LOCUS       sequence-299688-       10531 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Inducible expression of HO endonuclease in S. cerevisiae.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10531)
  AUTHORS   Gnugge R, Symington LS
  TITLE     Efficient DNA double-strand break formation at single or multiple 
            defined sites in the Saccharomyces cerevisiae genome.
  JOURNAL   Nucleic Acids Res. 2020 Oct 14. pii: 5923424. doi: 
            10.1093/nar/gkaa833.
  PUBMED    33053188
REFERENCE   2  (bases 1 to 10531)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic 
            Acids Res. 2020 Oct 14. pii: 5923424. doi: 10.1093/nar/gkaa833."
COMMENT     Sequence Label: pRG646_lexO-HO_LexA-TF_LEU2MX
FEATURES             Location/Qualifiers
     source          1..10531
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     2597..2619
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2611..2628
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2612..2628
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2635..2654
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2635..2653
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4220..4231
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             4835..4858
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     terminator      4908..5157
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     CDS             join(5827..6078,6079..6432)
                     /codon_start=1
                     /gene="lexA"
                     /product="E. coli DNA-binding protein that represses genes 
                     involved in the SOS response to DNA damage"
                     /label=LexA
                     /note="The N-terminal DNA-binding domain recognizes the 
                     16-bp palindromic sequence 5'-CTGTATATATATACAG-3'."
                     /translation="MKALTARQQEVFDLIRDHISQTGMPPTRAEIAQRLGFRSPNAAEE
                     HLKALARKGVIEIVSGASRGIRLLQEEEEGLPLVGRVAAGEPLLAQQHIEGHYQVDPSL
                     FKPNADFLLRVSGMSMKDIGIMDGDLLAVHKTQDVRNGQVVVARIDDEVTVKRLKKQGN
                     KVELLPENSEFKPIVVDLRQQSFTIEGLAVGVIRNGDWL"
     primer_bind     6370..6391
                     /label=LexA
                     /note="LexA DNA binding domain, forward primer"
     CDS             6451..7386
                     /codon_start=1
                     /product="mutated ligand-binding domain of the human 
                     estrogen receptor (Feil et al., 1997)"
                     /label=ERT2
                     /translation="AGDMRAANLWPSPLMIKRSKKNSLALSLTADQMVSALLDAEPPIL
                     YSEYDPTRPFSEASMMGLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEI
                     LMIGLVWRSMEHPVKLLFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEF
                     VCLKSIILLNSGVYTFLSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRL
                     AQLLLILSHIRHMSNKGMEHLYSMKCKNVVPLYDLLLEMLDAHRLHAPTSRGGASVEET
                     DQSHLATAGSTSSHSLQKYYITGEAEGFPAT"
     CDS             6499..7389
                     /codon_start=1
                     /gene="ESR1"
                     /product="estrogen nuclear receptor alpha ligand-binding 
                     domain"
                     /label=ER-LBD
                     /translation="KRSKKNSLALSLTADQMVSALLDAEPPILYSEYDPTRPFSEASMM
                     GLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEILMIGLVWRSMEHPVKL
                     LFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEFVCLKSIILLNSGVYTF
                     LSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRLAQLLLILSHIRHMSNK
                     GMEHLYSMKCKNVVPLYDLLLEMLDAHRLHAPTSRGGASVEETDQSHLATAGSTSSHSL
                     QKYYITGEAEGFPATV"
     terminator      7853..8102
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     promoter        complement(8265..8283)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(8304..8320)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8304..8320)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8317..8339)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8328..8344
                     /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)."
