LOCUS       sequence-439538-       16048 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses the bac-BGC in bacterial cells, produces bacillamide D.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 16048)
  AUTHORS   Hohmann M, Brunner V, Johannes W, Schum D, Carroll LM, Liu T, Sasaki
            D, Bosch J, Clavel T, Sieber SA, Zeller G, Tschurtschenthaler M, 
            Janssen KP, Gulder TAM
  TITLE     Bacillamide D produced by Bacillus cereus from the mouse intestinal 
            bacterial collection (miBC) is a potent cytotoxin in vitro.
  JOURNAL   Commun Biol. 2024 May 28;7(1):655. doi: 10.1038/s42003-024-06208-3.
  PUBMED    38806706
REFERENCE   2  (bases 1 to 16048)
  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: "Commun 
            Biol."; date: "2024-05-28"; volume: "7(1):655. doi"; pages: " 
            10.1038/s42003-024-06208-3"
COMMENT     Sequence Label: pET28b-ptetO::bac::gfp
FEATURES             Location/Qualifiers
     source          1..16048
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     173..190
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    205..344
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(270..289)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     430..452
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(446..637)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(1446..2528)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     2490..2509
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2529..2606)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     2812..2831
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     protein_bind    2918..2951
                     /label=lox71
                     /bound_moiety="Cre recombinase"
                     /note="Left element (LE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(2952..3578)
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MMSRLDKSKVISSALELLNEVGIEGLTTRKLAQRLGVEQPTLYWH
                     VKNKRALLDALAIEMLGRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGT
                     RPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPT
                     TDSMPPLLRQAIELFDHQGAEPAFLLGLELIICGLEKQLKCESGS"
     primer_bind     3483..3502
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     promoter        3594..3649
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    3600..3618
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     protein_bind    3630..3648
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             complement(10781..10792)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             join(13162..13164,13165..13167,13168..13878)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     terminator      13911..13958
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      13995..14450
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(14082..14101)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     14292..14313
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(14542..15357)
                     /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             complement(14542..15357)
                     /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     15265..15284
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     15349..15368
