LOCUS       sequence-462214-       11197 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid expressing Cas9 and gRNA GAAGTAACAAAGGAACCTAG which targets 
            the URA3 gene..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11197)
  TITLE     van Leeuwen Lab plasmids 2025
REFERENCE   2  (bases 1 to 11197)
  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: pML104-HygMx4-URA3-3
FEATURES             Location/Qualifiers
     source          1..11197
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1750..1770
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      1798..1985
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1853..1877)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     terminator      complement(2103..2122)
                     /label=SUP4 terminator
                     /note="transcription terminator for the S. cerevisiae SUP4 
                     tRNA gene"
     misc_RNA        complement(2127..2201)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(2222..2490)
                     /label=SNR52 promoter
                     /note="promoter for the S. cerevisiae small nucleolar RNA 
                     gene SNR52"
     primer_bind     complement(2521..2541)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2521..2539)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2560..2576)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2560..2576)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2573..2595)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2584..2600
                     /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(2608..2614,2615..2632,2633..2638))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2653..2674
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2791..2808)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2962..3550)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3042..3061)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3721..4512,4513..4581))
                     /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     4344..4363
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4582..4686)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4713..6055
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(5676..5723)
                     /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     6096..6114
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6152..6174)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6274..6293
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     gene            6482..8057
                     /label=hphMX6
                     /note="yeast selectable marker conferring hygromycin 
                     resistance"
     promoter        6482..6825
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             6826..7854
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEQFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     terminator      7860..8057
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     promoter        8133..8799
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     8669..8692
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     regulatory      8828..8837
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             8834..10552
                     /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"
ORIGIN
        1 aaggagctcg ggtcacagat ccttaaagag cacccggtgg aaaacaccca gcttcagaat
       61 gagaagctct atctgtacta ccttcaaaat ggacgcgata tgtatgtgga ccaagagctt
      121 gatatcaaca ggctctcaga ctacgacgtg gaccacatcg tccctcagag cttcctcaaa
      181 gacgactcaa ttgacaataa ggtgctgact cgctcagaca agaaccgggg aaagtcagat
      241 aacgtgccct cagaggaagt cgtgaaaaag atgaagaact attggcgcca gcttctgaac
      301 gcaaagctaa tcactcagcg gaagttcgac aatctcacta aggctgagag gggcggactg
      361 agcgaactgg acaaagcagg attcattaaa cggcaacttg tggagactcg gcagattact
      421 aaacatgtag cccaaatcct tgactcacgc atgaatacca agtacgacga aaacgacaaa
      481 cttatccgcg aggtgaaggt gattaccctg aagtccaagc tggtcagcga tttcagaaag
      541 gactttcaat tctacaaagt gcgggagatc aataactatc atcatgctca tgacgcatat
      601 ctgaatgccg tggtgggaac cgccctaatc aagaagtacc caaagctgga aagcgagttc
      661 gtgtacggag actacaaggt ctacgacgtg cgcaagatga ttgccaaatc tgagcaggag
