LOCUS       sequence-417150-       12081 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 12081)
  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 12081)
  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: pEK21
FEATURES             Location/Qualifiers
     source          1..12081
                     /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             1184..2209
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     CDS             1184..2209
                     /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             2225..2740
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     terminator      2762..2949
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2817..2841)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(2964..2980)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2964..2980)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2977..2999)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2988..3004
                     /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(3012..3018,3019..3036,3037..3042))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3057..3078
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_feature    3255..3279
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     primer_bind     complement(3561..3580)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3572..4387
                     /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             3572..4387
                     /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(3645..3664)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4482..5070
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4971..4990
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5224..5241
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5256..5396
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      5740..5934
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(6000..7073)
                     /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(7502..8131)
                     /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      9292..10634
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(10255..10302)
                     /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     10675..10693
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        10896..11111
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             11112..11915
                     /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 aagctttgct gtattctttc ttgctgtttt ataaatagct ctaatggatt atcagtccat
      301 tttataaact ggtttaaaat aaaggctttt tcatgataca agtcttttac atgtgttcgc
      361 caagaaggat agcgatacaa ggaaactcga aaagcaaatg ccagatattg tttgaatgac
      421 aatatgaatg tttgtaataa agcagcatca cgtgattggt tgggtttgtt atggcttatt
      481 cacgcacaac acatttgtat tcatcttgca gtgactatct tggaatatac aatagttgaa
      541 tatttgtttt gctttcctat tcaagattgg cagtttcaat aaacacaacg gtcattatat
      601 gatacacaat caaactatgg ctctaaactg atattgccct cccttgacat ggtcatacat
      661 tatgcaatct tcaagtttaa aattagatat tgaaatcgac tattttcaat acaattctat
      721 acaaacccct gtaccctaat ctcacagata tgtttgcgct ctagttcaag aaagcaatga
      781 ttgcatgtat tgggaataca agcgttgctt ccatgccata tccaatgctt ggtttgtaga
      841 gaaagataat tgaacaagtg tcaagaatgt agaatttatt gaagcagaga tgtgagcatt
      901 tagtcagctt tctatacaga aatgagcaag accagtcaac taaatagttt aaaagacaat
