LOCUS       sequence-417142-       11449 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Reporter gene expression in B. dendrobatidis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11449)
  AUTHORS   Kalinka E, Brody SM, Swafford AJM, Medina EM, Fritz-Laylin LK
  TITLE     Genetic transformation of the frog-killing chytrid fungus 
            Batrachochytrium dendrobatidis.
  JOURNAL   Proc Natl Acad Sci U S A. 2024 Jan 23;121(4):e2317928121. doi: 
            10.1073/pnas.2317928121. Epub 2024 Jan 18.
  PUBMED    38236738
REFERENCE   2  (bases 1 to 11449)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2024-01-23"; volume: "121(4):e2317928121. 
            doi"; pages: " 10.1073/pnas.2317928121. Epub 2024 Jan 18"
COMMENT     Sequence Label: pEK22
FEATURES             Location/Qualifiers
     source          1..11449
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..25
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     primer_bind     206..228
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     220..237
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     221..237
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             552..1577
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     CDS             552..1577
                     /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             1593..2108
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     terminator      2130..2317
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2185..2209)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(2332..2348)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2332..2348)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2345..2367)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2356..2372
                     /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(2380..2386,2387..2404,2405..2410))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2425..2446
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_feature    2623..2647
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     primer_bind     complement(2929..2948)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2940..3755
                     /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             2940..3755
                     /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(3013..3032)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3850..4438
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4339..4358
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4592..4609
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4624..4764
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      5108..5302
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(5368..6441)
                     /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(6870..7499)
                     /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      8660..10002
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(9623..9670)
                     /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     10043..10061
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        10264..10479
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             10480..11283
                     /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"
ORIGIN
        1 gtttacccgc caatatatcc tgtcaaacac tgatagttta aactgaaggc gggaaacgac
       61 aatctgatcc aagctcaagc tcattcgcca ttcaggctgc gcaactgttg ggaagggcga
