LOCUS       sequence-273752-       11636 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Agrobacterium T-DNA with Spun H2Bpr driving hph-mCerulean3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11636)
  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 11636)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: pGI3EM31
FEATURES             Location/Qualifiers
     source          1..11636
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1093..2435
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(2056..2103)
                     /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     2476..2494
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2697..2912
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             2913..3716
                     /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    3883..3907
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     primer_bind     4088..4110
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4102..4119
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4103..4119
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             4418..5443
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     CDS             4418..5443
                     /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             join(5459..5461,5462..5464,5465..6178)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAIHGNVYITADKQKNGIK
                     ANFGLNCNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5504..5525)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5765..5784)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6112..6133
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      6199..6386
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(6254..6278)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(6401..6417)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6401..6417)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6414..6436)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6425..6441
                     /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(6449..6455,6456..6473,6474..6479))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6494..6515
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_feature    6692..6716
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     primer_bind     complement(6998..7017)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             7009..7824
                     /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             7009..7824
                     /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(7082..7101)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      7919..8507
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8408..8427
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8661..8678
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    8693..8833
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      9177..9371
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(9437..10510)
                     /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(10939..11568)
                     /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 gttttactca aatacacatc acctttttag acggcggcgc tcggtttctt cagcggccaa
       61 gctggccggc caggccgcca gcttggcatc agacaaaccg gccaggattt catgcagccg
      121 cacggttgag acgtgcgcgg gcggctcgaa cacgtacccg gccgcgatca tctccgcctc
      181 gatctcttcg gtaatgaaaa acggttcgtc ctggccgtcc tggtgcggtt tcatgcttgt
