LOCUS       sequence-438349-       15649 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..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15649)
  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 15649)
  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: pVP141
FEATURES             Location/Qualifiers
     source          1..15649
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1313..1901
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1802..1821
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2055..2072
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2087..2227
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2153..2172)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2313..2335
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     rep_origin      2571..2765
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(2831..3904)
                     /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(4333..4962)
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     misc_feature    6293..6317
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             join(6636..6662,6663..6665,6666..9152,9153..9155,
                     9156..10739)
                     /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             10947..11012
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             11031..11096
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             11103..11726
                     /codon_start=1
                     /product="cytidine deaminase from the sea lamprey 
                     Petromyzon marinus (Rogozin et al., 2007)"
                     /label=PmCDA1
                     /translation="MTDAEYVRIHEKLDIYTFKKQFFNNKKSVSHRCYVLFELKRRGER
                     RACFWGYAVNKPQSGTERGIHAEIFSIRKVEEYLRDNPGQFTINWYSSWSPCADCAEKI
                     LEWYNQELRGNGHTLKIWACKLYYEKNARNQIGLWNLRDNGVGLNVMVSEHYQCCRKIF
                     IQSSHNQLNENRWLEKTLKRAEKRRSELSIMIQVKILHTTKSPAV"
     CDS             11733..11984
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="MTNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHT
                     AYDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     promoter        12056..12090
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     RBS             12246..12257
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             12264..12977
                     /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      12989..13060
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      13076..13103
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_RNA        13119..13194
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        13208..13653
                     /gene="sacR"
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             join(13654..13740,13741..15075)
                     /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"
     misc_feature    15234..15258
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     primer_bind     complement(15540..15559)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             join(15551..15649,1..717)
                     /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(15551..15649,1..717)