      121 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga
      181 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc
      241 aagctttgtg atagtagata gtagataata gagaactgtt gatgaggatt tgttgtgcaa
      301 gaaactgtgg tttatatatc gtacagagtt taaagatgcg gcgcagtagc ctgccccaaa
      361 accatgttcg attcaataga gctcgtggtt gtgccccacc ctgctgcgcc tcacccccat
      421 cataatccgc ccttgttttg cctataaatg aataggtgct tcccaccatt ctataactcg
      481 accatccagt accaactgct actttaacta ccacagttta cctactctta ctctatcatc
      541 tcaattaatt aatgggtaaa aagcctgaac tcaccgcgac gtctgtcgag aagtttctga
      601 tcgaaaagtt cgacagcgtc tccgacctga tgcagctctc ggagggcgaa gaatctcgtg
      661 ctttcagctt cgatgtagga gggcgtggat atgtcctgcg ggtaaatagc tgcgccgatg
      721 gtttctacaa agatcgttat gtttatcggc actttgcatc ggccgcgctc ccgattccgg
      781 aagtgcttga cattggggaa ttcagcgaga gcctgaccta ttgcatctcc cgccgtgcac
      841 agggtgtcac gttgcaagac ctgcctgaaa ccgaactgcc cgctgttctg cagccggtcg
      901 cggaggccat ggatgcgatc gctgcggccg atcttagcca gacgagcggg ttcggcccat
      961 tcggaccgca aggaatcggt caatacacta catggcgtga tttcatatgc gcgattgctg
     1021 atccccatgt gtatcactgg caaactgtga tggacgacac cgtcagtgcg tccgtcgcgc
     1081 aggctctcga tgagctgatg ctttgggccg aggactgccc cgaagtccgg cacctcgtgc
     1141 acgcggattt cggctccaac aatgtcctga cggacaatgg ccgcataaca gcggtcattg
     1201 actggagcga ggcgatgttc ggggattccc aatacgaggt cgccaacatc ttcttctgga
     1261 ggccgtggtt ggcttgtatg gagcagcaga cgcgctactt cgagcggagg catccggagc
     1321 ttgcaggatc gccgcggctc cgggcgtata tgctccgcat tggtcttgac caactctatc
     1381 agagcttggt tgacggcaat ttcgatgatg cagcttgggc gcagggtcga tgcgacgcaa
     1441 tcgtccgatc cggagccggg actgtcgggc gtacacaaat cgcccgcaga agcgcggccg
     1501 tctggaccga tggctgtgta gaagtactcg ccgatagtgg aaaccgacgc cccagcactc
     1561 gtccgagggc aaaggaagga ggaggaggat ccatggtctt cacactcgaa gatttcgttg
     1621 gggactggcg acagacagcc ggctacaacc tggaccaagt ccttgaacag ggaggtgtgt
     1681 ccagtttgtt tcagaatctc ggggtgtccg taactccgat ccaaaggatt gtcctgagcg
     1741 gtgaaaatgg gctgaagatc gacatccatg tcatcatccc gtatgaaggt ctgagcggcg
     1801 accaaatggg ccagatcgaa aaaattttta aggtggtgta ccctgtggat gatcatcact
     1861 ttaaggtgat cctgcactat ggcacactgg taatcgacgg ggttacgccg aacatgatcg
     1921 actatttcgg acggccgtat gaaggcatcg ccgtgttcga cggcaaaaag atcactgtaa
     1981 cagggaccct gtggaacggc aacaaaatta tcgacgagcg cctgatcaac cccgacggct
     2041 ccctgctgtt ccgagtaacc atcaacggag tgaccggctg gcggctgtgc gaacgcattc
     2101 tggcgtaaag gcgcgccact tctaaataag cgaatttctt atgatttatg atttttatta
     2161 ttaaataagt tataaaaaaa ataagtgtat acaaatttta aagtgactct taggttttaa
     2221 aacgaaaatt cttattcttg agtaactctt tcctgtaggt caggttgctt tctcaggtat
     2281 agtatgaggt cgctcttatt gaccacacct ctaccgggaa ttcgtaatca tgtcatagct
     2341 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     2401 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     2461 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     2521 cgcggggaga ggcggtttgc gtattggcta gagcaattcg gcgttaattc agtacattaa
     2581 aaacgtccgc aatgtgttat taagttgtct aagcgtcaat ttgtttacac cacaatatat
     2641 cctgccacca gccagccaac agctccccga ccggcagctc ggcacaaaat caccactcga
     2701 tacaggcagc ccatcagtcc gggacggcgt cagcgggaga gccgttgtaa ggcggcagac
     2761 tttgctcatg ttaccgatgc tattcggaag aacggcaact aagctgccgg gtttgaaaca
     2821 cggatgatct cgcggagggt agcatgttga ttgtaacgat gacagagcgt tgctgcctgt
     2881 gatccagatc atgaacaata aaactgtctg cttacataaa cagtaataca aggggtgtta
