat cgaagaaggt atcaaagaat taggaagtca gattcttaaa gagcatcctg
     8821 ttgaaaatac tcaattgcaa aatgaaaagc tctatctcta ttatctccaa aatggaagag
     8881 acatgtatgt ggaccaagaa ttagatatta atcgtttaag tgattatgat gtcgatgcca
     8941 ttgttccaca aagtttcctt aaagacgatt caatagacaa taaggtctta acgcgttctg
     9001 ataaaaatcg tggtaaatcg gataacgttc caagtgaaga agtagtcaaa aagatgaaaa
     9061 actattggag acaacttcta aacgccaagt taatcactca acgtaagttt gataatttaa
     9121 cgaaagctga acgtggaggt ttgagtgaac ttgataaagc tggttttatc aaacgccaat
     9181 tggttgaaac tcgccaaatc actaagcatg tggcacaaat tttggatagt cgcatgaata
     9241 ctaaatacga tgaaaatgat aaacttattc gagaggttaa agtgattacc ttaaaatcta
     9301 aattagtttc tgacttccga aaagatttcc aattctataa agtacgtgag attaacaatt
     9361 accatcatgc ccatgatgcg tatctaaatg ccgtcgttgg aactgctttg attaagaaat
     9421 atccaaaact tgaatcggag tttgtctatg gtgattataa agtttatgat gttcgtaaaa
     9481 tgattgctaa gtctgagcaa gaaataggca aagcaaccgc aaaatatttc ttttactcta
     9541 atatcatgaa cttcttcaaa acagaaatta cacttgcaaa tggagagatt cgcaaacgcc
     9601 ctctaatcga aactaatggg gaaactggag aaattgtctg ggataaaggg cgagattttg
     9661 ccacagtgcg caaagtattg tccatgcccc aagtcaatat tgtcaagaaa acagaagtac
     9721 agacaggcgg attctccaag gagtcaattt taccaaaaag aaattcggac aagcttattg
     9781 ctcgtaaaaa agactgggat ccaaaaaaat atggtggttt tgatagtcca acggtagctt
     9841 attcagtcct agtggttgct aaggtggaaa aagggaaatc gaagaagtta aaatccgtta
     9901 aagagttact agggatcaca attatggaaa gaagttcctt tgaaaaaaat ccgattgact
     9961 ttttagaagc taaaggatat aaggaagtta aaaaagactt aatcattaaa ctacctaaat
    10021 atagtctttt tgagttagaa aacggtcgta aacggatgct ggctagtgcc ggagaattac
    10081 aaaaaggaaa tgagctggct ctgccaagca aatatgtgaa ttttttatat ttagctagtc
    10141 attatgaaaa gttgaagggt agtccagaag ataacgaaca aaaacaattg tttgtggagc
    10201 agcataagca ttatttagat gagattattg agcaaatcag tgaattttct aagcgtgtta
    10261 ttttagcaga tgccaattta gataaagttc ttagtgcata taacaaacat agagacaaac
    10321 caatacgtga acaagcagaa aatattattc atttatttac gttgacgaat cttggagctc
    10381 ccgctgcttt taaatatttt gatacaacaa ttgatcgtaa acgatatacg tctacaaaag
    10441 aagttttaga tgccactctt atccatcaat ccatcactgg tctttatgaa acacgcattg
    10501 atttgagtca gctaggaggt gacggtggag gaggttctgg aggtggaggt tctgctgagt
    10561 atgtgcgagc cctctttgac tttaatggga atgatgaaga ggatcttccc tttaagaaag
    10621 gagacatcct gagaatccgg gataagcctg aggagcagtg gtggaatgca gaggacagcg
    10681 aaggaaagag ggggatgatt cctgtccctt acgtggagaa gtattccgga gactataagg
    10741 accacgacgg agactacaag gatcatgata ttgattacaa agacgatgac gataagtcta
    10801 ggctcgagtc cggagactat aaggaccacg acggagacta caaggatcat gatattgatt
