LOCUS       sequence-477068-       14913 bp    DNA     circular SYN 25-JUN-2025
DEFINITION  sfGFP linked with GFPuv via T2A expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14913)
  AUTHORS   Liu Y, Rajput R, Islam MT, Valle ID, Yao T, Agrawal R, Boone BA, 
            Eckert CA, Abraham PE, Chen JG, Tuskan GA, Yang X
  TITLE     A split ribozyme system for in vivo plant RNA imaging and genetic 
            engineering.
  JOURNAL   Plant Biotechnol J. 2025 May;23(5):1640-1649. doi: 
            10.1111/pbi.14612. Epub 2025 Feb 7.
  PUBMED    39919021
REFERENCE   2  (bases 1 to 14913)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 25, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1111/pbi.14612";
            journalName: "Plant Biotechnol J"; date: "2025-05"; volume: "23"; 
            issue: "5"; pages: "1640-1649"
COMMENT     Sequence Label: pXYB164
FEATURES             Location/Qualifiers
     source          1..14913
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(16..361)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     723..745
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     737..754
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     738..754
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1269..1614
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     CDS             join(1739..1741,1742..2452)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             1739..2452
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Nager et al., 2011)"
                     /label=superfolder GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(1769..1797)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2392..2412
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             2462..2518
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     terminator      3180..3428
                     /label=HSP terminator
                     /note="efficient transcription terminator from the 
                     Arabidopsis thaliana heat shock protein 18.2 gene (Nagaya 
                     et al., 2010)"
     primer_bind     complement(4118..4134)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4118..4134)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4131..4153)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4142..4158
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4166..4172,4173..4190,4191..4196))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4211..4232
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4423..5099
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     CDS             5167..5964
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MGIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR
                     PVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLL
                     SSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEH
                     QGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDI
                     ALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5224..5243)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5834..5853
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6004..6178
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    6256..6280
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             6705..7499
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      7586..8174
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8075..8094
