LOCUS       sequence-370803-       11127 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The CRISPR/Cas9-and-gRNA-expressing plasmid with a G418-resistent 
            selectable marker .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11127)
  TITLE     Vickers Lab plasmids
REFERENCE   2  (bases 1 to 11127)
  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"
COMMENT     Sequence Label: pJCKan
FEATURES             Location/Qualifiers
     source          1..11127
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    58..91
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     gene            146..1502
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        146..489
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             490..1299
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(557..576)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      1305..1502
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     protein_bind    1565..1598
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        1648..2314
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     2184..2207
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     regulatory      2343..2352
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2349..4067
                     /codon_start=1
                     /product="N-terminal portion of Streptococcus pyogenes Cas9
                     (Zetsche et al., 2015)"
                     /label=Cas9(N)
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIE"
     CDS             6462..6482
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      6510..6697
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(6565..6589)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     terminator      complement(6815..6834)
                     /label=SUP4 terminator
                     /note="transcription terminator for the S. cerevisiae SUP4 
                     tRNA gene"
     promoter        complement(6916..7184)
                     /label=SNR52 promoter
                     /note="promoter for the S. cerevisiae small nucleolar RNA 
                     gene SNR52"
     primer_bind     complement(7215..7235)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(7215..7233)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7254..7270)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7254..7270)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7267..7289)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7278..7294
                     /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(7302..7308,7309..7326,7327..7332))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7347..7368
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7485..7502)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7656..8244)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7736..7755)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8415..9206,9207..9275))
                     /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     9038..9057
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9276..9380)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      9407..10749
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(10370..10417)
                     /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     10790..10808
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(10846..10868)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     10968..10987
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gtatatatac gcatatgcag ctgaagcttc gtacgctgca ggtcgacaac ccttaatata
       61 acttcgtata atgtatgcta tacgaagtta ttaggtctag agatctgttt agcttgcctc
      121 gtccccgccg ggtcacccgg ccagcgacat ggaggcccag aataccctcc ttgacagtct
      181 tgacgtgcgc agctcagggg catgatgtga ctgtcgcccg tacatttagc ccatacatcc
      241 ccatgtataa tcatttgcat ccatacattt tgatggccgc acggcgcgaa gcaaaaatta
      301 cggctcctcg ctgcagacct gcgagcaggg aaacgctccc ctcacagacg cgttgaattg
      361 tccccacgcc gcgcccctgt agagaaatat aaaaggttag gatttgccac tgaggttctt
      421 ctttcatata cttcctttta aaatcttgct aggatacagt tctcacatca catccgaaca
      481 taaacaacca tgggtaagga aaagactcac gtttcgaggc cgcgattaaa ttccaacatg
      541 gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc aggtgcgaca
