LOCUS       sequence-208224-       10807 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid encoding iCas9, tracrRNA and crRNAs.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10807)
  AUTHORS   Bao Z, Xiao H, Liang J, Zhang L, Xiong X, Sun N, Si T, Zhao H
  TITLE     Homology-Integrated CRISPR-Cas (HI-CRISPR) System for One-Step 
            Multigene Disruption in Saccharomyces cerevisiae.
  JOURNAL   ACS Synth Biol. 2014 Sep 19.
  PUBMED    25207793
REFERENCE   2  (bases 1 to 10807)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "ACS Synth 
            Biol. 2014 Sep 19."
COMMENT     Sequence Label: pCRCT
FEATURES             Location/Qualifiers
     source          1..10807
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             54..857
                     /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"
     promoter        946..1214
                     /label=SNR52 promoter
                     /note="promoter for the S. cerevisiae small nucleolar RNA 
                     gene SNR52"
     CDS             complement(1257..1553)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSG"
     primer_bind     1403..1425
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1417..1434
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1418..1434
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1444..1463
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1444..1462
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1510..1530)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1510..1528)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1549..1565)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1573..1589
                     /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(1597..1603,1604..1621,1622..1627))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1642..1663
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     terminator      1736..1755
                     /label=SUP4 terminator
                     /note="transcription terminator for the S. cerevisiae SUP4 
                     tRNA gene"
     promoter        1757..2156
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             2168..2191
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2195..2215
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2216..6316
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTYKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSITHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             6320..6340
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      6344..6643
                     /label=ADH2 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 2 (ADH2) gene"
     promoter        6656..7062
                     /gene="RPR1"
                     /label=RPR1 promoter
                     /note="promoter for the S. cerevisiae RNase P RNA gene"
     misc_RNA        7063..7141
                     /label=tracrRNA
                     /note="trans-activating CRISPR RNA for the Streptococcus 
                     pyogenes CRISPR/Cas9 system"
     terminator      7149..7211
                     /label=RPR1 terminator
                     /note="transcription terminator for the S. cerevisiae RNase
                     P RNA gene"
     primer_bind     complement(7244..7260)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7268..7284
                     /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(7292..7298,7299..7316,7317..7322))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7337..7358
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7475..7492)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7646..8234)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7726..7745)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8405..9196,9197..9265))
                     /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     9028..9047
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9266..9370)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      9397..10739
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(10360..10407)
                     /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     10780..10798
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgaac ccaactgcac agaacaaaaa cctgcaggaa acgaagataa atcatgtcga
       61 aagctacata taaggaacgt gctgctactc atcctagtcc tgttgctgcc aagctattta