      241 tcctcttggc gttcattctc ggcggccgcc agggcgtcgg cctcggtcaa tgcgtcctca
      301 cggaaggcac cgcgccgcct ggcctcggtg ggcgtcactt cctcgctgcg ctcaagtgcg
      361 cggtacaggg tcgagcgatg cacgccaagc agtgcagccg cctctttcac ggtgcggcct
      421 tcctggtcga tcagctcgcg ggcgtgcgcg atctgtgccg gggtgagggt agggcggggg
      481 ccaaacttca cgcctcgggc cttggcggcc tcgcgcccgc tccgggtgcg gtcgatgatt
      541 agggaacgct cgaactcggc aatgccggcg aacacggtca acaccatgcg gccggccggc
      601 gtggtggtgt cggcccacgg ctctgccagg ctacgcaggc ccgcgccggc ctcctggatg
      661 cgctcggcaa tgtccagtag gtcgcgggtg ctgcgggcca ggcggtctag cctggtcact
      721 gtcacaacgt cgccagggcg taggtggtca agcatcctgg ccagctccgg gcggtcgcgc
      781 ctggtgccgg tgatcttctc ggaaaacagc ttggtgcagc cggccgcgtg cagttcggcc
      841 cgttggttgg tcaagtcctg gtcgtcggtg ctgacgcggg catagcccag caggccagcg
      901 gcggcgctct tgttcatggc gtaatgtctc cggttctagt cgcaagtatt ctactttatg
      961 cgactaaaac acgcgacaag aaaacgccag gaaaagggca gggcggcagc ctgtcgcgta
     1021 acttaggact tgtgcgacat gtcgttttca gaagacggct gcactgaacg tcagaaggaa
     1081 aagtgccacc tgaacgaagc atctgtgctt cattttgtag aacaaaaatg caacgcgaga
     1141 gcgctaattt ttcaaacaaa gaatctgagc tgcattttta cagaacagaa atgcaacgcg
     1201 aaagcgctat tttaccaacg aagaatctgt gcttcatttt tgtaaaacaa aaatgcaacg
     1261 cgagagcgct aatttttcaa acaaagaatc tgagctgcat ttttacagaa cagaaatgca
     1321 acgcgagagc gctattttac caacaaagaa tctatacttc ttttttgttc tacaaaaatg
     1381 catcccgaga gcgctatttt tctaacaaag catcttagat tacttttttt ctcctttgtg
     1441 cgctctataa tgcagtctct tgataacttt ttgcactgta ggtccgttaa ggttagaaga
     1501 aggctacttt ggtgtctatt ttctcttcca taaaaaaagc ctgactccac ttcccgcgtt
     1561 tactgattac tagcgaagct gcgggtgcat tttttcaaga taaaggcatc cccgattata
     1621 ttctataccg atgtggattg cgcatacttt gtgaacagaa agtgatagcg ttgatgattc
     1681 ttcattggtc agaaaattat gaacggtttc ttctattttg tctctatata ctacgtatag
     1741 gaaatgttta cattttcgta ttgttttcga ttcactctat gaatagttct tactacaatt
     1801 tttttgtcta aagagtaata ctagagataa acataaaaaa tgtagaggtc gagtttagat
     1861 gcaagttcaa ggagcgaaag gtggatgggt aggttatata gggatatagc acagagatat
     1921 atagcaaaga gatacttttg agcaatgttt gtggaagcgg tattcgcaat attttagtag
     1981 ctcgttacag tccggtgcgt ttttggtttt ttgaaagtgc gtcttcagag cgcttttggt
     2041 tttcaaaagc gctctgaagt tcctatactt tctagagaat aggaacttcg gaataggaac
     2101 ttcaaagcgt ttccgaaaac gagcgcttcc gaaaatgcaa cgcgagctgc gcacatacag
     2161 ctcactgttc acgtcgcacc tatatctgcg tgttgcctgt atatatatat acatgagaag
     2221 aacggcatag tgcgtgttta tgcttaaatg cgtacttata tgcgtctatt tatgtaggat
     2281 gaaaggtagt ctagtacctc ctgtgatatt atcccattcc atgcggggta tcgtatgctt
     2341 ccttcagcac taccctttag ctgttctata tgctgccact cctcaattgg attagtctca
     2401 tccttcaatg ctatcatttc ctttgatatt ggatcatact aagaaaccat tattatcatg
     2461 acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat
     2521 gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg
     2581 gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc
     2641 tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatac caccttttca
     2701 attcatcatt ttttttttat tctttttttt gatttcggtt tccttgaaat ttttttgatt
     2761 cggtaatctc cgaacagaag gaagaacgaa ggaaggagca cagacttaga ttggtatata
     2821 tacgcatatg tagtgttgaa gaaacatgaa attgcccagt attcttaacc caactgcaca
     2881 gaacaaaaac ctgcaggaaa cgaagataaa tcatgtcgaa agctacatat aaggaacgtg
     2941 ctgctactca tcctagtcct gttgctgcca agctatttaa tatcatgcac gaaaagcaaa
     3001 caaacttgtg tgcttcattg gatgttcgta ccaccaagga attactggag ttagttgaag