      601 atctatcgat tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt
      661 agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac ggaatttatg
      721 cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt actcaccact
      781 gcgatccccg gcaaaacagc attccaggta ttagaagaat atcctgattc aggtgaaaat
      841 attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt
      901 ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat gaataacggt
      961 ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga acaagtctgg
     1021 aaagaaatgc ataagctttt gccattctca ccggattcag tcgtcactca tggtgatttc
     1081 tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga tgttggacga
     1141 gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct cggtgagttt
     1201 tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc tgatatgaat
     1261 aaattgcagt ttcatttgat gctcgatgag tttttctaat cagtactgac aataaaaaga
     1321 ttcttgtttt caagaacttg tcatttgtat agttttttta tattgtagtt gttctatttt
     1381 aatcaaatgt tagcgtgatt tatatttttt ttcgcctcga catcatctgc ccagatgcga
     1441 agttaagtgc gcagaaagta atatcatgcg tcaatcgtat gtgaatgctg gtcgctatac
     1501 tgctgtcgat tcgatactaa cgccgccatc cagtgtcgaa aacgagctct cgagaaccct
     1561 taatataact tcgtataatg tatgctatac gaagttatta ggtgatatca gatccactag
     1621 tggcctatac tagtcagttc gagtttatca ttatcaatac tgccatttca aagaatacgt
     1681 aaataattaa tagtagtgat tttcctaact ttatttagtc aaaaaattag ccttttaatt
     1741 ctgctgtaac ccgtacatgc ccaaaatagg gggcgggtta cacagaatat ataacatcgt
     1801 aggtgtctgg gtgaacagtt tattcctggc atccactaaa tataatggag cccgcttttt
     1861 aagctggcat ccagaaaaaa aaagaatccc agcaccaaaa tattgttttc ttcaccaacc
     1921 atcagttcat aggtccattc tcttagcgca actacagaga acaggggcac aaacaggcaa
     1981 aaaacgggca caacctcaat ggagtgatgc aacctgcctg gagtaaatga tgacacaagg
     2041 caattgaccc acgcatgtat ctatctcatt ttcttacacc ttctattacc ttctgctctc
     2101 tctgatttgg aaaaagctga aaaaaaaggt tgaaaccagt tccctgaaat tattccccta
     2161 cttgactaat aagtatataa agacggtagg tattgattgt aattctgtaa atctatttct
     2221 taaacttctt aaattctact tttatagtta gtcttttttt tagttttaaa acaccaagaa
     2281 cttagtttcg aataaacaca cataaacaaa caaaagatct gatatggatc gaattagatc
     2341 tcgccaccat ggacaagaag tattctatcg gactggacat cgggactaat agcgtcgggt
     2401 gggccgtcat cactgacgag tacaaggtgc cctctaagaa gttcaaggtg ctcgggaaca
     2461 ccgaccggca ttccatcaag aaaaatctga tcggagctct cctctttgat tcaggggaga
     2521 ccgctgaagc aacccgcctc aagcggactg ctagacggcg gtacaccagg aggaagaacc
     2581 ggatttgtta ccttcaagag atattctcca acgaaatggc aaaggtcgac gacagcttct
     2641 tccataggct ggaagaatca ttcctcgtgg aagaggataa gaagcatgaa cggcatccca
     2701 tcttcggtaa tatcgtcgac gaggtggcct atcacgagaa atacccaacc atctaccatc
     2761 ttcgcaaaaa gctggtggac tcaaccgaca aggcagacct ccggcttatc tacctggccc
     2821 tggcccacat gattaagttc agaggccact tcctgatcga gggcgacctc aatcctgaca
     2881 atagcgatgt ggataaactg ttcatccagc tggtgcagac ttacaaccag ctctttgaag
     2941 agaaccccat caatgcaagc ggagtcgatg ccaaggccat tctgtcagcc cggctgtcaa
     3001 agagccgcag acttgagaat cttatcgctc agctgccggg tgaaaagaaa aatggactgt
     3061 tcgggaacct gattgctctt tcacttgggc tgactcccaa tttcaagtct aatttcgacc
     3121 tggcagagga tgccaagctg caactgtcca aggacaccta tgatgacgat ctcgacaacc
     3181 tcctggccca gatcggtgac caatacgccg accttttcct tgctgctaag aatctttctg
     3241 acgccatcct gctgtctgac attctccgcg tgaacactga aatcaccaag gcccctcttt
     3301 cagcttcaat gattaagcgg tatgatgagc accaccagga cctgaccctg cttaaggcac
     3361 tcgtccggca gcagcttccg gagaagtaca aggaaatctt ctttgaccag tcaaagaatg
     3421 gatacgccgg ctacatcgac ggaggtgcct cccaagagga attttataag tttatcaaac
     3481 ctatccttga gaagatggac ggcaccgaag agctcctcgt gaaactgaat cgggaggatc
     3541 tgctgcggaa gcagcgcact ttcgacaatg ggagcattcc ccaccagatc catcttgggg
     3601 agcttcacgc catccttcgg cgccaagagg acttctaccc ctttcttaag gacaacaggg
     3661 agaagattga gaaaattctc actttccgca tcccctacta cgtgggaccc ctcgccagag
     3721 gaaatagccg gtttgcttgg atgaccagaa agtcagaaga aactatcact ccctggaact
     3781 tcgaagaggt ggtggacaag ggagccagcg ctcagtcatt catcgaacgg atgactaact
     3841 tcgataagaa cctccccaat gagaaggtcc tgccgaaaca ttccctgctc tacgagtact
     3901 ttaccgtgta caacgagctg accaaggtga aatatgtcac cgaagggatg aggaagcccg
