LOCUS       sequence-273748-       13082 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Agrobacterium T-DNA with divergent Spun H2A/Bpr driving H2B-tdTomato
            and hph.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13082)
  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 13082)
  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: pGI3EM20C
FEATURES             Location/Qualifiers
     source          1..13082
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1652..2994
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(2615..2662)
                     /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     3035..3053
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3256..3471
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             3472..4275
                     /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    4442..4466
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     primer_bind     4647..4669
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4661..4678
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4662..4678
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      complement(4688..4875)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     CDS             complement(4897..5925)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     CDS             complement(4897..5925)
                     /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             6752..8182
                     /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      8204..8391
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(8406..8422)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8406..8422)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8419..8441)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8430..8446
                     /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(8454..8460,8461..8478,8479..8484))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8499..8520
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_feature    8697..8721
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     primer_bind     complement(9003..9022)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             9014..9829
                     /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             9014..9829
                     /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(9087..9106)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      9924..10512
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10413..10432
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10666..10683
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    10698..10838
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      11182..11376
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(11442..12515)
                     /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(join(12944..13082,1..491))
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
ORIGIN
        1 agaatgggca gctcgtaccc ggccagcgcc tcggcaacct caccgccgat gcgcgtgcct
       61 ttgatcgccc gcgacacgac aaaggccgct tgtagccttc catccgtgac ctcaatgcgc
      121 tgcttaacca gctccaccag gtcggcggtg gcccatatgt cgtaagggct tggctgcacc
      181 ggaatcagca cgaagtcggc tgccttgatc gcggacacag ccaagtccgc cgcctggggc
      241 gctccgtcga tcactacgaa gtcgcgccgg ccgatggcct tcacgtcgcg gtcaatcgtc
      301 gggcggtcga tgccgacaac ggttagcggt tgatcttccc gcacggccgc ccaatcgcgg
      361 gcactgccct ggggatcgga atcgactaac agaacatcgg ccccggcgag ttgcagggcg
      421 cgggctagat gggttgcgat ggtcgtcttg cctgacccgc ctttctggtt aagtacagcg
      481 ataaccttca tgcgttcccc ttgcgtattt gtttatttac tcatcgcatc atatacgcag
      541 cgaccgcatg acgcaagctg ttttactcaa atacacatca cctttttaga cggcggcgct
      601 cggtttcttc agcggccaag ctggccggcc aggccgccag cttggcatca gacaaaccgg