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8328..8345
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    8360..8500
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(8426..8445)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8586..8608
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     rep_origin      8844..9038
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(9104..10177)
                     /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(10606..11235)
                     /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    12536..12560
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     terminator      complement(12582..12834)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(12866..12883)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(join(12890..14707,14898..14912))
                     /codon_start=1
                     /product="beta-glucuronidase"
                     /label=GUSPlus(TM)
                     /note="codon-optimized Staphylococcus gusA gene with a 
                     catalase intron to ensure expression in plants but not 
                     bacteria"
                     /translation="MVDLRNRRTSLYPINTETRGVFDLNGVWNFKLDYGKGLEEKWYES
                     KLTDTISMAVPSSYNDIGVTKEIRNHIGYVWYEREFTVPAYLKDQRIVLRFGSATHKAI
                     VYVNGELVVEHKGGFLPFEAEINNSLRDGMNRVTVAVDNILDDSTLPVGLYSERHEEGL
                     GKVIRNKPNFDFFNYAGLHRPVKIYTTPFTYVEDISVVTDFNGPTGTVTYTVDFQGKAE
                     TVKVSVVDEEGKVVASTEGLSGNVEIPNVILWEPLNTYLYQIKVELVNDGLTIDVYEEP
                     FGVRTVEVNDGKFLINNKPFYFKGFGKHEDTPINGRGFNEASNVMDFNILKWIGANSFR
                     TAHYPYSEELMRLADREGLVVIDETPAVGVHLNFMATTGLGEGSERVSTWEKIRTFEHH
                     QDVLRELVSRDKNHPSVVMWSIANEAATEEEGAYEYFKPLVELTKELDPQKRPVTIVLF
                     VMATPETDKVAELIDVIALNRYNGWYFDGGDLEAAKVHLRQEFHAWNKRCPGKPIMITE
                     YGADTVAGFHDIDPVMFTEEYQVEYYQANHVVFDEFENFVGEQAWNFADFATSQGVMRV
                     QGNKKGVFTRDRKPKLAAHVFRERWTNIPDFGYKN"
     intron          14708..14897
                     /label=cat1 intron
                     /note="castor bean catalase intron, modified"
ORIGIN
        1 gtcaagagtc ccccgtgttc tctccaaatg aaatgaactt ccttatatag aggaagggtc
       61 ttgcgaagga tagtgggatt gtgcgtcatc ccttacgtca gtggagatat cacatcaatc
      121 cacttgcttt gaagacgtgg ttggaacgtc ttctttttcc acgatgctcc tcgtgggtgg
      181 gggtccatct ttgggaccac tgtcggcaga ggcatcttca acgatggcct ttcctttatc
      241 gcaatgatgg catttgtagg agccaccttc cttttccact atcttcacaa taaagtgaca
      301 gatagctggg caatggaatc cgaggaggtt tccggatatt accctttgtt gaaaagtctc
      361 aattgccctt tggtcttctg agactgtatc tttgatattt ttggagtaga caagtgtgtc
      421 gtgctccacc atgttgacga agattttctt cttgtcattg agtcgtaaga gactctgtat
      481 gaactgttcg ccagtcttta cggcgagttc tgttaggtcc tctatttgaa tctttgactc
      541 catgaagcta aactgaaggc gggaaacgac aatctgatcc aagctcaagc tgctctagca
      601 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
      661 acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt
      721 ttcccagtca cgacgttgta aaacgacggc cagtgccaag cttgcatgcc tgcaggtccc
      781 cagattagcc ttttcaattt cagaaagaat gctaacccac agatggttag agaggcttac
      841 gcagcaggtc tcatcaagac gatctacccg agcaataatc tccaggaaat caaatacctt
      901 cccaagaagg ttaaagatgc agtcaaaaga ttcaggacta actgcatcaa gaacacagag
      961 aaagatatat ttctcaagat cagaagtact attccagtat ggacgattca aggcttgctt