      721 atcggaaagg ccaccgcaaa gtacttcttc tacagcaaca tcatgaattt cttcaagacc
      781 gaaatcaccc ttgcaaacgg tgagatccgg aagaggccgc tcatcgagac taatggggag
      841 actggcgaaa tcgtgtggga caagggcaga gatttcgcta ccgtgcgcaa agtgctttct
      901 atgcctcaag tgaacatcgt gaagaaaacc gaggtgcaaa ccggaggctt ttctaaggaa
      961 tcaatcctcc ccaagcgcaa ctccgacaag ctcattgcaa ggaagaagga ttgggaccct
     1021 aagaagtacg gcggattcga ttcaccaact gtggcttatt ctgtcctggt cgtggctaag
     1081 gtggaaaaag gaaagtctaa gaagctcaag agcgtgaagg aactgctggg tatcaccatt
     1141 atggagcgca gctccttcga gaagaaccca attgactttc tcgaagccaa aggttacaag
     1201 gaagtcaaga aggaccttat catcaagctc ccaaagtata gcctgttcga actggagaat
     1261 gggcggaagc ggatgctcgc ctccgctggc gaacttcaga agggtaatga gctggctctc
     1321 ccctccaagt acgtgaattt cctctacctt gcaagccatt acgagaagct gaaggggagc
     1381 cccgaggaca acgagcaaaa gcaactgttt gtggagcagc ataagcatta tctggacgag
     1441 atcattgagc agatttccga gttttctaaa cgcgtcattc tcgctgatgc caacctcgat
     1501 aaagtcctta gcgcatacaa taagcacaga gacaaaccaa ttcgggagca ggctgagaat
     1561 atcatccacc tgttcaccct caccaatctt ggtgcccctg ccgcattcaa gtacttcgac
     1621 accaccatcg accggaaacg ctatacctcc accaaagaag tgctggacgc caccctcatc
     1681 caccagagca tcaccggact ttacgaaact cggattgacc tctcacagct cggaggggat
     1741 gagggagctc ccaagaaaaa gcgcaaggta ggtagttccg gatcctagac tctcgaggcg
     1801 aatttcttat gatttatgat ttttattatt aaataagtta taaaaaaaat aagtgtatac
     1861 aaattttaaa gtgactctta ggttttaaaa cgaaaattct tattcttgag taactctttc
     1921 ctgtaggtca ggttgctttc tcaggtatag catgaggtcg ctcttattga ccacacctct
     1981 accggcatgc cgagcaaatg cctgcaaatc gctccccatt tctctagagc ggccgtggta
     2041 tcgtttagat tggcaattac agtgtcttag ctcacatgct tataactaat tacatgactc
     2101 gaagacataa aaaacaaaaa aagcaccacc gactcggtgc cactttttca agttgataac
     2161 ggactagcct tattttaact tgctatttct agctctaaaa cctaggttcc tttgttactt
     2221 cgatcattta tctttcactg cggagaagtt tcgaacgccg aaacatgcgc accaactttc
     2281 acttctacag cgtttgacca aaatcttttg aacagaacat tgtagggtgt gaaaaaatgc
     2341 gcacctttac cgctagccca agagggcact acaaaatcta gagttgtact tcaaacgtac
     2401 atgtaatcac cttgtatata ctcgaaagaa aacatcaagt ttctgtataa atatgagtga
     2461 aagcataatc atacattatc ttttcaaaga ttgtccgcgg tggagctcca gcttttgttc
     2521 cctttagtga gggttaattg cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     2581 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     2641 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     2701 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     2761 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     2821 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     2881 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     2941 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     3001 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     3061 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     3121 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     3181 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     3241 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     3301 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     3361 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     3421 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     3481 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     3541 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     3601 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     3661 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     3721 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     3781 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     3841 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     3901 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     3961 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     4021 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     4081 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     4141 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     4201 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     4261 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     4321 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     4381 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     4441 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     4501 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     4561 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     4621 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     4681 tccgcgcaca tttccccgaa aagtgccacc tgaacgaagc atctgtgctt cattttgtag