      961 attgtccagt aagtttgatg gcatcattat ttcacgagat tcgctgttta ttatattttc
     1021 gcgcgttctc atgtctattg gcaccactgt aatttcctat aaatatggac tgaaatcgtt
     1081 tttctttcca ccatagtttt cccttttagc gtcttcacct ttaccgaagc cgcgaactga
     1141 ccaactactt ttcaactttc ttctactctt tttattttaa ttaatgggta aaaagcctga
     1201 actcaccgcg acgtctgtcg agaagtttct gatcgaaaag ttcgacagcg tctccgacct
     1261 gatgcagctc tcggagggcg aagaatctcg tgctttcagc ttcgatgtag gagggcgtgg
     1321 atatgtcctg cgggtaaata gctgcgccga tggtttctac aaagatcgtt atgtttatcg
     1381 gcactttgca tcggccgcgc tcccgattcc ggaagtgctt gacattgggg aattcagcga
     1441 gagcctgacc tattgcatct cccgccgtgc acagggtgtc acgttgcaag acctgcctga
     1501 aaccgaactg cccgctgttc tgcagccggt cgcggaggcc atggatgcga tcgctgcggc
     1561 cgatcttagc cagacgagcg ggttcggccc attcggaccg caaggaatcg gtcaatacac
     1621 tacatggcgt gatttcatat gcgcgattgc tgatccccat gtgtatcact ggcaaactgt
     1681 gatggacgac accgtcagtg cgtccgtcgc gcaggctctc gatgagctga tgctttgggc
     1741 cgaggactgc cccgaagtcc ggcacctcgt gcacgcggat ttcggctcca acaatgtcct
     1801 gacggacaat ggccgcataa cagcggtcat tgactggagc gaggcgatgt tcggggattc
     1861 ccaatacgag gtcgccaaca tcttcttctg gaggccgtgg ttggcttgta tggagcagca
     1921 gacgcgctac ttcgagcgga ggcatccgga gcttgcagga tcgccgcggc tccgggcgta
     1981 tatgctccgc attggtcttg accaactcta tcagagcttg gttgacggca atttcgatga
     2041 tgcagcttgg gcgcagggtc gatgcgacgc aatcgtccga tccggagccg ggactgtcgg
     2101 gcgtacacaa atcgcccgca gaagcgcggc cgtctggacc gatggctgtg tagaagtact
     2161 cgccgatagt ggaaaccgac gccccagcac tcgtccgagg gcaaaggaag gaggaggagg
     2221 atccatggtc ttcacactcg aagatttcgt tggggactgg cgacagacag ccggctacaa
     2281 cctggaccaa gtccttgaac agggaggtgt gtccagtttg tttcagaatc tcggggtgtc
     2341 cgtaactccg atccaaagga ttgtcctgag cggtgaaaat gggctgaaga tcgacatcca
     2401 tgtcatcatc ccgtatgaag gtctgagcgg cgaccaaatg ggccagatcg aaaaaatttt
     2461 taaggtggtg taccctgtgg atgatcatca ctttaaggtg atcctgcact atggcacact
     2521 ggtaatcgac ggggttacgc cgaacatgat cgactatttc ggacggccgt atgaaggcat
     2581 cgccgtgttc gacggcaaaa agatcactgt aacagggacc ctgtggaacg gcaacaaaat
     2641 tatcgacgag cgcctgatca accccgacgg ctccctgctg ttccgagtaa ccatcaacgg
     2701 agtgaccggc tggcggctgt gcgaacgcat tctggcgtaa aggcgcgcca cttctaaata
     2761 agcgaatttc ttatgattta tgatttttat tattaaataa gttataaaaa aaataagtgt
     2821 atacaaattt taaagtgact cttaggtttt aaaacgaaaa ttcttattct tgagtaactc
     2881 tttcctgtag gtcaggttgc tttctcaggt atagtatgag gtcgctctta ttgaccacac
     2941 ctctaccggg aattcgtaat catgtcatag ctgtttcctg tgtgaaattg ttatccgctc
     3001 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     3061 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     3121 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggc
     3181 tagagcaatt cggcgttaat tcagtacatt aaaaacgtcc gcaatgtgtt attaagttgt
     3241 ctaagcgtca atttgtttac accacaatat atcctgccac cagccagcca acagctcccc
     3301 gaccggcagc tcggcacaaa atcaccactc gatacaggca gcccatcagt ccgggacggc
     3361 gtcagcggga gagccgttgt aaggcggcag actttgctca tgttaccgat gctattcgga
     3421 agaacggcaa ctaagctgcc gggtttgaaa cacggatgat ctcgcggagg gtagcatgtt
     3481 gattgtaacg atgacagagc gttgctgcct gtgatccaga tcatgaacaa taaaactgtc
     3541 tgcttacata aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcttg
     3601 ctctaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg
     3661 cgataatgtc gggcaatcag gtgcgacaat ctatcgattg tatgggaagc ccgatgcgcc
