LOCUS       sequence-462144-       11258 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid expressing Cas9 and gRNA AAACCTTTTTACTCCACGCA which targets 
            the HIS3 gene..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11258)
  TITLE     van Leeuwen Lab plasmids 2025
REFERENCE   2  (bases 1 to 11258)
  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-HIS3-3
FEATURES             Location/Qualifiers
     source          1..11258
                     /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"
     promoter        6317..6532
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             6533..7336
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      complement(7467..7922)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7604..7625)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7816..7835
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8048..8070
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8062..8079
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8063..8079
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8089..8108
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        8089..8107
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     8132..8148
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     8133..8149
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        8194..8860
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     8730..8753
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     regulatory      8889..8898
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             8895..10613
                     /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 ctgcgtggag taaaaaggtt
     2221 tgatcattta 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 tacgcatatg tagtgttgaa gaaacatgaa attgcccagt
     6481 attcttaacc caactgcaca gaacaaaaac ctgcaggaaa cgaagataaa tcatgtcgaa
     6541 agctacatat aaggaacgtg ctgctactca tcctagtcct gttgctgcca agctatttaa
     6601 tatcatgcac gaaaagcaaa caaacttgtg tgcttcattg gatgttcgta ccaccaagga
     6661 attactggag ttagttgaag cattaggtcc caaaatttgt ttactaaaaa cacatgtgga
     6721 tatcttgact gatttttcca tggagggcac agttaagccg ctaaaggcat tatccgccaa
     6781 gtacaatttt ttactcttcg aagacagaaa atttgctgac attggtaata cagtcaaatt
     6841 gcagtactct gcgggtgtat acagaatagc agaatgggca gacattacga atgcacacgg
     6901 tgtggtgggc ccaggtattg ttagcggttt gaagcaggcg gcagaagaag taacaaagga
     6961 acctagaggc cttttgatgt tagcagaatt gtcatgcaag ggctccctat ctactggaga
     7021 atatactaag ggtactgttg acattgcgaa gagcgacaaa gattttgtta tcggctttat
     7081 tgctcaaaga gacatgggtg gaagagatga aggttacgat tggttgatta tgacacccgg
     7141 tgtgggttta gatgacaagg gagacgcatt gggtcaacag tatagaaccg tggatgatgt
     7201 ggtctctaca ggatctgaca ttattattgt tggaagagga ctatttgcaa agggaaggga
     7261 tgctaaggta gagggtgaac gttacagaaa agcaggctgg gaagcatatt tgagaagatg
     7321 cggccagcaa aactaaaaaa ctgtattata agtaaatgca tgtatactaa actcacaaat
     7381 tagagcttca atttaattat atcagttatt accctatgcg gtgtgaaata ccgcacagat
     7441 gcgtaaggag aaaataccgc atcaggaaat tgtaagcgtt aatattttgt taaaattcgc
     7501 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     7561 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     7621 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     7681 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     7741 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     7801 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     7861 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     7921 gtccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
     7981 ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca
     8041 gggttttccc agtcacgacg ttgtaaaacg acggccagtg agcgcgcgta atacgactca
     8101 ctatagggcg aattgggtac cgggcccccc ctcgaggtcg acggtatcga taagcttgat