      121 atatcatgca cgaaaagcaa acaaacttgt gtgcttcatt ggatgttcgt accaccaagg
      181 aattactgga gttagttgaa gcattaggtc ccaaaatttg tttactaaaa acacatgtgg
      241 atatcttgac tgatttttcc atggagggca cagttaagcc gctaaaggca ttatccgcca
      301 agtacaattt tttactcttc gaagacagaa aatttgctga cattggtaat acagtcaaat
      361 tgcagtactc tgcgggtgta tacagaatag cagaatgggc agacattacg aatgcacacg
      421 gtgtggtggg cccaggtatt gttagcggtt tgaagcaggc ggcagaagaa gtaacaaagg
      481 aacctagagg ccttttgatg ttagcagaat tgtcatgcaa gggctcccta tctactggag
      541 aatatactaa gggtactgtt gacattgcga agagcgacaa agattttgtt atcggcttta
      601 ttgctcaaag agacatgggt ggaagagatg aaggttacga ttggttgatt atgacacccg
      661 gtgtgggttt agatgacaag ggagacgcat tgggtcaaca gtatagaacc gtggatgatg
      721 tcgtttctac aggatctgac attattattg ttggaagagg actatttgca aagggaaggg
      781 atgctaaggt agagggtgaa cgttacagaa aagcaggctg ggaagcatat ttgagaagat
      841 gcggccagca aaactaaaaa actgtattat aagtaaatgc atgtatacta aactcacaaa
      901 ttagagcttc aatttaatta tatcagttat taccctgtgc tcgagtcttt gaaaagataa
      961 tgtatgatta tgctttcact catatttata cagaaacttg atgttttctt tcgagtatat
     1021 acaaggtgat tacatgtacg tttgaagtac aactctagat tttgtagtgc cctcttgggc
     1081 tagcggtaaa ggtgcgcatt ttttcacacc ctacaatgtt ctgttcaaaa gattttggtc
     1141 aaacgctgta gaagtgaaag ttggtgcgca tgtttcggcg ttcgaaactt ctccgcagtg
     1201 aaagataaat gatcgggttt tagagctatg ctgttttgaa tggtcccaaa gagacctcag
     1261 ccgctacagg gcgcgtccca ttcgccattc aggctgcgca actgttggga agggcgatcg
     1321 gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta
     1381 agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgagcgc
     1441 gcgtaatacg actcactata gggcgaattg ggtaccgggc cccccctcga ggtcctccag
     1501 cttttgttcc ctttagtgag ggttaattgc gcgcttggcg taatcatggt catagctgtt
     1561 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     1621 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     1681 gcccgctttc caccggtggt ctctagagct atgctgtttt gaatggtccc aaaacttttt
     1741 ttgtttttta tgtctgcata gcttcaaaat gtttctactc cttttttact cttccagatt
     1801 ttctcggact ccgcgcatcg ccgtaccact tcaaaacacc caagcacagc atactaaatt
     1861 tcccctcttt cttcctctag ggtgtcgtta attacccgta ctaaaggttt ggaaaagaaa
     1921 aaagaggccg cctcgtttct ttttcttcgt cgaaaaaggc aataaaaatt tttatcacgt
     1981 ttctttttct tgaaaatttt tttttttgat ttttttctct ttcgatgacc tcccattgat
     2041 atttaagtta ataaacggtc ttcaatttct caagtttcag tttcattttt cttgttctat
     2101 tacaactttt tttacttctt gctcattaga aagaaagcat agcaatctaa tctaagtttt
     2161 ctagatggat tataaagatg acgatgacaa acctccaaaa aagaagagaa aggtcgataa
     2221 gaaatactca ataggcttag atatcggcac aaatagcgtc ggatgggcgg tgatcactga
     2281 tgaatataag gttccgtcta aaaagttcaa ggttctggga aatacagacc gccacagtat
     2341 caaaaaaaat cttatagggg ctcttttatt tgacagtgga gagacagcgg aagcgactcg
     2401 tctcaaacgg acagctcgta gaaggtatac acgtcggaag aatcgtattt gttatctaca
     2461 ggagattttt tcaaatgaga tggcgaaagt agatgatagt ttctttcatc gacttgaaga
     2521 gtcttttttg gtggaagaag acaagaagca tgaacgtcat cctatttttg gaaatatagt
     2581 agatgaagtt gcttatcatg agaaatatcc aactatctat catctgcgaa aaaaattggt
     2641 agattctact tataaagcgg atttgcgctt aatctatttg gccttagcgc atatgattaa
     2701 gtttcgtggt cattttttga ttgagggaga tttaaatcct gataatagtg atgtggacaa
     2761 actatttatc cagttggtac aaacctacaa tcaattattt gaagaaaacc ctattaacgc
     2821 aagtggagta gatgctaaag cgattctttc tgcacgattg agtaaatcaa gacgattaga
     2881 aaatctcatt gctcagctcc ccggtgagaa gaaaaatggc ttatttggga atctcattgc
     2941 tttgtcattg ggtttgaccc ctaattttaa atcaaatttt gatttggcag aagatgctaa
     3001 attacagctt tcaaaagata cttacgatga tgatttagat aatttattgg cgcaaattgg
     3061 agatcaatat gctgatttgt ttttggcagc taagaattta tcagatgcta ttttactttc
     3121 agatatccta agagtaaata ctgaaataac taaggctccc ctatcagctt caatgattaa
     3181 acgctacgat gaacatcatc aagacttgac tcttttaaaa gctttagttc gacaacaact
     3241 tccagaaaag tataaagaaa tcttttttga tcaatcaaaa aacggatatg caggttatat
     3301 tgatggggga gctagccaag aagaatttta taaatttatc aaaccaattt tagaaaaaat
     3361 ggatggtact gaggaattat tggtgaaact aaatcgtgaa gatttgctgc gcaagcaacg
     3421 gacctttgac aacggctcta ttacccatca aattcacttg ggtgagctgc atgctatttt
     3481 gagaagacaa gaagactttt atccattttt aaaagacaat cgtgagaaga ttgaaaaaat
     3541 cttgactttt cgaattcctt attatgttgg tccattggcg cgtggcaata gtcgttttgc