     3721 agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt
     3781 cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac
     3841 tcctgatgat gcatggttac tcaccactgc gatccccggg aaaacagcat tccaggtatt
     3901 agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg
     3961 gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc
     4021 tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg
     4081 taatggctgg cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc
     4141 ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa
     4201 attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc
     4261 catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa
     4321 atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt
     4381 tttctaagaa ttaattcatg atcctgcatg accaaaatcc cttaacgtga gttttcgttc
     4441 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     4501 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     4561 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     4621 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     4681 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     4741 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     4801 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     4861 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     4921 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     4981 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     5041 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     5101 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     5161 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     5221 cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg
     5281 catctgtgcg gtatttcaca ccgcatatgg tgcactctca gtacaatctg ctctgatgcc
     5341 gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg gctgcgcccc
     5401 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     5461 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     5521 cgaaacgcgc gaggcagggt gccttgatgt gggcgccggc ggtcgagtgg cgacggcgcg
     5581 gcttgtccgc gccctggtag attgcctggc cgtaggccag ccatttttga gcggccagcg
     5641 gccgcgatag gccgacgcga agcggcgggg cgtagggagc gcagcgaccg aagggtaggc
     5701 gctttttgca gctcttcggc tgtgcgctgg ccagacagtt atgcacaggc caggcgggtt
     5761 ttaagagttt taataagttt taaagagttt taggcggaaa aatcgccttt tttctctttt
     5821 atatcagtca cttacatgtg tgaccggttc ccaatgtacg gctttgggtt cccaatgtac
     5881 gggttccggt tcccaatgta cggctttggg ttcccaatgt acgtgctatc cacaggaaag
     5941 agaccttttc gacctttttc ccctgctagg gcaatttgcc ctagcatctg ctccgtacat
     6001 taggaaccgg cggatgcttc gccctcgatc aggttgcggt agcgcatgac taggatcggg
     6061 ccagcctgcc ccgcctcctc cttcaaatcg tactccggca ggtcatttga cccgatcagc
     6121 ttgcgcacgg tgaaacagaa cttcttgaac tctccggcgc tgccactgcg ttcgtagatc
     6181 gtcttgaaca accatctggc ttctgccttg cctgcggcgc ggcgtgccag gcggtagaga
     6241 aaacggccga tgccgggatc gatcaaaaag taatcggggt gaaccgtcag cacgtccggg
     6301 ttcttgcctt ctgtgatctc gcggtacatc caatcagcta gctcgatctc gatgtactcc
     6361 ggccgcccgg tttcgctctt tacgatcttg tagcggctaa tcaaggcttc accctcggat
     6421 accgtcacca ggcggccgtt cttggccttc ttcgtacgct gcatggcaac gtgcgtggtg
     6481 tttaaccgaa tgcaggtttc taccaggtcg tctttctgct ttccgccatc ggctcgccgg
     6541 cagaacttga gtacgtccgc aacgtgtgga cggaacacgc ggccgggctt gtctcccttc
     6601 ccttcccggt atcggttcat ggattcggtt agatgggaaa ccgccatcag taccaggtcg