     3061 cattaggtcc caaaatttgt ttactaaaaa cacatgtgga tatcttgact gatttttcca
     3121 tggagggcac agttaagccg ctaaaggcat tatccgccaa gtacaatttt ttactcttcg
     3181 aagacagaaa atttgctgac attggtaata cagtcaaatt gcagtactct gcgggtgtat
     3241 acagaatagc agaatgggca gacattacga atgcacacgg tgtggtgggc ccaggtattg
     3301 ttagcggttt gaagcaggcg gcagaagaag taacaaagga acctagaggc cttttgatgc
     3361 tagcagaatt gtcatgcaag ggctccctat ctactggaga atatactaag ggtactgttg
     3421 acattgcgaa gagcgacaaa gattttgtta tcggctttat tgctcaaaga gacatgggtg
     3481 gaagagatga aggttacgat tggttgatta tgacacccgg tgtgggttta gatgacaagg
     3541 gagacgcatt gggtcaacag tatagaaccg tggatgatgt ggtctctaca ggatctgaca
     3601 ttattattgt tggaagagga ctatttgcaa agggaaggga tgctaaggta gagggtgaac
     3661 gttacagaaa agcaggctgg gaagcatatt tgagaagatg cggccagcaa aactaaaaaa
     3721 ctgtattata agtaaatgca tgtatactaa actcacaaat tagagcttca atttaattat
     3781 atcagttatt acctggcatg cacatacaaa tggacgaacg gataaacctt ttcacgccct
     3841 tttaaatatc cgattattct aataaacgct cttttctctt aggtttaccc gccaatatat
     3901 cctgtcaaac actgatagtt taaactgaag gcgggaaacg acaatctgat ccaagctcaa
     3961 gctcattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
     4021 ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca
     4081 gggttttccc agtcacgacg ttgtaaaacg acggccagtg ccaagctttt atgctccaag
     4141 cggagacgat gaggccgata ttacagtggc ttagtagcgt gtacaatatt tccataaaag
     4201 gctacatggt gagtttttca cgtgaccgta attagtccca attaggcttt attgtatatt
     4261 gctactaagc atcgttaaat gacctgctcg aagcttcgtc tataaaaggc gggcgtgtgt
     4321 gcgcttcccc cgctacgtct ttttcacaaa cactttgcga cttgaccaag acctttcacc
     4381 cgcaccactc tcatccatca cacacacaaa ttaattaatg ggtaaaaagc ctgaactcac
     4441 cgcgacgtct gtcgagaagt ttctgatcga aaagttcgac agcgtctccg acctgatgca
     4501 gctctcggag ggcgaagaat ctcgtgcttt cagcttcgat gtaggagggc gtggatatgt
     4561 cctgcgggta aatagctgcg ccgatggttt ctacaaagat cgttatgttt atcggcactt
     4621 tgcatcggcc gcgctcccga ttccggaagt gcttgacatt ggggaattca gcgagagcct
     4681 gacctattgc atctcccgcc gtgcacaggg tgtcacgttg caagacctgc ctgaaaccga
     4741 actgcccgct gttctgcagc cggtcgcgga ggccatggat gcgatcgctg cggccgatct
     4801 tagccagacg agcgggttcg gcccattcgg accgcaagga atcggtcaat acactacatg
     4861 gcgtgatttc atatgcgcga ttgctgatcc ccatgtgtat cactggcaaa ctgtgatgga
     4921 cgacaccgtc agtgcgtccg tcgcgcaggc tctcgatgag ctgatgcttt gggccgagga
     4981 ctgccccgaa gtccggcacc tcgtgcacgc ggatttcggc tccaacaatg tcctgacgga
     5041 caatggccgc ataacagcgg tcattgactg gagcgaggcg atgttcgggg attcccaata
     5101 cgaggtcgcc aacatcttct tctggaggcc gtggttggct tgtatggagc agcagacgcg
     5161 ctacttcgag cggaggcatc cggagcttgc aggatcgccg cggctccggg cgtatatgct
     5221 ccgcattggt cttgaccaac tctatcagag cttggttgac ggcaatttcg atgatgcagc
     5281 ttgggcgcag ggtcgatgcg acgcaatcgt ccgatccgga gccgggactg tcgggcgtac
     5341 acaaatcgcc cgcagaagcg cggccgtctg gaccgatggc tgtgtagaag tactcgccga
     5401 tagtggaaac cgacgcccca gcactcgtcc gagggcaaag gaaggaggag gaggatccat
     5461 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
     5521 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
     5581 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
     5641 cgtgaccacc ctgagctggg gcgtgcagtg cttcgcccgc taccccgacc acatgaagca
     5701 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
     5761 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     5821 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     5881 gctggagtac aacgccatcc acggcaacgt ctatatcacc gccgacaagc agaagaacgg
