EEB;ID EU855177; SV 1; circular; other DNA; STD; SYN; 9953 BP. AC EU855177; DT 03-AUG-2008 (Rel. 96, Created) DT 03-AUG-2008 (Rel. 96, Last updated, Version 1) DE Cloning vector pAW8_1XFLAG, complete sequence. KW . OS Cloning vector pAW8_1XFLAG OC other sequences; artificial sequences; vectors. RN [1] RP 1-9953 RX DOI; 10.1016/j.gene.2007.09.024. RX PUBMED; 18054176. RA Watson A.T., Garcia V., Bone N., Carr A.M., Armstrong J.; RT "Gene tagging and gene replacement using recombinase-mediated cassette RT exchange in Schizosaccharomyces pombe"; RL Gene 407(1-2):63-74(2008). RN [2] RP 1-9953 RA Watson A.T.; RT ; RL Submitted (26-JUN-2008) to the EMBL/GenBank/DDBJ databases. RL Genome Damage and Stability Centre, University of Sussex, Falmer, Brighton, RL East Sussex BN1 9RQ, UK FH Key Location/Qualifiers FT source 1..9953 FT /organism="Cloning vector pAW8_1XFLAG" FT /mol_type="other DNA" FT /db_xref="taxon:550439" FT misc_feature 1..3873 FT /note="Cre-expression plasmid pAW8 backbone" FT misc_recomb 3874..3907 FT /note="loxP" FT misc_feature 3916..3954 FT /note="1 X FLAG epitope tag" FT terminator 3963..4191 FT /note="Saccharomyces cerevisiae ADH1" FT gene 4581..5390 FT /gene="kanR" FT CDS 4581..5390 FT /codon_start=1 FT /transl_table=11 FT /gene="kanR" FT /product="kanamycin and geneticin G418 resistance protein" FT /protein_id="ACF75940.1" FT /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP FT DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA FT FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD FT FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD FT RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" FT misc_recomb 5638..5671 FT /note="loxM3" FT misc_feature 5672..9953 FT /note="Cre-expression plasmid pAW8 backbone" SQ Sequence 9953 BP; 2800 A; 2134 C; 2080 G; 2939 T; 0 other; caaaaagtta ggtgtaacag aaaatcatga aactatgatt tctaatttat atattggagg 60 attttctcta aaaaaaaaaa tacaacaaat aaaaaacact caatgacctg accatttgat 120 ggagtttaag tcaatacctt cttgaaccat ttcccataat ggtgaaagtt ccctcaagaa 180 ttttactctg tcagaaacgg ccttaacgac gtagtcgatc cggacgatgg ccaaccttcc 240 aattcattaa atcgagtcct ccatcagatg atttacaagg ttttgcatag aatccctaag 300 taagagaagt tgttagtgat ttcttcaagt ttactaaaat ttacatacaa atggatgatc 360 tcttctccat tgtttccttt cttcttgcaa acgcgtttta caaagagatg acatgctatt 420 ttctgattat ttttttctat agttttctat tttgtacgtc cttgggagtc cgaaatgtaa 480 aatcggggta tggttagtag cggcagcgtt cattatggta tataaaagat aaaaagtagg 540 ggcaagcgag acagaatggg atacaagggc atcgtctata attatagcta aaaattgtat 600 tttaatttgt attttttgta attttatttc tcattgtttt acttaaaatg aatgcgaatt 660 agagaaaact tcaacgaaat gtcaaaataa gctcagcaaa atatacaatt ttagggaaag 720 cgatcagcaa cttattctcc ggtgtagatt ctttttcttc ataaattcca acaatgtaac 780 tgccaattga tttttctcta gatgctaata aattgaaact aatgaatata ataacacagg 840 ttttgataaa ttgtaaactc ttactcaaaa aaagaatgtt aaaactaatt taaccttttt 900 ataaaggctc atggcgttaa agaggatagt aaaaattcta caatttttta atttaatgtc 960 acaattgtct tgcgaatata attctattaa ttttcgctaa ctgctgtttt agtactatta 1020 ctttcaatat gccttatatg caactttaat tcatgaagaa tgcaatattt ttgcaatctt 1080 tcttgtactg agcgcttttt atcatttagt actcattatt taatttttca gtaaagggca 1140 ataaataaat ttgtagaaga tgcaatgtaa tctcctctat ccttttgctc atatgtttat 1200 gagtatacct agtctagaaa ggcttgtatt taaaatatta ttcaataaaa attcacaatt 1260 tttacgacat gtgctaccat tcacttaact tcctgattat aaaattggtt cgtttatact 1320 aattacttaa gtacctttaa ctaaacaaaa tgcctatata tatattaatt tacaatgagt 1380 gtcagataag tcactatgtc cgagtggtta aggagttaga ctcgccatcg atggtatggt 1440 gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa 1500 