ÆP’ÆP’ÿ}8ID FJ205735; SV 1; circular; other DNA; STD; SYN; 8252 BP. AC FJ205735; DT 14-OCT-2008 (Rel. 97, Created) DT 16-OCT-2008 (Rel. 97, Last updated, Version 2) DE Cre-expression plasmid pAW8_Xho, complete sequence. KW . OS Cre-expression plasmid pAW8_Xho OC other sequences; artificial sequences; vectors. RN [1] RP 1-8252 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 1-8252 RA Watson A.T.; RT ; RL Submitted (08-SEP-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..8252 FT /organism="Cre-expression plasmid pAW8_Xho" FT /mol_type="other DNA" FT /db_xref="taxon:563989" FT rep_origin 231..1424 FT /note="fission yeast ars1" FT misc_feature 1425..3860 FT /note="pUC19 vector backbone" FT misc_recomb 3874..3907 FT /note="loxP" FT misc_feature 3908..3939 FT /note="MCS polylinker; XhoI-SacI-SalI-SpeI" FT misc_recomb 3940..3972 FT /note="loxM3" FT terminator complement(4213..4769) FT /note="nmt1" FT gene complement(4775..5855) FT /gene="cre" FT CDS complement(join(4775..5767,5817..5855)) FT /codon_start=1 FT /transl_table=11 FT /gene="cre" FT /product="bacteriophage P1 recombinase" FT /protein_id="ACI42000.1" FT /translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML FT LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL FT PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF FT LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE FT RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ FT RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE FT DGD" FT intron complement(5768..5816) FT /gene="cre" FT /number=1 FT /note="Schizosaccharomyces pombe rad50" FT promoter complement(5856..6462) FT /note="nmt41" FT gene 6481..8252 FT /gene="LEU2" FT promoter 6481..6900 FT /gene="LEU2" FT CDS 6901..7995 FT /codon_start=1 FT /transl_table=11 FT /gene="LEU2" FT /product="beta-isopropyl-malate dehydrogenase" FT /note="involved in leucine biosynthesis" FT /protein_id="ACI42001.1" FT /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL FT IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA FT NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT FT VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ FT LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF FT GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG FT DLGGSNSTTEVGDAVAEEVKKILA" FT terminator 7996..8252 FT /gene="LEU2" SQ Sequence 8252 BP; 2344 A; 1773 C; 1715 G; 2420 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 aagttatctc gagggagagc tcggagtcga cgaactagta taacttcgta tatggtatta 3960 tatacgaagt tatgcggccg cgcgaattca ctggccgtcg ttttacaacg tcgtgactgg 4020 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg 4080 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc 4140 gaatggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata 4200 ccatcgtccg gatcaatgaa gtagatgagt ttactacctg tatatacttt