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      join(15479..16048,1..19)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     15968..15987
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
       61 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
      121 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
      181 agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg
      241 tatttcacac cgcaatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
      301 cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga cacccgccaa
      361 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
      421 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
      481 ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt
      541 catccgcgtc cagctcgttg agtttctcca gaagcgttaa tgtctggctt ctgataaagc
      601 gggccatgtt aagggcggtt ttttcctgtt tggtcactga tgcctccgtg taagggggat
      661 ttctgttcat gggggtaatg ataccgatga aacgagagag gatgctcacg atacgggtta
      721 ctgatgatga acatgcccgg ttactggaac gttgtgaggg taaacaactg gcggtatgga
      781 tgcggcggga ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt aatacagatg
      841 taggtgttcc acagggtagc cagcagcatc ctgcgatgca gatccggaac ataatggtgc
      901 agggcgctga cttccgcgtt tccagacttt acgaaacacg gaaaccgaag accattcatg
      961 ttgttgctca ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg
     1021 gtgattcatt ctgctaacca gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca
     1081 ggagcacgat catgcgcacc cgtggggccg ccatgccggc gataatggcc tgcttctcgc
     1141 cgaaacgttt ggtggcggga ccagtgacga aggcttgagc gagggcgtgc aagattccga
     1201 ataccgcaag cgacaggccg atcatcgtcg cgctccagcg aaagcggtcc tcgccgaaaa
     1261 tgacccagag cgctgccggc acctgtccta cgagttgcat gataaagaag acagtcataa
     1321 gtgcggcgac gatagtcatg ccccgcgccc accggaagga gctgactggg ttgaaggctc
     1381 tcaagggcat cggtcgagat cccggtgcct aatgagtgag ctaacttaca ttaattgcgt
     1441 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     1501 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc
     1561 agtgagacgg gcaacagctg attgcccttc accgcctggc cctgagagag ttgcagcaag
     1621 cggtccacgc tggtttgccc cagcaggcga aaatcctgtt tgatggtggt taacggcggg
     1681 atataacatg agctgtcttc ggtatcgtcg tatcccacta ccgagatatc cgcaccaacg
     1741 cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc
     1801 agcatcgcag tgggaacgat gccctcattc agcatttgca tggtttgttg aaaaccggac
     1861 atggcactcc agtcgccttc ccgttccgct atcggctgaa tttgattgcg agtgagatat
     1921 ttatgccagc cagccagacg cagacgcgcc gagacagaac ttaatgggcc cgctaacagc
     1981 gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca
     2041 tgggagaaaa taatactgtt gatgggtgtc tggtcagaga catcaagaaa taacgccgga
     2101 acattagtgc aggcagcttc cacagcaatg gcatcctggt catccagcgg atagttaatg
     2161 atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg
     2221 ccgcttcgtt ctaccatcga caccaccacg ctggcaccca gttgatcggc gcgagattta
     2281 atcgccgcga caatttgcga cggcgcgtgc agggccagac tggaggtggc aacgccaatc
     2341 agcaacgact gtttgcccgc cagttgttgt gccacgcggt tgggaatgta attcagctcc
     2401 gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc
     2461 acgcgggaaa cggtctgata agagacaccg gcatactctg cgacatcgta taacgttact
     2521 ggtttcacat tcaccaccct gaattgactc tcttccgggc gctatcatgc cataccgcga
     2581 aaggttttgc gccattcgat ggtgtccggg atctcgacgc tctcccttat gcgactcctg