                     /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(15624..15643)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 gggcaatcag gtgcgacaat ctatcgattg tatgggaagc ccgatgcgcc agagttgttt
       61 ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt cagactaaac
      121 tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac tcctgatgat
      181 gcatggttac tcaccactgc gatccccggg aaaacagcat tccaggtatt agaagaatat
      241 cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg
      301 attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa
      361 tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg taatggctgg
      421 cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc ggattcagtc
      481 gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa attaataggt
      541 tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc catcctatgg
      601 aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa atatggtatt
      661 gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt tttctaacag
      721 cggcggagga actctttgat ccggttcctg aacaggatct atttgaggcg ctaaatgaaa
      781 ccttaacgct atggaactcg ccgcccgact gggctggcga tgagcgaaat gtagtgctta
      841 cgttgtcccg catttggtac agcgcagtaa ccggcaaaat cgcgccgaag gatgtcgctg
      901 ccgactgggc aatggagcgc ctgccggccc agtatcagcc cgtcatactt gaagctagac
      961 aggcttatct tggacaagaa gaagatcgct tggcctcgcg cgcagatcag ttggaagaat
     1021 ttgtccacta cgtgaaaggc gagatcacca aggtagtcgg caaataatgt ctagctagaa
     1081 attcgttcaa gccgacgccg cttcgcggcg cggcttaact caagcgttag atgcactaag
     1141 cacataattg ctcacagcca aactatcagg tcaagtctgc ttttattatt tttaagcgtg
     1201 cataataagc cctacacaaa ttgggagata tatcatgcat gaccaaaatc ccttaacgtg
     1261 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     1321 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     1381 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     1441 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
     1501 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     1561 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     1621 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     1681 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     1741 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     1801 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     1861 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     1921 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     1981 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     2041 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt
     2101 ttctccttac gcatctgtgc ggtatttcac accgcatatg gtgcactctc agtacaatct
     2161 gctctgatgc cgcatagtta agccagtata cactccgcta tcgctacgtg actgggtcat
     2221 ggctgcgccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     2281 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     2341 accgtcatca ccgaaacgcg cgaggcaggg tgccttgatg tgggcgccgg cggtcgagtg
     2401 gcgacggcgc ggcttgtccg cgccctggta gattgcctgg ccgtaggcca gccatttttg
     2461 agcggccagc ggccgcgata ggccgacgcg aagcggcggg gcgtagggag cgcagcgacc
     2521 gaagggtagg cgctttttgc agctcttcgg ctgtgcgctg gccagacagt tatgcacagg
     2581 ccaggcgggt tttaagagtt ttaataagtt ttaaagagtt ttaggcggaa aaatcgcctt
     2641 ttttctcttt tatatcagtc acttacatgt gtgaccggtt cccaatgtac ggctttgggt