     6661 taatcccaca cactggccat gccggccggc cctgcggaaa cctctacgtg cccgtctgga
     6721 agctcgtagc ggatcacctc gccagctcgt cggtcacgct tcgacagacg gaaaacggcc
     6781 acgtccatga tgctgcgact atcgcgggtg cccacgtcat agagcatcgg aacgaaaaaa
     6841 tctggttgct cgtcgccctt gggcggcttc ctaatcgacg gcgcaccggc tgccggcggt
     6901 tgccgggatt ctttgcggat tcgatcagcg gccgcttgcc acgattcacc ggggcgtgct
     6961 tctgcctcga tgcgttgccg ctgggcggcc tgcgcggcct tcaacttctc caccaggtca
     7021 tcacccagcg ccgcgccgat ttgtaccggg ccggatggtt tgcgaccgct cacgccgatt
     7081 cctcgggctt gggggttcca gtgccattgc agggccggca gacaacccag ccgcttacgc
     7141 ctggccaacc gcccgttcct ccacacatgg ggcattccac ggcgtcggtg cctggttgtt
     7201 cttgattttc catgccgcct cctttagccg ctaaaattca tctactcatt tattcatttg
     7261 ctcatttact ctggtagctg cgcgatgtat tcagatagca gctcggtaat ggtcttgcct
     7321 tggcgtaccg cgtacatctt cagcttggtg tgatcctccg ccggcaactg aaagttgacc
     7381 cgcttcatgg ctggcgtgtc tgccaggctg gccaacgttg cagccttgct gctgcgtgcg
     7441 ctcggacggc cggcacttag cgtgtttgtg cttttgctca ttttctcttt acctcattaa
     7501 ctcaaatgag ttttgattta atttcagcgg ccagcgcctg gacctcgcgg gcagcgtcgc
     7561 cctcgggttc tgattcaaga acggttgtgc cggcggcggc agtgcctggg tagctcacgc
     7621 gctgcgtgat acgggactca agaatgggca gctcgtaccc ggccagcgcc tcggcaacct
     7681 caccgccgat gcgcgtgcct ttgatcgccc gcgacacgac aaaggccgct tgtagccttc
     7741 catccgtgac ctcaatgcgc tgcttaacca gctccaccag gtcggcggtg gcccatatgt
     7801 cgtaagggct tggctgcacc ggaatcagca cgaagtcggc tgccttgatc gcggacacag
     7861 ccaagtccgc cgcctggggc gctccgtcga tcactacgaa gtcgcgccgg ccgatggcct
     7921 tcacgtcgcg gtcaatcgtc gggcggtcga tgccgacaac ggttagcggt tgatcttccc
     7981 gcacggccgc ccaatcgcgg gcactgccct ggggatcgga atcgactaac agaacatcgg
     8041 ccccggcgag ttgcagggcg cgggctagat gggttgcgat ggtcgtcttg cctgacccgc
     8101 ctttctggtt aagtacagcg ataaccttca tgcgttcccc ttgcgtattt gtttatttac
     8161 tcatcgcatc atatacgcag cgaccgcatg acgcaagctg ttttactcaa atacacatca
     8221 cctttttaga cggcggcgct cggtttcttc agcggccaag ctggccggcc aggccgccag
     8281 cttggcatca gacaaaccgg ccaggatttc atgcagccgc acggttgaga cgtgcgcggg
     8341 cggctcgaac acgtacccgg ccgcgatcat ctccgcctcg atctcttcgg taatgaaaaa
     8401 cggttcgtcc tggccgtcct ggtgcggttt catgcttgtt cctcttggcg ttcattctcg
     8461 gcggccgcca gggcgtcggc ctcggtcaat gcgtcctcac ggaaggcacc gcgccgcctg
     8521 gcctcggtgg gcgtcacttc ctcgctgcgc tcaagtgcgc ggtacagggt cgagcgatgc
     8581 acgccaagca gtgcagccgc ctctttcacg gtgcggcctt cctggtcgat cagctcgcgg
     8641 gcgtgcgcga tctgtgccgg ggtgagggta gggcgggggc caaacttcac gcctcgggcc
     8701 ttggcggcct cgcgcccgct ccgggtgcgg tcgatgatta gggaacgctc gaactcggca
     8761 atgccggcga acacggtcaa caccatgcgg ccggccggcg tggtggtgtc ggcccacggc
     8821 tctgccaggc tacgcaggcc cgcgccggcc tcctggatgc gctcggcaat gtccagtagg
     8881 tcgcgggtgc tgcgggccag gcggtctagc ctggtcactg tcacaacgtc gccagggcgt
     8941 aggtggtcaa gcatcctggc cagctccggg cggtcgcgcc tggtgccggt gatcttctcg
     9001 gaaaacagct tggtgcagcc ggccgcgtgc agttcggccc gttggttggt caagtcctgg
     9061 tcgtcggtgc tgacgcgggc atagcccagc aggccagcgg cggcgctctt gttcatggcg
     9121 taatgtctcc ggttctagtc gcaagtattc tactttatgc gactaaaaca cgcgacaaga
     9181 aaacgccagg aaaagggcag ggcggcagcc tgtcgcgtaa cttaggactt gtgcgacatg
     9241 tcgttttcag aagacggctg cactgaacgt cagaaggaaa agtgccacct gaacgaagca
     9301 tctgtgcttc attttgtaga acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag
     9361 aatctgagct gcatttttac agaacagaaa tgcaacgcga aagcgctatt ttaccaacga
     9421 agaatctgtg cttcattttt gtaaaacaaa aatgcaacgc gagagcgcta atttttcaaa
     9481 caaagaatct gagctgcatt tttacagaac agaaatgcaa cgcgagagcg ctattttacc
     9541 aacaaagaat ctatacttct tttttgttct acaaaaatgc atcccgagag cgctattttt
     9601 ctaacaaagc atcttagatt actttttttc tcctttgtgc gctctataat gcagtctctt
     9661 gataactttt tgcactgtag gtccgttaag gttagaagaa ggctactttg gtgtctattt
     9721 tctcttccat aaaaaaagcc tgactccact tcccgcgttt actgattact agcgaagctg
     9781 cgggtgcatt ttttcaagat aaaggcatcc ccgattatat tctataccga tgtggattgc
     9841 gcatactttg tgaacagaaa gtgatagcgt tgatgattct tcattggtca gaaaattatg
     9901 aacggtttct tctattttgt ctctatatac tacgtatagg aaatgtttac attttcgtat
     9961 tgttttcgat tcactctatg aatagttctt actacaattt ttttgtctaa agagtaatac
    10021 tagagataaa cataaaaaat gtagaggtcg agtttagatg caagttcaag gagcgaaagg
    10081 tggatgggta ggttatatag ggatatagca cagagatata tagcaaagag atacttttga
    10141 gcaatgtttg tggaagcggt attcgcaata ttttagtagc tcgttacagt ccggtgcgtt
    10201 tttggttttt tgaaagtgcg tcttcagagc gcttttggtt ttcaaaagcg ctctgaagtt
    10261 cctatacttt ctagagaata ggaacttcgg aataggaact tcaaagcgtt tccgaaaacg
    10321 agcgcttccg aaaatgcaac gcgagctgcg cacatacagc tcactgttca cgtcgcacct
    10381 atatctgcgt gttgcctgta tatatatata catgagaaga acggcatagt gcgtgtttat
    10441 gcttaaatgc gtacttatat gcgtctattt atgtaggatg aaaggtagtc tagtacctcc
    10501 tgtgatatta tcccattcca tgcggggtat cgtatgcttc cttcagcact accctttagc
    10561 tgttctatat gctgccactc ctcaattgga ttagtctcat ccttcaatgc tatcatttcc
    10621 tttgatattg gatcatacta agaaaccatt attatcatga cattaaccta taaaaatagg
    10681 cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
    10741 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
    10801 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca
    10861 gagcagattg tactgagagt gcaccatacc accttttcaa ttcatcattt tttttttatt
    10921 cttttttttg atttcggttt ccttgaaatt tttttgattc ggtaatctcc gaacagaagg
    10981 aagaacgaag gaaggagcac agacttagat tggtatatat acgcatatgt agtgttgaag
    11041 aaacatgaaa ttgcccagta ttcttaaccc aactgcacag aacaaaaacc tgcaggaaac
    11101 gaagataaat catgtcgaaa gctacatata aggaacgtgc tgctactcat cctagtcctg
    11161 ttgctgccaa gctatttaat atcatgcacg aaaagcaaac aaacttgtgt gcttcattgg
    11221 atgttcgtac caccaaggaa ttactggagt tagttgaagc attaggtccc aaaatttgtt
    11281 tactaaaaac acatgtggat atcttgactg atttttccat ggagggcaca gttaagccgc
    11341 taaaggcatt atccgccaag tacaattttt tactcttcga agacagaaaa tttgctgaca
    11401 ttggtaatac agtcaaattg cagtactctg cgggtgtata cagaatagca gaatgggcag
    11461 acattacgaa tgcacacggt gtggtgggcc caggtattgt tagcggtttg aagcaggcgg
    11521 cagaagaagt aacaaaggaa cctagaggcc ttttgatgct agcagaattg tcatgcaagg
    11581 gctccctatc tactggagaa tatactaagg gtactgttga cattgcgaag agcgacaaag
    11641 attttgttat cggctttatt gctcaaagag acatgggtgg aagagatgaa ggttacgatt
    11701 ggttgattat gacacccggt gtgggtttag atgacaaggg agacgcattg ggtcaacagt
    11761 atagaaccgt ggatgatgtg gtctctacag gatctgacat tattattgtt ggaagaggac
    11821 tatttgcaaa gggaagggat gctaaggtag agggtgaacg ttacagaaaa gcaggctggg
    11881 aagcatattt gagaagatgc ggccagcaaa actaaaaaac tgtattataa gtaaatgcat
    11941 gtatactaaa ctcacaaatt agagcttcaa tttaattata tcagttatta cctggcatgc
    12001 acatacaaat ggacgaacgg ataaaccttt tcacgccctt ttaaatatcc gattattcta
    12061 ataaacgctc ttttctctta g
//