     2641 cattaggaag cagcccagta gtaggttgag gccgttgagc accgccgccg caaggaatgg
     2701 tgcatgcaag gagatggcgc ccaacagtcc cccggccacg gggcctgcca ccatacccac
     2761 gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga tcttccccat cggtgatgtc
     2821 ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc
     2881 ggcgtagagg atcgagatct caattcgttc aagccgaata acttcgtata atgtatgcta
     2941 tacgaacggt attaagaccc actttcacat ttaagttgtt tttctaatcc gcatatgatc
     3001 aattcaaggc cgagtaagaa ggctggctct gcaccttggt gatcaaataa ttcgatagct
     3061 tgtcgtaata atggcggcat actatcagta gtaggtgttt ccctttcttc tttagcgact
     3121 tgatgctctt gatcttccaa tacgcaacct aaagtaaaat gccccacagc gctgagtgca
     3181 tataatgcat tctctagtga aaaaccttgt tggcataaaa aggctaattg attttcgaga
     3241 gtttcatact gtttttctgt aggccgtgta cctaaatgta cttttgctcc atcgcgatga
     3301 cttagtaaag cacatctaaa acttttagcg ttattacgta aaaaatcttg ccagctttcc
     3361 ccttctaaag ggcaaaagtg agtatggtgc ctacctaaca tctcaatggc taaggcgtcg
     3421 agcaaagccc gcttattttt tacatgccaa tacaatgtag gctgctctac acctagcctc
     3481 tgggcgagtt tacgggttgt taaaccttcg attccgacct cattaagcag ctctaatgcg
     3541 ctgctaatca ctttactttt atctaatcta gacatcatta attcctaatt tttgttgaca
     3601 ctctatcatt gatagagtta ttttaccact ccctatcagt gatagagaag aggatcgacc
     3661 atgagcgaga aaatttatta ctggtctccg attaagcatt gggagaagtt gcataatgaa
     3721 gttttgatag gggaaatgag atttacaggt attctgtctg agtggtttcc agatttttat
     3781 tttatggctc aaaaaggtgt gaaaattagt gaactagtag agcgcttttc gttaggaaat
     3841 gtagaagaga cacaaaaaaa catagagctc atgataaaaa atcgtgtgtt agtaagtaat
     3901 atattacacc caagagaagt attttcttca caagaaaaga ttttttcaaa tccatatagc
     3961 aaccagattc gtttttctaa agaaaagtta gataagtata tgagtaaaca attaaatcgc
     4021 acgcacgttg ctgcacggtc gaccgaaatt caacttgaaa caacaaatga gctacctact
     4081 ataatcaagg aacgtaggtc ttgtcgtcaa tttgatatgg aaaaacatat tagtttttcg
     4141 gaattttccc gattcttatc aactctaaaa caggttaggg aagaaaaaat atactatcat
     4201 tacgcaagtg caggcgggct ctatcccatt gatatctttg tttatgtaaa accaaaacgt
     4261 atagaggata tgaaagctgg attttattat tataacccat caaaaaatag tcttgtgatt
     4321 gtaaacaaca ttgatcaagt aatcaaaagt gatcatgagt taattaatca ggatttattt
     4381 actcaatctg ccttctcagt ttatttggtt tataatgcga acgcttccat cccgaaatac
     4441 ggttcagatg gatatttatt tgcttgtatt gagtcaggca ttatcactgc taccttaaat
     4501 atggttgctg agacattaaa tttaggggtt tgttcagtcg gtcatatgaa atttgaagat
     4561 attcagcaat ttttatgttt ggataatcat caagtgtttc ttcatgggct tgaagttggc
     4621 ttgaaaatta acgagtagaa gaaatgaata ttagggggat aaagataaaa atgcacatta
     4681 aagatcgaag ttcgattcat gaaaataaag aagtagaagc gttttggaag gagcaattat
     4741 cttccgagat tctcgatttt tctgtattta aatttaataa cgattaccaa ctaaataaag
     4801 aaacaccttg tgaatacgtt caagtcatag tgaatgtaga atttagtaaa agtatacaaa
     4861 aaataagtaa gtctcaagat ttattattat ttagctggtt tcaagccgct ttaagggttt
     4921 gtttggcaca ttatacagat caagaaagaa ttactattgg tattcctata attgaaaaga
     4981 atcttgaaca aagtgagaat cttaataaat tgattccttt aggaagtgaa atccagccgc
     5041 atcacacctt taaacaatta gtaaatgaga ttgaacaaac gactttattc gggtatgaaa
     5101 atcaggagta tccgctctct gaattattag ttaaacacaa tttaactcct tttcaaattg
     5161 taatcgggat ggaaggatta cataatgaaa attcgctaga agaatacgca gagaataatt
     5221 tagccctatg gatacgaaga aaatggaatg aaatacataa tgaatttgaa tttcaaattt
