LOCUS       sequence-273750-       12737 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Agrobacterium T-DNA with divergent H2A/Bpr driving LifeAct-tdTomato 
            and hph.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12737)
  AUTHORS   Medina EM, Robinson KA, Bellingham-Johnstun K, Ianiri G, Laplante C,
            Fritz-Laylin LK, Buchler NE
  TITLE     Genetic transformation of Spizellomyces punctatus, a resource for 
            studying chytrid biology and evolutionary cell biology.
  JOURNAL   Elife. 2020 May 11;9. pii: 52741. doi: 10.7554/eLife.52741.
  PUBMED    32392127
REFERENCE   2  (bases 1 to 12737)
  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: "Elife."; 
            date: "2020-05-11"; volume: "9. pii: 52741. doi"; pages: " 
            10.7554/eLife.52741"
COMMENT     Sequence Label: pGI3EM22C
FEATURES             Location/Qualifiers
     source          1..12737
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(687..1316)
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     rep_origin      2477..3819
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(3440..3487)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     primer_bind     3860..3878
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4081..4296
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             4297..5100
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     misc_feature    5267..5291
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     primer_bind     5472..5494
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5486..5503
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5487..5503
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      complement(5513..5700)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     CDS             complement(5722..6750)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     CDS             complement(5722..6750)
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     CDS             7166..7216
                     /codon_start=1
                     /product="peptide that binds filamentous actin (Riedl et 
                     al., 2008)"
                     /label=Lifeact
                     /note="first 17 amino acids of budding yeast Abp140"
                     /translation="MGVADLIKKFESISKEE"
     CDS             7232..8662
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     terminator      8684..8871
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(8886..8902)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8886..8902)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8899..8921)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8910..8926
                     /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(8934..8940,8941..8958,8959..8964))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8979..9000
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_feature    9177..9201
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     primer_bind     complement(9483..9502)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             9494..10309
                     /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             9494..10309
                     /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(9567..9586)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      10404..10992
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10893..10912
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     11146..11163
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    11178..11318
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      11662..11856
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(join(11922..12737,1..258))
                     /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"
ORIGIN
        1 gtcatagagc atcggaacga aaaaatctgg ttgctcgtcg cccttgggcg gcttcctaat