      661 ccaggatttc atgcagccgc acggttgaga cgtgcgcggg cggctcgaac acgtacccgg
      721 ccgcgatcat ctccgcctcg atctcttcgg taatgaaaaa cggttcgtcc tggccgtcct
      781 ggtgcggttt catgcttgtt cctcttggcg ttcattctcg gcggccgcca gggcgtcggc
      841 ctcggtcaat gcgtcctcac ggaaggcacc gcgccgcctg gcctcggtgg gcgtcacttc
      901 ctcgctgcgc tcaagtgcgc ggtacagggt cgagcgatgc acgccaagca gtgcagccgc
      961 ctctttcacg gtgcggcctt cctggtcgat cagctcgcgg gcgtgcgcga tctgtgccgg
     1021 ggtgagggta gggcgggggc caaacttcac gcctcgggcc ttggcggcct cgcgcccgct
     1081 ccgggtgcgg tcgatgatta gggaacgctc gaactcggca atgccggcga acacggtcaa
     1141 caccatgcgg ccggccggcg tggtggtgtc ggcccacggc tctgccaggc tacgcaggcc
     1201 cgcgccggcc tcctggatgc gctcggcaat gtccagtagg tcgcgggtgc tgcgggccag
     1261 gcggtctagc ctggtcactg tcacaacgtc gccagggcgt aggtggtcaa gcatcctggc
     1321 cagctccggg cggtcgcgcc tggtgccggt gatcttctcg gaaaacagct tggtgcagcc
     1381 ggccgcgtgc agttcggccc gttggttggt caagtcctgg tcgtcggtgc tgacgcgggc
     1441 atagcccagc aggccagcgg cggcgctctt gttcatggcg taatgtctcc ggttctagtc
     1501 gcaagtattc tactttatgc gactaaaaca cgcgacaaga aaacgccagg aaaagggcag
     1561 ggcggcagcc tgtcgcgtaa cttaggactt gtgcgacatg tcgttttcag aagacggctg
     1621 cactgaacgt cagaaggaaa agtgccacct gaacgaagca tctgtgcttc attttgtaga
     1681 acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag aatctgagct gcatttttac
     1741 agaacagaaa tgcaacgcga aagcgctatt ttaccaacga agaatctgtg cttcattttt
     1801 gtaaaacaaa aatgcaacgc gagagcgcta atttttcaaa caaagaatct gagctgcatt
     1861 tttacagaac agaaatgcaa cgcgagagcg ctattttacc aacaaagaat ctatacttct
     1921 tttttgttct acaaaaatgc atcccgagag cgctattttt ctaacaaagc atcttagatt
     1981 actttttttc tcctttgtgc gctctataat gcagtctctt gataactttt tgcactgtag
     2041 gtccgttaag gttagaagaa ggctactttg gtgtctattt tctcttccat aaaaaaagcc
     2101 tgactccact tcccgcgttt actgattact agcgaagctg cgggtgcatt ttttcaagat
     2161 aaaggcatcc ccgattatat tctataccga tgtggattgc gcatactttg tgaacagaaa
     2221 gtgatagcgt tgatgattct tcattggtca gaaaattatg aacggtttct tctattttgt
     2281 ctctatatac tacgtatagg aaatgtttac attttcgtat tgttttcgat tcactctatg
     2341 aatagttctt actacaattt ttttgtctaa agagtaatac tagagataaa cataaaaaat
     2401 gtagaggtcg agtttagatg caagttcaag gagcgaaagg tggatgggta ggttatatag
     2461 ggatatagca cagagatata tagcaaagag atacttttga gcaatgtttg tggaagcggt
     2521 attcgcaata ttttagtagc tcgttacagt ccggtgcgtt tttggttttt tgaaagtgcg
     2581 tcttcagagc gcttttggtt ttcaaaagcg ctctgaagtt cctatacttt ctagagaata
     2641 ggaacttcgg aataggaact tcaaagcgtt tccgaaaacg agcgcttccg aaaatgcaac
     2701 gcgagctgcg cacatacagc tcactgttca cgtcgcacct atatctgcgt gttgcctgta
     2761 tatatatata catgagaaga acggcatagt gcgtgtttat gcttaaatgc gtacttatat
     2821 gcgtctattt atgtaggatg aaaggtagtc tagtacctcc tgtgatatta tcccattcca
     2881 tgcggggtat cgtatgcttc cttcagcact accctttagc tgttctatat gctgccactc
     2941 ctcaattgga ttagtctcat ccttcaatgc tatcatttcc tttgatattg gatcatacta
     3001 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     3061 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     3121 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     3181 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
     3241 gcaccatacc accttttcaa ttcatcattt tttttttatt cttttttttg atttcggttt
     3301 ccttgaaatt tttttgattc ggtaatctcc gaacagaagg aagaacgaag gaaggagcac
     3361 agacttagat tggtatatat acgcatatgt agtgttgaag aaacatgaaa ttgcccagta
     3421 ttcttaaccc aactgcacag aacaaaaacc tgcaggaaac gaagataaat catgtcgaaa
     3481 gctacatata aggaacgtgc tgctactcat cctagtcctg ttgctgccaa gctatttaat
     3541 atcatgcacg aaaagcaaac aaacttgtgt gcttcattgg atgttcgtac caccaaggaa
     3601 ttactggagt tagttgaagc attaggtccc aaaatttgtt tactaaaaac acatgtggat
     3661 atcttgactg atttttccat ggagggcaca gttaagccgc taaaggcatt atccgccaag