     2701 tcccaatgta cgggttccgg ttcccaatgt acggctttgg gttcccaatg tacgtgctat
     2761 ccacaggaaa gagagctttt cgaccttttt cccctgctag ggcaatttgc cctagcatct
     2821 gctccgtaca ttaggaaccg gcggatgctt cgccctcgat caggttgcgg tagcgcatga
     2881 ctaggatcgg gccagcctgc cccgcctcct ccttcaaatc gtactccggc aggtcatttg
     2941 acccgatcag cttgcgcacg gtgaaacaga acttcttgaa ctctccggcg ctgccactgc
     3001 gttcgtagat cgtcttgaac aaccatctgg cttctgcctt gcctgcggcg cggcgtgcca
     3061 ggcggtagag aaaacggccg atgccgggat cgatcaaaaa gtaatcgggg tgaaccgtca
     3121 gcacgtccgg gttcttgcct tctgtgatct cgcggtacat ccaatcagct agctcgatct
     3181 cgatgtactc cggccgcccg gtttcgctct ttacgatctt gtagcggcta atcaaggctt
     3241 caccctcgga taccgtcacc aggcggccgt tcttggcctt cttcgtacgc tgcatggcaa
     3301 cgtgcgtggt gtttaaccga atgcaggttt ctaccaggtc gtctttctgc tttccgccat
     3361 cggctcgccg gcagaacttg agtacgtccg caacgtgtgg acggaacacg cggccgggct
     3421 tgtctccctt cccttcccgg tatcggttca tggattcggt tagatgggaa accgccatca
     3481 gtaccaggtc gtaatcccac acactggcca tgccggccgg ccctgcggaa acctctacgt
     3541 gcccgtctgg aagctcgtag cggatcacct cgccagctcg tcggtcacgc ttcgacagac
     3601 ggaaaacggc cacgtccatg atgctgcgac tatcgcgggt gcccacgtca tagagcatcg
     3661 gaacgaaaaa atctggttgc tcgtcgccct tgggcggctt cctaatcgac ggcgcaccgg
     3721 ctgccggcgg ttgccgggat tctttgcgga ttcgatcagc ggccgcttgc cacgattcac
     3781 cggggcgtgc ttctgcctcg atgcgttgcc gctgggcggc ctgcgcggcc ttcaacttct
     3841 ccaccaggtc atcacccagc gccgcgccga tttgtaccgg gccggatggt ttgcgaccgc
     3901 tcacgccgat tcctcgggct tgggggttcc agtgccattg cagggccggc agacaaccca
     3961 gccgcttacg cctggccaac cgcccgttcc tccacacatg gggcattcca cggcgtcggt
     4021 gcctggttgt tcttgatttt ccatgccgcc tcctttagcc gctaaaattc atctactcat
     4081 ttattcattt gctcatttac tctggtagct gcgcgatgta ttcagatagc agctcggtaa
     4141 tggtcttgcc ttggcgtacc gcgtacatct tcagcttggt gtgatcctcc gccggcaact
     4201 gaaagttgac ccgcttcatg gctggcgtgt ctgccaggct ggccaacgtt gcagccttgc
     4261 tgctgcgtgc gctcggacgg ccggcactta gcgtgtttgt gcttttgctc attttctctt
     4321 tacctcatta actcaaatga gttttgattt aatttcagcg gccagcgcct ggacctcgcg
     4381 ggcagcgtcg ccctcgggtt ctgattcaag aacggttgtg ccggcggcgg cagtgcctgg
     4441 gtagctcacg cgctgcgtga tacgggactc aagaatgggc agctcgtacc cggccagcgc
     4501 ctcggcaacc tcaccgccga tgcgcgtgcc tttgatcgcc cgcgacacga caaaggccgc
     4561 ttgtagcctt ccatccgtga cctcaatgcg ctgcttaacc agctccacca ggtcggcggt
     4621 ggcccatatg tcgtaagggc ttggctgcac cggaatcagc acgaagtcgg ctgccttgat
     4681 cgcggacaca gccaagtccg ccgcctgggg cgctccgtcg atcactacga agtcgcgccg
     4741 gccgatggcc ttcacgtcgc ggtcaatcgt cgggcggtcg atgccgacaa cggttagcgg
     4801 ttgatcttcc cgcacggccg cccaatcgcg ggcactgccc tggggatcgg aatcgactaa
     4861 cagaacatcg gccccggcga gttgcagggc gcgggctaga tgggttgcga tggtcgtctt
     4921 gcctgacccg cctttctggt taagtacagc gataaccttc atgcgttccc cttgcgtatt
     4981 tgtttattta ctcatcgcat catatacgca gcgaccgcat gacgcaagct gttttactca
     5041 aatacacatc acctttttag acggcggcgc tcggtttctt cagcggccaa gctggccggc
     5101 caggccgcca gcttggcatc agacaaaccg gccaggattt catgcagccg cacggttgag
     5161 acgtgcgcgg gcggctcgaa cacgtacccg gccgcgatca tctccgcctc gatctcttcg
     5221 gtaatgaaaa acggttcgtc ctggccgtcc tggtgcggtt tcatgcttgt tcctcttggc