     primer_bind     complement(8603..8620)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8774..9362)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8854..8873)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9541..10332,10333..10401))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     10164..10183
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10402..10506)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacgtcgaaa gaacgcggca ttagcaacat ttgcgaagcg aacgattttg cgaactgcta
       61 ttaattaact ctggcgcgcc taactttttg tgtggtgccc tcctccttgt caatattaat
      121 gttaaagtgc aattcttttt ccttatcacg ttgagccatt agtatcaatt tgcttacctg
      181 tattccttta catcctcctt tttctccttc ttgataaatg tatgtagatt gcgtatatag
      241 tttcgtctac cctatgaaca tattccattt tgtaatttcg tgtcgcatat gatccgcagg
      301 ctaaccggaa cctgtattat ttagtttatg ctacgttaaa taaagacctt tcgttcacat
      361 aactgaatgt gtaatggcct tgagatttca agcataccaa gttggtggag acggggtcgt
      421 tacaaaagac tctttaagcc aagatttcct tgacagcctt ggcgatagca tcgccaacct
      481 cagtggtaga gttagaacca ccaaggtcac cggttctgac acctgcatcc aagacatttc
      541 taacagcttc ttcaagagcc ctaccttctt caaccaaatc caaggataac ttcaacatca
      601 tagctgcaga taagatggta gcaattgggt taaccttgtt tgctggtaaa tctggggcag
      661 aaccatgaca tggttcgtac aaaccgaatg ccttgttagt gtcaggtagg gaagctagag
      721 atgcagaagg taataaaccc aaagaacctg gaataacaga ggcttcatcg gagataatat
      781 caccaaacat gttgttggta ataacaacac cgtttagctt agttggtgat ttaaccaaaa
      841 tcatagcagc agagtcgatc aattggtgct gaacagttaa ttgtgggaac tcagtcttga
      901 tggtttcttc aacagtcttt ctccacaatc tggaagaggc aagcacgtta gccttgtcaa
      961 gagaccagat tggtaatggt gggttttgtt gcaatgccaa gaaagcagcc attcttgtaa
     1021 ttctttgaac ttcaggaaca ctgtatttct cagagtccca agcaactccg tcaccttcat
     1081 cttcttttct ttcaccaaag tagataccac caaccaattc tctaacgacg acgaaatcag
     1141 taccctttgc atattcaggc ttcaaaggag aaagatctag taaagaatca gaagcaaagt
     1201 tacatggtct caagttggcg tatagaccca attcctttct gatcttcaat agaccttgtt
     1261 ctggtctaac tgcgcccgta ccccattttg gaccaccaac agcacctagt aagacagcat
     1321 cggctttctt agaggcttca agagcttcat ctggtaaagg agtgccagtg gcatcgatgg
     1381 cagcaccccc gatcaagtga tgttggaaat tgaacttaat ttctggacgg acttcagcaa
     1441 tggcattcaa gaccttaata gcttcgtcag taacttcttt accgacgtga tcacccggta
     1501 ggacaacgat attcttagac atggtataat ctgtgtagtg tgggatactt tttacttctt
     1561 caaataggta tcaacaataa aaataaatcg aagcaaatgt aggaatgcgt taaagcagat
     1621 gtacttactt agagtacata gatatattta tataattcaa tatataaaag tatatgaagc
     1681 atctgatgtt gaacctgtca tgactctaaa ctggaatggg tagttatgct tctgaaggtt
     1741 ttccgttaga aaatgagtgc agggatcagg cccggaaccg gttttagcct gaaaggaaaa
     1801 gaaaaaaatg cggtgattac tagcacgtga ctgcgctgaa ttggaatcta ccacaagtac
     1861 agcgaatggc aagagggaag gggaatatat tgtgcctggc ggtagcgtct gctatatcaa
     1921 atgttaaaca taactaaatg cagtagtgaa tagagtaatg tatttctaca tctgtgggta
     1981 gggccaatgg gagcccgatg tctattgtac agatattttc cttttatagt taatcaattc
     2041 aacttggaaa tcattgagtc gatgtctact tggataactg gagccgtggc agcgtttgaa
     2101 atttgacgag tcttcttgtt ttgttcgcgc attttcagtt tattcaaacg ggagatggca
     2161 ccgaatccac tgacttctct gttcaatttg aatggtaata ggaagaaagt acatttcgag
     2221 aaattttcgc ccaacgtcga aatctcatgg atgatttcct cgatacaaat gacaccttca