     1021 cacaaaccaa ggcaagtaat agagattgga gtctctaaaa aggtagttcc cactgaatca
     1081 aaggccatgg agtcaaagat tcaaatagag gacctaacag aactcgccgt aaagactggc
     1141 gaacagttca tacagagtct cttacgactc aatgacaaga agaaaatctt cgtcaacatg
     1201 gtggagcacg acacacttgt ctactccaaa aatatcaaag atacagtctc agaagaccaa
     1261 agggcaattg agacttttca acaaagggta atatccggaa acctcctcgg attccattgc
     1321 ccagctatct gtcactttat tgtgaagata gtggaaaagg aaggtggctc ctacaaatgc
     1381 catcattgcg ataaaggaaa ggccatcgtt gaagatgcct ctgccgacag tggtcccaaa
     1441 gatggacccc cacccacgag gagcatcgtg gaaaaagaag acgttccaac cacgtcttca
     1501 aagcaagtgg attgatgtga tatctccact gacgtaaggg atgacgcaca atcccactat
     1561 ccttcgcaag acccttcctc tatataagga agttcatttc atttggagag aacacggggg
     1621 actcaaacat tactcattca caaaaccatc ttaaagcaac tacacaagtc ttgaaatttt
     1681 ctcatatttt ctatttacta tataaacttt taatcaaatc aagattaaag ttaattaaat
     1741 gagcaaagga gaagaacttt tcactggagt tgtcccaatt cttgttgaat tagatggtga
     1801 tgttaatggg cacaaatttt ctgtccgtgg agagggtgaa ggtgatgcta caaacggaaa
     1861 actcaccctt aaatttattt gcactactgg aaaactacct gttccgtggc caacacttgt
     1921 cactactctg acctatggtg ttcaatgctt ttcccgttat ccggatcaca tgaaacggca
     1981 tgactttttc aagagtgcca tgcccgaagg ttatgtacag gaacgcacta tatctttcaa
     2041 agatgacggg acctacaaga cgcgtgctga agtcaagttt gaaggtgata cccttgttaa
     2101 tcgtatcgag ttaaagggta ttgattttaa agaagatgga aacattcttg gacacaaact
     2161 cgagtacaac tttaactcac acaatgtata catcacggca gacaaacaaa agaatggaat
     2221 caaagctaac ttcaaaattc gccacaacgt tgaagatggt tccgttcaac tagcagacca
     2281 ttatcaacaa aatactccaa ttggcgatgg ccctgtcctt ttaccagaca accattacct
     2341 gtcgacacaa tctgtccttt cgaaagatcc caacgaaaag cgtgaccaca tggtccttct
     2401 tgagtttgta actgctgctg ggattacaca tggcatggat gagctctaca aaggaagcgg
     2461 agctactaac ttcagcctgc tgaagcaggc tggagacgtg gaggagaacc ctggacctat
     2521 gacaaccttc aaaatcgagt cccggatcca cggcaacctc aacggggaga agttcgagtt
     2581 ggttggaggt ggagtaggtg aggagggtcg cctcgagatt gagatgaaga ctaaagataa
     2641 accactggca ttctctccct tcctgctgac cacttgcatg ggttacgggt tctaccactt
     2701 cgccagcttc ccaaagggga ttaagaacat ctatcttcat gctgcaacga acggaggtta
     2761 caccaacacc aggaaggaga tctatgaaga cggcggcatc ttggaggtca acttccgtta
     2821 cacttacgag ttcaacaaga tcatcggtga cgtcgagtgc attggacatg gattcccaag
     2881 tcagagtccg atcttcaagg acacgatcgt gaagagttgt cccacggtgg acctgatgtt
     2941 gccaatgtcc gggaacatca tcgccagctc ctacgcttac gccttccaac tgaaggacgg
     3001 ctctttctac acggcagaag tcaagaacaa catagacttc aagaatccaa tccacgagtc
     3061 cttctcgaag tcagggccca tgttcaccca cagacgtgtc gaggagactc tcaccaagga
     3121 gaaccttgcc atagtggagt accagcaggt tttcaacagc gccccaagag acatgtagat
     3181 atgaagatga agatgaaata tttggtgtgt caaataaaaa gcttgtgtgc ttaagtttgt
     3241 gtttttttct tggcttgttg tgttatgaat ttgtggcttt ttctaatatt aaatgaatgt
     3301 aagatctcat tataatgaat aaacaaatgt ttctataatc cattgtgaat gttttgttgg
     3361 atctcttctg cagcatataa ctactgtatg tgctatggta tggactatgg aatatgatta
     3421 aagataagat gggctcatag agtaaaacga ggcgagggac ctataaacct cccttcatca
     3481 tgctatttca tgatctattt tataaaataa agatgtagaa aaaagtaagc gtaataaccg
     3541 caaaacaaat gatttaaaac atggcacata atgaggagat taagttcggt ttacgtttat
     3601 tttagtacta attgtaacgt gagactacgt atcgggaatc gcctaattaa agcattaatg
     3661 cgaacctgat tagattcacc gaccctccta tcgtgtcgac ctttctgttt cttagaattt