     2941 tgagccatat tcaacgggaa acgtcttgct ctaggccgcg attaaattcc aacatggatg
     3001 ctgatttata tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct
     3061 atcgattgta tgggaagccc gatgcgccag agttgtttct gaaacatggc aaaggtagcg
     3121 ttgccaatga tgttacagat gagatggtca gactaaactg gctgacggaa tttatgcctc
     3181 ttccgaccat caagcatttt atccgtactc ctgatgatgc atggttactc accactgcga
     3241 tccccgggaa aacagcattc caggtattag aagaatatcc tgattcaggt gaaaatattg
     3301 ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt
     3361 ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg
     3421 ttgatgcgag tgattttgat gacgagcgta atggctggcc tgttgaacaa gtctggaaag
     3481 aaatgcataa acttttgcca ttctcaccgg attcagtcgt cactcatggt gatttctcac
     3541 ttgataacct tatttttgac gaggggaaat taataggttg tattgatgtt ggacgagtcg
     3601 gaatcgcaga ccgataccag gatcttgcca tcctatggaa ctgcctcggt gagttttctc
     3661 cttcattaca gaaacggctt tttcaaaaat atggtattga taatcctgat atgaataaat
     3721 tgcagtttca tttgatgctc gatgagtttt tctaagaatt aattcatgat cctgcatgac
     3781 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     3841 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     3901 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     3961 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
     4021 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     4081 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     4141 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     4201 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     4261 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     4321 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     4381 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     4441 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     4501 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     4561 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     4621 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatggtg
     4681 cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg
     4741 ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga
     4801 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     4861 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagggtgc cttgatgtgg
     4921 gcgccggcgg tcgagtggcg acggcgcggc ttgtccgcgc cctggtagat tgcctggccg
     4981 taggccagcc atttttgagc ggccagcggc cgcgataggc cgacgcgaag cggcggggcg
     5041 tagggagcgc agcgaccgaa gggtaggcgc tttttgcagc tcttcggctg tgcgctggcc
     5101 agacagttat gcacaggcca ggcgggtttt aagagtttta ataagtttta aagagtttta
     5161 ggcggaaaaa tcgccttttt tctcttttat atcagtcact tacatgtgtg accggttccc
     5221 aatgtacggc tttgggttcc caatgtacgg gttccggttc ccaatgtacg gctttgggtt
     5281 cccaatgtac gtgctatcca caggaaagag accttttcga cctttttccc ctgctagggc
     5341 aatttgccct agcatctgct ccgtacatta ggaaccggcg gatgcttcgc cctcgatcag
     5401 gttgcggtag cgcatgacta ggatcgggcc agcctgcccc gcctcctcct tcaaatcgta
     5461 ctccggcagg tcatttgacc cgatcagctt gcgcacggtg aaacagaact tcttgaactc
     5521 tccggcgctg ccactgcgtt cgtagatcgt cttgaacaac catctggctt ctgccttgcc
     5581 tgcggcgcgg cgtgccaggc ggtagagaaa acggccgatg ccgggatcga tcaaaaagta
     5641 atcggggtga accgtcagca cgtccgggtt cttgccttct gtgatctcgc ggtacatcca
     5701 atcagctagc tcgatctcga tgtactccgg ccgcccggtt tcgctcttta cgatcttgta
     5761 gcggctaatc aaggcttcac cctcggatac cgtcaccagg cggccgttct tggccttctt
     5821 cgtacgctgc atggcaacgt gcgtggtgtt taaccgaatg caggtttcta ccaggtcgtc