     5281 gttcattctc ggcggccgcc agggcgtcgg cctcggtcaa tgcgtcctca cggaaggcac
     5341 cgcgccgcct ggcctcggtg ggcgtcactt cctcgctgcg ctcaagtgcg cggtacaggg
     5401 tcgagcgatg cacgccaagc agtgcagccg cctctttcac ggtgcggcct tcctggtcga
     5461 tcagctcgcg ggcgtgcgcg atctgtgccg gggtgagggt agggcggggg ccaaacttca
     5521 cgcctcgggc cttggcggcc tcgcgcccgc tccgggtgcg gtcgatgatt agggaacgct
     5581 cgaactcggc aatgccggcg aacacggtca acaccatgcg gccggccggc gtggtggtgt
     5641 cggcccacgg ctctgccagg ctacgcaggc ccgcgccggc ctcctggatg cgctcggcaa
     5701 tgtccagtag gtcgcgggtg ctgcgggcca ggcggtctag cctggtcact gtcacaacgt
     5761 cgccagggcg taggtggtca agcatcctgg ccagctccgg gcggtcgcgc ctggtgccgg
     5821 tgatcttctc ggaaaacagc ttggtgcagc cggccgcgtg cagttcggcc cgttggttgg
     5881 tcaagtcctg gtcgtcggtg ctgacgcggg catagcccag caggccagcg gcggcgctct
     5941 tgttcatggc gtaatgtctc cggttctagt cgcaagtatt ctactttatg cgactaaaac
     6001 acgcgacaag aaaacgccag gaaaagggca gggcggcagc ctgtcgcgta acttaggact
     6061 tgtgcgacat gtcgttttca gaagagggct gcactgaacg tcagaagccg actgcactat
     6121 agcagcggag gggttggatc aaagtacttt aaagtacttt aaagtacttt aaagtacttt
     6181 gatcccgagg ggaaccctgt ggttggcatg cacatacaaa tggacgaacg gataaacctt
     6241 ttcacgccct tttaaatatc cgattattct aataaacgct cttttctctt aggtttaccc
     6301 gccaatatat cctgtcaaac actgatagtt taaactgaag gcgggaaacg acaatctgat
     6361 ccaagctcaa gctaagcttc gagcattttc gttctgagcc tgacctgacc aactgcggcc
     6421 tgaataggtc gatccggttg cttagtcatg gatgcgcggt tctcggtcca tgttgcgttc
     6481 caagacgccg ggcgaggttt ctcgcttcaa ttgaaatcat aaagaagcaa ttgaaaattt
     6541 tcgagtaacc gaccctcccg ataatcttca acataaaaca acgcacttct tccaacggga
     6601 gaggcggtgt tagttgcgag ctaaggagag gctccatgga taagaaatac tcaataggct
     6661 tagctatcgg cacaaatagc gtcggatggg cggtgatcac tgatgaatat aaggttccgt
     6721 ctaaaaagtt caaggttctg ggaaatacag accgccacag tatcaaaaaa aatcttatag
     6781 gggctctttt atttgacagt ggagagacag cggaagcgac tcgtctcaaa cggacagctc
     6841 gtagaaggta tacacgtcgg aagaatcgta tttgttatct acaggagatt ttttcaaatg
     6901 agatggcgaa agtagatgat agtttctttc atcgacttga agagtctttt ttggtggaag
     6961 aagacaagaa gcatgaacgt catcctattt ttggaaatat agtagatgaa gttgcttatc
     7021 atgagaaata tccaactatc tatcatctgc gaaaaaaatt ggtagattct actgataaag
     7081 cggatttgcg cttaatctat ttggccttag cgcatatgat taagtttcgt ggtcattttt
     7141 tgattgaggg agatttaaat cctgataata gtgatgtgga caaactattt atccagttgg
     7201 tacaaaccta caatcaatta tttgaagaaa accctattaa cgcaagtgga gtagatgcta
     7261 aagcgattct ttctgcacga ttgagtaaat caagacgatt agaaaatctc attgctcagc
     7321 tccccggtga gaagaaaaat ggcttatttg ggaatctcat tgctttgtca ttgggtttga
     7381 cccctaattt taaatcaaat tttgatttgg cagaagatgc taaattacag ctttcaaaag
     7441 atacttacga tgatgattta gataatttat tggcgcaaat tggagatcaa tatgctgatt
     7501 tgtttttggc agctaagaat ttatcagatg ctattttact ttcagatatc ctaagagtaa
     7561 atactgaaat aactaaggct cccctatcag cttcaatgat taaacgctac gatgaacatc
     7621 atcaagactt gactctttta aaagctttag ttcgacaaca acttccagaa aagtataaag
     7681 aaatcttttt tgatcaatca aaaaacggat atgcaggtta tattgatggg ggagctagcc
     7741 aagaagaatt ttataaattt atcaaaccaa ttttagaaaa aatggatggt actgaggaat
     7801 tattggtgaa actaaatcgt gaagatttgc tgcgcaagca acggaccttt gacaacggct