       61 cgacggcgca ccggctgccg gcggttgccg ggattctttg cggattcgat cagcggccgc
      121 ttgccacgat tcaccggggc gtgcttctgc ctcgatgcgt tgccgctggg cggcctgcgc
      181 ggccttcaac ttctccacca ggtcatcacc cagcgccgcg ccgatttgta ccgggccgga
      241 tggtttgcga ccgctcacgc cgattcctcg ggcttggggg ttccagtgcc attgcagggc
      301 cggcagacaa cccagccgct tacgcctggc caaccgcccg ttcctccaca catggggcat
      361 tccacggcgt cggtgcctgg ttgttcttga ttttccatgc cgcctccttt agccgctaaa
      421 attcatctac tcatttattc atttgctcat ttactctggt agctgcgcga tgtattcaga
      481 tagcagctcg gtaatggtct tgccttggcg taccgcgtac atcttcagct tggtgtgatc
      541 ctccgccggc aactgaaagt tgacccgctt catggctggc gtgtctgcca ggctggccaa
      601 cgttgcagcc ttgctgctgc gtgcgctcgg acggccggca cttagcgtgt ttgtgctttt
      661 gctcattttc tctttacctc attaactcaa atgagttttg atttaatttc agcggccagc
      721 gcctggacct cgcgggcagc gtcgccctcg ggttctgatt caagaacggt tgtgccggcg
      781 gcggcagtgc ctgggtagct cacgcgctgc gtgatacggg actcaagaat gggcagctcg
      841 tacccggcca gcgcctcggc aacctcaccg ccgatgcgcg tgcctttgat cgcccgcgac
      901 acgacaaagg ccgcttgtag ccttccatcc gtgacctcaa tgcgctgctt aaccagctcc
      961 accaggtcgg cggtggccca tatgtcgtaa gggcttggct gcaccggaat cagcacgaag
     1021 tcggctgcct tgatcgcgga cacagccaag tccgccgcct ggggcgctcc gtcgatcact
     1081 acgaagtcgc gccggccgat ggccttcacg tcgcggtcaa tcgtcgggcg gtcgatgccg
     1141 acaacggtta gcggttgatc ttcccgcacg gccgcccaat cgcgggcact gccctgggga
     1201 tcggaatcga ctaacagaac atcggccccg gcgagttgca gggcgcgggc tagatgggtt
     1261 gcgatggtcg tcttgcctga cccgcctttc tggttaagta cagcgataac cttcatgcgt
     1321 tccccttgcg tatttgttta tttactcatc gcatcatata cgcagcgacc gcatgacgca
     1381 agctgtttta ctcaaataca catcaccttt ttagacggcg gcgctcggtt tcttcagcgg
     1441 ccaagctggc cggccaggcc gccagcttgg catcagacaa accggccagg atttcatgca
     1501 gccgcacggt tgagacgtgc gcgggcggct cgaacacgta cccggccgcg atcatctccg
     1561 cctcgatctc ttcggtaatg aaaaacggtt cgtcctggcc gtcctggtgc ggtttcatgc
     1621 ttgttcctct tggcgttcat tctcggcggc cgccagggcg tcggcctcgg tcaatgcgtc
     1681 ctcacggaag gcaccgcgcc gcctggcctc ggtgggcgtc acttcctcgc tgcgctcaag
     1741 tgcgcggtac agggtcgagc gatgcacgcc aagcagtgca gccgcctctt tcacggtgcg
     1801 gccttcctgg tcgatcagct cgcgggcgtg cgcgatctgt gccggggtga gggtagggcg
     1861 ggggccaaac ttcacgcctc gggccttggc ggcctcgcgc ccgctccggg tgcggtcgat
     1921 gattagggaa cgctcgaact cggcaatgcc ggcgaacacg gtcaacacca tgcggccggc
     1981 cggcgtggtg gtgtcggccc acggctctgc caggctacgc aggcccgcgc cggcctcctg
     2041 gatgcgctcg gcaatgtcca gtaggtcgcg ggtgctgcgg gccaggcggt ctagcctggt
     2101 cactgtcaca acgtcgccag ggcgtaggtg gtcaagcatc ctggccagct ccgggcggtc
     2161 gcgcctggtg ccggtgatct tctcggaaaa cagcttggtg cagccggccg cgtgcagttc
     2221 ggcccgttgg ttggtcaagt cctggtcgtc ggtgctgacg cgggcatagc ccagcaggcc
     2281 agcggcggcg ctcttgttca tggcgtaatg tctccggttc tagtcgcaag tattctactt
     2341 tatgcgacta aaacacgcga caagaaaacg ccaggaaaag ggcagggcgg cagcctgtcg
     2401 cgtaacttag gacttgtgcg acatgtcgtt ttcagaagac ggctgcactg aacgtcagaa
     2461 ggaaaagtgc cacctgaacg aagcatctgt gcttcatttt gtagaacaaa aatgcaacgc
     2521 gagagcgcta atttttcaaa caaagaatct gagctgcatt tttacagaac agaaatgcaa
     2581 cgcgaaagcg ctattttacc aacgaagaat ctgtgcttca tttttgtaaa acaaaaatgc
     2641 aacgcgagag cgctaatttt tcaaacaaag aatctgagct gcatttttac agaacagaaa
     2701 tgcaacgcga gagcgctatt ttaccaacaa agaatctata cttctttttt gttctacaaa
     2761 aatgcatccc gagagcgcta tttttctaac aaagcatctt agattacttt ttttctcctt
     2821 tgtgcgctct ataatgcagt ctcttgataa ctttttgcac tgtaggtccg ttaaggttag
     2881 aagaaggcta ctttggtgtc tattttctct tccataaaaa aagcctgact ccacttcccg
     2941 cgtttactga ttactagcga agctgcgggt gcattttttc aagataaagg catccccgat
     3001 tatattctat accgatgtgg attgcgcata ctttgtgaac agaaagtgat agcgttgatg
     3061 attcttcatt ggtcagaaaa ttatgaacgg tttcttctat tttgtctcta tatactacgt
     3121 ataggaaatg tttacatttt cgtattgttt tcgattcact ctatgaatag ttcttactac