     3721 tacaattttt tactcttcga agacagaaaa tttgctgaca ttggtaatac agtcaaattg
     3781 cagtactctg cgggtgtata cagaatagca gaatgggcag acattacgaa tgcacacggt
     3841 gtggtgggcc caggtattgt tagcggtttg aagcaggcgg cagaagaagt aacaaaggaa
     3901 cctagaggcc ttttgatgct agcagaattg tcatgcaagg gctccctatc tactggagaa
     3961 tatactaagg gtactgttga cattgcgaag agcgacaaag attttgttat cggctttatt
     4021 gctcaaagag acatgggtgg aagagatgaa ggttacgatt ggttgattat gacacccggt
     4081 gtgggtttag atgacaaggg agacgcattg ggtcaacagt atagaaccgt ggatgatgtg
     4141 gtctctacag gatctgacat tattattgtt ggaagaggac tatttgcaaa gggaagggat
     4201 gctaaggtag agggtgaacg ttacagaaaa gcaggctggg aagcatattt gagaagatgc
     4261 ggccagcaaa actaaaaaac tgtattataa gtaaatgcat gtatactaaa ctcacaaatt
     4321 agagcttcaa tttaattata tcagttatta cctggcatgc acatacaaat ggacgaacgg
     4381 ataaaccttt tcacgccctt ttaaatatcc gattattcta ataaacgctc ttttctctta
     4441 ggtttacccg ccaatatatc ctgtcaaaca ctgatagttt aaactgaagg cgggaaacga
     4501 caatctgatc caagctcaag ctcattcgcc attcaggctg cgcaactgtt gggaagggcg
     4561 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg
     4621 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtgc
     4681 caagcttccg gtagaggtgt ggtcaataag agcgacctca tactatacct gagaaagcaa
     4741 cctgacctac aggaaagagt tactcaagaa taagaatttt cgttttaaaa cctaagagtc
     4801 actttaaaat ttgtatacac ttattttttt tataacttat ttaataataa aaatcataaa
     4861 tcataagaaa ttcgcttatt tagaagtggc gcgccattat tcctttgccc tcggacgagt
     4921 gctggggcgt cggtttccac tatcggcgag tacttctaca cagccatcgg tccagacggc
     4981 cgcgcttctg cgggcgattt gtgtacgccc gacagtcccg gctccggatc ggacgattgc
     5041 gtcgcatcga ccctgcgccc aagctgcatc atcgaaattg ccgtcaacca agctctgata
     5101 gagttggtca agaccaatgc ggagcatata cgcccggagc cgcggcgatc ctgcaagctc
     5161 cggatgcctc cgctcgaagt agcgcgtctg ctgctccata caagccaacc acggcctcca
     5221 gaagaagatg ttggcgacct cgtattggga atccccgaac atcgcctcgc tccagtcaat
     5281 gaccgctgtt atgcggccat tgtccgtcag gacattgttg gagccgaaat ccgcgtgcac
     5341 gaggtgccgg acttcggggc agtcctcggc ccaaagcatc agctcatcga gagcctgcgc
     5401 gacggacgca ctgacggtgt cgtccatcac agtttgccag tgatacacat ggggatcagc
     5461 aatcgcgcat atgaaatcac gccatgtagt gtattgaccg attccttgcg gtccgaatgg
     5521 gccgaacccg ctcgtctggc taagatcggc cgcagcgatc gcatccatgg cctccgcgac
     5581 cggctgcaga acagcgggca gttcggtttc aggcaggtct tgcaacgtga caccctgtgc
     5641 acggcgggag atgcaatagg tcaggctctc gctgaattcc ccaatgtcaa gcacttccgg
     5701 aatcgggagc gcggccgatg caaagtgccg ataaacataa cgatctttgt agaaaccatc
     5761 ggcgcagcta tttacccgca ggacatatcc acgccctcct acatcgaagc tgaaagcacg
     5821 agattcttcg ccctccgaga gctgcatcag gtcggagacg ctgtcgaact tttcgatcag
     5881 aaacttctcg acagacgtcg cggtgagttc aggcttttta cccatttaat taatgctgtg
     5941 taaggtgtgc gtgagagaga gggtcgtgag tgagtcttga agagaaacgt caaagcttga
     6001 agcgtagcag aatacggaaa acgacttggg ggaatggcga ggagcgtcgc cttttatgct
     6061 ccaagcggag acgatgaggc cgatattaca gtggcttagt agcgtgtaca atatttccat
     6121 aaaaggctac atggtgagtt tttcacgtga ccgtaattag tcccaattag gctttattgt
     6181 atattgctac taagcatcgt taaatgacct gctcgaagct tcgtctataa aaggcgggcg
     6241 tgtgtgcgct tcccccgcta cgtctttttc acaaacactt tgcgacttga ccaagacctt
     6301 tcacccgcac cactctcatc catcacacac acaagtcgac atggcaccca aggaagctcc
     6361 cgctgctgca gcccctactg gtggcaagaa gcccgctgga aaggcacccg caaagcccgc
     6421 aacagagaag aaggaaaagt caaagaggaa gactatccgt cgtgagacct actctacata
     6481 catctacaag gtgttgaagc aagtgcaccc cgacactggt atctccaaca aggccatgtc
     6541 catcatgaac tctttcgtca acgacatctt tgagcgtatt gcgggagagg catccaagct
     6601 cgctgcctac aacaagcgct ccaccatcag ctcacgtgag attcaaacct cagtgcgtct
     6661 tattcttccc ggtgaacttg ccaagcacgc tgtttctgag ggtaccaagg ctgtgacgaa
     6721 gtaccagtct gctaaatcct cctccggatc catggtgagc aagggcgagg aggtcatcaa