     7861 ctattcccca tcaaattcac ttgggtgagc tgcatgctat tttgagaaga caagaagact
     7921 tttatccatt tttaaaagac aatcgtgaga agattgaaaa aatcttgact tttcgaattc
     7981 cttattatgt tggtccattg gcgcgtggca atagtcgttt tgcatggatg actcggaagt
     8041 ctgaagaaac aattacccca tggaattttg aagaagttgt cgataaaggt gcttcagctc
     8101 aatcatttat tgaacgcatg acaaactttg ataaaaatct tccaaatgaa aaagtactac
     8161 caaaacatag tttgctttat gagtatttta cggtttataa cgaattgaca aaggtcaaat
     8221 atgttactga aggaatgcga aaaccagcat ttctttcagg tgaacagaag aaagccattg
     8281 ttgatttact cttcaaaaca aatcgaaaag taaccgttaa gcaattaaaa gaagattatt
     8341 tcaaaaaaat agaatgtttt gatagtgttg aaatttcagg agttgaagat agatttaatg
     8401 cttcattagg tacctaccat gatttgctaa aaattattaa agataaagat tttttggata
     8461 atgaagaaaa tgaagatatc ttagaggata ttgttttaac attgacctta tttgaagata
     8521 gggagatgat tgaggaaaga cttaaaacat atgctcacct ctttgatgat aaggtgatga
     8581 aacagcttaa acgtcgccgt tatactggtt ggggacgttt gtctcgaaaa ttgattaatg
     8641 gtattaggga taagcaatct ggcaaaacaa tattagattt tttgaaatca gatggttttg
     8701 ccaatcgcaa ttttatgcag ctgatccatg atgatagttt gacatttaaa gaagacattc
     8761 aaaaagcaca agtgtctgga caaggcgata gtttacatga acatattgca aatttagctg
     8821 gtagccctgc tattaaaaaa ggtattttac agactgtaaa agttgttgat gaattggtca
     8881 aagtaatggg gcggcataag ccagaaaata tcgttattga aatggcacgt gaaaatcaga
     8941 caactcaaaa gggccagaaa aattcgcgag agcgtatgaa acgaatcgaa gaaggtatca
     9001 aagaattagg aagtcagatt cttaaagagc atcctgttga aaatactcaa ttgcaaaatg
     9061 aaaagctcta tctctattat ctccaaaatg gaagagacat gtatgtggac caagaattag
     9121 atattaatcg tttaagtgat tatgatgtcg atgccattgt tccacaaagt ttccttaaag
     9181 acgattcaat agacaataag gtcttaacgc gttctgataa aaatcgtggt aaatcggata
     9241 acgttccaag tgaagaagta gtcaaaaaga tgaaaaacta ttggagacaa cttctaaacg
     9301 ccaagttaat cactcaacgt aagtttgata atttaacgaa agctgaacgt ggaggtttga
     9361 gtgaacttga taaagctggt tttatcaaac gccaattggt tgaaactcgc caaatcacta
     9421 agcatgtggc acaaattttg gatagtcgca tgaatactaa atacgatgaa aatgataaac
     9481 ttattcgaga ggttaaagtg attaccttaa aatctaaatt agtttctgac ttccgaaaag
     9541 atttccaatt ctataaagta cgtgagatta acaattacca tcatgcccat gatgcgtatc
     9601 taaatgccgt cgttggaact gctttgatta agaaatatcc aaaacttgaa tcggagtttg
     9661 tctatggtga ttataaagtt tatgatgttc gtaaaatgat tgctaagtct gagcaagaaa
     9721 taggcaaagc aaccgcaaaa tatttctttt actctaatat catgaacttc ttcaaaacag
     9781 aaattacact tgcaaatgga gagattcgca aacgccctct aatcgaaact aatggggaaa
     9841 ctggagaaat tgtctgggat aaagggcgag attttgccac agtgcgcaaa gtattgtcca
     9901 tgccccaagt caatattgtc aagaaaacag aagtacagac aggcggattc tccaaggagt
     9961 caattttacc aaaaagaaat tcggacaagc ttattgctcg taaaaaagac tgggatccaa
    10021 aaaaatatgg tggttttgat agtccaacgg tagcttattc agtcctagtg gttgctaagg
    10081 tggaaaaagg gaaatcgaag aagttaaaat ccgttaaaga gttactaggg atcacaatta
    10141 tggaaagaag ttcctttgaa aaaaatccga ttgacttttt agaagctaaa ggatataagg
    10201 aagttaaaaa agacttaatc attaaactac ctaaatatag tctttttgag ttagaaaacg
    10261 gtcgtaaacg gatgctggct agtgccggag aattacaaaa aggaaatgag ctggctctgc
    10321 caagcaaata tgtgaatttt ttatatttag ctagtcatta tgaaaagttg aagggtagtc
    10381 cagaagataa cgaacaaaaa caattgtttg tggagcagca taagcattat ttagatgaga