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg 1560 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga 1620 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt 1680 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt 1740 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat 1800 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt 1860 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg 1920 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga 1980 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc 2040 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac 2100 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg 2160 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca 2220 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg 2280 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg 2340 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg 2400 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag 2460 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg 2520 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct 2580 cccgtatcgt agttatctac acgacgggga gtcaggccac tatggatgaa cgaaatagac 2640 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact 2700 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga 2760 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt 2820 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct 2880 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc 2940 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc 3000 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc 3060 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg 3120 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt 3180 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg 3240 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg 3300 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt 3360 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag 3420 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt 3480 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta 3540 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt 3600 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc 3660 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca 3720 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc 3780 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg 3840 accatgatta cgccaagctt gcatggcggc cgcataactt cgtatagcat acattatacg 3900 aagttatgca tgctcactag tgactacaag gacgacgatg acaagtctag atgaggcgcg 3960 ccacttctaa ataagcgaat ttcttatgat ttatgatttt tattattaaa taagttataa 4020 aaaaaataag tgtatacaaa ttttaaagtg actcttaggt tttaaaacga aaattcttat 4080 tcttgagtaa ctctttcctg taggtcaggt tgctttctca ggtatagtat gaggtcgctc 4140 ttattgacca cacctctacc ggcagatccg ctagggataa cagggtaata tagatctgtt 4200 tagcttgcct cgtccccgcc gggtcacccg gccagcgaca tggaggccca gaataccctc 4260 cttgacagtc ttgacgtgcg cagctcaggg gcatgatgtg actgtcgccc gtacatttag 4320 cccatacatc cccatgtata atcatttgca tccatacatt ttgatggccg cacggcgcga 4380 agcaaaaatt acggctcctc gctgcagacc tgcgagcagg gaaacgctcc cctcacagac 4440 gcgttgaatt gtccccacgc cgcgcccctg tagagaaata taaaaggtta ggatttgcca 4500 ctgaggttct tctttcatat acttcctttt aaaatcttgc taggatacag ttctcacatc 4560 acatccgaac ataaacaacc atgggtaagg aaaagactca cgtttcgagg ccgcgattaa 4620 attccaacat ggatgctgat ttatatgggt ataaatgggc tcgcgataat gtcgggcaat 4680 caggtgcgac aatctatcga ttgtatggga agcccgatgc gccagagttg tttctgaaac 4740 atggcaaagg tagcgttgcc aatgatgtta cagatgagat ggtcagacta aactggctga 4800 cggaatttat gcctcttccg accatcaagc attttatccg tactcctgat gatgcatggt 4860 tactcaccac tgcgatcccc ggcaaaacag