     5281 cttgcaagtc ccgttcattc tcttctttgc aacagtttgc caattcgtat tgttatgttt
     5341 taaagcaagt ttcacaaaat ccgaatttgg cattgaaaga tatttgtatt attgcagaag
     5401 aagaaaaaga gttgttagaa ggattgaata cttttcaaac gggcatagac ttgtttcaat
     5461 catttcaaga tttgtttgaa gcgcaagtat taaaaactcc agatcaaatt gctgttgttt
     5521 gtaatgatca atctttgaca tatagagaat tgaataagaa agcgaatcaa ttggcttcta
     5581 ttcttcaaag taaaggtgtt tcaaaagaaa gtattattgg cgttatggta gatcggtcat
     5641 tagaaatgat tattggtatg atggggattt taaaagcagg agcagcgtat ttaccaatcg
     5701 accctaacta cccaactgaa cgaattgaat atatgttaca agatagtcaa gcaaaatatt
     5761 tacttagtaa gcgaacagag gaagtattgc cgcaatttgc gggtgaagta ctgtatcttg
     5821 ataaagaata tcttttccaa ggtgaagaaa gtaatttagt acgagaacat aatcctaatg
     5881 atttggctta tgttatttac acatcaggat cgacaggaaa tccaaaaggg gtaatgatag
     5941 aacaaaaatc gctagttcat tttttaaatg taatgagaga aaaagtaaaa gctaatcaat
     6001 cattcttgtt tcttgctagt gtatcttttg atatttcctt gttagaaata tgtcttcctt
     6061 taacgcaagg ttcaaaagta gtaattgcta cagaggaaca gtttgcaaca aaagaattgg
     6121 ctcatttggt caaaaaacat aaggtggatt tatgggaatc aacgccatcg cgaatggaag
     6181 tgattttgag tgatccagaa ggggcaaatt ttctaaaaga tttaaaatct attttattag
     6241 caggagaagc attttccatt gatttggttg aaaaaattcg ttgtatcagt gaagcaacaa
     6301 ttttaaatat ttatggtccc acagaaacaa ccatttgcgc aacagtaaaa gacttgagca
     6361 cttccaaaga agttacgatc ggcaaaccaa atccgaatta tcattcctat atattgaata
     6421 agtatggcca attaaaacca tttggaattc ccggtgaact ctgcattgct ggagtagcgg
     6481 tagcacgagg gtatcttggt aaaagtaaac tgacagatga aaaatttgta cagagtcctt
     6541 ttgcggcagg ggaaatgatg tatcatacag gtgatctcgt tcgttggttg cctaatggag
     6601 aacttgagta tttaggtcga atggatcatc aagtgaaaat tcgagggtat cggattgaat
     6661 taagagaaat cgaaactcaa ttaagggaat atccagaaat aaaccaggct atcgttgttg
     6721 atcaagtgta tggaaatagg aaattattag ctgcctacta tgtatcggat aacaaggtgt
     6781 cttttggtga aatcagaaaa tatttaagtg ataaattacc tgagtttatg attcctgaaa
     6841 aaatgattca ggtagaagag attccattga atccaaatgg aaaagtggat agaaaaaggt
     6901 tattggatat aacccaaacg gattatgtat caattccata cgttgccccg cgcaatgaaa
     6961 ttgaacaaaa attggtgcgt gcgtgggaaa aagttatgga gattgaggga atcggtattc
     7021 atgataattt ttttgcatta aaaggagaat ccattaaagc acttcaggta attaatttat
     7081 taagaaaaga ctgcttaaaa ataagtacaa cagatttttt taagcaccct acaatcgctc
     7141 aactgagttc ttgtgtaaag ttagagcata aagaagaaaa ctttgaaagt aaaccaactc
     7201 atgttaaaga tttgtctaat ccattctcct taacagaagt tcagacagcg tatatgttag
     7261 gaaggaattc acagtttgaa ttaagtggta tttcccctca aacatatttt gaatatgaaa
     7321 cagcattgga tatcaatcgt ctttcaaaaa gttttcaaaa ggtgatccat cgtcatccaa
     7381 tgttgcgagc tgtaatttta ccagagggaa aacagcaaat attacaaagc gttcctgatt
     7441 acgagattga aatagtgagt ctcatagatt tagatgatag gaatcaacat gctcgcttac
     7501 aagaagagcg ttctcgaatg actaaccatg tattcccttt ggggcaatgg cctttgtttg
     7561 aattgaaagc ttttctttta aaagaagaca cgtatttatt atgttttcga tacgatgctt
     7621 tattaatgga cggagcaagc atgaacattg ttgggcagga tttgttgcat tattattata
     7681 aaccaaatca aaaattagaa tctttatctt ttgattttca agattatatg tttatctatg
     7741 atgaaatgga acaaagtgaa gaatataaaa cagcaaaaga ttattggaca agtaagcttc
     7801 ctgactttcc gtttgcacct tctttgctgt taaaaaagga tccagcagag attgctactc
     7861 ctaaattcca atcacttaca aaaatcctgg ataataaaaa atggacaaag ctacgaaagt