    10441 ttattgagca aatcagtgaa ttttctaagc gtgttatttt agcagatgcc aatttagata
    10501 aagttcttag tgcatataac aaacatagag acaaaccaat acgtgaacaa gcagaaaata
    10561 ttattcattt atttacgttg acgaatcttg gagctcccgc tgcttttaaa tattttgata
    10621 caacaattga tcgtaaacga tatacgtcta caaaagaagt tttagatgcc actcttatcc
    10681 atcaatccat cactggtctt tatgaaacac gcattgattt gagtcagcta ggaggtgacg
    10741 gtggaggagg ttctggaggt ggaggttctg ctgagtatgt gcgagccctc tttgacttta
    10801 atgggaatga tgaagaggat cttcccttta agaaaggaga catcctgaga atccgggata
    10861 agcctgagga gcagtggtgg aatgcagagg acagcgaagg aaagaggggg atgattcctg
    10921 tcccttacgt ggagaagtat tccggagact ataaggacca cgacggagac tacaaggatc
    10981 atgatattga ttacaaagac gatgacgata agtctaggct cgagtccgga gactataagg
    11041 accacgacgg agactacaag gatcatgata ttgattacaa agacgatgac gataagtcta
    11101 ggatgaccga cgctgagtac gtgagaatcc atgagaagtt ggacatctac acgtttaaga
    11161 aacagttttt caacaacaaa aaatccgtgt cgcatagatg ctacgttctc tttgaattaa
    11221 aacgacgggg tgaacgtaga gcgtgttttt ggggctatgc tgtgaataaa ccacagagcg
    11281 ggacagaacg tggcattcac gccgaaatct ttagcattag aaaagtcgaa gaatacctgc
    11341 gcgacaaccc cggacaattc acgataaatt ggtactcatc ctggagtcct tgtgcagatt
    11401 gcgctgaaaa gatcttagaa tggtataacc aggagctgcg ggggaacggc cacactttga
    11461 aaatctgggc ttgcaaactc tattacgaga aaaatgcgag gaatcaaatt gggctgtgga
    11521 acctcagaga taacggggtt gggttgaatg taatggtaag tgaacactac caatgttgca
    11581 ggaaaatatt catccaatcg tcgcacaatc aattgaatga gaatagatgg cttgagaaga
    11641 ctttgaagcg agctgaaaaa cgacggagcg agttgtccat tatgattcag gtaaaaatac
    11701 tccacaccac taagagtcct gctgtttcta gaatgaccaa cctttccgac atcatagaga
    11761 aggaaacagg caaacagttg gtcatccaag agtcgatact catgcttcct gaagaagttg
    11821 aggaggtcat tgggaataag ccggaaagtg acattctcgt acacactgcg tatgatgaga
    11881 gcaccgatga gaacgtgatg ctgctcacgt cagatgcccc agagtacaaa ccctgggctc
    11941 tggtgattca ggactctaat ggagagaaca agatcaagat gctaggactg gttgcataat
    12001 cagaataagc aaaccccttg ggttccctct ttaggagtct gaggggtttt ttgcattgac
    12061 agctagctca gtcctaggta taatgctagc ggagaccgaa agtgaaacgt gatttcatgc
    12121 gtcattttga acattttgta aatcttattt aataatgtgt gcggcaattc acatttaatt
    12181 tatgaatgtt ttcttaacat cgcggcaact caagaaacgg caggttcgga tcttagctac
    12241 tagagaaaga ggagaaatac tagatgcgta aaggcgaaga gctgttcact ggtgtcgtcc
    12301 ctattctggt ggaactggat ggtgatgtca acggtcataa gttttccgtg cgtggcgagg
    12361 gtgaaggtga cgcaactaat ggtaaactga cgctgaagtt catctgtact actggtaaac
    12421 tgccggttcc ttggccgact ctggtaacga cgctgactta tggtgttcag tgctttgctc
    12481 gttatccgga ccatatgaag cagcatgact tcttcaagtc cgccatgccg gaaggctatg
    12541 tgcaggaacg cacgatttcc tttaaggatg acggcacgta caaaacgcgt gcggaagtga
    12601 aatttgaagg cgataccctg gtaaaccgca ttgagctgaa aggcattgac tttaaagagg
    12661 acggcaatat cctgggccat aagctggaat acaattttaa cagccacaat gtttacatca
    12721 ccgccgataa acaaaaaaat ggcattaaag cgaattttaa aattcgccac aacgtggagg
    12781 atggcagcgt gcagctggct gatcactacc agcaaaacac tccaatcggt gatggtcctg
    12841 ttctgctgcc agacaatcac tatctgagca cgcaaagcgt tctgtctaaa gatccgaacg
    12901 agaaacgcga tcatatggtt ctgctggagt tcgtaaccgc agcgggcatc acgcatggta
    12961 tggatgaact gtacaaatga ccaggcatca aataaaacga aaggctcagt cgaaagactg
    13021 ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc tactagagtc acactggctc