cattccaggt attagaagaa tatcctgatt 4920 caggtgaaaa tattgttgat gcgctggcag tgttcctgcg ccggttgcat tcgattcctg 4980 tttgtaattg tccttttaac agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa 5040 tgaataacgg tttggttgat gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg 5100 aacaagtctg gaaagaaatg cataagcttt tgccattctc accggattca gtcgtcactc 5160 atggtgattt ctcacttgat aaccttattt ttgacgaggg gaaattaata ggttgtattg 5220 atgttggacg agtcggaatc gcagaccgat accaggatct tgccatccta tggaactgcc 5280 tcggtgagtt ttctccttca ttacagaaac ggctttttca aaaatatggt attgataatc 5340 ctgatatgaa taaattgcag tttcatttga tgctcgatga gtttttctaa tcagtactga 5400 caataaaaag attcttgttt tcaagaactt gtcatttgta tagttttttt atattgtagt 5460 tgttctattt taatcaaatg ttagcgtgat ttatattttt tttcgcctcg acatcatctg 5520 cccagatgcg aagttaagtg cgcagaaagt aatatcatgc gtcaatcgta tgtgaatgct 5580 ggtcgctata ctgctgtcga ttcgatacta acgccgccat ccagtcgacg aactagtata 5640 acttcgtata tggtattata tacgaagtta tgcggccgct gaattcactg gccgtcgttt 5700 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc 5760 cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt 5820 tgcgcagcct gaatggcgaa tggcgcctga tgcggtattt tctccttacg catctgtgcg 5880 gtatttcaca ccgcatacca tcgtccggat caatgaagta gatgagttta ctacctgtat 5940 atactttttt ttttgtcaaa gtcactttta tggcatttcc atgttgaagg attagccaca 6000 gcactaaatt ttcagcgtat gattgctttt aaatatttaa ttttcatcgt tttttaatta 6060 atattcaaaa cgatttaatg ccttctacat ctgaagtcaa gctcatagac tcgggttggg 6120 tagctaaaac tacatctaac acgaaggggt atagccttct taaagtatac tcatgtataa 6180 acgttttggg ttcatatttt tgggccacgt ggtttataaa aattcttaac tacaccactc 6240 ggtatcccgc acccgtctac gtttctacga tttcgagcta atattgagta aagtgattag 6300 caaaaaaaat aaaagtactc gttgtcggag atcaagaatt tttctattat ctcatctaaa 6360 ccactttcta aaagcgaaaa acaaaatcgt aatatgcagc ttgaatgggc ttccatagtt 6420 tgaaagaaaa accctagcag tactggcaag ggagacattc cttttacctg gctaatcgcc 6480 atcttccagc aggcgcacca ttgcccctgt ttcactatcc aggttacgga tatagttcat 6540 gacaatattt acattggtcc agccaccagc ttgcatgatc tccggtattg aaactccagc 6600 gcgggccata tctcgcgcgg ctccgacacg ggcactgtgt ccagaccagg ccaggtatct 6660 ctgaccagag tcatccttag cgccgtaaat caatcgatga gttgcttcaa aaatcccttc 6720 cagggcgcga gttgatagct ggctggtggc agatggcgcg gcaacaccat tttttctgac 6780 ccggcaaaac aggtagttat tcggatcatc agctacacca gagacggaaa tccatcgctc 6840 gaccagttta gttaccccca ggctaagtgc cttctctaca cctgcggtgc taaccagcgt 6900 tttcgttctg ccaatatgga ttaacattct cccaccgtca gtacgtgaga tatctttaac 6960 cctgatcctg gcaatttcgg ctatacgtaa cagggtgtta taagcaatcc ccagaaatgc 7020 cagattacgt atatcctggc agcgatcgct attttccatg agtgaacgaa cctggtcgaa 7080 atcagtgcgt tcgaacgcta gagcctgttt tgcacgttca ccggcatcaa cgttttcttt 7140 tcggatccgc cgcataacca gtgaaacagc attgctgtca cttggtcgtg gcagcccgga 7200 ccgacgatga agcatgttta gctggcccaa atgttgctgg atagttttta ctgccagacc 7260 gcgcgcctga agatatagaa gataatcgcg aacatcttca ggttctgcgg gaaaccattt 7320 ccggttattc aacttgcacc atgccgccca cgaccggcaa acggacagaa gcattttcca 7380 ggtatgctca gaaaacgcct ggcgatccct gaacatgtcc atcaggttct tgcgaacctc 7440 atcactcgtt gcatcgaccg gtaactttta gattaacatt tgtttaaata tgaaattagg 7500 caatgtaaca tactgcaggc aaattttggt gtacggtaag taaattggac atatgattta 7560 acaaagcgac tataagtcag aaagtgagaa tgagattgaa ataattaatt caacttattc 7620 aattgatgat atgccaggat tcctcttcct ttatatatct tcaatgtttc gattaacagg 7680 tccattatcc ggtttttagt ttcgctgcac attgccgaat gacaatcgga ttcctttaag 7740 ctttgctttt gccctgtttt ctctccccag taatcgttgg gtgactgaac catccagtag 7800 ccgagagtta cgtttttcct gatttaataa atatccatat ctatttgtct cgtggtgttt 7860 tacagcaact aaccgaaatt actattttgc atcatccaac cataaagaga caacaataat 7920 tctagaaaac