     5881 tttctgcttt ccgccatcgg ctcgccggca gaacttgagt acgtccgcaa cgtgtggacg
     5941 gaacacgcgg ccgggcttgt ctcccttccc ttcccggtat cggttcatgg attcggttag
     6001 atgggaaacc gccatcagta ccaggtcgta atcccacaca ctggccatgc cggccggccc
     6061 tgcggaaacc tctacgtgcc cgtctggaag ctcgtagcgg atcacctcgc cagctcgtcg
     6121 gtcacgcttc gacagacgga aaacggccac gtccatgatg ctgcgactat cgcgggtgcc
     6181 cacgtcatag agcatcggaa cgaaaaaatc tggttgctcg tcgcccttgg gcggcttcct
     6241 aatcgacggc gcaccggctg ccggcggttg ccgggattct ttgcggattc gatcagcggc
     6301 cgcttgccac gattcaccgg ggcgtgcttc tgcctcgatg cgttgccgct gggcggcctg
     6361 cgcggccttc aacttctcca ccaggtcatc acccagcgcc gcgccgattt gtaccgggcc
     6421 ggatggtttg cgaccgctca cgccgattcc tcgggcttgg gggttccagt gccattgcag
     6481 ggccggcaga caacccagcc gcttacgcct ggccaaccgc ccgttcctcc acacatgggg
     6541 cattccacgg cgtcggtgcc tggttgttct tgattttcca tgccgcctcc tttagccgct
     6601 aaaattcatc tactcattta ttcatttgct catttactct ggtagctgcg cgatgtattc
     6661 agatagcagc tcggtaatgg tcttgccttg gcgtaccgcg tacatcttca gcttggtgtg
     6721 atcctccgcc ggcaactgaa agttgacccg cttcatggct ggcgtgtctg ccaggctggc
     6781 caacgttgca gccttgctgc tgcgtgcgct cggacggccg gcacttagcg tgtttgtgct
     6841 tttgctcatt ttctctttac ctcattaact caaatgagtt ttgatttaat ttcagcggcc
     6901 agcgcctgga cctcgcgggc agcgtcgccc tcgggttctg attcaagaac ggttgtgccg
     6961 gcggcggcag tgcctgggta gctcacgcgc tgcgtgatac gggactcaag aatgggcagc
     7021 tcgtacccgg ccagcgcctc ggcaacctca ccgccgatgc gcgtgccttt gatcgcccgc
     7081 gacacgacaa aggccgcttg tagccttcca tccgtgacct caatgcgctg cttaaccagc
     7141 tccaccaggt cggcggtggc ccatatgtcg taagggcttg gctgcaccgg aatcagcacg
     7201 aagtcggctg ccttgatcgc ggacacagcc aagtccgccg cctggggcgc tccgtcgatc
     7261 actacgaagt cgcgccggcc gatggccttc acgtcgcggt caatcgtcgg gcggtcgatg
     7321 ccgacaacgg ttagcggttg atcttcccgc acggccgccc aatcgcgggc actgccctgg
     7381 ggatcggaat cgactaacag aacatcggcc ccggcgagtt gcagggcgcg ggctagatgg
     7441 gttgcgatgg tcgtcttgcc tgacccgcct ttctggttaa gtacagcgat aaccttcatg
     7501 cgttcccctt gcgtatttgt ttatttactc atcgcatcat atacgcagcg accgcatgac
     7561 gcaagctgtt ttactcaaat acacatcacc tttttagacg gcggcgctcg gtttcttcag
     7621 cggccaagct ggccggccag gccgccagct tggcatcaga caaaccggcc aggatttcat
     7681 gcagccgcac ggttgagacg tgcgcgggcg gctcgaacac gtacccggcc gcgatcatct
     7741 ccgcctcgat ctcttcggta atgaaaaacg gttcgtcctg gccgtcctgg tgcggtttca
     7801 tgcttgttcc tcttggcgtt cattctcggc ggccgccagg gcgtcggcct cggtcaatgc
     7861 gtcctcacgg aaggcaccgc gccgcctggc ctcggtgggc gtcacttcct cgctgcgctc
     7921 aagtgcgcgg tacagggtcg agcgatgcac gccaagcagt gcagccgcct ctttcacggt
     7981 gcggccttcc tggtcgatca gctcgcgggc gtgcgcgatc tgtgccgggg tgagggtagg
     8041 gcgggggcca aacttcacgc ctcgggcctt ggcggcctcg cgcccgctcc gggtgcggtc
     8101 gatgattagg gaacgctcga actcggcaat gccggcgaac acggtcaaca ccatgcggcc
     8161 ggccggcgtg gtggtgtcgg cccacggctc tgccaggcta cgcaggcccg cgccggcctc
     8221 ctggatgcgc tcggcaatgt ccagtaggtc gcgggtgctg cgggccaggc ggtctagcct
     8281 ggtcactgtc acaacgtcgc cagggcgtag gtggtcaagc atcctggcca gctccgggcg
     8341 gtcgcgcctg gtgccggtga tcttctcgga aaacagcttg gtgcagccgg ccgcgtgcag
     8401 ttcggcccgt tggttggtca agtcctggtc gtcggtgctg acgcgggcat agcccagcag
     8461 gccagcggcg gcgctcttgt tcatggcgta atgtctccgg ttctagtcgc aagtattcta
     8521 ctttatgcga ctaaaacacg cgacaagaaa acgccaggaa aagggcaggg cggcagcctg
     8581 tcgcgtaact taggacttgt gcgacatgtc gttttcagaa gacggctgca ctgaacgtca