     3181 aatttttttg tctaaagagt aatactagag ataaacataa aaaatgtaga ggtcgagttt
     3241 agatgcaagt tcaaggagcg aaaggtggat gggtaggtta tatagggata tagcacagag
     3301 atatatagca aagagatact tttgagcaat gtttgtggaa gcggtattcg caatatttta
     3361 gtagctcgtt acagtccggt gcgtttttgg ttttttgaaa gtgcgtcttc agagcgcttt
     3421 tggttttcaa aagcgctctg aagttcctat actttctaga gaataggaac ttcggaatag
     3481 gaacttcaaa gcgtttccga aaacgagcgc ttccgaaaat gcaacgcgag ctgcgcacat
     3541 acagctcact gttcacgtcg cacctatatc tgcgtgttgc ctgtatatat atatacatga
     3601 gaagaacggc atagtgcgtg tttatgctta aatgcgtact tatatgcgtc tatttatgta
     3661 ggatgaaagg tagtctagta cctcctgtga tattatccca ttccatgcgg ggtatcgtat
     3721 gcttccttca gcactaccct ttagctgttc tatatgctgc cactcctcaa ttggattagt
     3781 ctcatccttc aatgctatca tttcctttga tattggatca tactaagaaa ccattattat
     3841 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg
     3901 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     3961 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     4021 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc ataccacctt
     4081 ttcaattcat catttttttt ttattctttt ttttgatttc ggtttccttg aaattttttt
     4141 gattcggtaa tctccgaaca gaaggaagaa cgaaggaagg agcacagact tagattggta
     4201 tatatacgca tatgtagtgt tgaagaaaca tgaaattgcc cagtattctt aacccaactg
     4261 cacagaacaa aaacctgcag gaaacgaaga taaatcatgt cgaaagctac atataaggaa
     4321 cgtgctgcta ctcatcctag tcctgttgct gccaagctat ttaatatcat gcacgaaaag
     4381 caaacaaact tgtgtgcttc attggatgtt cgtaccacca aggaattact ggagttagtt
     4441 gaagcattag gtcccaaaat ttgtttacta aaaacacatg tggatatctt gactgatttt
     4501 tccatggagg gcacagttaa gccgctaaag gcattatccg ccaagtacaa ttttttactc
     4561 ttcgaagaca gaaaatttgc tgacattggt aatacagtca aattgcagta ctctgcgggt
     4621 gtatacagaa tagcagaatg ggcagacatt acgaatgcac acggtgtggt gggcccaggt
     4681 attgttagcg gtttgaagca ggcggcagaa gaagtaacaa aggaacctag aggccttttg
     4741 atgctagcag aattgtcatg caagggctcc ctatctactg gagaatatac taagggtact
     4801 gttgacattg cgaagagcga caaagatttt gttatcggct ttattgctca aagagacatg
     4861 ggtggaagag atgaaggtta cgattggttg attatgacac ccggtgtggg tttagatgac
     4921 aagggagacg cattgggtca acagtataga accgtggatg atgtggtctc tacaggatct
     4981 gacattatta ttgttggaag aggactattt gcaaagggaa gggatgctaa ggtagagggt
     5041 gaacgttaca gaaaagcagg ctgggaagca tatttgagaa gatgcggcca gcaaaactaa
     5101 aaaactgtat tataagtaaa tgcatgtata ctaaactcac aaattagagc ttcaatttaa
     5161 ttatatcagt tattacctgg catgcacata caaatggacg aacggataaa ccttttcacg
     5221 cccttttaaa tatccgatta ttctaataaa cgctcttttc tcttaggttt acccgccaat
     5281 atatcctgtc aaacactgat agtttaaact gaaggcggga aacgacaatc tgatccaagc
     5341 tcaagctcat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
     5401 ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac
     5461 gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgccaagc ttccggtaga
     5521 ggtgtggtca ataagagcga cctcatacta tacctgagaa agcaacctga cctacaggaa
     5581 agagttactc aagaataaga attttcgttt taaaacctaa gagtcacttt aaaatttgta
     5641 tacacttatt ttttttataa cttatttaat aataaaaatc ataaatcata agaaattcgc
     5701 ttatttagaa gtggcgcgcc attattcctt tgccctcgga cgagtgctgg ggcgtcggtt
     5761 tccactatcg gcgagtactt ctacacagcc atcggtccag acggccgcgc ttctgcgggc
     5821 gatttgtgta cgcccgacag tcccggctcc ggatcggacg attgcgtcgc atcgaccctg
     5881 cgcccaagct gcatcatcga aattgccgtc aaccaagctc tgatagagtt ggtcaagacc
     5941 aatgcggagc atatacgccc ggagccgcgg cgatcctgca agctccggat gcctccgctc
     6001 gaagtagcgc gtctgctgct ccatacaagc caaccacggc ctccagaaga agatgttggc
     6061 gacctcgtat tgggaatccc cgaacatcgc ctcgctccag tcaatgaccg ctgttatgcg
     6121 gccattgtcc gtcaggacat tgttggagcc gaaatccgcg tgcacgaggt gccggacttc
     6181 ggggcagtcc tcggcccaaa gcatcagctc atcgagagcc tgcgcgacgg acgcactgac
     6241 ggtgtcgtcc atcacagttt gccagtgata cacatgggga tcagcaatcg cgcatatgaa