     6781 agagttcatg cgcttcaagg tgcgcatgga gggctccatg aacggccacg agttcgagat
     6841 cgagggcgag ggcgagggcc gcccctacga gggcacccag accgccaagc tgaaggtgac
     6901 caagggcggc cccctgccct tcgcctggga catcctgtcc ccccagttca tgtacggctc
     6961 caaggcgtac gtgaagcacc ccgccgacat ccccgattac aagaagctgt ccttccccga
     7021 gggcttcaag tgggagcgcg tgatgaactt cgaggacggc ggtctggtga ccgtgaccca
     7081 ggactcctcc ctgcaggacg gcacgctgat ctacaaggtg aagatgcgcg gcaccaactt
     7141 cccccccgac ggccccgtaa tgcagaagaa gaccatgggc tgggaggcct ccaccgagcg
     7201 cctgtacccc cgcgacggcg tgctgaaggg cgagatccac caggccctga agctgaagga
     7261 cggcggccac tacctggtgg agttcaagac catctacatg gccaagaagc ccgtgcaact
     7321 gcccggctac tactacgtgg acaccaagct ggacatcacc tcccacaacg aggactacac
     7381 catcgtggaa cagtacgagc gctccgaggg ccgccaccac ctgttcctgg ggcatggcac
     7441 cggcagcacc ggcagcggca gctccggcac cgcctcctcc gaggacaaca acatggccgt
     7501 catcaaagag ttcatgcgct tcaaggtgcg catggagggc tccatgaacg gccacgagtt
     7561 cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg ccaagctgaa
     7621 ggtgaccaag ggcggccccc tgcccttcgc ctgggacatc ctgtcccccc agttcatgta
     7681 cggctccaag gcgtacgtga agcaccccgc cgacatcccc gattacaaga agctgtcctt
     7741 ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggtc tggtgaccgt
     7801 gacccaggac tcctccctgc aggacggcac gctgatctac aaggtgaaga tgcgcggcac
     7861 caacttcccc cccgacggcc ccgtaatgca gaagaagacc atgggctggg aggcctccac
     7921 cgagcgcctg tacccccgcg acggcgtgct gaagggcgag atccaccagg ccctgaagct
     7981 gaaggacggc ggccactacc tggtggagtt caagaccatc tacatggcca agaagcccgt
     8041 gcaactgccc ggctactact acgtggacac caagctggac atcacctccc acaacgagga
     8101 ctacaccatc gtggaacagt acgagcgctc cgagggccgc caccacctgt tcctgtacgg
     8161 catggacgag ctgtacaagt aaaggcgcgc cacttctaaa taagcgaatt tcttatgatt
     8221 tatgattttt attattaaat aagttataaa aaaaataagt gtatacaaat tttaaagtga
     8281 ctcttaggtt ttaaaacgaa aattcttatt cttgagtaac tctttcctgt aggtcaggtt
     8341 gctttctcag gtatagtatg aggtcgctct tattgaccac acctctaccg ggaattcgta
     8401 atcatgtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     8461 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     8521 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     8581 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg gctagagcaa ttcggcgtta
     8641 attcagtaca ttaaaaacgt ccgcaatgtg ttattaagtt gtctaagcgt caatttgttt
     8701 acaccacaat atatcctgcc accagccagc caacagctcc ccgaccggca gctcggcaca
     8761 aaatcaccac tcgatacagg cagcccatca gtccgggacg gcgtcagcgg gagagccgtt
     8821 gtaaggcggc agactttgct catgttaccg atgctattcg gaagaacggc aactaagctg
     8881 ccgggtttga aacacggatg atctcgcgga gggtagcatg ttgattgtaa cgatgacaga
     8941 gcgttgctgc ctgtgatcca gatcatgaac aataaaactg tctgcttaca taaacagtaa
     9001 tacaaggggt gttatgagcc atattcaacg ggaaacgtct tgctctaggc cgcgattaaa
     9061 ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc
     9121 aggtgcgaca atctatcgat tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca
     9181 tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac
     9241 ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt
     9301 actcaccact gcgatccccg ggaaaacagc attccaggta ttagaagaat atcctgattc
     9361 aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt
     9421 ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat
     9481 gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga
     9541 acaagtctgg aaagaaatgc ataaactttt gccattctca ccggattcag tcgtcactca
     9601 tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga
     9661 tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct
     9721 cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc
     9781 tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttctaag aattaattca