     4741 aacaaaaatg caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc tgcattttta
     4801 cagaacagaa atgcaacgcg aaagcgctat tttaccaacg aagaatctgt gcttcatttt
     4861 tgtaaaacaa aaatgcaacg cgagagcgct aatttttcaa acaaagaatc tgagctgcat
     4921 ttttacagaa cagaaatgca acgcgagagc gctattttac caacaaagaa tctatacttc
     4981 ttttttgttc tacaaaaatg catcccgaga gcgctatttt tctaacaaag catcttagat
     5041 tacttttttt ctcctttgtg cgctctataa tgcagtctct tgataacttt ttgcactgta
     5101 ggtccgttaa ggttagaaga aggctacttt ggtgtctatt ttctcttcca taaaaaaagc
     5161 ctgactccac ttcccgcgtt tactgattac tagcgaagct gcgggtgcat tttttcaaga
     5221 taaaggcatc cccgattata ttctataccg atgtggattg cgcatacttt gtgaacagaa
     5281 agtgatagcg ttgatgattc ttcattggtc agaaaattat gaacggtttc ttctattttg
     5341 tctctatata ctacgtatag gaaatgttta cattttcgta ttgttttcga ttcactctat
     5401 gaatagttct tactacaatt tttttgtcta aagagtaata ctagagataa acataaaaaa
     5461 tgtagaggtc gagtttagat gcaagttcaa ggagcgaaag gtggatgggt aggttatata
     5521 gggatatagc acagagatat atagcaaaga gatacttttg agcaatgttt gtggaagcgg
     5581 tattcgcaat attttagtag ctcgttacag tccggtgcgt ttttggtttt ttgaaagtgc
     5641 gtcttcagag cgcttttggt tttcaaaagc gctctgaagt tcctatactt tctagagaat
     5701 aggaacttcg gaataggaac ttcaaagcgt ttccgaaaac gagcgcttcc gaaaatgcaa
     5761 cgcgagctgc gcacatacag ctcactgttc acgtcgcacc tatatctgcg tgttgcctgt
     5821 atatatatat acatgagaag aacggcatag tgcgtgttta tgcttaaatg cgtacttata
     5881 tgcgtctatt tatgtaggat gaaaggtagt ctagtacctc ctgtgatatt atcccattcc
     5941 atgcggggta tcgtatgctt ccttcagcac taccctttag ctgttctata tgctgccact
     6001 cctcaattgg attagtctca tccttcaatg ctatcatttc ctttgatatt ggatcatact
     6061 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     6121 gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg
     6181 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg
     6241 gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag
     6301 tgcaccatac caccttttca attcatcatt ttttttttat tctttttttt gatttcggtt
     6361 tccttgaaat ttttttgatt cggtaatctc cgaacagaag gaagaacgaa ggaaggagca
     6421 cagacttaga ttggtatata tacgcaagct tgcctcgtcc ccgccgggtc acccggccag
     6481 cgacatggag gcccagaata ccctccttga cagtcttgac gtgcgcagct caggggcatg
     6541 atgtgactgt cgcccgtaca tctagcccat acatccccat gtataatcat ttgcatccat
     6601 acattttgat ggccgcacgg cgcgaagcaa aaattacggc ccctcgctgc ggacctgcga
     6661 gcagggaaac gctcccctca cagacgcgtt gaattgtccc cacgccgcgc ccctgtagag
     6721 aaatataaaa ggttaggatt tgccactgag gttcttcttt catatacttc cttttaaaat
     6781 cttgctagga tacagttctc acatcacatc cgaacataaa caaccatggg taaaaagcct
     6841 gaactcaccg cgacgtctgt cgagaagttt ctgatcgaac agttcgacag cgtctccgac
     6901 ctgatgcagc tctcggaggg cgaagaatct cgtgctttca gcttcgatgt aggagggcgt
     6961 ggatatgtcc tgcgggtaaa tagctgcgcc gatggtttct acaaagatcg ttatgtttat
     7021 cggcactttg catcggccgc gctcccgatt ccggaagtgc ttgacattgg ggaattcagc
     7081 gagagcctga cctattgcat ctcccgccgt gcacagggtg tcacgttgca agacctgcct
     7141 gaaaccgaac tgcccgctgt tctgcagccg gtcgcggagg ccatggatgc gatcgctgcg
     7201 gccgatctta gccagacgag cgggttcggc ccattcggac cgcaaggaat cggtcaatac
     7261 actacatggc gtgatttcat atgcgcgatt gctgatcccc atgtgtatca ctggcaaact
     7321 gtgatggacg acaccgtcag tgcgtccgtc gcgcaggctc tcgatgagct gatgctttgg
     7381 gccgaggact gccccgaagt ccggcacctc gtgcacgcgg atttcggctc caacaatgtc
     7441 ctgacggaca atggccgcat aacagcggtc attgactgga gcgaggcgat gttcggggat
     7501 tcccaatacg aggtcgccaa catcttcttc tggaggccgt ggttggcttg tatggagcag
     7561 cagacgcgct acttcgagcg gaggcatccg gagcttgcag gatcgccgcg gctccgggcg
     7621 tatatgctcc gcattggtct tgaccaactc tatcagagct tggttgacgg caatttcgat
     7681 gatgcagctt gggcgcaggg tcgatgcgac gcaatcgtcc gatccggagc cgggactgtc
     7741 gggcgtacac aaatcgcccg cagaagcgcg gccgtctgga ccgatggctg tgtagaagta