     3721 tttggtagtc tatgtactaa taatgtcagc ttcgtattta tttcataagc aatttgcatt
     3781 tgcaatttgt tttttacttt tatttttatt gtattgtgga atgtggactc gtaccaacat
     3841 gaagttatat accaccaaaa aaattacagt tagtcaaaag attcacgagt gagagctact
     3901 tatgattgtc ttttacgtat atgtctaatt gtctatttgc tcaataatct ttgtactttc
     3961 ttttgtcgtt gataaaatca caaagttcca aaagtaatcg aatgatttgc ttttaagaaa
     4021 agaagagctc aataattcaa catatatctg tacacaatta ccgcctttga gtgagctcta
     4081 gatgccacct gacgtctaag aagaattcgt aatcatggtc atagctgttt cctgtgtgaa
     4141 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     4201 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     4261 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     4321 gtttgcgtat tggctagagc agcttgccaa catggtggag cacgacactc tcgtctactc
     4381 caagaatatc aaagatacag tctcagaaga ccaaagggct attgagactt ttcaacaaag
     4441 ggtaatatcg ggaaacctcc tcggattcca ttgcccagct atctgtcact tcatcaaaag
     4501 gacagtagaa aaggaaggtg gcacctacaa atgccatcat tgcgataaag gaaaggctat
     4561 cgttcaagat gcctctgccg acagtggtcc caaagatgga cccccaccca cgaggagcat
     4621 cgtggaaaaa gaagacgttc caaccacgtc ttcaaagcaa gtggattgat gtgaacatgg
     4681 tggagcacga cactctcgtc tactccaaga atatcaaaga tacagtctca gaagaccaaa
     4741 gggctattga gacttttcaa caaagggtaa tatcgggaaa cctcctcgga ttccattgcc
     4801 cagctatctg tcacttcatc aaaaggacag tagaaaagga aggtggcacc tacaaatgcc
     4861 atcattgcga taaaggaaag gctatcgttc aagatgcctc tgccgacagt ggtcccaaag
     4921 atggaccccc acccacgagg agcatcgtgg aaaaagaaga cgttccaacc acgtcttcaa
     4981 agcaagtgga ttgatgtgat atctccactg acgtaaggga tgacgcacaa tcccactatc
     5041 cttcgcaaga cccttcctct atataaggaa gttcatttca tttggagagg acacgctgaa
     5101 atcaccagtc tctctctaca aatctatctc tctcgagctt tcgcagatcc cggggggcaa
     5161 tgagatatgg ggattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     5221 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     5281 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     5341 aatgaactcc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     5401 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     5461 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     5521 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     5581 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     5641 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     5701 atgcccgacg gcgaggatct cgtcgtgaca catggcgatg cctgcttgcc gaatatcatg
     5761 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     5821 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     5881 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     5941 cgccttcttg acgagttctt ctgaagtaga tgccgaccgg atctgtcgat cgacaagctc
     6001 gagtttctcc ataataatgt gtgagtagtt cccagataag ggaattaggg ttcctatagg
     6061 gtttcgctca tgtgttgagc atataagaaa cccttagtat gtatttgtat ttgtaaaata
     6121 cttctatcaa taaaatttct aattcctaaa accaaaatcc agtactaaaa tccagatccc
     6181 ccgaattaat tcggcgttaa ttcagtacat taaaaacgtc cgcaatgtgt tattaagttg
     6241 tctaagcgtc aatttgttta caccacaata tatcctgcca ccagccagcc aacagctccc
     6301 cgaccggcag ctcggcacaa aatcaccact cgatacaggc agcccatcag tccgggacgg
     6361 cgtcagcggg agagccgttg taaggcggca gactttgctc atgttaccga tgctattcgg
     6421 aagaacggca actaagctgc cgggtttgaa acacggatga tctcgcggag ggtagcatgt
     6481 tgattgtaac gatgacagag cgttgctgcc tgtgatcacc gcggtttcaa aatcggctcc
     6541 gtcgatacta tgttatacgc caactttgaa aacaactttg aaaaagctgt tttctggtat