     7921 tagcacaaga gaaagaggta acgccttccg ctctactttg tacaatatat ggtgatgtgt
     7981 tggcatattg gagtaatcaa cgtcgattag caattaactt gactgtattt aaccgttatc
     8041 ctgttcatga tgaagtagaa cagattgttg gagatttcac atcacttatt ttattggatg
     8101 ttgatgtaaa accagagcaa actttcttta ctagagtaaa agaaacgcaa gctactttat
     8161 tagatggcct tgaacatcgt cattacgatg gagttaactt tattcgagat tttactcgat
     8221 atcatcaaat gacgccaaaa gcagttatgc caatcgtttt cacctcgatg ttagcaggtg
     8281 caggtgcttt ctcttgggaa caacttggtt ctcttcgcta tattcatgct cggactccac
     8341 aagtgtattt ggataatgta gtaattgaaa aaaatggaga gttattaatt agctggaact
     8401 atgttgaaga gttgtttgat gttgatgtaa tagaagcaat gttttcccaa tttgttgact
     8461 tattggaaca attggtgaaa caaagcgata taacatcttt gcagatgaaa gaatcggatc
     8521 aaactttaat taagcaatac aatgaaacaa cagaaaaaat accttcaact actctgtatc
     8581 aattatttac agaccaagta aaacgtacac cggatgaagt agcagtggta tttgaacaag
     8641 aatggttaac atactcggaa cttcataaac gttctaatca aattgcccac ttcttaaaag
     8701 agcaaggtat tggtctaggt gacaaagtgg gtcttttagc aaaaaggcga gttgacacaa
     8761 ttgttaatat gttaggtatt ttaaaatcag gtgccgctta tgttccgatt gatcccgatc
     8821 atcctttaga tcgccaaaca tatattttaa aaaatagctc atgcaagctt ttattagaac
     8881 ctagtcttta tgaagaaaat catttgtcat tttatacaac agaagatatg cccgctattg
     8941 ctggtccaga agatattgcc tatattatat acacttcagg aagtacagga aaaccaaaag
     9001 gagtaattat tactcaccaa gcggttacaa atactattca agacataaat caaaaatatg
     9061 aggtgaatga agatgatcgt attattggta tctcttccat gtgttttgac ctatctgtat
     9121 acgatatttt cggagcttta agcacaggtg caatgttagt aatgattcgt gatccgagag
     9181 acatgcaaga attaatccgc actgtcgaac gtagagggat tacaatttgg aatacagtgc
     9241 ctgcaatcat ggatttggca ttggagcaag taggttctca ttttgaatat ccttctttac
     9301 gtttagtttt acatagtgga gattggattc cgctttcttt accagaaaaa ataaaacgac
     9361 attttccgat tgcagaagta gtctcactag gaggggcaac agaagcatcg atttggtcaa
     9421 tttattatcc ggtcaaacag gtggaatcac attggaatag tattccatac gggatgccct
     9481 tggcaaacca aacgtattat gtgttgaact atgaaaagaa aatgtgccct gttggtgtaa
     9541 ttggagattt atatatcggg ggagtgggcc ttgctaaagg atatgtaaat gatgaaaaga
     9601 aaacaaatga agcgtttgtg tcacatcctg attttggtct tatatataaa acaggagatt
     9661 gtggaaaaat gcattctgag ggatatattg aatttttagg ccgccaagat taccaagtga
     9721 aaattcaggg atatcgagta gagttagaag aaatctctca ttgtcttcta acttataaac
     9781 aagtcgaaca tgctgtcgta attgatcaaa cagatgagaa cggaataaaa tttttagttg
     9841 catatgtggt aactgagcaa aatattagta caacagaact acgaaagcat ttgcgagatc
     9901 acttgccaga ttatatgatt ccttcttatt ttgtttattt ggaccaatta cccctgactc
     9961 caaacgggaa attagataga aaagcgctcc ctactccgga gaaacaaaaa aataaagtat
    10021 ttattgcccc aaaaactgaa atggaaaaaa tattaactag tatttggcaa gaagtactca
    10081 aaatagatca agttggagta aatgatcatt tcttcgcctt aggtggagat tcgataaaag
    10141 cgattcaagt gtccgtcaga ctgtatcgac aaggattcga gcttgaacca aaaaatatat
    10201 tttcatttcc tatattaagg gacatggttc aatttgttaa aaagctagac cagttttcaa
    10261 gttcacaaac aaatgatgta aacataatga acgataataa agctacatta ctaaatgtta
    10321 aagataaaca acttaatcaa catacactta acaaaataca agaaaaaatg ccggatataa
    10381 aaatagaacg tatttacccg ttagcacctt tccaggaagt tatttatgag catgctacga
    10441 atgaacctga tacaaatgca tattttgaac aaacaatttg ggcacttgag ggggagctgg
    10501 atattaaact ctttattcag tgtttccaac aattagttga tcgtcatgat tctttaagaa
    10561 caataattgt gcaagatgaa caagcaaaac cttggcaagc tgttctatat gatgttcaaa