     3961 cattcctgtc aggcgaacaa aagaaggcaa ttgtggacct tctgttcaag accaatagaa
     4021 aggtgaccgt gaagcagctg aaggaggact atttcaagaa aattgaatgc ttcgactctg
     4081 tggagattag cggggtcgaa gatcggttca acgcaagcct gggtacctac catgatctgc
     4141 ttaagatcat caaggacaag gattttctgg acaatgagga gaaagaggac atccttgagg
     4201 acattgtcct gactctcact ctgttcgagg accgggaaat gatcgaggag aggcttaaga
     4261 cctacgccca tctgttcgac gataaagtga tgaagcaact taaacggaga agatataccg
     4321 gatggggacg ccttagccgc aaactcatca acggaatccg ggacaaacag agcggaaaga
     4381 ccattcttga tttccttaag agcgacggat tcgctaatcg caacttcatg caacttatcc
     4441 atgatgattc cctgaccttt aaggaggaca tccagaaggc ccaagtgtct ggacaaggtg
     4501 actcactgca cgagcatatc gcaaatctgg ctggttcacc cgctattaag aagggtattc
     4561 tccagaccgt gaaagtcgtg gacgagctgg tcaaggtgat gggtcgccat aaaccagaga
     4621 acattgtcat cgagatggcc agggaaaacc agactaccca gaagggacag aagaacagca
     4681 gggagcggat gaaaagaatt gaggaaggga ttaaggagct cgggtcacag atccttaaag
     4741 agcacccggt ggaaaacacc cagcttcaga atgagaagct ctatctgtac taccttcaaa
     4801 atggacgcga tatgtatgtg gaccaagagc ttgatatcaa caggctctca gactacgacg
     4861 tggaccacat cgtccctcag agcttcctca aagacgactc aattgacaat aaggtgctga
     4921 ctcgctcaga caagaaccgg ggaaagtcag ataacgtgcc ctcagaggaa gtcgtgaaaa
     4981 agatgaagaa ctattggcgc cagcttctga acgcaaagct aatcactcag cggaagttcg
     5041 acaatctcac taaggctgag aggggcggac tgagcgaact ggacaaagca ggattcatta
     5101 aacggcaact tgtggagact cggcagatta ctaaacatgt agcccaaatc cttgactcac
     5161 gcatgaatac caagtacgac gaaaacgaca aacttatccg cgaggtgaag gtgattaccc
     5221 tgaagtccaa gctggtcagc gatttcagaa aggactttca attctacaaa gtgcgggaga
     5281 tcaataacta tcatcatgct catgacgcat atctgaatgc cgtggtggga accgccctaa
     5341 tcaagaagta cccaaagctg gaaagcgagt tcgtgtacgg agactacaag gtctacgacg
     5401 tgcgcaagat gattgccaaa tctgagcagg agatcggaaa ggccaccgca aagtacttct
     5461 tctacagcaa catcatgaat ttcttcaaga ccgaaatcac ccttgcaaac ggtgagatcc
     5521 ggaagaggcc gctcatcgag actaatgggg agactggcga aatcgtgtgg gacaagggca
     5581 gagatttcgc taccgtgcgc aaagtgcttt ctatgcctca agtgaacatc gtgaagaaaa
     5641 ccgaggtgca aaccggaggc ttttctaagg aatcaatcct ccccaagcgc aactccgaca
     5701 agctcattgc aaggaagaag gattgggacc ctaagaagta cggcggattc gattcaccaa
     5761 ctgtggctta ttctgtcctg gtcgtggcta aggtggaaaa aggaaagtct aagaagctca
     5821 agagcgtgaa ggaactgctg ggtatcacca ttatggagcg cagctccttc gagaagaacc
     5881 caattgactt tctcgaagcc aaaggttaca aggaagtcaa gaaggacctt atcatcaagc
     5941 tcccaaagta tagcctgttc gaactggaga atgggcggaa gcggatgctc gcctccgctg
     6001 gcgaacttca gaagggtaat gagctggctc tcccctccaa gtacgtgaat ttcctctacc
     6061 ttgcaagcca ttacgagaag ctgaagggga gccccgagga caacgagcaa aagcaactgt
     6121 ttgtggagca gcataagcat tatctggacg agatcattga gcagatttcc gagttttcta
     6181 aacgcgtcat tctcgctgat gccaacctcg ataaagtcct tagcgcatac aataagcaca
     6241 gagacaaacc aattcgggag caggctgaga atatcatcca cctgttcacc ctcaccaatc
     6301 ttggtgcccc tgccgcattc aagtacttcg acaccaccat cgaccggaaa cgctatacct
     6361 ccaccaaaga agtgctggac gccaccctca tccaccagag catcaccgga ctttacgaaa
     6421 ctcggattga cctctcacag ctcggagggg atgagggagc tcccaagaaa aagcgcaagg
     6481 taggtagttc cggatcctag actctcgagg cgaatttctt atgatttatg atttttatta
     6541 ttaaataagt tataaaaaaa ataagtgtat acaaatttta aagtgactct taggttttaa
     6601 aacgaaaatt cttattcttg agtaactctt tcctgtaggt caggttgctt tctcaggtat
     6661 agcatgaggt cgctcttatt gaccacacct ctaccggcat gccgagcaaa tgcctgcaaa
     6721 tcgctcccca tttctctaga gcggccgtgg tatcgtttag attggcaatt acagtgtctt
     6781 agctcacatg cttataacta attacatgac tcgaagacat aaaaaacaaa aaaagcacca
     6841 ccgactcggt gccacttttt caagttgata acggactagc cttattttaa cttgctattt
     6901 aaatgcatcg attctgatca tttatctttc actgcggaga agtttcgaac gccgaaacat
     6961 gcgcaccaac tttcacttct acagcgtttg accaaaatct tttgaacaga acattgtagg
     7021 gtgtgaaaaa atgcgcacct ttaccgctag cccaagaggg cactacaaaa tctagagttg
     7081 tacttcaaac gtacatgtaa tcaccttgta tatactcgaa agaaaacatc aagtttctgt
     7141 ataaatatga gtgaaagcat aatcatacat tatcttttca aagattgtcc gcggtggagc
     7201 tccagctttt gttcccttta gtgagggtta attgcgcgct tggcgtaatc atggtcatag