     6601 ttaaggtttt agaatgcaag gaacagtgaa ttggagttcg tcttgttata attagcttct
     6661 tggggtatct ttaaatactg tagaaaagag gaaggaaata ataaatggct aaaatgagaa
     6721 tatcaccgga attgaaaaaa ctgatcgaaa aataccgctg cgtaaaagat acggaaggaa
     6781 tgtctcctgc taaggtatat aagctggtgg gagaaaatga aaacctatat ttaaaaatga
     6841 cggacagccg gtataaaggg accacctatg atgtggaacg ggaaaaggac atgatgctat
     6901 ggctggaagg aaagctgcct gttccaaagg tcctgcactt tgaacggcat gatggctgga
     6961 gcaatctgct catgagtgag gccgatggcg tcctttgctc ggaagagtat gaagatgaac
     7021 aaagccctga aaagattatc gagctgtatg cggagtgcat caggctcttt cactccatcg
     7081 acatatcgga ttgtccctat acgaatagct tagacagccg cttagccgaa ttggattact
     7141 tactgaataa cgatctggcc gatgtggatt gcgaaaactg ggaagaagac actccattta
     7201 aagatccgcg cgagctgtat gattttttaa agacggaaaa gcccgaagag gaacttgtct
     7261 tttcccacgg cgacctggga gacagcaaca tctttgtgaa agatggcaaa gtaagtggct
     7321 ttattgatct tgggagaagc ggcagggcgg acaagtggta tgacattgcc ttctgcgtcc
     7381 ggtcgatcag ggaggatatc ggggaagaac agtatgtcga gctatttttt gacttactgg
     7441 ggatcaagcc tgattgggag aaaataaaat attatatttt actggatgaa ttgttttagt
     7501 acctagaatg catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     7561 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     7621 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     7681 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     7741 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     7801 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     7861 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     7921 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     7981 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     8041 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     8101 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     8161 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     8221 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     8281 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     8341 gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     8401 cacaccgcat atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt
     8461 atacactccg ctatcgctac gtgactgggt catggctgcg ccccgacacc cgccaacacc
     8521 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     8581 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca
     8641 gggtgccttg atgtgggcgc cggcggtcga gtggcgacgg cgcggcttgt ccgcgccctg
     8701 gtagattgcc tggccgtagg ccagccattt ttgagcggcc agcggccgcg ataggccgac
     8761 gcgaagcggc ggggcgtagg gagcgcagcg accgaagggt aggcgctttt tgcagctctt
     8821 cggctgtgcg ctggccagac agttatgcac aggccaggcg ggttttaaga gttttaataa
     8881 gttttaaaga gttttaggcg gaaaaatcgc cttttttctc ttttatatca gtcacttaca
     8941 tgtgtgaccg gttcccaatg tacggctttg ggttcccaat gtacgggttc cggttcccaa
     9001 tgtacggctt tgggttccca atgtacgtgc tatccacagg aaagagacct tttcgacctt
     9061 tttcccctgc tagggcaatt tgccctagca tctgctccgt acattaggaa ccggcggatg
     9121 cttcgccctc gatcaggttg cggtagcgca tgactaggat cgggccagcc tgccccgcct
     9181 cctccttcaa atcgtactcc ggcaggtcat ttgacccgat cagcttgcgc acggtgaaac
     9241 agaacttctt gaactctccg gcgctgccac tgcgttcgta gatcgtcttg aacaaccatc
     9301 tggcttctgc cttgcctgcg gcgcggcgtg ccaggcggta gagaaaacgg ccgatgccgg
     9361 gatcgatcaa aaagtaatcg gggtgaaccg tcagcacgtc cgggttcttg ccttctgtga
     9421 tctcgcggta catccaatca gctagctcga tctcgatgta ctccggccgc ccggtttcgc