    10621 taaattcaga aataaaaaac ttgattgaga tgacgaaaca agaacagcaa gaattcctaa
    10681 aacagtggat gaatgaaaat ttaagtcgag gatttaaaaa tgatgaactt atgacacgtc
    10741 tctgtatatt ccaactaggt aataacgagc ataaagtaat ttggagttcc catcatctat
    10801 tatgtgatgg ttggagcgtc agcattttga tagacgaatt attccagctc tacgaaacta
    10861 caaatgccgg aactacggcg gggttaccca tggtaaaacc ttttgaaaca tttatagatt
    10921 ggactttggc acaagatcat gagaaagcac gtcatttttg gcgagattat ttatcaggat
    10981 acaatcagaa aattagtatt cctaaaaaga aagtcccatc taaaagtgaa gggttaaact
    11041 tcgcttttat ggatttaaca ttagacaagg atttgactga tcagttacag acatttgtaa
    11101 ctaaaaataa tataactatg aatacggcat tactagctgt atggggaata ttgcttggca
    11161 aattcaacgg aacgagagaa gtagtattgc ctaatctagt atcagtcaga cctcccgaag
    11221 tagaaggaat tgaaaatatg ttcggtctgt tcacaaatgt tttgcctatc cgattagcat
    11281 ggacagaaac gcaatctttt gtggacttcc tgcaaaaggt acaactagag acattgaaat
    11341 gtaatgagta tgcatactat tcatttgttg atatacaaaa gcaaagtgaa ttaaaaactg
    11401 agttgactga ccatctttgg gtttttgaaa actatccaac taatccagca atcttttctt
    11461 caaatgaaaa cagacatttt gctatcacgg attacgaagt tgttgatgag ccacatgtaa
    11521 agtatggaat tatgtgcttc cctgaagaaa gacttacaat aaagtttgga tatgatcaaa
    11581 cggtatatga aacagagaaa atacaggaaa tattaaatca catccagggg ttaatcaaca
    11641 cgattcttca aaatccaata caaacaattt gtgattacaa attataaatg acaaaaaaat
    11701 tacagttatc agccgaagaa atgcgtcagt taggatatca agcggttgac ttaattgttg
    11761 atcatatgaa tcatttaaaa agtaaaccag tttcagaaac gattgatagt aatattttta
    11821 gagacaagct aattgaaacg attccggaaa acggatctaa tccaaaagaa ttacttcatt
    11881 ttcttaataa caatgtgttt aatcaaatta ctcatgtgga tcatcctcat tttatggctt
    11941 ttgtgccagg tcctaataat tatgttggtg tattagcaga ttttttagca agtgggttta
    12001 atgtatttcc tacagcttgg atagtaggtg caggtgccga acaaattgaa ttaacgacaa
    12061 ttaactggtt aaaatcaatg ctaggattcc ctgattcagc tgaaggatta tttgtcagtg
    12121 gtggttccat ggctaatttg actgcactta ctgtagcgag acaggtcaag ctgaataatg
    12181 acatagaaaa tgcgattgtt tatttttcta accaaactca tttttctgta gatcgagcac
    12241 ttaaagtatt aggttttaaa cagcatcaaa tttgccgaat tgagacagat gaagacttaa
    12301 aaatttcagt tagtacttta agaaaacaaa taaaagaaga tcgattaaaa gggaaaaaac
    12361 cattctgtgt tattgctaat gcagggacaa cgaattgcgg agcggtggac tccctcgatg
    12421 aattggctga tgtatgtggt gatgaagata tatggctaca cgtggatggg gcttatgggg
    12481 ctgcagcaat tctcagtgaa aaaggaaggg agttattaag tggaattcat cgtgccgatt
    12541 ctttaacgtt agacccgcat aaatggcttt ttcagcctta tgatgtcggt tgtgtactta
    12601 ttcgaaatag ccaatattta agtgaaacat tccgtatgat cccagagtat attaaggata
    12661 cagaaactaa tatagaagaa aaagtgaatt ttggggagcg tggaattgaa ctttctcgta
    12721 gatttagagc attaaaggta tggctttctt ttaaagcatt cggagttaca gcttttcgtg
    12781 aggcaattga tcatggcatt atgctagctg aacaagttga agagttttta aggaaagaaa
    12841 aagattggga agtagtaaca cctgcccaat taggaattgt tacttttcgt tatattccat
    12901 gtgagctaac atctacagat acaattcatg aaataaacaa aaaattggtg gaagaaatta
    12961 atcaaagagg atttgctatg ctaagtacga ctaaattaaa agaaaaagta gtaattcgac
    13021 tttgctctat caacccaaga acaacaaaag aagaaatact tcaaatcatg atgaacataa
    13081 aagcattagc agaggaaata aacacatcgc ctaaacatct tatatctgtt tctcaacctt
    13141 gatatattag gcggaggcct catggttagc aaaggtgaag aactgtttac cggcgttgtg
    13201 ccgattctgg tggaactgga tggtgatgtg aatggccata aatttagcgt tcgtggcgaa