     2281 tcacctaatt tctcttcaac gatattgttg tcattcaaga ttatctcttt tggttcttta
     2341 tcttctggcc tttgccacat tattctagtt ctcttttgga ttaaagaacg gatagatgca
     2401 agtgaaggtt gtccgataac aatatatggt gatacaagtt tcaacaatgg gtacacgtcc
     2461 ttggtaagtt tgatgaaaac accagtgttc aactcaacaa gtctcaaaag agataacact
     2521 ttgtaagcct ttgctgggat cttcacagcg atcggatgtg ctgcaaggcg attaagttgg
     2581 gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga attgtaatac
     2641 gactcactat agggcgaatt ggagaaacag aataatatat aaacctgtat aatataacct
     2701 tgaagactat atttcttttc gcggccgcct atagggcgaa ttggagctcc ctaggtgctg
     2761 tatatactca cagcataact gtatatacac ccagggtcta ggtgctgtat atactcacag
     2821 cataactgta tatacaccca gggtctaggt gctgtatata ctcacagcat aactgtatat
     2881 acacccaggg tctaggtgct gtatatactc acagcataac tgtatataca cccagggtct
     2941 agagcatgtg ctctgtatgt atataaaact cttgttttct tcttttctct tctttcctta
     3001 tacattagga cctttgcagc ataaattact atacttctat actagtggat cccccgggct
     3061 gcaggaattc atgctttctg aaaacacgac tattctgatg gctaacggtg aaattaaaga
     3121 catcgcaaac gtcacggcta actcttacgt tatgtgcgca gatggctccg ctgcccgcgt
     3181 cataaatgtc acacagggct atcagaaaat ctataatata cagcaaaaaa ccaaacacag
     3241 agcttttgaa ggtgaacctg gtaggttaga tcccaggcgt agaacagttt atcagcgtct
     3301 tgcattacaa tgtactgcag gtcataaatt gtcagtcagg gtccctacca aaccactgtt
     3361 ggaaaaaagt ggtagaaatg ccaccaaata taaagtgaga tggagaaatc tgcagcaatg
     3421 tcagacgctt gatggtagga taataataat tccaaaaaac catcataaga cattcccaat
     3481 gacagttgaa ggtgagtttg ccgcaaaacg cttcatagaa gaaatggagc gctctaaagg
     3541 agaatatttc aactttgaca ttgaagttag agatttggat tatcttgatg ctcaattgag
     3601 aatttctagc tgcataagat ttggtccagt actcacagga aatggtgttt tatctaaatt
     3661 tctcactgga cgtagtgacc ttgtaactcc tgctgtaaaa agtatggctt ggatgcttgg
     3721 tctgtggtta ggtgacggta caacaaaaga gccagaaatc tcagtagata gcttggatcc
     3781 taagctaatg gagagtttaa gagaaaatgc gaaaatctgg ggtctctacc ttacggtttg
     3841 tgacgatcac gttccgctac gtgccaaaca tgtaaggctt cattatggag atggtccaga
     3901 tgaaaacagg aagacaagga atttgaggaa aaataatcca ttctggaaag ctgtcacaat
     3961 tttaaagttt aaaagggatc ttgatggaga gaagcaaatc cctgaattta tgtacggcga
     4021 gcatatagaa gttcgtgaag cattcttagc cggcttgatc gactcagatg ggtacgttgt
     4081 gaaaaagggc gaaggccctg aatcttataa aatagcaatt caaactgttt attcatccat
     4141 tatggacgga attgtccata tttcaagatc tcttggtatg tcagctactg tgacgaccag
     4201 gtcagctagg gaggaaatca ttgaaggaag aaaagtccaa tgtcaattta catacgactg
     4261 taatgttgct gggggaacaa ctttacagaa tgttttgtca tattgtcgaa gtggtcacaa
     4321 aacaagagaa gttccgccaa ttataaaaag ggaacccgta tatttcagct tcacggatga
     4381 tttccagggt gagagtactg tatatgggct tacgatagaa ggccataaaa atttcttgct
     4441 tggcaacaaa atagaagtga aatcatgtcg aggctgctgt gtgggagaac agcataaaat
     4501 atcacaaaaa aagaatctaa aacactgtgt tgcttgtccc agaaagggaa tcaagtattt
     4561 ttataaagat tggagtggta aaaatcgagt atgtgctaga tgctatggaa gatacaaatt
     4621 cagcggtcat cactgtataa attgcaagta tgtaccagaa gcacgtgaag tgaaaaaggc
     4681 aaaagacaaa ggcgaaaaat tgggcattac gcccgaaggt ttgccagtta aaggaccaga
     4741 gtgtataaaa tgtggcggaa tcttacagtt tgatgctgtc cgcgggcctc ataagagttg
     4801 tggtaacaac gcaggtgcgc gcatctgcga tatcgactac aaggatgacg acgacaagaa
     4861 gctttaacta actaacctcg agacaggccc cttttccttt gtctatatca tgtaattagt