    10861 acaaagacga tgacgataag tctaggatga ccgacgctga gtacgtgaga atccatgaga
    10921 agttggacat ctacacgttt aagaaacagt ttttcaacaa caaaaaatcc gtgtcgcata
    10981 gatgctacgt tctctttgaa ttaaaacgac ggggtgaacg tagagcgtgt ttttggggct
    11041 atgctgtgaa taaaccacag agcgggacag aacgtggcat tcacgccgaa atctttagca
    11101 ttagaaaagt cgaagaatac ctgcgcgaca accccggaca attcacgata aattggtact
    11161 catcctggag tccttgtgca gattgcgctg aaaagatctt agaatggtat aaccaggagc
    11221 tgcgggggaa cggccacact ttgaaaatct gggcttgcaa actctattac gagaaaaatg
    11281 cgaggaatca aattgggctg tggaacctca gagataacgg ggttgggttg aatgtaatgg
    11341 taagtgaaca ctaccaatgt tgcaggaaaa tattcatcca atcgtcgcac aatcaattga
    11401 atgagaatag atggcttgag aagactttga agcgagctga aaaacgacgg agcgagttgt
    11461 ccattatgat tcaggtaaaa atactccaca ccactaagag tcctgctgtt tctagaatga
    11521 ccaacctttc cgacatcata gagaaggaaa caggcaaaca gttggtcatc caagagtcga
    11581 tactcatgct tcctgaagaa gttgaggagg tcattgggaa taagccggaa agtgacattc
    11641 tcgtacacac tgcgtatgat gagagcaccg atgagaacgt gatgctgctc acgtcagatg
    11701 ccccagagta caaaccctgg gctctggtga ttcaggactc taatggagag aacaagatca
    11761 agatgctagg actggttgca taatcagaat aagcaaaccc cttgggttcc ctctttagga
    11821 gtctgagggg ttttttgcat tgacagctag ctcagtccta ggtataatgc tagcggagac
    11881 cgaaagtgaa acgtgatttc atgcgtcatt ttgaacattt tgtaaatctt atttaataat
    11941 gtgtgcggca attcacattt aatttatgaa tgttttctta acatcgcggc aactcaagaa
    12001 acggcaggtt cggatcttag ctactagaga aagaggagaa atactagatg cgtaaaggcg
    12061 aagagctgtt cactggtgtc gtccctattc tggtggaact ggatggtgat gtcaacggtc
    12121 ataagttttc cgtgcgtggc gagggtgaag gtgacgcaac taatggtaaa ctgacgctga
    12181 agttcatctg tactactggt aaactgccgg ttccttggcc gactctggta acgacgctga
    12241 cttatggtgt tcagtgcttt gctcgttatc cggaccatat gaagcagcat gacttcttca
    12301 agtccgccat gccggaaggc tatgtgcagg aacgcacgat ttcctttaag gatgacggca
    12361 cgtacaaaac gcgtgcggaa gtgaaatttg aaggcgatac cctggtaaac cgcattgagc
    12421 tgaaaggcat tgactttaaa gaggacggca atatcctggg ccataagctg gaatacaatt
    12481 ttaacagcca caatgtttac atcaccgccg ataaacaaaa aaatggcatt aaagcgaatt
    12541 ttaaaattcg ccacaacgtg gaggatggca gcgtgcagct ggctgatcac taccagcaaa
    12601 acactccaat cggtgatggt cctgttctgc tgccagacaa tcactatctg agcacgcaaa
    12661 gcgttctgtc taaagatccg aacgagaaac gcgatcatat ggttctgctg gagttcgtaa
    12721 ccgcagcggg catcacgcat ggtatggatg aactgtacaa atgaccaggc atcaaataaa
    12781 acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc
    12841 tctctactag agtcacactg gctcaccttc gggtgggcct ttctgcgttt atagaggtct
    12901 ctgttttaga gctagaaata gcaagttaaa ataaggctag tccgttatca acttgaaaaa
    12961 gtggcaccga gtcggtgctt tttttcaggt gcacatatac ctgccgttca ctattattta
    13021 gtgaaatgag atattatgat attttctgaa ttgtgattaa aaaggcaact ttatgcccat
    13081 gcaacagaaa ctataaaaaa tacagagaat gaaaagaaac agatagattt tttagttctt
    13141 taggcccgta gtctgcaaat ccttttatga ttttctatca aacaaaagag gaaaatagac
    13201 cagttgcaat ccaaacgaga gtctaataga atgaggtcga aaagtaaatc gcgcgggttt
    13261 gttactgata aagcaggcaa gacctaaaat gtgtaaaggg caaagtgtat actttggcgt
    13321 caccccttac atattttagg tcttttttta ttgtgcgtaa ctaacttgcc atcttcaaac
    13381 aggagggctg gaagaagcag accgctaaca cagtacataa aaaaggagac atgaacgatg
    13441 aacatcaaaa agtttgcaaa acaagcaaca gtattaacct ttactaccgc actgctggca
    13501 ggaggcgcaa ctcaagcgtt tgcgaaagaa acgaaccaaa agccatataa ggaaacatac
    13561 ggcatttccc atattacacg ccatgatatg ctgcaaatcc ctgaacagca aaaaaatgaa
    13621 aaatatcaag ttcctgagtt cgattcgtcc acaattaaaa atatctcttc tgcaaaaggc
    13681 ctggacgttt gggacagctg gccattacaa aacgctgacg gcactgtcgc aaactatcac
    13741 ggctaccaca tcgtctttgc attagccgga gatcctaaaa atgcggatga cacatcgatt
    13801 tacatgttct atcaaaaagt cggcgaaact tctattgaca gctggaaaaa cgctggccgc
    13861 gtctttaaag acagcgacaa attcgatgca aatgattcta tcctaaaaga ccaaacacaa
    13921 gaatggtcag gttcagccac atttacatct gacggaaaaa tccgtttatt ctacactgat
    13981 ttctccggta aacattacgg caaacaaaca ctgacaactg cacaagttaa cgtatcagca
    14041 tcagacagct ctttgaacat caacggtgta gaggattata aatcaatctt tgacggtgac
    14101 ggaaaaacgt atcaaaatgt acagcagttc atcgatgaag gcaactacag ctcaggcgac
    14161 aaccatacgc tgagagatcc tcactacgta gaagataaag gccacaaata cttagtattt
    14221 gaagcaaaca ctggaactga agatggctac caaggcgaag aatctttatt taacaaagca
    14281 tactatggca aaagcacatc attcttccgt caagaaagtc aaaaacttct gcaaagcgat
    14341 aaaaaacgca cggctgagtt agcaaacggc gctctcggta tgattgagct aaacgatgat
    14401 tacacactga aaaaagtgat gaaaccgctg attgcatcta acacagtaac agatgaaatt
    14461 gaacgcgcga acgtctttaa aatgaacggc aaatggtacc tgttcactga ctcccgcgga
    14521 tcaaaaatga cgattgacgg cattacgtct aacgatattt acatgcttgg ttatgtttct
    14581 aattctttaa ctggcccata caagccgctg aacaaaactg gccttgtgtt aaaaatggat
    14641 cttgatccta acgatgtaac ctttacttac tcacacttcg ctgtacctca agcgaaagga
    14701 aacaatgtcg tgattacaag ctatatgaca aacagaggat tctacgcaga caaacaatca
    14761 acgtttgcgc cgagcttcct gctgaacatc aaaggcaaga aaacatctgt tgtcaaagac
    14821 agcatccttg aacaaggaca attaacagtt aacaaataac acctgcagct gtatctagaa
    14881 tcgcgtatgc gagtgtggca cgaaccttgc tgcctgtgat ccagatcatg aacaataaaa
    14941 ctgtctgctt acataaacag taatacaagg ggtgttatga gccatattca acgggaaacg
    15001 tcttgctcta ggccgcgatt aaattccaac atggatgctg atttatatgg gtataaatgg
    15061 gctcgcgata atgtcgggca atcaggtgcg acaatctat
//