     6301 atcacgccat gtagtgtatt gaccgattcc ttgcggtccg aatgggccga acccgctcgt
     6361 ctggctaaga tcggccgcag cgatcgcatc catggcctcc gcgaccggct gcagaacagc
     6421 gggcagttcg gtttcaggca ggtcttgcaa cgtgacaccc tgtgcacggc gggagatgca
     6481 ataggtcagg ctctcgctga attccccaat gtcaagcact tccggaatcg ggagcgcggc
     6541 cgatgcaaag tgccgataaa cataacgatc tttgtagaaa ccatcggcgc agctatttac
     6601 ccgcaggaca tatccacgcc ctcctacatc gaagctgaaa gcacgagatt cttcgccctc
     6661 cgagagctgc atcaggtcgg agacgctgtc gaacttttcg atcagaaact tctcgacaga
     6721 cgtcgcggtg agttcaggct ttttacccat ttaattaatg ctgtgtaagg tgtgcgtgag
     6781 agagagggtc gtgagtgagt cttgaagaga aacgtcaaag cttgaagcgt agcagaatac
     6841 ggaaaacgac ttgggggaat ggcgaggagc gtcgcctttt atgctccaag cggagacgat
     6901 gaggccgata ttacagtggc ttagtagcgt gtacaatatt tccataaaag gctacatggt
     6961 gagtttttca cgtgaccgta attagtccca attaggcttt attgtatatt gctactaagc
     7021 atcgttaaat gacctgctcg aagcttcgtc tataaaaggc gggcgtgtgt gcgcttcccc
     7081 cgctacgtct ttttcacaaa cactttgcga cttgaccaag acctttcacc cgcaccactc
     7141 tcatccatca cacacacaag tcgacatggg cgtggccgac ttgatcaaga agttcgagtc
     7201 catctccaag gaggagggag gaggaggatc catggtgagc aagggcgagg aggtcatcaa
     7261 agagttcatg cgcttcaagg tgcgcatgga gggctccatg aacggccacg agttcgagat
     7321 cgagggcgag ggcgagggcc gcccctacga gggcacccag accgccaagc tgaaggtgac
     7381 caagggcggc cccctgccct tcgcctggga catcctgtcc ccccagttca tgtacggctc
     7441 caaggcgtac gtgaagcacc ccgccgacat ccccgattac aagaagctgt ccttccccga
     7501 gggcttcaag tgggagcgcg tgatgaactt cgaggacggc ggtctggtga ccgtgaccca
     7561 ggactcctcc ctgcaggacg gcacgctgat ctacaaggtg aagatgcgcg gcaccaactt
     7621 cccccccgac ggccccgtaa tgcagaagaa gaccatgggc tgggaggcct ccaccgagcg
     7681 cctgtacccc cgcgacggcg tgctgaaggg cgagatccac caggccctga agctgaagga
     7741 cggcggccac tacctggtgg agttcaagac catctacatg gccaagaagc ccgtgcaact
     7801 gcccggctac tactacgtgg acaccaagct ggacatcacc tcccacaacg aggactacac
     7861 catcgtggaa cagtacgagc gctccgaggg ccgccaccac ctgttcctgg ggcatggcac
     7921 cggcagcacc ggcagcggca gctccggcac cgcctcctcc gaggacaaca acatggccgt
     7981 catcaaagag ttcatgcgct tcaaggtgcg catggagggc tccatgaacg gccacgagtt
     8041 cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg ccaagctgaa
     8101 ggtgaccaag ggcggccccc tgcccttcgc ctgggacatc ctgtcccccc agttcatgta
     8161 cggctccaag gcgtacgtga agcaccccgc cgacatcccc gattacaaga agctgtcctt
     8221 ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggtc tggtgaccgt
     8281 gacccaggac tcctccctgc aggacggcac gctgatctac aaggtgaaga tgcgcggcac
     8341 caacttcccc cccgacggcc ccgtaatgca gaagaagacc atgggctggg aggcctccac
     8401 cgagcgcctg tacccccgcg acggcgtgct gaagggcgag atccaccagg ccctgaagct
     8461 gaaggacggc ggccactacc tggtggagtt caagaccatc tacatggcca agaagcccgt
     8521 gcaactgccc ggctactact acgtggacac caagctggac atcacctccc acaacgagga
     8581 ctacaccatc gtggaacagt acgagcgctc cgagggccgc caccacctgt tcctgtacgg
     8641 catggacgag ctgtacaagt aaaggcgcgc cacttctaaa taagcgaatt tcttatgatt
     8701 tatgattttt attattaaat aagttataaa aaaaataagt gtatacaaat tttaaagtga
     8761 ctcttaggtt ttaaaacgaa aattcttatt cttgagtaac tctttcctgt aggtcaggtt
     8821 gctttctcag gtatagtatg aggtcgctct tattgaccac acctctaccg ggaattcgta
     8881 atcatgtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     8941 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     9001 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     9061 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg gctagagcaa ttcggcgtta
     9121 attcagtaca ttaaaaacgt ccgcaatgtg ttattaagtt gtctaagcgt caatttgttt
     9181 acaccacaat atatcctgcc accagccagc caacagctcc ccgaccggca gctcggcaca
     9241 aaatcaccac tcgatacagg cagcccatca gtccgggacg gcgtcagcgg gagagccgtt
     9301 gtaaggcggc agactttgct catgttaccg atgctattcg gaagaacggc aactaagctg
     9361 ccgggtttga aacacggatg atctcgcgga gggtagcatg ttgattgtaa cgatgacaga