     4921 tatgtcacgc ttacattcac gccctcctcc cacatccgct ctaaccgaaa aggaaggagt
     4981 tagacaacct gaagtctagg tccctattta ttttttttaa tagttatgtt agtattaaga
     5041 acgttattta tatttcaaat ttttcttttt tttctgtaca aacgcgtgta cgcatgtaac
     5101 attatactga aaaccttgct tgagaaggtt ttgggacgct cgaaggcttt aatttgcaag
     5161 gttcgcagtt tacactctca tcgtcgctct catcatcgct tccgttgttg ttttccttag
     5221 tagcgtctgc ttggtaccca gcttcggccg ctctagagaa gcgggtaagc tgccacagca
     5281 attaatgcac aacatttaac ctacattctt ccttatcgga tcctcaaaac ccttaaaaac
     5341 atatgcctca ccctaacata ttttccaatt aaccctcaat atttctctgt cacccggcct
     5401 ctattttcca ttttcttctt tacccgccac gcgttttttt ctttcaaatt tttttcttcc
     5461 ttcttctttt tcttccacgt cctcttgcat aaataaataa accgttttga aaccaaactc
     5521 gcctctctct ctcctttttg aaatattttt gggtttgttt gatcctttcc ttcccaatct
     5581 ctcttgttta atatatattc atttatatca cgctctcttt ttatcttcct ttttttcctc
     5641 tctcttgtat tcttccttcc cctttctact caaaccaaga agaaaaagaa aaggtcaatc
     5701 tttgttaaag aataggatct tctactacat cagcttttag atttttcacg cttactgctt
     5761 ttttcttccc aagatcgaaa atttactgaa ttaacaacta gtggatcccc cgggctgcag
     5821 gaattcatga aagcgttaac ggccaggcaa caagaggtgt ttgatctcat ccgtgatcac
     5881 atcagccaga caggtatgcc gccgacgcgt gcggaaatcg cgcagcgttt ggggttccgt
     5941 tccccaaacg cggctgaaga acatctgaag gcgctggcac gcaaaggcgt tattgaaatt
     6001 gtttccggcg catcacgcgg gattcgtctg ttgcaggaag aggaagaagg gttgccgctg
     6061 gtaggtcgtg tggctgccgg tgaaccactt ctggcgcaac agcatattga aggtcattat
     6121 caggtcgatc cttccttatt caagccgaat gctgatttcc tgctgcgcgt cagcgggatg
     6181 tcgatgaaag acatcggcat tatggatggt gacttgctgg cagtgcataa aactcaggat
     6241 gtacgtaacg gtcaggtcgt tgtcgcacgt attgatgacg aggttaccgt taagcgcctg
     6301 aaaaaacagg gcaataaagt cgaactgttg ccagaaaata gcgagtttaa accaattgtc
     6361 gtagatcttc gtcagcagag cttcaccatt gaagggctgg cggttggggt tattcgcaac
     6421 ggcgactggc tgggttctaa ggatatctct gctggagaca tgagagctgc caacctttgg
     6481 ccaagcccgc tcatgatcaa acgctctaag aagaacagcc tggccttgtc cctgacggcc
     6541 gaccagatgg tcagtgcctt gttggatgct gagcccccca tactctattc cgagtatgat
     6601 cctaccagac ccttcagtga agcttcgatg atgggcttac tgaccaacct ggcagacagg
     6661 gagctggttc acatgatcaa ctgggcgaag agggtgccag gctttgtgga tttgaccctc
     6721 catgatcagg tccaccttct agaatgtgcc tggttagaga tcctgatgat tggtctcgtc
     6781 tggcgctcca tggagcaccc agtgaagcta ctgtttgctc ctaacttgct cttggacagg
     6841 aaccagggaa aatgtgtaga gggcatggtg gagatcttcg acatgctgct ggctacatca
     6901 tctcggttcc gcatgatgaa tctgcaggga gaggagtttg tgtgcctcaa atctattatt
     6961 ttgcttaatt ctggagtgta cacatttctg tccagcaccc tgaagtctct ggaagagaag
     7021 gaccatatcc accgagtcct ggacaagatc acagacactt tgatccacct gatggccaag
     7081 gcaggcctga ccctgcagca gcagcaccag cggctggccc agctcctcct catcctctcc
     7141 cacatcaggc acatgagtaa caaaggcatg gagcatctgt acagcatgaa gtgcaagaac
     7201 gtggtgcccc tctatgacct gctgctggag atgctggacg cccaccgcct acatgcgccc
     7261 actagccgtg gaggggcatc cgtggaggag acggaccaaa gccacttggc cactgcgggc
     7321 tctacttcat cgcattcctt gcaaaagtat tacatcacgg gggaggcaga gggtttccct
     7381 gccacagtcg atatcatgca ggatctgccg ggcaacgata acagcaccgc gggcgaattt
     7441 ccaggtatta ctttgagaat ccaagaaact gatatgttgt ataaaggtga tactttgtat
     7501 ttggattggt tggaagatgg tattgctgaa ttggttttcg atgctccagg ttctgttaat
     7561 aaattggata ctgctgttgc ttctttgggt gaagctattg gtgttttgga acaacaatct