     9481 tctttacgat cttgtagcgg ctaatcaagg cttcaccctc ggataccgtc accaggcggc
     9541 cgttcttggc cttcttcgta cgctgcatgg caacgtgcgt ggtgtttaac cgaatgcagg
     9601 tttctaccag gtcgtctttc tgctttccgc catcggctcg ccggcagaac ttgagtacgt
     9661 ccgcaacgtg tggacggaac acgcggccgg gcttgtctcc cttcccttcc cggtatcggt
     9721 tcatggattc ggttagatgg gaaaccgcca tcagtaccag gtcgtaatcc cacacactgg
     9781 ccatgccggc cggccctgcg gaaacctcta cgtgcccgtc tggaagctcg tagcggatca
     9841 cctcgccagc tcgtcggtca cgcttcgaca gacggaaaac ggccacgtcc atgatgctgc
     9901 gactatcgcg ggtgcccacg tcatagagca tcggaacgaa aaaatctggt tgctcgtcgc
     9961 ccttgggcgg cttcctaatc gacggcgcac cggctgccgg cggttgccgg gattctttgc
    10021 ggattcgatc agcggccgct tgccacgatt caccggggcg tgcttctgcc tcgatgcgtt
    10081 gccgctgggc ggcctgcgcg gccttcaact tctccaccag gtcatcaccc agcgccgcgc
    10141 cgatttgtac cgggccggat ggtttgcgac cgctcacgcc gattcctcgg gcttgggggt
    10201 tccagtgcca ttgcagggcc ggcaggcaac ccagccgctt acgcctggcc aaccgcccgt
    10261 tcctccacac atggggcatt ccacggcgtc ggtgcctggt tgttcttgat tttccatgcc
    10321 gcctccttta gccgctaaaa ttcatctact catttattca tttgctcatt tactctggta
    10381 gctgcgcgat gtattcagat agcagctcgg taatggtctt gccttggcgt accgcgtaca
    10441 tcttcagctt ggtgtgatcc tccgccggca actgaaagtt gacccgcttc atggctggcg
    10501 tgtctgccag gctggccaac gttgcagcct tgctgctgcg tgcgctcgga cggccggcac
    10561 ttagcgtgtt tgtgcttttg ctcattttct ctttacctca ttaactcaaa tgagttttga
    10621 tttaatttca gcggccagcg cctggacctc gcgggcagcg tcgccctcgg gttctgattc
    10681 aagaacggtt gtgccggcgg cggcagtgcc tgggtagctc acgcgctgcg tgatacggga
    10741 ctcaagaatg ggcagctcgt acccggccag cgcctcggca acctcaccgc cgatgcgcgt
    10801 gcctttgatc gcccgcgaca cgacaaaggc cgcttgtagc cttccatccg tgacctcaat
    10861 gcgctgctta accagctcca ccaggtcggc ggtggcccat atgtcgtaag ggcttggctg
    10921 caccggaatc agcacgaagt cggctgcctt gatcgcggac acagccaagt ccgccgcctg
    10981 gggcgctccg tcgatcacta cgaagtcgcg ccggccgatg gccttcacgt cgcggtcaat
    11041 cgtcgggcgg tcgatgccga caacggttag cggttgatct tcccgcacgg ccgcccaatc
    11101 gcgggcactg ccctggggat cggaatcgac taacagaaca tcggccccgg cgagttgcag
    11161 ggcgcgggct agatgggttg cgatggtcgt cttgcctgac ccgcctttct ggttaagtac
    11221 agcgataacc ttcatgcgtt ccccttgcgt atttgtttat ttactcatcg catcatatac
    11281 gcagcgaccg catgacgcaa gctgttttac tcaaatacac atcacctttt tagacggcgg
    11341 cgctcggttt cttcagcggc caagctggcc ggccaggccg ccagcttggc atcagacaaa
    11401 ccggccagga tttcatgcag ccgcacggtt gagacgtgcg cgggcggctc gaacacgtac
    11461 ccggccgcga tcatctccgc ctcgatctct tcggtaatga aaaacggttc gtcctggccg
    11521 tcctggtgcg gtttcatgct tgttcctctt ggcgttcatt ctcggcggcc gccagggcgt
    11581 cggcctcggt caatgcgtcc tcacggaagg caccgcgccg cctggcctcg gtgggcgtca
    11641 cttcctcgct gcgctcaagt gcgcggtaca gggtcgagcg atgcacgcca agcagtgcag
    11701 ccgcctcttt cacggtgcgg ccttcctggt cgatcagctc gcgggcgtgc gcgatctgtg
    11761 ccggggtgag ggtagggcgg gggccaaact tcacgcctcg ggccttggcg gcctcgcgcc
    11821 cgctccgggt gcggtcgatg attagggaac gctcgaactc ggcaatgccg gcgaacacgg
    11881 tcaacaccat gcggccggcc ggcgtggtgg tgtcggccca cggctctgcc aggctacgca
    11941 ggcccgcgcc ggcctcctgg atgcgctcgg caatgtccag taggtcgcgg gtgctgcggg
    12001 ccaggcggtc tagcctggtc actgtcacaa cgtcgccagg gcgtaggtgg tcaagcatcc
    12061 tggccagctc cgggcggtcg cgcctggtgc cggtgatctt ctcggaaaac agcttggtgc
    12121 agccggccgc gtgcagttcg gcccgttggt tggtcaagtc ctggtcgtcg gtgctgacgc
