I^I^K;ID EU855179; SV 1; circular; other DNA; STD; SYN; 9932 BP. AC EU855179; DT 03-AUG-2008 (Rel. 96, Created) DT 03-AUG-2008 (Rel. 96, Last updated, Version 1) DE Cloning vector pAW8_6XHis, complete sequence. KW . OS Cloning vector pAW8_6XHis OC other sequences; artificial sequences; vectors. RN [1] RP 1-9932 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 1-9932 RA Watson A.T.; RT ; RL Submitted (27-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..9932 FT /organism="Cloning vector pAW8_6XHis" FT /mol_type="other DNA" FT /db_xref="taxon:550441" FT misc_feature 1..3873 FT /note="Cre-expression plasmid pAW8 backbone" FT misc_recomb 3874..3907 FT /note="loxP" FT misc_feature 3916..3933 FT /note="6 X histidine (His) affinity tag" FT terminator 3942..4170 FT /note="Saccharomyces cerevisiae ADH1" FT gene 4560..5369 FT /gene="kanR" FT CDS 4560..5369 FT /codon_start=1 FT /transl_table=11 FT /gene="kanR" FT /product="kanamycin and geneticin G418 resistance protein" FT /protein_id="ACF75942.1" FT /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP FT DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA FT FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD FT FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD FT RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" FT misc_recomb 5617..5650 FT /note="loxM3" FT misc_feature 5651..9932 FT /note="Cre-expression plasmid pAW8 backbone" SQ Sequence 9932 BP; 2793 A; 2132 C; 2070 G; 2937 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 tgcatcatca ccatcatcat taaggcgcgc cacttctaaa taagcgaatt 3960 tcttatgatt tatgattttt attattaaat aagttataaa aaaaataagt gtatacaaat 4020 tttaaagtga ctcttaggtt ttaaaacgaa aattcttatt cttgagtaac tctttcctgt 4080 aggtcaggtt gctttctcag gtatagtatg aggtcgctct tattgaccac acctctaccg 4140 gcagatccgc tagggataac agggtaatat agatctgttt agcttgcctc gtccccgccg 4200 ggtcacccgg ccagcgacat ggaggcccag aataccctcc ttgacagtct tgacgtgcgc 4260 agctcagggg catgatgtga ctgtcgcccg tacatttagc ccatacatcc ccatgtataa 4320 tcatttgcat ccatacattt tgatggccgc acggcgcgaa gcaaaaatta cggctcctcg 4380 ctgcagacct gcgagcaggg aaacgctccc ctcacagacg cgttgaattg tccccacgcc 4440 gcgcccctgt agagaaatat aaaaggttag gatttgccac tgaggttctt ctttcatata 4500 cttcctttta aaatcttgct aggatacagt tctcacatca catccgaaca taaacaacca 4560 tgggtaagga aaagactcac gtttcgaggc cgcgattaaa ttccaacatg gatgctgatt 4620 tatatgggta taaatgggct cgcgataatg tcgggcaatc aggtgcgaca atctatcgat 4680 tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca 4740 atgatgttac agatgagatg gtcagactaa actggctgac ggaatttatg cctcttccga 4800 ccatcaagca ttttatccgt actcctgatg atgcatggtt actcaccact gcgatccccg 4860 gcaaaacagc attccaggta ttagaagaat atcctgattc aggtgaaaat attgttgatg 4920 cgctggcagt gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt ccttttaaca 4980 gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat gaataacggt ttggttgatg 5040 cgagtgattt tgatgacgag cgtaatggct ggcctgttga acaagtctgg aaagaaatgc 5100 ataagctttt gccattctca ccggattcag tcgtcactca tggtgatttc tcacttgata 5160 accttatttt tgacgagggg aaattaatag gttgtattga tgttggacga gtcggaatcg 5220 cagaccgata ccaggatctt gccatcctat ggaactgcct cggtgagttt tctccttcat 5280 tacagaaacg gctttttcaa aaatatggta ttgataatcc tgatatgaat aaattgcagt 5340 ttcatttgat gctcgatgag tttttctaat cagtactgac aataaaaaga ttcttgtttt 