     7621 gatttgattt gggaaacttt gactgttaaa gatgctaaag ttaatttcga ttctggtttg
     7681 gaaaaattcg aagaagctat tccatctgct gatgatttcg atccagttgc tgaaagaaga
     7741 tcctctggtg aatttagagc tgaaagacat tctggtggta ctgatttgtg tttcaagctt
     7801 taaacgcgtt taaagcttat cctcgagaca ggcccctttt cctttgtcta tatcatgtaa
     7861 ttagttatgt cacgcttaca ttcacgccct cctcccacat ccgctctaac cgaaaaggaa
     7921 ggagttagac aacctgaagt ctaggtccct atttattttt tttaatagtt atgttagtat
     7981 taagaacgtt atttatattt caaatttttc ttttttttct gtacaaacgc gtgtacgcat
     8041 gtaacattat actgaaaacc ttgcttgaga aggttttggg acgctcgaag gctttaattt
     8101 gcaaggttcg cagtttacac tctcatcgtc gctctcatca tcgcttccgt tgttgttttc
     8161 cttagtagcg tctgcttggt acccagcttc ggccgtattt acagataacg atgtcgttat
     8221 tatatatata tatatatatt gtcacttgta cccagctttt gttcccttta gtgagggtta
     8281 atttcgagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     8341 acaacagctg taatatatag tagtaacctg aaaaatagtc aatgcccgtg gttcatttgc
     8401 caatagaaca gtcgatgtga tcaatgtaca cgaaagtcaa tataaaatat ccaaatagcc
     8461 aatggtgtca ggacggctaa ggcaaatgca actttgaaat acttattcgg tttgcatctt
     8521 ggcgcgcctc ttcctgcagg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     8581 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     8641 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     8701 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     8761 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     8821 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     8881 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     8941 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     9001 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     9061 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     9121 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     9181 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     9241 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     9301 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     9361 aagaagatcc tttgatcttt tgtttaaacc tacggggtct gacgctcagt ggaacgaaaa
     9421 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     9481 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     9541 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     9601 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     9661 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     9721 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     9781 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     9841 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     9901 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     9961 ggttagctcc ttcggtcctc cgatggttgt cagaagtaag ttggccgcag tgttatcact
    10021 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
    10081 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
    10141 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
    10201 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
    10261 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
    10321 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
    10381 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
    10441 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
    10501 tccgcgcaca tttccccgaa aagtgccacc t
//