    13081 accttcgggt gggcctttct gcgtttatag aggtctctgt tttagagcta gaaatagcaa
    13141 gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg gtgctttttt
    13201 tcaggtgcac atatacctgc cgttcactat tatttagtga aatgagatat tatgatattt
    13261 tctgaattgt gattaaaaag gcaactttat gcccatgcaa cagaaactat aaaaaataca
    13321 gagaatgaaa agaaacagat agatttttta gttctttagg cccgtagtct gcaaatcctt
    13381 ttatgatttt ctatcaaaca aaagaggaaa atagaccagt tgcaatccaa acgagagtct
    13441 aatagaatga ggtcgaaaag taaatcgcgc gggtttgtta ctgataaagc aggcaagacc
    13501 taaaatgtgt aaagggcaaa gtgtatactt tggcgtcacc ccttacatat tttaggtctt
    13561 tttttattgt gcgtaactaa cttgccatct tcaaacagga gggctggaag aagcagaccg
    13621 ctaacacagt acataaaaaa ggagacatga acgatgaaca tcaaaaagtt tgcaaaacaa
    13681 gcaacagtat taacctttac taccgcactg ctggcaggag gcgcaactca agcgtttgcg
    13741 aaagaaacga accaaaagcc atataaggaa acatacggca tttcccatat tacacgccat
    13801 gatatgctgc aaatccctga acagcaaaaa aatgaaaaat atcaagttcc tgagttcgat
    13861 tcgtccacaa ttaaaaatat ctcttctgca aaaggcctgg acgtttggga cagctggcca
    13921 ttacaaaacg ctgacggcac tgtcgcaaac tatcacggct accacatcgt ctttgcatta
    13981 gccggagatc ctaaaaatgc ggatgacaca tcgatttaca tgttctatca aaaagtcggc
    14041 gaaacttcta ttgacagctg gaaaaacgct ggccgcgtct ttaaagacag cgacaaattc
    14101 gatgcaaatg attctatcct aaaagaccaa acacaagaat ggtcaggttc agccacattt
    14161 acatctgacg gaaaaatccg tttattctac actgatttct ccggtaaaca ttacggcaaa
    14221 caaacactga caactgcaca agttaacgta tcagcatcag acagctcttt gaacatcaac
    14281 ggtgtagagg attataaatc aatctttgac ggtgacggaa aaacgtatca aaatgtacag
    14341 cagttcatcg atgaaggcaa ctacagctca ggcgacaacc atacgctgag agatcctcac
    14401 tacgtagaag ataaaggcca caaatactta gtatttgaag caaacactgg aactgaagat
    14461 ggctaccaag gcgaagaatc tttatttaac aaagcatact atggcaaaag cacatcattc
    14521 ttccgtcaag aaagtcaaaa acttctgcaa agcgataaaa aacgcacggc tgagttagca
    14581 aacggcgctc tcggtatgat tgagctaaac gatgattaca cactgaaaaa agtgatgaaa
    14641 ccgctgattg catctaacac agtaacagat gaaattgaac gcgcgaacgt ctttaaaatg
    14701 aacggcaaat ggtacctgtt cactgactcc cgcggatcaa aaatgacgat tgacggcatt
    14761 acgtctaacg atatttacat gcttggttat gtttctaatt ctttaactgg cccatacaag
    14821 ccgctgaaca aaactggcct tgtgttaaaa atggatcttg atcctaacga tgtaaccttt
    14881 acttactcac acttcgctgt acctcaagcg aaaggaaaca atgtcgtgat tacaagctat
    14941 atgacaaaca gaggattcta cgcagacaaa caatcaacgt ttgcgccgag cttcctgctg
    15001 aacatcaaag gcaagaaaac atctgttgtc aaagacagca tccttgaaca aggacaatta
    15061 acagttaaca aataacacct gcagctgtat ctagaatcgc gtatgcgagt gtctgcaggc
    15121 atgcgacgtc gggccctcta gaggatcccc gggtaccgcg aattcaattc ggcgttaatt
    15181 cagtacatta aaaacgtccg caatgtgtta ttaagttgtc taagcgtcaa tttgtttaca
    15241 ccacaatata tcctgccacc agccagccaa cagctccccg accggcagct cggcacaaaa
    15301 tcaccactcg atacaggcag cccatcagtc cgggacggcg tcagcgggag agccgttgta
    15361 aggcggcaga ctttgctcat gttaccgatg ctattcggaa gaacggcaac taagctgccg
    15421 ggtttgaaac acggatgatc tcgcggaggg tagcatgttg attgtaacga tgacagagcg
    15481 ttgctgcctg tgatccagat catgaacaat aaaactgtct gcttacataa acagtaatac
    15541 aaggggtgtt atgagccata ttcaacggga aacgtcttgc tctaggccgc gattaaattc
    15601 caacatggat gctgatttat atgggtataa atgggctcgc gataatgtc
//