     3601 atggatgact cggaagtctg aagaaacaat taccccatgg aattttgaag aagttgtcga
     3661 taaaggtgct tcagctcaat catttattga acgcatgaca aactttgata aaaatcttcc
     3721 aaatgaaaaa gtactaccaa aacatagttt gctttatgag tattttacgg tttataacga
     3781 attgacaaag gtcaaatatg ttactgaagg aatgcgaaaa ccagcatttc tttcaggtga
     3841 acagaagaaa gccattgttg atttactctt caaaacaaat cgaaaagtaa ccgttaagca
     3901 attaaaagaa gattatttca aaaaaataga atgttttgat agtgttgaaa tttcaggagt
     3961 tgaagataga tttaatgctt cattaggtac ctaccatgat ttgctaaaaa ttattaaaga
     4021 taaagatttt ttggataatg aagaaaatga agatatctta gaggatattg ttttaacatt
     4081 gaccttattt gaagataggg agatgattga ggaaagactt aaaacatatg ctcacctctt
     4141 tgatgataag gtgatgaaac agcttaaacg tcgccgttat actggttggg gacgtttgtc
     4201 tcgaaaattg attaatggta ttagggataa gcaatctggc aaaacaatat tagatttttt
     4261 gaaatcagat ggttttgcca atcgcaattt tatgcagctg atccatgatg atagtttgac
     4321 atttaaagaa gacattcaaa aagcacaagt gtctggacaa ggcgatagtt tacatgaaca
     4381 tattgcaaat ttagctggta gccctgctat taaaaaaggt attttacaga ctgtaaaagt
     4441 tgttgatgaa ttggtcaaag taatggggcg gcataagcca gaaaatatcg ttattgaaat
     4501 ggcacgtgaa aatcagacaa ctcaaaaggg ccagaaaaat tcgcgagagc gtatgaaacg
     4561 aatcgaagaa ggtatcaaag aattaggaag tcagattctt aaagagcatc ctgttgaaaa
     4621 tactcaattg caaaatgaaa agctctatct ctattatctc caaaatggaa gagacatgta
     4681 tgtggaccaa gaattagata ttaatcgttt aagtgattat gatgtcgatc acattgttcc
     4741 acaaagtttc cttaaagacg attcaataga caataaggtc ttaacgcgtt ctgataaaaa
     4801 tcgtggtaaa tcggataacg ttccaagtga agaagtagtc aaaaagatga aaaactattg
     4861 gagacaactt ctaaacgcca agttaatcac tcaacgtaag tttgataatt taacgaaagc
     4921 tgaacgtgga ggtttgagtg aacttgataa agctggtttt atcaaacgcc aattggttga
     4981 aactcgccaa atcactaagc atgtggcaca aattttggat agtcgcatga atactaaata
     5041 cgatgaaaat gataaactta ttcgagaggt taaagtgatt accttaaaat ctaaattagt
     5101 ttctgacttc cgaaaagatt tccaattcta taaagtacgt gagattaaca attaccatca
     5161 tgcccatgat gcgtatctaa atgccgtcgt tggaactgct ttgattaaga aatatccaaa
     5221 acttgaatcg gagtttgtct atggtgatta taaagtttat gatgttcgta aaatgattgc
     5281 taagtctgag caagaaatag gcaaagcaac cgcaaaatat ttcttttact ctaatatcat
     5341 gaacttcttc aaaacagaaa ttacacttgc aaatggagag attcgcaaac gccctctaat
     5401 cgaaactaat ggggaaactg gagaaattgt ctgggataaa gggcgagatt ttgccacagt
     5461 gcgcaaagta ttgtccatgc cccaagtcaa tattgtcaag aaaacagaag tacagacagg
     5521 cggattctcc aaggagtcaa ttttaccaaa aagaaattcg gacaagctta ttgctcgtaa
     5581 aaaagactgg gatccaaaaa aatatggtgg ttttgatagt ccaacggtag cttattcagt
     5641 cctagtggtt gctaaggtgg aaaaagggaa atcgaagaag ttaaaatccg ttaaagagtt
     5701 actagggatc acaattatgg aaagaagttc ctttgaaaaa aatccgattg actttttaga
     5761 agctaaagga tataaggaag ttaaaaaaga cttaatcatt aaactaccta aatatagtct
     5821 ttttgagtta gaaaacggtc gtaaacggat gctggctagt gccggagaat tacaaaaagg
     5881 aaatgagctg gctctgccaa gcaaatatgt gaatttttta tatttagcta gtcattatga
     5941 aaagttgaag ggtagtccag aagataacga acaaaaacaa ttgtttgtgg agcagcataa
     6001 gcattattta gatgagatta ttgagcaaat cagtgaattt tctaagcgtg ttattttagc
     6061 agatgccaat ttagataaag ttcttagtgc atataacaaa catagagaca aaccaatacg
     6121 tgaacaagca gaaaatatta ttcatttatt tacgttgacg aatcttggag ctcccgctgc
     6181 ttttaaatat tttgatacaa caattgatcg taaacgatat acgtctacaa aagaagtttt
     6241 agatgccact cttatccatc aatccatcac tggtctttat gaaacacgca ttgatttgag
     6301 tcagctagga ggtgaccctc caaaaaagaa gagaaaggtc tgagcggatc tcttatgtct
     6361 ttacgattta tagttttcat tatcaagtat gcctatatta gtatatagca tctttagatg
     6421 acagtgttcg aagtttcacg aataaaagat aatattctac tttttgctcc caccgcgttt
     6481 gctagcacga gtgaacacca tccctcgcct gtgagttgta cccattcctc taaactgtag
     6541 acatggtagc ttcagcagtg ttcgttatgt acggcatcct ccaacaaaca gtcggttata
     6601 gtttgtcctg ctcctctgaa tcgagtccct cgatatttct catactagtt ctagagatct
     6661 gccaattgaa cataacatgg tagttacata tactagtaat atggttcggc acacattaaa
     6721 agtataaaaa ctatctgaat tacgaattac atatattggt cataaaaatc aatcaatcat
     6781 cgtgtgtttt atatgtctct tatctaagta taagaatatc catagttaat attcacttac
     6841 gctacctttt aacctgtaat cattgtcaac aggatatgtt aacgacccac attgataaac
     6901 gctagtattt ctttttcctc ttcttattgg ccggctgtct ctatactccc ctatagtctg
     6961 tttcttttcg tttcgattgt tttacgtttg aggcctcgtg gcgcacatgg tacgctgtgg