    12181 gggcatagcc cagcaggcca gcggcggcgc tcttgttcat ggcgtaatgt ctccggttct
    12241 agtcgcaagt attctacttt atgcgactaa aacacgcgac aagaaaacgc caggaaaagg
    12301 gcagggcggc agcctgtcgc gtaacttagg acttgtgcga catgtcgttt tcagaagacg
    12361 gctgcactga acgtcagaag ccgactgcac tatagcagcg gaggggttgg atcaaagtac
    12421 tttgatcccg aggggaaccc tgtggttggc atgcacatac aaatggacga acggataaac
    12481 cttttcacgc ccttttaaat atccgattat tctaataaac gctcttttct cttaggttta
    12541 cccgccaata tatcctgtca aacactgata gtttaattcc cgatctagta acatagatga
    12601 caccgcgcgc gataatttat cctagtttgc gcgctatatt ttgttttcta tcgcgtatta
    12661 aatgtataat tgcgggactc taatcataaa aacccatctc ataaataacg tcatgcatta
    12721 catgttaatt attacatgct taacgtaatt caacagaaat tatatgataa tcatcgcaag
    12781 accggcaaca ggattcaatc ttaagaaact ttattgccaa atgtttgaac gatcggggaa
    12841 attcgagctg gtcaccaatt cacacgtgat ggtgatggtg atggctagcg ttcttgtagc
    12901 cgaaatctgg aatgttggtc cagcgctcgc gaaagacgtg cgcggcgagc ttcggcttgc
    12961 ggtcacgagt gaacacgccc ttcttgtttc cttggacgcg catcacgccc tgagaggtcg
    13021 cgaagtccgc gaagttccac gcttgctcac ccacgaagtt ctcaaactca tcgaacacga
    13081 cgtggttcgc ctggtagtac tcgacttgat attcctcggt gaacatcact ggatcaatgt
    13141 cgtgaaagcc cgcaacggtg tctgcgccgt actcagtgat catgatcggc tttcctgggc
    13201 aacgcttgtt ccacgcgtga aattcctggc ggagatggac tttggccgct tcgagatcac
    13261 cgccatcgaa gtaccatccg ttatagcgat tgagcgcgat gacgtcaatc agttcggcga
    13321 ctttgtccgt ctccggggta gccatcacaa acagcacgat cgtgaccgga cgcttctgtg
    13381 ggtcgagttc cttggtcagc tccaccaacg gcttgaagta ctcgtacgcg ccctcttcct
    13441 cagtcgccgc ctcgttggcg atgctccaca tcacgacgct tggatggttc ttgtcacgag
    13501 acaccagttc acggagaacg tcttgatggt gctcaaacgt ccgaatcttc tcccaggtac
    13561 tgacgcgctc gctgccttcg ccgagtcccg tggtggccat gaagttgagg tgcacgccaa
    13621 ctgccggagt ctcgtcgatc acgaccagac cctcgcgatc cgcaagacgc atcaactctt
    13681 cagagtacgg atagtgtgcg gtccggaagc tgttggcgcc gatccatttg aggatattga
    13741 aatccatcac attgctcgct tcgttaaagc cacggccgtt gataggagtg tcctcatgtt
    13801 tgccaaagcc cttgaagtag aacggtttgt tgttgatgag gaacttgccg tcgttgactt
    13861 ccacggtccg cacgccgaac ggctcttcat agacatcgat ggtcagtccg tcgttcacca
    13921 gttccacttt gatctggtag agatacgtgt tcagtggttc ccagaggatg acattcggaa
    13981 tctccacgtt accgctcagg ccctcggtgc ttgcgaccac tttgccttcc tcatccacga
    14041 ccgacacttt cacggtctcg gctttgcctt gaaagtccac cgtataggtc acagtcccgg
    14101 ttgggccatt gaagtcggtc acaaccgaga tgtcctcgac gtacgtaaac ggggtcgtgt
    14161 agattttcac cggacggtgc aggcctgcat agttgaagaa gtcgaagttc ggcttgttac
    14221 gaatgacttt tccgaggccc tcttcgtggc gctcgctgta cagccccacc gggagggtgc
    14281 tatcgtcgag gatgttgtcc acggcgacgg tgacgcgatt catgccatca cgcagcgagt
    14341 tgttgatttc cgcttcgaat ggcaggaatc cgcccttgtg ctccacgacc agctcaccat
    14401 tgacatagac aattgctttg tgagttgcag agccgaagcg gagcacgata cgctgatcct
    14461 tcagataggc cggcaccgtg aactcacgtt cgtaccagac atatccgata tggttgcgga
    14521 tttccttggt cacgccaatg tcattgtaac tgcttgggac ggccatacta atagtgtcgg
    14581 tcagcttgct ttcgtaccac ttctcttcca gtcctttccc gtagtccagc ttgaagttcc
    14641 agacgccatt gaggtcgaag acgccacggg tctcggtgtt gatcgggtac agactagttc
    14701 gtcggttctg taactatcat catcatcata gacacacgaa ataaagtaat cagattatca
    14761 gttaaagcta tgtaatattt acaccataac caatcaatta aaaaatagat cagtttaaag
    14821 aaagatcaaa gctcaaaaaa ataaaaagag aaaagggtcc taaccaagaa aatgaaggag
    14881 aaaaactaga aatttaccct cagatctacc atg
//