     9841 tgatcctgca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     9901 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     9961 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
    10021 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
    10081 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
    10141 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
    10201 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
    10261 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
    10321 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
    10381 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
    10441 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
    10501 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
    10561 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
    10621 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
    10681 gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
    10741 caccgcatat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
    10801 acactccgct atcgctacgt gactgggtca tggctgcgcc ccgacacccg ccaacacccg
    10861 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
    10921 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgaggcagg
    10981 gtgccttgat gtgggcgccg gcggtcgagt ggcgacggcg cggcttgtcc gcgccctggt
    11041 agattgcctg gccgtaggcc agccattttt gagcggccag cggccgcgat aggccgacgc
    11101 gaagcggcgg ggcgtaggga gcgcagcgac cgaagggtag gcgctttttg cagctcttcg
    11161 gctgtgcgct ggccagacag ttatgcacag gccaggcggg ttttaagagt tttaataagt
    11221 tttaaagagt tttaggcgga aaaatcgcct tttttctctt ttatatcagt cacttacatg
    11281 tgtgaccggt tcccaatgta cggctttggg ttcccaatgt acgggttccg gttcccaatg
    11341 tacggctttg ggttcccaat gtacgtgcta tccacaggaa agagaccttt tcgacctttt
    11401 tcccctgcta gggcaatttg ccctagcatc tgctccgtac attaggaacc ggcggatgct
    11461 tcgccctcga tcaggttgcg gtagcgcatg actaggatcg ggccagcctg ccccgcctcc
    11521 tccttcaaat cgtactccgg caggtcattt gacccgatca gcttgcgcac ggtgaaacag
    11581 aacttcttga actctccggc gctgccactg cgttcgtaga tcgtcttgaa caaccatctg
    11641 gcttctgcct tgcctgcggc gcggcgtgcc aggcggtaga gaaaacggcc gatgccggga
    11701 tcgatcaaaa agtaatcggg gtgaaccgtc agcacgtccg ggttcttgcc ttctgtgatc
    11761 tcgcggtaca tccaatcagc tagctcgatc tcgatgtact ccggccgccc ggtttcgctc
    11821 tttacgatct tgtagcggct aatcaaggct tcaccctcgg ataccgtcac caggcggccg
    11881 ttcttggcct tcttcgtacg ctgcatggca acgtgcgtgg tgtttaaccg aatgcaggtt
    11941 tctaccaggt cgtctttctg ctttccgcca tcggctcgcc ggcagaactt gagtacgtcc
    12001 gcaacgtgtg gacggaacac gcggccgggc ttgtctccct tcccttcccg gtatcggttc
    12061 atggattcgg ttagatggga aaccgccatc agtaccaggt cgtaatccca cacactggcc
    12121 atgccggccg gccctgcgga aacctctacg tgcccgtctg gaagctcgta gcggatcacc
    12181 tcgccagctc gtcggtcacg cttcgacaga cggaaaacgg ccacgtccat gatgctgcga
    12241 ctatcgcggg tgcccacgtc atagagcatc ggaacgaaaa aatctggttg ctcgtcgccc
    12301 ttgggcggct tcctaatcga cggcgcaccg gctgccggcg gttgccggga ttctttgcgg
    12361 attcgatcag cggccgcttg ccacgattca ccggggcgtg cttctgcctc gatgcgttgc
    12421 cgctgggcgg cctgcgcggc cttcaacttc tccaccaggt catcacccag cgccgcgccg
    12481 atttgtaccg ggccggatgg tttgcgaccg ctcacgccga ttcctcgggc ttgggggttc
    12541 cagtgccatt gcagggccgg cagacaaccc agccgcttac gcctggccaa ccgcccgttc
    12601 ctccacacat ggggcattcc acggcgtcgg tgcctggttg ttcttgattt tccatgccgc
    12661 ctcctttagc cgctaaaatt catctactca tttattcatt tgctcattta ctctggtagc
    12721 tgcgcgatgt attcagatag cagctcggta atggtcttgc cttggcgtac cgcgtacatc
    12781 ttcagcttgg tgtgatcctc cgccggcaac tgaaagttga cccgcttcat ggctggcgtg
    12841 tctgccaggc tggccaacgt tgcagccttg ctgctgcgtg cgctcggacg gccggcactt
    12901 agcgtgtttg tgcttttgct cattttctct ttacctcatt aactcaaatg agttttgatt
    12961 taatttcagc ggccagcgcc tggacctcgc gggcagcgtc gccctcgggt tctgattcaa
    13021 gaacggttgt gccggcggcg gcagtgcctg ggtagctcac gcgctgcgtg atacgggact
    13081 ca
//