5400 caagaacttg tcatttgtat agttttttta tattgtagtt gttctatttt aatcaaatgt 5460 tagcgtgatt tatatttttt ttcgcctcga catcatctgc ccagatgcga agttaagtgc 5520 gcagaaagta atatcatgcg tcaatcgtat gtgaatgctg gtcgctatac tgctgtcgat 5580 tcgatactaa cgccgccatc cagtcgacga actagtataa cttcgtatat ggtattatat 5640 acgaagttat gcggccgctg aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa 5700 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta 5760 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat 5820 ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcataccat 5880 cgtccggatc aatgaagtag atgagtttac tacctgtata tacttttttt tttgtcaaag 5940 tcacttttat ggcatttcca tgttgaagga ttagccacag cactaaattt tcagcgtatg 6000 attgctttta aatatttaat tttcatcgtt ttttaattaa tattcaaaac gatttaatgc 6060 cttctacatc tgaagtcaag ctcatagact cgggttgggt agctaaaact acatctaaca 6120 cgaaggggta tagccttctt aaagtatact catgtataaa cgttttgggt tcatattttt 6180 gggccacgtg gtttataaaa attcttaact acaccactcg gtatcccgca cccgtctacg 6240 tttctacgat ttcgagctaa tattgagtaa agtgattagc aaaaaaaata aaagtactcg 6300 ttgtcggaga tcaagaattt ttctattatc tcatctaaac cactttctaa aagcgaaaaa 6360 caaaatcgta atatgcagct tgaatgggct tccatagttt gaaagaaaaa ccctagcagt 6420 actggcaagg gagacattcc ttttacctgg ctaatcgcca tcttccagca ggcgcaccat 6480 tgcccctgtt tcactatcca ggttacggat atagttcatg acaatattta cattggtcca 6540 gccaccagct tgcatgatct ccggtattga aactccagcg cgggccatat ctcgcgcggc 6600 tccgacacgg gcactgtgtc cagaccaggc caggtatctc tgaccagagt catccttagc 6660 gccgtaaatc aatcgatgag ttgcttcaaa aatcccttcc agggcgcgag ttgatagctg 6720 gctggtggca gatggcgcgg caacaccatt ttttctgacc cggcaaaaca ggtagttatt 6780 cggatcatca gctacaccag agacggaaat ccatcgctcg accagtttag ttacccccag 6840 gctaagtgcc ttctctacac ctgcggtgct aaccagcgtt ttcgttctgc caatatggat 6900 taacattctc ccaccgtcag tacgtgagat atctttaacc ctgatcctgg caatttcggc 6960 tatacgtaac agggtgttat aagcaatccc cagaaatgcc agattacgta tatcctggca 7020 gcgatcgcta ttttccatga gtgaacgaac ctggtcgaaa tcagtgcgtt cgaacgctag 7080 agcctgtttt gcacgttcac cggcatcaac gttttctttt cggatccgcc gcataaccag 7140 tgaaacagca ttgctgtcac ttggtcgtgg cagcccggac cgacgatgaa gcatgtttag 7200 ctggcccaaa tgttgctgga tagtttttac tgccagaccg cgcgcctgaa gatatagaag 7260 ataatcgcga acatcttcag gttctgcggg aaaccatttc cggttattca acttgcacca 7320 tgccgcccac gaccggcaaa cggacagaag cattttccag gtatgctcag aaaacgcctg 7380 gcgatccctg aacatgtcca tcaggttctt gcgaacctca tcactcgttg catcgaccgg 7440 taacttttag attaacattt gtttaaatat gaaattaggc aatgtaacat actgcaggca 7500 aattttggtg tacggtaagt aaattggaca tatgatttaa caaagcgact ataagtcaga 7560 aagtgagaat gagattgaaa taattaattc aacttattca attgatgata tgccaggatt 7620 cctcttcctt tatatatctt caatgtttcg attaacaggt ccattatccg gtttttagtt 7680 tcgctgcaca ttgccgaatg acaatcggat tcctttaagc tttgcttttg ccctgttttc 7740 tctccccagt aatcgttggg tgactgaacc atccagtagc cgagagttac gtttttcctg 7800 atttaataaa tatccatatc tatttgtctc gtggtgtttt acagcaacta accgaaatta 7860 ctattttgca tcatccaacc ataaagagac aacaataatt ctagaaaaca tataacaagt 7920 gtttttttct atttacgtct catacgcaaa gtccacttta ccatgttttg gaaacttgta 7980 cccaaccatt catttttcga ttttgtaatg aggctctttt cttctaacaa tcattgaggt 8040 cattatcatg tgtcctcacc tattgtttaa attgaggctt gcgctattcc caaaattgtg 8100 ggtattattt tcatatatca gtatctgcaa tttcggttca actccggaag ctcggtaccc 8160 aactgtggga atactcaggt atcgtaagat gcaagagttc