     8161 atcgaattca gtctactagt cagttcgagt ttatcattat caatactgcc atttcaaaga
     8221 atacgtaaat aattaatagt agtgattttc ctaactttat ttagtcaaaa aattagcctt
     8281 ttaattctgc tgtaacccgt acatgcccaa aatagggggc gggttacaca gaatatataa
     8341 catcgtaggt gtctgggtga acagtttatt cctggcatcc actaaatata atggagcccg
     8401 ctttttaagc tggcatccag aaaaaaaaag aatcccagca ccaaaatatt gttttcttca
     8461 ccaaccatca gttcataggt ccattctctt agcgcaacta cagagaacag gggcacaaac
     8521 aggcaaaaaa cgggcacaac ctcaatggag tgatgcaacc tgcctggagt aaatgatgac
     8581 acaaggcaat tgacccacgc atgtatctat ctcattttct tacaccttct attaccttct
     8641 gctctctctg atttggaaaa agctgaaaaa aaaggttgaa accagttccc tgaaattatt
     8701 cccctacttg actaataagt atataaagac ggtaggtatt gattgtaatt ctgtaaatct
     8761 atttcttaaa cttcttaaat tctactttta tagttagtct tttttttagt tttaaaacac
     8821 caagaactta gtttcgaata aacacacata aacaaacaaa agatctgata tggatcgaat
     8881 tagatctcgc caccatggac aagaagtatt ctatcggact ggacatcggg actaatagcg
     8941 tcgggtgggc cgtcatcact gacgagtaca aggtgccctc taagaagttc aaggtgctcg
     9001 ggaacaccga ccggcattcc atcaagaaaa atctgatcgg agctctcctc tttgattcag
     9061 gggagaccgc tgaagcaacc cgcctcaagc ggactgctag acggcggtac accaggagga
     9121 agaaccggat ttgttacctt caagagatat tctccaacga aatggcaaag gtcgacgaca
     9181 gcttcttcca taggctggaa gaatcattcc tcgtggaaga ggataagaag catgaacggc
     9241 atcccatctt cggtaatatc gtcgacgagg tggcctatca cgagaaatac ccaaccatct
     9301 accatcttcg caaaaagctg gtggactcaa ccgacaaggc agacctccgg cttatctacc
     9361 tggccctggc ccacatgatt aagttcagag gccacttcct gatcgagggc gacctcaatc
     9421 ctgacaatag cgatgtggat aaactgttca tccagctggt gcagacttac aaccagctct
     9481 ttgaagagaa ccccatcaat gcaagcggag tcgatgccaa ggccattctg tcagcccggc
     9541 tgtcaaagag ccgcagactt gagaatctta tcgctcagct gccgggtgaa aagaaaaatg
     9601 gactgttcgg gaacctgatt gctctttcac ttgggctgac tcccaatttc aagtctaatt
     9661 tcgacctggc agaggatgcc aagctgcaac tgtccaagga cacctatgat gacgatctcg
     9721 acaacctcct ggcccagatc ggtgaccaat acgccgacct tttccttgct gctaagaatc
     9781 tttctgacgc catcctgctg tctgacattc tccgcgtgaa cactgaaatc accaaggccc
     9841 ctctttcagc ttcaatgatt aagcggtatg atgagcacca ccaggacctg accctgctta
     9901 aggcactcgt ccggcagcag cttccggaga agtacaagga aatcttcttt gaccagtcaa
     9961 agaatggata cgccggctac atcgacggag gtgcctccca agaggaattt tataagttta
    10021 tcaaacctat ccttgagaag atggacggca ccgaagagct cctcgtgaaa ctgaatcggg
    10081 aggatctgct gcggaagcag cgcactttcg acaatgggag cattccccac cagatccatc
    10141 ttggggagct tcacgccatc cttcggcgcc aagaggactt ctaccccttt cttaaggaca
    10201 acagggagaa gattgagaaa attctcactt tccgcatccc ctactacgtg ggacccctcg
    10261 ccagaggaaa tagccggttt gcttggatga ccagaaagtc agaagaaact atcactccct
    10321 ggaacttcga agaggtggtg gacaagggag ccagcgctca gtcattcatc gaacggatga
    10381 ctaacttcga taagaacctc cccaatgaga aggtcctgcc gaaacattcc ctgctctacg
    10441 agtactttac cgtgtacaac gagctgacca aggtgaaata tgtcaccgaa gggatgagga
    10501 agcccgcatt cctgtcaggc gaacaaaaga aggcaattgt ggaccttctg ttcaagacca
    10561 atagaaaggt gaccgtgaag cagctgaagg aggactattt caagaaaatt gaatgcttcg
    10621 actctgtgga gattagcggg gtcgaagatc ggttcaacgc aagcctgggt acctaccatg
    10681 atctgcttaa gatcatcaag gacaaggatt ttctggacaa tgaggagaaa gaggacatcc
    10741 ttgaggacat tgtcctgact ctcactctgt tcgaggaccg ggaaatgatc gaggagaggc
    10801 ttaagaccta cgcccatctg ttcgacgata aagtgatgaa gcaacttaaa cggagaagat
    10861 ataccggatg gggacgcctt agccgcaaac tcatcaacgg aatccgggac aaacagagcg
    10921 gaaagaccat tcttgatttc cttaagagcg acggattcgc taatcgcaac ttcatgcaac
    10981 ttatccatga tgattccctg acctttaagg aggacatcca gaaggcccaa gtgtctggac
    11041 aaggtgactc actgcacgag catatcgcaa atctggctgg ttcacccgct attaagaagg
    11101 gtattctcca gaccgtgaaa gtcgtggacg agctggtcaa ggtgatgggt cgccataaac
    11161 cagagaacat tgtcatcgag atggccaggg aaaaccagac tacccagaag ggacagaaga
    11221 acagcaggga gcggatgaaa agaattgagg aagggatt
//