tttttttgtc 4260 aaagtcactt ttatggcatt tccatgttga aggattagcc acagcactaa attttcagcg 4320 tatgattgct tttaaatatt taattttcat cgttttttaa ttaatattca aaacgattta 4380 atgccttcta catctgaagt caagctcata gactcgggtt gggtagctaa aactacatct 4440 aacacgaagg ggtatagcct tcttaaagta tactcatgta taaacgtttt gggttcatat 4500 ttttgggcca cgtggtttat aaaaattctt aactacacca ctcggtatcc cgcacccgtc 4560 tacgtttcta cgatttcgag ctaatattga gtaaagtgat tagcaaaaaa aataaaagta 4620 ctcgttgtcg gagatcaaga atttttctat tatctcatct aaaccacttt ctaaaagcga 4680 aaaacaaaat cgtaatatgc agcttgaatg ggcttccata gtttgaaaga aaaaccctag 4740 cagtactggc aagggagaca ttccttttac ctggctaatc gccatcttcc agcaggcgca 4800 ccattgcccc tgtttcacta tccaggttac ggatatagtt catgacaata tttacattgg 4860 tccagccacc agcttgcatg atctccggta ttgaaactcc agcgcgggcc atatctcgcg 4920 cggctccgac acgggcactg tgtccagacc aggccaggta tctctgacca gagtcatcct 4980 tagcgccgta aatcaatcga tgagttgctt caaaaatccc ttccagggcg cgagttgata 5040 gctggctggt ggcagatggc gcggcaacac cattttttct gacccggcaa aacaggtagt 5100 tattcggatc atcagctaca ccagagacgg aaatccatcg ctcgaccagt ttagttaccc 5160 ccaggctaag tgccttctct acacctgcgg tgctaaccag cgttttcgtt ctgccaatat 5220 ggattaacat tctcccaccg tcagtacgtg agatatcttt aaccctgatc ctggcaattt 5280 cggctatacg taacagggtg ttataagcaa tccccagaaa tgccagatta cgtatatcct 5340 ggcagcgatc gctattttcc atgagtgaac gaacctggtc gaaatcagtg cgttcgaacg 5400 ctagagcctg ttttgcacgt tcaccggcat caacgttttc ttttcggatc cgccgcataa 5460 ccagtgaaac agcattgctg tcacttggtc gtggcagccc ggaccgacga tgaagcatgt 5520 ttagctggcc caaatgttgc tggatagttt ttactgccag accgcgcgcc tgaagatata 5580 gaagataatc gcgaacatct tcaggttctg cgggaaacca tttccggtta ttcaacttgc 5640 accatgccgc ccacgaccgg caaacggaca gaagcatttt ccaggtatgc tcagaaaacg 5700 cctggcgatc cctgaacatg tccatcaggt tcttgcgaac ctcatcactc gttgcatcga 5760 ccggtaactt ttagattaac atttgtttaa atatgaaatt aggcaatgta acatactgca 5820 ggcaaatttt ggtgtacggt aagtaaattg gacatatgat ttaacaaagc gactataagt 5880 cagaaagtga gaatgagatt gaaataatta attcaactta ttcaattgat gatatgccag 5940 gattcctctt cctttatctt caatgtttcg attaacaggt ccattatccg gtttttagtt 6000 tcgctgcaca ttgccgaatg acaatcggat tcctttaagc tttgcttttg ccctgttttc 6060 tctccccagt aatcgttggg tgactgaacc atccagtagc cgagagttac gtttttcctg 6120 atttaataaa tatccatatc tatttgtctc gtggtgtttt acagcaacta accgaaatta 6180 ctattttgca tcatccaacc ataaagagac aacaataatt ctagaaaaca tataacaagt 6240 gtttttttct atttacgtct catacgcaaa gtccacttta ccatgttttg gaaacttgta 6300 cccaaccatt catttttcga ttttgtaatg aggctctttt cttctaacaa tcattgaggt 6360 cattatcatg tgtcctcacc tattgtttaa attgaggctt gcgctattcc caaaattgtg 6420 ggtattattt tcatatatca gtatctgcaa tttcggttca actccggaag ctcggtaccc 6480 aactgtggga atactcaggt atcgtaagat gcaagagttc gaatctctta gcaaccatta 6540 ttttttttct ctcaacataa cgagaacaca caggggcgct atcgcacaga atcaaattcg 6600 atgactggaa attttttgtt aatttcagag gtcgcctgac gcatatacct