gaatctctta gcaaccatta 8220 ttttttttct ctcaacataa cgagaacaca caggggcgct atcgcacaga atcaaattcg 8280 atgactggaa attttttgtt aatttcagag gtcgcctgac gcatatacct ttttcaactg 8340 aaaaattggg agaaaaagga aaggtgagag cgccggaacc ggcttttcat atagaataga 8400 gaagcgttca tgactaaatg cttgcatcac aatacttgaa gttgacaata ttatttaagg 8460 acctattgtt ttttccaata ggtggttagc aatcgtctta ctttctaact tttcttacct 8520 tttacatttc agcaatatat atatatatat atatatatat ttcaaggata taccattgta 8580 atgtctgccc ctaagaagat cgtcgttttg ccaggtgacc acgttggtca agaaatcaca 8640 gccgaagcca ttaaggttct taaagctatt tctgatgttc gttccaatgt caagttcgat 8700 ttcgaaaatc atttaattgg tggtgctgct atcgatgcta caggtgttcc acttccagat 8760 gaggcgctgg aagcctccaa gaaggctgat gccgttttgt taggtgctgt gggtggtcct 8820 aaatggggta ccggtagtgt tagacctgaa caaggtttac taaaaatccg taaagaactt 8880 caattgtacg ccaacttaag accatgtaac tttgcatccg actctctttt agacttatct 8940 ccaatcaagc cacaatttgc taaaggtact gacttcgttg ttgttagaga attagtggga 9000 ggtatttact ttggtaagag aaaggaagac gatggtgatg gtgtcgcttg ggatagtgaa 9060 caatacaccg ttccagaagt gcaaagaatc acaagaatgg ccgctttcat ggccctacaa 9120 catgagccac cattgcctat ttggtccttg gataaagcta atgttttggc ctcttcaaga 9180 ttatggagaa aaactgtgga ggaaaccatc aagaacgaat ttcctacatt gaaagttcaa 9240 catcaattga ttgattctgc cgccatgatc ctagttaaga acccaaccca cctaaatggt 9300 attataatca ccagcaacat gtttggtgat atcatctccg atgaagcctc cgttatccca 9360 ggctccttgg gtttgttgcc atctgcgtcc ttggcctctt tgccagacaa gaacaccgca 9420 tttggtttgt acgaaccatg ccatggttcc gctccagatt tgccaaagaa taaggtcaac 9480 cctatcgcca ctatcttgtc tgctgcaatg atgttgaaat tgtcattgaa cttgcctgaa 9540 gaaggtaaag ccattgaaga tgcagttaaa aaggttttgg atgcaggtat cagaactggt 9600 gatttaggtg gttccaacag taccaccgaa gtcggtgatg ctgtcgccga agaagttaag 9660 aaaatccttg cttaaaaaga ttctcttttt ttgtgatatt tgtacataaa ctttataaat 9720 gaaattcata atagaaacga cacgaaatta caaaatggaa tatgttcata gggtagacga 9780 aactatatac gcaatctaca tacatttatc aagaaggaga aaaaggagga tgtaaaggaa 9840 tacaggtaag caaattgata ctaatggctc aacgtgataa ggaaaaagaa ttgcacttta 9900 acattaatat tgacaaggag gagggcatca ca 9932 & pAW8_6XHis"Times New Roman I^MluI(392),XbaI(798),ScaI(1014) Arial&c[RR I^+ScaI(1111),NdeI(1192),XbaI(1214),ClaI(1429) Arial&!w!GXw I^ScaI ArialP&Xajj I^FPvuII(3677),HindIII(3856),NotI(3868),EagI(3868),AscI(3936),BglII(4172) Arial & W5 I^6PstI(4386),MluI(4419),NcoI(4559),NruI(4643),ClaI(4677) Arial& ^ I^\HindIII(5104),ScaI(5375),SalI(5604),SpeI(5612),NotI(5653),EagI(5653),EcoRI(5661),PvuII(5753) Arial H&$   I^+PacI(6038),ScaI(6296),ScaI(6421),ClaI(6674) Arial& I^DEcoRV(6931),SexAI(7050),BamHI(7123),NruI(7268),PstI(7497),NdeI(7531) Arial&^`bC> I^#HindIII(7718),XbaI(7901),KpnI(8159) Arial&H I^!ClaI(8733),KpnI(8832),EcoRV(9331) Arial"&~woKo I^NcoI Arial$&0T-J)@4C@I^C&"8 pGEX_3_primer"Times New RomanI^CɆ& AmpR_promoter"Times New RomanI^C&  Ampicillin"Times New RomanI^CɆ&y ORF 2"Times New RomanI^&   pBR322_ori"Times New RomanI^& lac_promoter"Times New RomanI^C&M13_pUC_rev_primer"Times New RomanI^CɆ&M13_reverse_primer"Times New RomanI^C&"CloxP"Times New RomanI^&pHybLex_rev_primer"Times New RomanI^C&kanR"Times New RomanI^C&"2M13_forward20_primer"Times New RomanI^&+AM13_pUC_fwd_primer"Times New RomanI^C&3ORF 1"Times New RomanI^C&!%ORF 1"Times New Roman I^test"Times New RomanB&^F:V I^test"Times New Roman&C  I^test"Times New Romanj&