     5941 catcaaggcc aacttcggcc tcaactgcaa catcgaggac ggcagcgtgc agctcgccga
     6001 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     6061 cctgagcacc cagtccaagc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     6121 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaagg
     6181 cgcgccactt ctaaataagc gaatttctta tgatttatga tttttattat taaataagtt
     6241 ataaaaaaaa taagtgtata caaattttaa agtgactctt aggttttaaa acgaaaattc
     6301 ttattcttga gtaactcttt cctgtaggtc aggttgcttt ctcaggtata gtatgaggtc
     6361 gctcttattg accacacctc taccgggaat tcgtaatcat gtcatagctg tttcctgtgt
     6421 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     6481 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     6541 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     6601 gcggtttgcg tattggctag agcaattcgg cgttaattca gtacattaaa aacgtccgca
     6661 atgtgttatt aagttgtcta agcgtcaatt tgtttacacc acaatatatc ctgccaccag
     6721 ccagccaaca gctccccgac cggcagctcg gcacaaaatc accactcgat acaggcagcc
     6781 catcagtccg ggacggcgtc agcgggagag ccgttgtaag gcggcagact ttgctcatgt
     6841 taccgatgct attcggaaga acggcaacta agctgccggg tttgaaacac ggatgatctc
     6901 gcggagggta gcatgttgat tgtaacgatg acagagcgtt gctgcctgtg atccagatca
     6961 tgaacaataa aactgtctgc ttacataaac agtaatacaa ggggtgttat gagccatatt
     7021 caacgggaaa cgtcttgctc taggccgcga ttaaattcca acatggatgc tgatttatat
     7081 gggtataaat gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgattgtat
     7141 gggaagcccg atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat
     7201 gttacagatg agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc
     7261 aagcatttta tccgtactcc tgatgatgca tggttactca ccactgcgat ccccgggaaa
     7321 acagcattcc aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg
     7381 gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagcgat
     7441 cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt
     7501 gattttgatg acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataaa
     7561 cttttgccat tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt
     7621 atttttgacg aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac
     7681 cgataccagg atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag
     7741 aaacggcttt ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat
     7801 ttgatgctcg atgagttttt ctaagaatta attcatgatc ctgcatgacc aaaatccctt
     7861 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     7921 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     7981 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     8041 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
     8101 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     8161 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     8221 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     8281 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     8341 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     8401 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     8461 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     8521 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     8581 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     8641 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc
     8701 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catatggtgc actctcagta
     8761 caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg
     8821 ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     8881 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     8941 gttttcaccg tcatcaccga aacgcgcgag gcagggtgcc ttgatgtggg cgccggcggt
     9001 cgagtggcga cggcgcggct tgtccgcgcc ctggtagatt gcctggccgt aggccagcca
     9061 tttttgagcg gccagcggcc gcgataggcc gacgcgaagc ggcggggcgt agggagcgca
     9121 gcgaccgaag ggtaggcgct ttttgcagct cttcggctgt gcgctggcca gacagttatg
     9181 cacaggccag gcgggtttta agagttttaa taagttttaa agagttttag gcggaaaaat
     9241 cgcctttttt ctcttttata tcagtcactt acatgtgtga ccggttccca atgtacggct
     9301 ttgggttccc aatgtacggg ttccggttcc caatgtacgg ctttgggttc ccaatgtacg
     9361 tgctatccac aggaaagaga ccttttcgac ctttttcccc tgctagggca atttgcccta
     9421 gcatctgctc cgtacattag gaaccggcgg atgcttcgcc ctcgatcagg ttgcggtagc
     9481 gcatgactag gatcgggcca gcctgccccg cctcctcctt caaatcgtac tccggcaggt
     9541 catttgaccc gatcagcttg cgcacggtga aacagaactt cttgaactct ccggcgctgc
     9601 cactgcgttc gtagatcgtc ttgaacaacc atctggcttc tgccttgcct gcggcgcggc
     9661 gtgccaggcg gtagagaaaa cggccgatgc cgggatcgat caaaaagtaa tcggggtgaa
     9721 ccgtcagcac gtccgggttc ttgccttctg tgatctcgcg gtacatccaa tcagctagct
     9781 cgatctcgat gtactccggc cgcccggttt cgctctttac gatcttgtag cggctaatca
     9841 aggcttcacc ctcggatacc gtcaccaggc ggccgttctt ggccttcttc gtacgctgca
     9901 tggcaacgtg cgtggtgttt aaccgaatgc aggtttctac caggtcgtct ttctgctttc
     9961 cgccatcggc tcgccggcag aacttgagta cgtccgcaac gtgtggacgg aacacgcggc
    10021 cgggcttgtc tcccttccct tcccggtatc ggttcatgga ttcggttaga tgggaaaccg
    10081 ccatcagtac caggtcgtaa tcccacacac tggccatgcc ggccggccct gcggaaacct
    10141 ctacgtgccc gtctggaagc tcgtagcgga tcacctcgcc agctcgtcgg tcacgcttcg
    10201 acagacggaa aacggccacg tccatgatgc tgcgactatc gcgggtgccc acgtcataga
    10261 gcatcggaac gaaaaaatct ggttgctcgt cgcccttggg cggcttccta atcgacggcg
    10321 caccggctgc cggcggttgc cgggattctt tgcggattcg atcagcggcc gcttgccacg
    10381 attcaccggg gcgtgcttct gcctcgatgc gttgccgctg ggcggcctgc gcggccttca
    10441 acttctccac caggtcatca cccagcgccg cgccgatttg taccgggccg gatggtttgc
    10501 gaccgctcac gccgattcct cgggcttggg ggttccagtg ccattgcagg gccggcagac
    10561 aacccagccg cttacgcctg gccaaccgcc cgttcctcca cacatggggc attccacggc
    10621 gtcggtgcct ggttgttctt gattttccat gccgcctcct ttagccgcta aaattcatct
    10681 actcatttat tcatttgctc atttactctg gtagctgcgc gatgtattca gatagcagct
    10741 cggtaatggt cttgccttgg cgtaccgcgt acatcttcag cttggtgtga tcctccgccg
    10801 gcaactgaaa gttgacccgc ttcatggctg gcgtgtctgc caggctggcc aacgttgcag
    10861 ccttgctgct gcgtgcgctc ggacggccgg cacttagcgt gtttgtgctt ttgctcattt
    10921 tctctttacc tcattaactc aaatgagttt tgatttaatt tcagcggcca gcgcctggac
    10981 ctcgcgggca gcgtcgccct cgggttctga ttcaagaacg gttgtgccgg cggcggcagt
    11041 gcctgggtag ctcacgcgct gcgtgatacg ggactcaaga atgggcagct cgtacccggc
    11101 cagcgcctcg gcaacctcac cgccgatgcg cgtgcctttg atcgcccgcg acacgacaaa
    11161 ggccgcttgt agccttccat ccgtgacctc aatgcgctgc ttaaccagct ccaccaggtc
    11221 ggcggtggcc catatgtcgt aagggcttgg ctgcaccgga atcagcacga agtcggctgc
    11281 cttgatcgcg gacacagcca agtccgccgc ctggggcgct ccgtcgatca ctacgaagtc
    11341 gcgccggccg atggccttca cgtcgcggtc aatcgtcggg cggtcgatgc cgacaacggt
    11401 tagcggttga tcttcccgca cggccgccca atcgcgggca ctgccctggg gatcggaatc
    11461 gactaacaga acatcggccc cggcgagttg cagggcgcgg gctagatggg ttgcgatggt
    11521 cgtcttgcct gacccgcctt tctggttaag tacagcgata accttcatgc gttccccttg
    11581 cgtatttgtt tatttactca tcgcatcata tacgcagcga ccgcatgacg caagct
//