ttttcaactg 6660 aaaaattggg agaaaaagga aaggtgagag cgccggaacc ggcttttcat atagaataga 6720 gaagcgttca tgactaaatg cttgcatcac aatacttgaa gttgacaata ttatttaagg 6780 acctattgtt ttttccaata ggtggttagc aatcgtctta ctttctaact tttcttacct 6840 tttacatttc agcaatatat atatatatat atatatatat ttcaaggata taccattgta 6900 atgtctgccc ctaagaagat cgtcgttttg ccaggtgacc acgttggtca agaaatcaca 6960 gccgaagcca ttaaggttct taaagctatt tctgatgttc gttccaatgt caagttcgat 7020 ttcgaaaatc atttaattgg tggtgctgct atcgatgcta caggtgttcc acttccagat 7080 gaggcgctgg aagcctccaa gaaggctgat gccgttttgt taggtgctgt gggtggtcct 7140 aaatggggta ccggtagtgt tagacctgaa caaggtttac taaaaatccg taaagaactt 7200 caattgtacg ccaacttaag accatgtaac tttgcatccg actctctttt agacttatct 7260 ccaatcaagc cacaatttgc taaaggtact gacttcgttg ttgttagaga attagtggga 7320 ggtatttact ttggtaagag aaaggaagac gatggtgatg gtgtcgcttg ggatagtgaa 7380 caatacaccg ttccagaagt gcaaagaatc acaagaatgg ccgctttcat ggccctacaa 7440 catgagccac cattgcctat ttggtccttg gataaagcta atgttttggc ctcttcaaga 7500 ttatggagaa aaactgtgga ggaaaccatc aagaacgaat ttcctacatt gaaagttcaa 7560 catcaattga ttgattctgc cgccatgatc ctagttaaga acccaaccca cctaaatggt 7620 attataatca ccagcaacat gtttggtgat atcatctccg atgaagcctc cgttatccca 7680 ggctccttgg gtttgttgcc atctgcgtcc ttggcctctt tgccagacaa gaacaccgca 7740 tttggtttgt acgaaccatg ccatggttcc gctccagatt tgccaaagaa taaggtcaac 7800 cctatcgcca ctatcttgtc tgctgcaatg atgttgaaat tgtcattgaa cttgcctgaa 7860 gaaggtaaag ccattgaaga tgcagttaaa aaggttttgg atgcaggtat cagaactggt 7920 gatttaggtg gttccaacag taccaccgaa gtcggtgatg ctgtcgccga agaagttaag 7980 aaaatccttg cttaaaaaga ttctcttttt ttgtgatatt tgtacataaa ctttataaat 8040 gaaattcata atagaaacga cacgaaatta caaaatggaa tatgttcata gggtagacga 8100 aactatatac gcaatctaca tacatttatc aagaaggaga aaaaggagga tgtaaaggaa 8160 tacaggtaag caaattgata ctaatggctc aacgtgataa ggaaaaagaa ttgcacttta 8220 acattaatat tgacaaggag gagggcatca ca 8252 < pAW8_Xhoðÿÿÿ"Times New Roman ÍÍÍÍÆP’ÍÍÍÍMluI(392),XbaI(798) Arialˆ< ïZòPôFç F ÍÍÍÍÆP’ÍÍÍÍ6ScaI(1014),ScaI(1111),NdeI(1192),XbaI(1214),ClaI(1429) Arial¨< a¨i¢p›p_§› ÍÍÍÍÆP’ÍÍÍÍScaI ArialP< ‘>›>¤>¤8×D ÍÍÍÍÆP’ÍÍÍ͉PvuII(3677),HindIII(3856),NotI(3868),EagI(3868),XhoI(3909),SacI(3922),SalI(3927),SpeI(3935),NotI(3976),EagI(3976),EcoRI(3985),PvuII(4077) ArialR< Ïä7ÑM’MÑÝ ÍÍÍÍÆP’ÍÍÍÍ PacI(4362),ScaI(4620),ScaI(4745) Arial < Y UQ>d] ÍÍÍÍÆP’ÍÍÍÍ.ClaI(4998),EcoRV(5255),SexAI(5374),BamHI(5447) Arial‡< þÌöÓîÙ®Ùî  ÍÍÍÍÆP’ÍÍÍÍ.NruI(5592),PstI(5821),NdeI(5855),HindIII(6038) Arial½< ÐyÆ|¼~}f¼– ÍÍÍÍÆP’ÍÍÍÍXbaI(6221),KpnI(6479) ArialM< Æ1¼1²0|$²< ÍÍÍÍÆP’ÍÍÍÍClaI(7053),KpnI(7152) Arial< ô«ì¥äž®†äž ÍÍÍÍÆP’ÍÍÍÍEcoRV(7651),NcoI(7762) Arialã< Ci?`:Wú?:WÍÍÍÍÆP’ÉCÍÍÍÍ< "8Ž pGEX_3_primerõÿÿÿ"Times New RomanèÍÍÍÍÆP’CɆÍÍÍÍ< ×óŽ AmpR_promoterõÿÿÿ"Times New RomanèÍÍÍÍÆP’ÿCÍÍÍÍ<  Ž Ampicillinõÿÿÿ"Times New RomanèÍÍÍÍÆP’CɆÍÍÍÍ< y ŽORF 2õÿÿÿ"Times New RomanèