LOCUS       sequence-438346-       14906 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 eforRED for plasmid eviction..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 13701 to 13735)
  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 14906)
  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: pVP144
FEATURES             Location/Qualifiers
     source          1..14906
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             4183..4248
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             4267..4332
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             4339..4962
                     /codon_start=1
                     /product="cytidine deaminase from the sea lamprey 
                     Petromyzon marinus (Rogozin et al., 2007)"
                     /label=PmCDA1
                     /translation="MTDAEYVRIHEKLDIYTFKKQFFNNKKSVSHRCYVLFELKRRGER
                     RACFWGYAVNKPQSGTERGIHAEIFSIRKVEEYLRDNPGQFTINWYSSWSPCADCAEKI
                     LEWYNQELRGNGHTLKIWACKLYYEKNARNQIGLWNLRDNGVGLNVMVSEHYQCCRKIF
                     IQSSHNQLNENRWLEKTLKRAEKRRSELSIMIQVKILHTTKSPAV"
     CDS             4969..5220
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="MTNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHT
                     AYDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     promoter        5292..5326
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     RBS             5482..5493
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             5500..6213
                     /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      6225..6296
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      6312..6339
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_RNA        6355..6430
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        6444..6889
                     /gene="sacR"
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             join(6890..6976,6977..8311)
                     /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"
     CDS             8626..9417
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      9663..10250
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10151..10170
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10404..10421
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    10436..10576
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(10502..10521)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     10662..10684
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     rep_origin      10920..11114
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(11180..12253)
                     /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(12682..13311)
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     promoter        13701..13735
                     /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             13747..13758
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      14465..14511
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             join(14778..14804,14805..14807,14808..14906,1..2388,
                     2389..2391,2392..3975)
                     /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"
ORIGIN
        1 aaaaaaaatc ttataggggc tcttttattt gacagtggag agacagcgga agcgactcgt
       61 ctcaaacgga cagctcgtag aaggtataca cgtcggaaga atcgtatttg ttatctacag
      121 gagatttttt caaatgagat ggcgaaagta gatgatagtt tctttcatcg acttgaagag
      181 tcttttttgg tggaagaaga caagaagcat gaacgtcatc ctatttttgg aaatatagta
      241 gatgaagttg cttatcatga gaaatatcca actatctatc atctgcgaaa aaaattggta
      301 gattctactg ataaagcgga tttgcgctta atctatttgg ccttagcgca tatgattaag
      361 tttcgtggtc attttttgat tgagggagat ttaaatcctg ataatagtga tgtggacaaa
      421 ctatttatcc agttggtaca aacctacaat caattatttg aagaaaaccc tattaacgca
      481 agtggagtag atgctaaagc gattctttct gcacgattga gtaaatcaag acgattagaa
      541 aatctcattg ctcagctccc cggtgagaag aaaaatggct tatttgggaa tctcattgct
      601 ttgtcattgg gtttgacccc taattttaaa tcaaattttg atttggcaga agatgctaaa
      661 ttacagcttt caaaagatac ttacgatgat gatttagata atttattggc gcaaattgga
      721 gatcaatatg ctgatttgtt tttggcagct aagaatttat cagatgctat tttactttca
      781 gatatcctaa gagtaaatac tgaaataact aaggctcccc tatcagcttc aatgattaaa
      841 cgctacgatg aacatcatca agacttgact cttttaaaag ctttagttcg acaacaactt
      901 ccagaaaagt ataaagaaat cttttttgat caatcaaaaa acggatatgc aggttatatt
      961 gatgggggag ctagccaaga agaattttat aaatttatca aaccaatttt agaaaaaatg
     1021 gatggtactg aggaattatt ggtgaaacta aatcgtgaag atttgctgcg caagcaacgg
     1081 acctttgaca acggctctat tccccatcaa attcacttgg gtgagctgca tgctattttg
     1141 agaagacaag aagactttta tccattttta aaagacaatc gtgagaagat tgaaaaaatc
     1201 ttgacttttc gaattcctta ttatgttggt ccattggcgc gtggcaatag tcgttttgca
     1261 tggatgactc ggaagtctga agaaacaatt accccatgga attttgaaga agttgtcgat
     1321 aaaggtgctt cagctcaatc atttattgaa cgcatgacaa actttgataa aaatcttcca
     1381 aatgaaaaag tactaccaaa acatagtttg ctttatgagt attttacggt ttataacgaa
     1441 ttgacaaagg tcaaatatgt tactgaagga atgcgaaaac cagcatttct ttcaggtgaa
     1501 cagaagaaag ccattgttga tttactcttc aaaacaaatc gaaaagtaac cgttaagcaa
     1561 ttaaaagaag attatttcaa aaaaatagaa tgttttgata gtgttgaaat ttcaggagtt
     1621 gaagatagat ttaatgcttc attaggtacc taccatgatt tgctaaaaat tattaaagat
     1681 aaagattttt tggataatga agaaaatgaa gatatcttag aggatattgt tttaacattg
     1741 accttatttg aagataggga gatgattgag gaaagactta aaacatatgc tcacctcttt
     1801 gatgataagg tgatgaaaca gcttaaacgt cgccgttata ctggttgggg acgtttgtct
     1861 cgaaaattga ttaatggtat tagggataag caatctggca aaacaatatt agattttttg
     1921 aaatcagatg gttttgccaa tcgcaatttt atgcagctga tccatgatga tagtttgaca
     1981 tttaaagaag acattcaaaa agcacaagtg tctggacaag gcgatagttt acatgaacat
     2041 attgcaaatt tagctggtag ccctgctatt aaaaaaggta ttttacagac tgtaaaagtt
     2101 gttgatgaat tggtcaaagt aatggggcgg cataagccag aaaatatcgt tattgaaatg
     2161 gcacgtgaaa atcagacaac tcaaaagggc cagaaaaatt cgcgagagcg tatgaaacga
     2221 atcgaagaag gtatcaaaga attaggaagt cagattctta aagagcatcc tgttgaaaat
     2281 actcaattgc aaaatgaaaa gctctatctc tattatctcc aaaatggaag agacatgtat
     2341 gtggaccaag aattagatat taatcgttta agtgattatg atgtcgatgc cattgttcca
     2401 caaagtttcc ttaaagacga ttcaatagac aataaggtct taacgcgttc tgataaaaat
     2461 cgtggtaaat cggataacgt tccaagtgaa gaagtagtca aaaagatgaa aaactattgg
     2521 agacaacttc taaacgccaa gttaatcact caacgtaagt ttgataattt aacgaaagct
     2581 gaacgtggag gtttgagtga acttgataaa gctggtttta tcaaacgcca attggttgaa
     2641 actcgccaaa tcactaagca tgtggcacaa attttggata gtcgcatgaa tactaaatac
     2701 gatgaaaatg ataaacttat tcgagaggtt aaagtgatta ccttaaaatc taaattagtt
     2761 tctgacttcc gaaaagattt ccaattctat aaagtacgtg agattaacaa ttaccatcat
     2821 gcccatgatg cgtatctaaa tgccgtcgtt ggaactgctt tgattaagaa atatccaaaa
     2881 cttgaatcgg agtttgtcta tggtgattat aaagtttatg atgttcgtaa aatgattgct
     2941 aagtctgagc aagaaatagg caaagcaacc gcaaaatatt tcttttactc taatatcatg
     3001 aacttcttca aaacagaaat tacacttgca aatggagaga ttcgcaaacg ccctctaatc
     3061 gaaactaatg gggaaactgg agaaattgtc tgggataaag ggcgagattt tgccacagtg
     3121 cgcaaagtat tgtccatgcc ccaagtcaat attgtcaaga aaacagaagt acagacaggc
     3181 ggattctcca aggagtcaat tttaccaaaa agaaattcgg acaagcttat tgctcgtaaa
     3241 aaagactggg atccaaaaaa atatggtggt tttgatagtc caacggtagc ttattcagtc
     3301 ctagtggttg ctaaggtgga aaaagggaaa tcgaagaagt taaaatccgt taaagagtta
     3361 ctagggatca caattatgga aagaagttcc tttgaaaaaa atccgattga ctttttagaa
     3421 gctaaaggat ataaggaagt taaaaaagac ttaatcatta aactacctaa atatagtctt
     3481 tttgagttag aaaacggtcg taaacggatg ctggctagtg ccggagaatt acaaaaagga
     3541 aatgagctgg ctctgccaag caaatatgtg aattttttat atttagctag tcattatgaa
     3601 aagttgaagg gtagtccaga agataacgaa caaaaacaat tgtttgtgga gcagcataag
     3661 cattatttag atgagattat tgagcaaatc agtgaatttt ctaagcgtgt tattttagca
     3721 gatgccaatt tagataaagt tcttagtgca tataacaaac atagagacaa accaatacgt
     3781 gaacaagcag aaaatattat tcatttattt acgttgacga atcttggagc tcccgctgct
     3841 tttaaatatt ttgatacaac aattgatcgt aaacgatata cgtctacaaa agaagtttta
     3901 gatgccactc ttatccatca atccatcact ggtctttatg aaacacgcat tgatttgagt
     3961 cagctaggag gtgacggtgg aggaggttct ggaggtggag gttctgctga gtatgtgcga
     4021 gccctctttg actttaatgg gaatgatgaa gaggatcttc cctttaagaa aggagacatc
     4081 ctgagaatcc gggataagcc tgaggagcag tggtggaatg cagaggacag cgaaggaaag
     4141 agggggatga ttcctgtccc ttacgtggag aagtattccg gagactataa ggaccacgac
     4201 ggagactaca aggatcatga tattgattac aaagacgatg acgataagtc taggctcgag
     4261 tccggagact ataaggacca cgacggagac tacaaggatc atgatattga ttacaaagac
     4321 gatgacgata agtctaggat gaccgacgct gagtacgtga gaatccatga gaagttggac
     4381 atctacacgt ttaagaaaca gtttttcaac aacaaaaaat ccgtgtcgca tagatgctac
     4441 gttctctttg aattaaaacg acggggtgaa cgtagagcgt gtttttgggg ctatgctgtg
     4501 aataaaccac agagcgggac agaacgtggc attcacgccg aaatctttag cattagaaaa
     4561 gtcgaagaat acctgcgcga caaccccgga caattcacga taaattggta ctcatcctgg
     4621 agtccttgtg cagattgcgc tgaaaagatc ttagaatggt ataaccagga gctgcggggg
     4681 aacggccaca ctttgaaaat ctgggcttgc aaactctatt acgagaaaaa tgcgaggaat
     4741 caaattgggc tgtggaacct cagagataac ggggttgggt tgaatgtaat ggtaagtgaa
     4801 cactaccaat gttgcaggaa aatattcatc caatcgtcgc acaatcaatt gaatgagaat
     4861 agatggcttg agaagacttt gaagcgagct gaaaaacgac ggagcgagtt gtccattatg
     4921 attcaggtaa aaatactcca caccactaag agtcctgctg tttctagaat gaccaacctt
     4981 tccgacatca tagagaagga aacaggcaaa cagttggtca tccaagagtc gatactcatg
     5041 cttcctgaag aagttgagga ggtcattggg aataagccgg aaagtgacat tctcgtacac
     5101 actgcgtatg atgagagcac cgatgagaac gtgatgctgc tcacgtcaga tgccccagag
     5161 tacaaaccct gggctctggt gattcaggac tctaatggag agaacaagat caagatgcta
     5221 ggactggttg cataatcaga ataagcaaac cccttgggtt ccctctttag gagtctgagg
     5281 ggttttttgc attgacagct agctcagtcc taggtataat gctagcggag accgaaagtg
     5341 aaacgtgatt tcatgcgtca ttttgaacat tttgtaaatc ttatttaata atgtgtgcgg
     5401 caattcacat ttaatttatg aatgttttct taacatcgcg gcaactcaag aaacggcagg
     5461 ttcggatctt agctactaga gaaagaggag aaatactaga tgcgtaaagg cgaagagctg
     5521 ttcactggtg tcgtccctat tctggtggaa ctggatggtg atgtcaacgg tcataagttt
     5581 tccgtgcgtg gcgagggtga aggtgacgca actaatggta aactgacgct gaagttcatc
     5641 tgtactactg gtaaactgcc ggttccttgg ccgactctgg taacgacgct gacttatggt
     5701 gttcagtgct ttgctcgtta tccggaccat atgaagcagc atgacttctt caagtccgcc
     5761 atgccggaag gctatgtgca ggaacgcacg atttccttta aggatgacgg cacgtacaaa
     5821 acgcgtgcgg aagtgaaatt tgaaggcgat accctggtaa accgcattga gctgaaaggc
     5881 attgacttta aagaggacgg caatatcctg ggccataagc tggaatacaa ttttaacagc
     5941 cacaatgttt acatcaccgc cgataaacaa aaaaatggca ttaaagcgaa ttttaaaatt
     6001 cgccacaacg tggaggatgg cagcgtgcag ctggctgatc actaccagca aaacactcca
     6061 atcggtgatg gtcctgttct gctgccagac aatcactatc tgagcacgca aagcgttctg
     6121 tctaaagatc cgaacgagaa acgcgatcat atggttctgc tggagttcgt aaccgcagcg
     6181 ggcatcacgc atggtatgga tgaactgtac aaatgaccag gcatcaaata aaacgaaagg
     6241 ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctctact
     6301 agagtcacac tggctcacct tcgggtgggc ctttctgcgt ttatagaggt ctctgtttta
     6361 gagctagaaa tagcaagtta aaataaggct agtccgttat caacttgaaa aagtggcacc
     6421 gagtcggtgc tttttttcag gtgcacatat acctgccgtt cactattatt tagtgaaatg
     6481 agatattatg atattttctg aattgtgatt aaaaaggcaa ctttatgccc atgcaacaga
     6541 aactataaaa aatacagaga atgaaaagaa acagatagat tttttagttc tttaggcccg
     6601 tagtctgcaa atccttttat gattttctat caaacaaaag aggaaaatag accagttgca
     6661 atccaaacga gagtctaata gaatgaggtc gaaaagtaaa tcgcgcgggt ttgttactga
     6721 taaagcaggc aagacctaaa atgtgtaaag ggcaaagtgt atactttggc gtcacccctt
     6781 acatatttta ggtctttttt tattgtgcgt aactaacttg ccatcttcaa acaggagggc
     6841 tggaagaagc agaccgctaa cacagtacat aaaaaaggag acatgaacga tgaacatcaa
     6901 aaagtttgca aaacaagcaa cagtattaac ctttactacc gcactgctgg caggaggcgc
     6961 aactcaagcg tttgcgaaag aaacgaacca aaagccatat aaggaaacat acggcatttc
     7021 ccatattaca cgccatgata tgctgcaaat ccctgaacag caaaaaaatg aaaaatatca
     7081 agttcctgag ttcgattcgt ccacaattaa aaatatctct tctgcaaaag gcctggacgt
     7141 ttgggacagc tggccattac aaaacgctga cggcactgtc gcaaactatc acggctacca
     7201 catcgtcttt gcattagccg gagatcctaa aaatgcggat gacacatcga tttacatgtt
     7261 ctatcaaaaa gtcggcgaaa cttctattga cagctggaaa aacgctggcc gcgtctttaa
     7321 agacagcgac aaattcgatg caaatgattc tatcctaaaa gaccaaacac aagaatggtc
     7381 aggttcagcc acatttacat ctgacggaaa aatccgttta ttctacactg atttctccgg
     7441 taaacattac ggcaaacaaa cactgacaac tgcacaagtt aacgtatcag catcagacag
     7501 ctctttgaac atcaacggtg tagaggatta taaatcaatc tttgacggtg acggaaaaac
     7561 gtatcaaaat gtacagcagt tcatcgatga aggcaactac agctcaggcg acaaccatac
     7621 gctgagagat cctcactacg tagaagataa aggccacaaa tacttagtat ttgaagcaaa
     7681 cactggaact gaagatggct accaaggcga agaatcttta tttaacaaag catactatgg
     7741 caaaagcaca tcattcttcc gtcaagaaag tcaaaaactt ctgcaaagcg ataaaaaacg
     7801 cacggctgag ttagcaaacg gcgctctcgg tatgattgag ctaaacgatg attacacact
     7861 gaaaaaagtg atgaaaccgc tgattgcatc taacacagta acagatgaaa ttgaacgcgc
     7921 gaacgtcttt aaaatgaacg gcaaatggta cctgttcact gactcccgcg gatcaaaaat
     7981 gacgattgac ggcattacgt ctaacgatat ttacatgctt ggttatgttt ctaattcttt
     8041 aactggccca tacaagccgc tgaacaaaac tggccttgtg ttaaaaatgg atcttgatcc
     8101 taacgatgta acctttactt actcacactt cgctgtacct caagcgaaag gaaacaatgt
     8161 cgtgattaca agctatatga caaacagagg attctacgca gacaaacaat caacgtttgc
     8221 gccgagcttc ctgctgaaca tcaaaggcaa gaaaacatct gttgtcaaag acagcatcct
     8281 tgaacaagga caattaacag ttaacaaata acacctgcag ctgtatctag aatcgcgtat
     8341 gcgagtgtgg cacgaaccca gtggacataa gcctgttcgg ttcgtaagct gtaatgcaag
     8401 tagcgtatgc gctcacgcaa ctggtccaga accttgaccg aacgcagcgg tggtaacggc
     8461 gcagtggcgg ttttcatggc ttgttatgac tgtttttttg gggtacagtc tatgcctcgg
     8521 gcatccaagc agcaagcgcg ttacgccgtg ggtcgatgtt tgatgttatg gagcagcaac
     8581 gatgttacgc agcagggcag tcgccctaaa acaaagttaa acatcatggg ggaagcggtg
     8641 atcgccgaag tatcgactca actatcagag gtagttggcg tcatcgagcg ccatctcgaa
     8701 ccgacgttgc tggccgtaca tttgtacggc tccgcagtgg atggcggcct gaagccacac
     8761 agtgatattg atttgctggt tacggtgacc gtaaggcttg atgaaacaac gcggcgagct
     8821 ttgatcaacg accttttgga aacttcggct tcccctggag agagcgagat tctccgcgct
     8881 gtagaagtca ccattgttgt gcacgacgac atcattccgt ggcgttatcc agctaagcgc
     8941 gaactgcaat ttggagaatg gcagcgcaat gacattcttg caggtatctt cgagccagcc
     9001 acgatcgaca ttgatctggc tatcttgctg acaaaagcaa gagaacatag cgttgccttg
     9061 gtaggtccag cggcggagga actctttgat ccggttcctg aacaggatct atttgaggcg
     9121 ctaaatgaaa ccttaacgct atggaactcg ccgcccgact gggctggcga tgagcgaaat
     9181 gtagtgctta cgttgtcccg catttggtac agcgcagtaa ccggcaaaat cgcgccgaag
     9241 gatgtcgctg ccgactgggc aatggagcgc ctgccggccc agtatcagcc cgtcatactt
     9301 gaagctagac aggcttatct tggacaagaa gaagatcgct tggcctcgcg cgcagatcag
     9361 ttggaagaat ttgtccacta cgtgaaaggc gagatcacca aggtagtcgg caaataatgt
     9421 ctagctagaa attcgttcaa gccgacgccg cttcgcggcg cggcttaact caagcgttag
     9481 atgcactaag cacataattg ctcacagcca aactatcagg tcaagtctgc ttttattatt
     9541 tttaagcgtg cataataagc cctacacaaa ttgggagata tatcatgcat gaccaaaatc
     9601 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     9661 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa ccaccgctac
     9721 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     9781 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     9841 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     9901 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     9961 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
    10021 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
    10081 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
    10141 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
    10201 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
    10261 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
    10321 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
    10381 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga
    10441 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatatgg tgcactctca
    10501 gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat cgctacgtga
    10561 ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
    10621 tctgctcccg gcatccgctt acagacaagc tgtgaccggc tccgggagct gcatgtgtca
    10681 gaggttttca ccgtcatcac cgaaacgcgc gaggcagggt gccttgatgt gggcgccggc
    10741 ggtcgagtgg cgacggcgcg gcttgtccgc gccctggtag attgcctggc cgtaggccag
    10801 ccatttttga gcggccagcg gccgcgatag gccgacgcga agcggcgggg cgtagggagc
    10861 gcagcgaccg aagggtaggc gctttttgca gctcttcggc tgtgcgctgg ccagacagtt
    10921 atgcacaggc caggcgggtt ttaagagttt taataagttt taaagagttt taggcggaaa
    10981 aatcgccttt tttctctttt atatcagtca cttacatgtg tgaccggttc ccaatgtacg
    11041 gctttgggtt cccaatgtac gggttccggt tcccaatgta cggctttggg ttcccaatgt
    11101 acgtgctatc cacaggaaag agagcttttc gacctttttc ccctgctagg gcaatttgcc
    11161 ctagcatctg ctccgtacat taggaaccgg cggatgcttc gccctcgatc aggttgcggt
    11221 agcgcatgac taggatcggg ccagcctgcc ccgcctcctc cttcaaatcg tactccggca
    11281 ggtcatttga cccgatcagc ttgcgcacgg tgaaacagaa cttcttgaac tctccggcgc
    11341 tgccactgcg ttcgtagatc gtcttgaaca accatctggc ttctgccttg cctgcggcgc
    11401 ggcgtgccag gcggtagaga aaacggccga tgccgggatc gatcaaaaag taatcggggt
    11461 gaaccgtcag cacgtccggg ttcttgcctt ctgtgatctc gcggtacatc caatcagcta
    11521 gctcgatctc gatgtactcc ggccgcccgg tttcgctctt tacgatcttg tagcggctaa
    11581 tcaaggcttc accctcggat accgtcacca ggcggccgtt cttggccttc ttcgtacgct
    11641 gcatggcaac gtgcgtggtg tttaaccgaa tgcaggtttc taccaggtcg tctttctgct
    11701 ttccgccatc ggctcgccgg cagaacttga gtacgtccgc aacgtgtgga cggaacacgc
    11761 ggccgggctt gtctcccttc ccttcccggt atcggttcat ggattcggtt agatgggaaa
    11821 ccgccatcag taccaggtcg taatcccaca cactggccat gccggccggc cctgcggaaa
    11881 cctctacgtg cccgtctgga agctcgtagc ggatcacctc gccagctcgt cggtcacgct
    11941 tcgacagacg gaaaacggcc acgtccatga tgctgcgact atcgcgggtg cccacgtcat
    12001 agagcatcgg aacgaaaaaa tctggttgct cgtcgccctt gggcggcttc ctaatcgacg
    12061 gcgcaccggc tgccggcggt tgccgggatt ctttgcggat tcgatcagcg gccgcttgcc
    12121 acgattcacc ggggcgtgct tctgcctcga tgcgttgccg ctgggcggcc tgcgcggcct
    12181 tcaacttctc caccaggtca tcacccagcg ccgcgccgat ttgtaccggg ccggatggtt
    12241 tgcgaccgct cacgccgatt cctcgggctt gggggttcca gtgccattgc agggccggca
    12301 gacaacccag ccgcttacgc ctggccaacc gcccgttcct ccacacatgg ggcattccac
    12361 ggcgtcggtg cctggttgtt cttgattttc catgccgcct cctttagccg ctaaaattca
    12421 tctactcatt tattcatttg ctcatttact ctggtagctg cgcgatgtat tcagatagca
    12481 gctcggtaat ggtcttgcct tggcgtaccg cgtacatctt cagcttggtg tgatcctccg
    12541 ccggcaactg aaagttgacc cgcttcatgg ctggcgtgtc tgccaggctg gccaacgttg
    12601 cagccttgct gctgcgtgcg ctcggacggc cggcacttag cgtgtttgtg cttttgctca
    12661 ttttctcttt acctcattaa ctcaaatgag ttttgattta atttcagcgg ccagcgcctg
    12721 gacctcgcgg gcagcgtcgc cctcgggttc tgattcaaga acggttgtgc cggcggcggc
    12781 agtgcctggg tagctcacgc gctgcgtgat acgggactca agaatgggca gctcgtaccc
    12841 ggccagcgcc tcggcaacct caccgccgat gcgcgtgcct ttgatcgccc gcgacacgac
    12901 aaaggccgct tgtagccttc catccgtgac ctcaatgcgc tgcttaacca gctccaccag
    12961 gtcggcggtg gcccatatgt cgtaagggct tggctgcacc ggaatcagca cgaagtcggc
    13021 tgccttgatc gcggacacag ccaagtccgc cgcctggggc gctccgtcga tcactacgaa
    13081 gtcgcgccgg ccgatggcct tcacgtcgcg gtcaatcgtc gggcggtcga tgccgacaac
    13141 ggttagcggt tgatcttccc gcacggccgc ccaatcgcgg gcactgccct ggggatcgga
    13201 atcgactaac agaacatcgg ccccggcgag ttgcagggcg cgggctagat gggttgcgat
    13261 ggtcgtcttg cctgacccgc ctttctggtt aagtacagcg ataaccttca tgcgttcccc
    13321 ttgcgtattt gtttatttac tcatcgcatc atatacgcag cgaccgcatg acgcaagctg
    13381 ttttactcaa atacacatca cctttttaga cggcggcgct cggtttcttc agcggccaag
    13441 ctggccggcc aggccgccag cttggcatca gacaaaccgg ccaggatttc atgcagccgc
    13501 acggttgata cgtgcgcggg cggctcgaac acgtacccgg ccgcgatcat ctccgcctcg
    13561 atctcttcgg taatgaaaaa cggttcgtcc tggccgtcct ggtgcggttt catgcttgtt
    13621 cctcttggcg ttcattctcg gcggccgcca gggcgtcggc ctcggtcaat gcgtcctcac
    13681 ggaaggcacc gcgccgcctg ttgacggcta gctcagtcct aggtacagtg ctagctacta
    13741 gagattaaag aggagaaaga gtcccatggc cgtcatcaag cagatcatga agaccaagct
    13801 gcacctggaa ggcaccgtca acggccacga cttcaccatc gaaggcaagg gcgaaggcaa
    13861 gccgtacgaa ggcctgcagc acatgaagat gaccgtcacc aagggcgccc cgctgccgtt
    13921 ctccgtccac atcctgaccc cgtcccacat gtacggctcc aagccgttca acaagtaccc
    13981 ggccgacatc ccggactacc acaagcagtc cttcccggaa ggcatgtcct gggaacgctc
    14041 catgatcttc gaagacggcg gcgtctgcac cgcctccaac cactcctcca tcaacctgca
    14101 ggaaaactgc ttcatctacg acgtcaagtt ccacggcgtc aacctgccgc cggacggccc
    14161 ggtcatgcag aagaccatcg ccggctggga accgtccgtc gaaaccctgt acgtccgcga
    14221 cggcatgctg aagtccgaca ccgccatggt cttcaagctg aagggcggcg gccaccaccg
    14281 cgtcgacttc aagaccacct acaaggccaa gaagccggtc aagctgccgg aattccactt
    14341 cgtcgaacac cgcctggaac tgaccaagca cgacaaggac ttcaccacct gggaccagca
    14401 ggaagccgcc gaaggccact tctccccgct gccgaaggcc ctgccgtaat agagctcagg
    14461 acatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt
    14521 acgagcattt tcgttctgag cctgacctga ccaactgcgg cctgaatagg tcgatccggt
    14581 tgcttagtca tggatgcgcg gttctcggtc catgttgcgt tccaagacgc cgggcgaggt
    14641 ttctcgcttc aattgaaatc ataaagaagc aattgaaaat tttcgagtaa ccgaccctcc
    14701 cgataatctt caacataaaa caacgcactt cttccaacgg gagaggcggt gttagttgcg
    14761 agctaaggag aggctccatg gataagaaat actcaatagg cttagctatc ggcacaaata
    14821 gcgtcggatg ggcggtgatc actgatgaat ataaggttcc gtctaaaaag ttcaaggttc
    14881 tgggaaatac agaccgccac agtatc
//