     7801 ctcgccgata gtggaaaccg acgccccagc actcgtccga gggcaaagga ataatcagta
     7861 ctgacaataa aaagattctt gttttcaaga acttgtcatt tgtatagttt ttttatattg
     7921 tagttgttct attttaatca aatgttagcg tgatttatat tttttttcgc ctcgacatca
     7981 tctgcccaga tgcgaagtta agtgcgcaga aagtaatatc atgcgtcaat cgtatgtgaa
     8041 tgctggtcgc tatactgctg tcgattcgat actaacgccg ccatccagtg tcgaatgata
     8101 tcgaattcag tctactagtc agttcgagtt tatcattatc aatactgcca tttcaaagaa
     8161 tacgtaaata attaatagta gtgattttcc taactttatt tagtcaaaaa attagccttt
     8221 taattctgct gtaacccgta catgcccaaa atagggggcg ggttacacag aatatataac
     8281 atcgtaggtg tctgggtgaa cagtttattc ctggcatcca ctaaatataa tggagcccgc
     8341 tttttaagct ggcatccaga aaaaaaaaga atcccagcac caaaatattg ttttcttcac
     8401 caaccatcag ttcataggtc cattctctta gcgcaactac agagaacagg ggcacaaaca
     8461 ggcaaaaaac gggcacaacc tcaatggagt gatgcaacct gcctggagta aatgatgaca
     8521 caaggcaatt gacccacgca tgtatctatc tcattttctt acaccttcta ttaccttctg
     8581 ctctctctga tttggaaaaa gctgaaaaaa aaggttgaaa ccagttccct gaaattattc
     8641 ccctacttga ctaataagta tataaagacg gtaggtattg attgtaattc tgtaaatcta
     8701 tttcttaaac ttcttaaatt ctacttttat agttagtctt ttttttagtt ttaaaacacc
     8761 aagaacttag tttcgaataa acacacataa acaaacaaaa gatctgatat ggatcgaatt
     8821 agatctcgcc accatggaca agaagtattc tatcggactg gacatcggga ctaatagcgt
     8881 cgggtgggcc gtcatcactg acgagtacaa ggtgccctct aagaagttca aggtgctcgg
     8941 gaacaccgac cggcattcca tcaagaaaaa tctgatcgga gctctcctct ttgattcagg
     9001 ggagaccgct gaagcaaccc gcctcaagcg gactgctaga cggcggtaca ccaggaggaa
     9061 gaaccggatt tgttaccttc aagagatatt ctccaacgaa atggcaaagg tcgacgacag
     9121 cttcttccat aggctggaag aatcattcct cgtggaagag gataagaagc atgaacggca
     9181 tcccatcttc ggtaatatcg tcgacgaggt ggcctatcac gagaaatacc caaccatcta
     9241 ccatcttcgc aaaaagctgg tggactcaac cgacaaggca gacctccggc ttatctacct
     9301 ggccctggcc cacatgatta agttcagagg ccacttcctg atcgagggcg acctcaatcc
     9361 tgacaatagc gatgtggata aactgttcat ccagctggtg cagacttaca accagctctt
     9421 tgaagagaac cccatcaatg caagcggagt cgatgccaag gccattctgt cagcccggct
     9481 gtcaaagagc cgcagacttg agaatcttat cgctcagctg ccgggtgaaa agaaaaatgg
     9541 actgttcggg aacctgattg ctctttcact tgggctgact cccaatttca agtctaattt
     9601 cgacctggca gaggatgcca agctgcaact gtccaaggac acctatgatg acgatctcga
     9661 caacctcctg gcccagatcg gtgaccaata cgccgacctt ttccttgctg ctaagaatct
     9721 ttctgacgcc atcctgctgt ctgacattct ccgcgtgaac actgaaatca ccaaggcccc
     9781 tctttcagct tcaatgatta agcggtatga tgagcaccac caggacctga ccctgcttaa
     9841 ggcactcgtc cggcagcagc ttccggagaa gtacaaggaa atcttctttg accagtcaaa
     9901 gaatggatac gccggctaca tcgacggagg tgcctcccaa gaggaatttt ataagtttat
     9961 caaacctatc cttgagaaga tggacggcac cgaagagctc ctcgtgaaac tgaatcggga
    10021 ggatctgctg cggaagcagc gcactttcga caatgggagc attccccacc agatccatct
    10081 tggggagctt cacgccatcc ttcggcgcca agaggacttc tacccctttc ttaaggacaa
    10141 cagggagaag attgagaaaa ttctcacttt ccgcatcccc tactacgtgg gacccctcgc
    10201 cagaggaaat agccggtttg cttggatgac cagaaagtca gaagaaacta tcactccctg
    10261 gaacttcgaa gaggtggtgg acaagggagc cagcgctcag tcattcatcg aacggatgac
    10321 taacttcgat aagaacctcc ccaatgagaa ggtcctgccg aaacattccc tgctctacga
    10381 gtactttacc gtgtacaacg agctgaccaa ggtgaaatat gtcaccgaag ggatgaggaa
    10441 gcccgcattc ctgtcaggcg aacaaaagaa ggcaattgtg gaccttctgt tcaagaccaa
    10501 tagaaaggtg accgtgaagc agctgaagga ggactatttc aagaaaattg aatgcttcga
    10561 ctctgtggag attagcgggg tcgaagatcg gttcaacgca agcctgggta cctaccatga
    10621 tctgcttaag atcatcaagg acaaggattt tctggacaat gaggagaaag aggacatcct
    10681 tgaggacatt gtcctgactc tcactctgtt cgaggaccgg gaaatgatcg aggagaggct
    10741 taagacctac gcccatctgt tcgacgataa agtgatgaag caacttaaac ggagaagata
    10801 taccggatgg ggacgcctta gccgcaaact catcaacgga atccgggaca aacagagcgg
    10861 aaagaccatt cttgatttcc ttaagagcga cggattcgct aatcgcaact tcatgcaact
    10921 tatccatgat gattccctga cctttaagga ggacatccag aaggcccaag tgtctggaca
    10981 aggtgactca ctgcacgagc atatcgcaaa tctggctggt tcacccgcta ttaagaaggg
    11041 tattctccag accgtgaaag tcgtggacga gctggtcaag gtgatgggtc gccataaacc
    11101 agagaacatt gtcatcgaga tggccaggga aaaccagact acccagaagg gacagaagaa
    11161 cagcagggag cggatgaaaa gaattgagga agggatt
//