atataacaag tgtttttttc tatttacgtc tcatacgcaa agtccacttt 7980 accatgtttt ggaaacttgt acccaaccat tcatttttcg attttgtaat gaggctcttt 8040 tcttctaaca atcattgagg tcattatcat gtgtcctcac ctattgttta aattgaggct 8100 tgcgctattc ccaaaattgt gggtattatt ttcatatatc agtatctgca atttcggttc 8160 aactccggaa gctcggtacc caactgtggg aatactcagg tatcgtaaga tgcaagagtt 8220 cgaatctctt agcaaccatt attttttttc tctcaacata acgagaacac acaggggcgc 8280 tatcgcacag aatcaaattc gatgactgga aattttttgt taatttcaga ggtcgcctga 8340 cgcatatacc tttttcaact gaaaaattgg gagaaaaagg aaaggtgaga gcgccggaac 8400 cggcttttca tatagaatag agaagcgttc atgactaaat gcttgcatca caatacttga 8460 agttgacaat attatttaag gacctattgt tttttccaat aggtggttag caatcgtctt 8520 actttctaac ttttcttacc ttttacattt cagcaatata tatatatata tatatatata 8580 tttcaaggat ataccattgt aatgtctgcc cctaagaaga tcgtcgtttt gccaggtgac 8640 cacgttggtc aagaaatcac agccgaagcc attaaggttc ttaaagctat ttctgatgtt 8700 cgttccaatg tcaagttcga tttcgaaaat catttaattg gtggtgctgc tatcgatgct 8760 acaggtgttc cacttccaga tgaggcgctg gaagcctcca agaaggctga tgccgttttg 8820 ttaggtgctg tgggtggtcc taaatggggt accggtagtg ttagacctga acaaggttta 8880 ctaaaaatcc gtaaagaact tcaattgtac gccaacttaa gaccatgtaa ctttgcatcc 8940 gactctcttt tagacttatc tccaatcaag ccacaatttg ctaaaggtac tgacttcgtt 9000 gttgttagag aattagtggg aggtatttac tttggtaaga gaaaggaaga cgatggtgat 9060 ggtgtcgctt gggatagtga acaatacacc gttccagaag tgcaaagaat cacaagaatg 9120 gccgctttca tggccctaca acatgagcca ccattgccta tttggtcctt ggataaagct 9180 aatgttttgg cctcttcaag attatggaga aaaactgtgg aggaaaccat caagaacgaa 9240 tttcctacat tgaaagttca acatcaattg attgattctg ccgccatgat cctagttaag 9300 aacccaaccc acctaaatgg tattataatc accagcaaca tgtttggtga tatcatctcc 9360 gatgaagcct ccgttatccc aggctccttg ggtttgttgc catctgcgtc cttggcctct 9420 ttgccagaca agaacaccgc atttggtttg tacgaaccat gccatggttc cgctccagat 9480 ttgccaaaga ataaggtcaa ccctatcgcc actatcttgt ctgctgcaat gatgttgaaa 9540 ttgtcattga acttgcctga agaaggtaaa gccattgaag atgcagttaa aaaggttttg 9600 gatgcaggta tcagaactgg tgatttaggt ggttccaaca gtaccaccga agtcggtgat 9660 gctgtcgccg aagaagttaa gaaaatcctt gcttaaaaag attctctttt tttgtgatat 9720 ttgtacataa actttataaa tgaaattcat aatagaaacg acacgaaatt acaaaatgga 9780 atatgttcat agggtagacg aaactatata cgcaatctac atacatttat caagaaggag 9840 aaaaaggagg atgtaaagga atacaggtaa gcaaattgat actaatggct caacgtgata 9900 aggaaaaaga attgcacttt aacattaata ttgacaagga ggagggcatc aca 9953 & pAW8_1XFLAG"Times New Roman EMluI(392),XbaI(798),ScaI(1014) Arial&_ WN#N E+ScaI(1111),NdeI(1192),XbaI(1214),ClaI(1429) Arial&8?{EsEC|s EScaI ArialP&{ E\PvuII(3677),HindIII(3856),NotI(3868),EagI(3868),SpeI(3917),XbaI(3947),AscI(3957),BglII(4193) Arial &29??~G E6PstI(4407),MluI(4440),NcoI(4580),NruI(4664),ClaI(4698) Arial& "-9K E\HindIII(5125),ScaI(5396),SalI(5625),SpeI(5633),NotI(5674),EagI(5674),EcoRI(5682),PvuII(5774) Arial&B>y E+PacI(6059),ScaI(6317),ScaI(6442),ClaI(6695) Arial& EDEcoRV(6952),SexAI(7071),BamHI(7144),NruI(7289),PstI(7518),NdeI(7552) Arial&Z\^g: E#HindIII(7739),XbaI(7922),KpnI(8180) Arial&l E!ClaI(8754),KpnI(8853),EcoRV(9352) Arial"&zskGk ENcoI Arial$&TPQFM<30g<EC&"8 pGEX_3_primer"Times New RomanECɆ& AmpR_promoter"Times New RomanEC&  Ampicillin"Times New Roman(E&y ORF 2"Times New RomanE&   pBR322_ori"Times New RomanE& lac_promoter"Times New RomanEC&M13_pUC_rev_primer"Times New RomanECɆ&M13_reverse_primer"Times New RomanEC&"CloxP"Times New RomanE&pHybLex_rev_primer"Times New RomanE&kanR"Times New RomanEC&7GM13_forward20_primer"Times New RomanECɆ&@VM13_pUC_fwd_primer"Times New RomanE&HORF 1"Times New RomanEC&!%ORF 1"Times New Roman Etest"Times New RomanU&,#?5