     9421 gcgttgctgc ctgtgatcca gatcatgaac aataaaactg tctgcttaca taaacagtaa
     9481 tacaaggggt gttatgagcc atattcaacg ggaaacgtct tgctctaggc cgcgattaaa
     9541 ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc
     9601 aggtgcgaca atctatcgat tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca
     9661 tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac
     9721 ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt
     9781 actcaccact gcgatccccg ggaaaacagc attccaggta ttagaagaat atcctgattc
     9841 aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt
     9901 ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat
     9961 gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga
    10021 acaagtctgg aaagaaatgc ataaactttt gccattctca ccggattcag tcgtcactca
    10081 tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga
    10141 tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct
    10201 cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc
    10261 tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttctaag aattaattca
    10321 tgatcctgca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
    10381 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
    10441 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
    10501 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
    10561 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
    10621 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
    10681 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
    10741 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
    10801 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
    10861 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
    10921 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
    10981 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
    11041 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
    11101 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
    11161 gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
    11221 caccgcatat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
    11281 acactccgct atcgctacgt gactgggtca tggctgcgcc ccgacacccg ccaacacccg
    11341 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
    11401 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgaggcagg
    11461 gtgccttgat gtgggcgccg gcggtcgagt ggcgacggcg cggcttgtcc gcgccctggt
    11521 agattgcctg gccgtaggcc agccattttt gagcggccag cggccgcgat aggccgacgc
    11581 gaagcggcgg ggcgtaggga gcgcagcgac cgaagggtag gcgctttttg cagctcttcg
    11641 gctgtgcgct ggccagacag ttatgcacag gccaggcggg ttttaagagt tttaataagt
    11701 tttaaagagt tttaggcgga aaaatcgcct tttttctctt ttatatcagt cacttacatg
    11761 tgtgaccggt tcccaatgta cggctttggg ttcccaatgt acgggttccg gttcccaatg
    11821 tacggctttg ggttcccaat gtacgtgcta tccacaggaa agagaccttt tcgacctttt
    11881 tcccctgcta gggcaatttg ccctagcatc tgctccgtac attaggaacc ggcggatgct
    11941 tcgccctcga tcaggttgcg gtagcgcatg actaggatcg ggccagcctg ccccgcctcc
    12001 tccttcaaat cgtactccgg caggtcattt gacccgatca gcttgcgcac ggtgaaacag
    12061 aacttcttga actctccggc gctgccactg cgttcgtaga tcgtcttgaa caaccatctg
    12121 gcttctgcct tgcctgcggc gcggcgtgcc aggcggtaga gaaaacggcc gatgccggga
    12181 tcgatcaaaa agtaatcggg gtgaaccgtc agcacgtccg ggttcttgcc ttctgtgatc
    12241 tcgcggtaca tccaatcagc tagctcgatc tcgatgtact ccggccgccc ggtttcgctc
    12301 tttacgatct tgtagcggct aatcaaggct tcaccctcgg ataccgtcac caggcggccg
    12361 ttcttggcct tcttcgtacg ctgcatggca acgtgcgtgg tgtttaaccg aatgcaggtt
    12421 tctaccaggt cgtctttctg ctttccgcca tcggctcgcc ggcagaactt gagtacgtcc
    12481 gcaacgtgtg gacggaacac gcggccgggc ttgtctccct tcccttcccg gtatcggttc
    12541 atggattcgg ttagatggga aaccgccatc agtaccaggt cgtaatccca cacactggcc
    12601 atgccggccg gccctgcgga aacctctacg tgcccgtctg gaagctcgta gcggatcacc
    12661 tcgccagctc gtcggtcacg cttcgacaga cggaaaacgg ccacgtccat gatgctgcga
    12721 ctatcgcggg tgcccac
//