    13261 ggcgaaggtg atgcgaccaa cggtaaactg accctgaaat ttatttgcac caccggtaaa
    13321 ctgccggttc cgtggccgac cctggtgacc accctgacct atggcgttca gtgctttagc
    13381 cgctatccgg atcatatgaa acgccatgat ttctttaaaa gcgcgatgcc ggaaggctat
    13441 gtgcaggaac gtaccattag cttcaaagat gatggcacct ataaaacccg tgcggaagtt
    13501 aaatttgaag gcgataccct ggtgaaccgc attgaactga aaggtattga ttttaaagaa
    13561 gatggcaaca ttctgggtca taaactggaa tataatttca acagccataa tgtgtatatt
    13621 accgccgata aacagaaaaa tggcatcaaa gcgaacttta aaatccgtca caacgtggaa
    13681 gatggtagcg tgcagctggc ggatcattat cagcagaata ccccgattgg tgatggcccg
    13741 gtgctgctgc cggataatca ttatctgagc acccagagcg ttctgagcaa agatccgaat
    13801 gaaaaacgtg atcatatggt gctgctggaa tttgttaccg ccgcgggcat tacccacggt
    13861 atggatgaac tgtataaagg cagctaaggc agagtggtaa aatgcaataa ctagcataac
    13921 cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaaaggagga actatatccg
    13981 gattggcgaa tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
    14041 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
    14101 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
    14161 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
    14221 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
    14281 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
    14341 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
    14401 ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt aggtggcact
    14461 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
    14521 tatccgctca tgaattaatt cttagaaaaa ctcatcgagc atcaaatgaa actgcaattt
    14581 attcatatca ggattatcaa taccatattt ttgaaaaagc cgtttctgta atgaaggaga
    14641 aaactcaccg aggcagttcc ataggatggc aagatcctgg tatcggtctg cgattccgac
    14701 tcgtccaaca tcaatacaac ctattaattt cccctcgtca aaaataaggt tatcaagtga
    14761 gaaatcacca tgagtgacga ctgaatccgg tgagaatggc aaaagtttat gcatttcttt
    14821 ccagacttgt tcaacaggcc agccattacg ctcgtcatca aaatcactcg catcaaccaa
    14881 accgttattc attcgtgatt gcgcctgagc gagacgaaat acgcgatcgc tgttaaaagg
    14941 acaattacaa acaggaatcg aatgcaaccg gcgcaggaac actgccagcg catcaacaat
    15001 attttcacct gaatcaggat attcttctaa tacctggaat gctgttttcc cggggatcgc
    15061 agtggtgagt aaccatgcat catcaggagt acggataaaa tgcttgatgg tcggaagagg
    15121 cataaattcc gtcagccagt ttagtctgac catctcatct gtaacatcat tggcaacgct
    15181 acctttgcca tgtttcagaa acaactctgg cgcatcgggc ttcccataca atcgatagat
    15241 tgtcgcacct gattgcccga cattatcgcg agcccattta tacccatata aatcagcatc
    15301 catgttggaa tttaatcgcg gcctagagca agacgtttcc cgttgaatat ggctcataac
    15361 accccttgta ttactgttta tgtaagcaga cagttttatt gttcatgacc aaaatccctt
    15421 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
    15481 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
    15541 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
    15601 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
    15661 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
    15721 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
    15781 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
    15841 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
    15901 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
    15961 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
    16021 agcgtcgatt tttgtgatgc tcgtcagg
//