     8641 gaaggaaaag tgccacctga acgaagcatc tgtgcttcat tttgtagaac aaaaatgcaa
     8701 cgcgagagcg ctaatttttc aaacaaagaa tctgagctgc atttttacag aacagaaatg
     8761 caacgcgaaa gcgctatttt accaacgaag aatctgtgct tcatttttgt aaaacaaaaa
     8821 tgcaacgcga gagcgctaat ttttcaaaca aagaatctga gctgcatttt tacagaacag
     8881 aaatgcaacg cgagagcgct attttaccaa caaagaatct atacttcttt tttgttctac
     8941 aaaaatgcat cccgagagcg ctatttttct aacaaagcat cttagattac tttttttctc
     9001 ctttgtgcgc tctataatgc agtctcttga taactttttg cactgtaggt ccgttaaggt
     9061 tagaagaagg ctactttggt gtctattttc tcttccataa aaaaagcctg actccacttc
     9121 ccgcgtttac tgattactag cgaagctgcg ggtgcatttt ttcaagataa aggcatcccc
     9181 gattatattc tataccgatg tggattgcgc atactttgtg aacagaaagt gatagcgttg
     9241 atgattcttc attggtcaga aaattatgaa cggtttcttc tattttgtct ctatatacta
     9301 cgtataggaa atgtttacat tttcgtattg ttttcgattc actctatgaa tagttcttac
     9361 tacaattttt ttgtctaaag agtaatacta gagataaaca taaaaaatgt agaggtcgag
     9421 tttagatgca agttcaagga gcgaaaggtg gatgggtagg ttatataggg atatagcaca
     9481 gagatatata gcaaagagat acttttgagc aatgtttgtg gaagcggtat tcgcaatatt
     9541 ttagtagctc gttacagtcc ggtgcgtttt tggttttttg aaagtgcgtc ttcagagcgc
     9601 ttttggtttt caaaagcgct ctgaagttcc tatactttct agagaatagg aacttcggaa
     9661 taggaacttc aaagcgtttc cgaaaacgag cgcttccgaa aatgcaacgc gagctgcgca
     9721 catacagctc actgttcacg tcgcacctat atctgcgtgt tgcctgtata tatatataca
     9781 tgagaagaac ggcatagtgc gtgtttatgc ttaaatgcgt acttatatgc gtctatttat
     9841 gtaggatgaa aggtagtcta gtacctcctg tgatattatc ccattccatg cggggtatcg
     9901 tatgcttcct tcagcactac cctttagctg ttctatatgc tgccactcct caattggatt
     9961 agtctcatcc ttcaatgcta tcatttcctt tgatattgga tcatactaag aaaccattat
    10021 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt
    10081 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
    10141 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
    10201 tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accataccac
    10261 cttttcaatt catcattttt tttttattct tttttttgat ttcggtttcc ttgaaatttt
    10321 tttgattcgg taatctccga acagaaggaa gaacgaagga aggagcacag acttagattg
    10381 gtatatatac gcatatgtag tgttgaagaa acatgaaatt gcccagtatt cttaacccaa
    10441 ctgcacagaa caaaaacctg caggaaacga agataaatca tgtcgaaagc tacatataag
    10501 gaacgtgctg ctactcatcc tagtcctgtt gctgccaagc tatttaatat catgcacgaa
    10561 aagcaaacaa acttgtgtgc ttcattggat gttcgtacca ccaaggaatt actggagtta
    10621 gttgaagcat taggtcccaa aatttgttta ctaaaaacac atgtggatat cttgactgat
    10681 ttttccatgg agggcacagt taagccgcta aaggcattat ccgccaagta caatttttta
    10741 ctcttcgaag acagaaaatt tgctgacatt ggtaatacag tcaaattgca gtactctgcg
    10801 ggtgtataca gaatagcaga atgggcagac attacgaatg cacacggtgt ggtgggccca
    10861 ggtattgtta gcggtttgaa gcaggcggca gaagaagtaa caaaggaacc tagaggcctt
    10921 ttgatgctag cagaattgtc atgcaagggc tccctatcta ctggagaata tactaagggt
    10981 actgttgaca ttgcgaagag cgacaaagat tttgttatcg gctttattgc tcaaagagac
    11041 atgggtggaa gagatgaagg ttacgattgg ttgattatga cacccggtgt gggtttagat
    11101 gacaagggag acgcattggg tcaacagtat agaaccgtgg atgatgtggt ctctacagga
    11161 tctgacatta ttattgttgg aagaggacta tttgcaaagg gaagggatgc taaggtagag
    11221 ggtgaacgtt acagaaaagc aggctgggaa gcatatttga gaagatgcgg ccagcaaaac
    11281 taaaaaactg tattataagt aaatgcatgt atactaaact cacaaattag agcttcaatt
    11341 taattatatc agttattacc tggcatgcac atacaaatgg acgaacggat aaaccttttc
    11401 acgccctttt aaatatccga ttattctaat aaacgctctt ttctcttag
//