     7261 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     7321 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     7381 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     7441 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     7501 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     7561 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     7621 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     7681 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     7741 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     7801 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     7861 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     7921 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     7981 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     8041 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     8101 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     8161 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     8221 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     8281 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     8341 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     8401 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     8461 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     8521 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     8581 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     8641 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     8701 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     8761 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     8821 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     8881 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     8941 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     9001 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     9061 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     9121 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     9181 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     9241 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     9301 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     9361 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgaacg aagcatctgt
     9421 gcttcatttt gtagaacaaa aatgcaacgc gagagcgcta atttttcaaa caaagaatct
     9481 gagctgcatt tttacagaac agaaatgcaa cgcgaaagcg ctattttacc aacgaagaat
     9541 ctgtgcttca tttttgtaaa acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag
     9601 aatctgagct gcatttttac agaacagaaa tgcaacgcga gagcgctatt ttaccaacaa
     9661 agaatctata cttctttttt gttctacaaa aatgcatccc gagagcgcta tttttctaac
     9721 aaagcatctt agattacttt ttttctcctt tgtgcgctct ataatgcagt ctcttgataa
     9781 ctttttgcac tgtaggtccg ttaaggttag aagaaggcta ctttggtgtc tattttctct
     9841 tccataaaaa aagcctgact ccacttcccg cgtttactga ttactagcga agctgcgggt
     9901 gcattttttc aagataaagg catccccgat tatattctat accgatgtgg attgcgcata
     9961 ctttgtgaac agaaagtgat agcgttgatg attcttcatt ggtcagaaaa ttatgaacgg
    10021 tttcttctat tttgtctcta tatactacgt ataggaaatg tttacatttt cgtattgttt
    10081 tcgattcact ctatgaatag ttcttactac aatttttttg tctaaagagt aatactagag
    10141 ataaacataa aaaatgtaga ggtcgagttt agatgcaagt tcaaggagcg aaaggtggat
    10201 gggtaggtta tatagggata tagcacagag atatatagca aagagatact tttgagcaat
    10261 gtttgtggaa gcggtattcg caatatttta gtagctcgtt acagtccggt gcgtttttgg
    10321 ttttttgaaa gtgcgtcttc agagcgcttt tggttttcaa aagcgctctg aagttcctat
    10381 actttctaga gaataggaac ttcggaatag gaacttcaaa gcgtttccga aaacgagcgc
    10441 ttccgaaaat gcaacgcgag ctgcgcacat acagctcact gttcacgtcg cacctatatc
    10501 tgcgtgttgc ctgtatatat atatacatga gaagaacggc atagtgcgtg tttatgctta
    10561 aatgcgtact tatatgcgtc tatttatgta ggatgaaagg tagtctagta cctcctgtga
    10621 tattatccca ttccatgcgg ggtatcgtat gcttccttca gcactaccct ttagctgttc
    10681 tatatgctgc cactcctcaa ttggattagt ctcatccttc aatgctatca tttcctttga
    10741 tattggatca tactaagaaa ccattattat catgacatta acctataaaa ataggcgtat
    10801 cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
    10861 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
    10921 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
    10981 gattgtactg agagtgcacc ataccacctt ttcaattcat catttttttt ttattctttt
    11041 ttttgatttc ggtttccttg aaattttttt gattcggtaa tctccgaaca gaaggaagaa
    11101 cgaaggaagg agcacagact tagattg
//