     7021 tgctcgcggc tgggaacgaa actctgggag ctgcgattgg caggaaccat tcaaaacagc
     7081 atagcaagtt aaaataaggc tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg
     7141 ctttttttcc atatccaact tccaatttaa tctttctttt ttaattttca cttatttgcg
     7201 atacagaaag accctgcagg catgcaagct tggcgtaatc atggtcatag ctgtttcctg
     7261 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     7321 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     7381 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     7441 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     7501 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     7561 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     7621 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     7681 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     7741 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     7801 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     7861 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     7921 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     7981 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     8041 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     8101 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     8161 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     8221 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     8281 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     8341 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     8401 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     8461 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     8521 gccccagtgc tgcaatgata ccgcggctcc cacgctcacc ggctccagat ttatcagcaa
     8581 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     8641 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     8701 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     8761 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     8821 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     8881 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     8941 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     9001 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     9061 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     9121 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     9181 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     9241 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     9301 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     9361 gggttccgcg cacatttccc cgaaaagtgc cacctgaacg aagcatctgt gcttcatttt
     9421 gtagaacaaa aatgcaacgc gagagcgcta atttttcaaa caaagaatct gagctgcatt
     9481 tttacagaac agaaatgcaa cgcgaaagcg ctattttacc aacgaagaat ctgtgcttca
     9541 tttttgtaaa acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag aatctgagct
     9601 gcatttttac agaacagaaa tgcaacgcga gagcgctatt ttaccaacaa agaatctata
     9661 cttctttttt gttctacaaa aatgcatccc gagagcgcta tttttctaac aaagcatctt
     9721 agattacttt ttttctcctt tgtgcgctct ataatgcagt ctcttgataa ctttttgcac
     9781 tgtaggtccg ttaaggttag aagaaggcta ctttggtgtc tattttctct tccataaaaa
     9841 aagcctgact ccacttcccg cgtttactga ttactagcga agctgcgggt gcattttttc
     9901 aagataaagg catccccgat tatattctat accgatgtgg attgcgcata ctttgtgaac
     9961 agaaagtgat agcgttgatg attcttcatt ggtcagaaaa ttatgaacgg tttcttctat
    10021 tttgtctcta tatactacgt ataggaaatg tttacatttt cgtattgttt tcgattcact
    10081 ctatgaatag ttcttactac aatttttttg tctaaagagt aatactagag ataaacataa
    10141 aaaatgtaga ggtcgagttt agatgcaagt tcaaggagcg aaaggtggat gggtaggtta
    10201 tatagggata tagcacagag atatatagca aagagatact tttgagcaat gtttgtggaa
    10261 gcggtattcg caatatttta gtagctcgtt acagtccggt gcgtttttgg ttttttgaaa
    10321 gtgcgtcttc agagcgcttt tggttttcaa aagcgctctg aagttcctat actttctaga
    10381 gaataggaac ttcggaatag gaacttcaaa gcgtttccga aaacgagcgc ttccgaaaat
    10441 gcaacgcgag ctgcgcacat acagctcact gttcacgtcg cacctatatc tgcgtgttgc
    10501 ctgtatatat atatacatga gaagaacggc atagtgcgtg tttatgctta aatgcgtact
    10561 tatatgcgtc tatttatgta ggatgaaagg tagtctagta cctcctgtga tattatccca
    10621 ttccatgcgg ggtatcgtat gcttccttca gcactaccct ttagctgttc tatatgctgc
    10681 cactcctcaa ttggattagt ctcatccttc aatgctatca tttcctttga tattggatca
    10741 tactaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
    10801 tttcgtc
//
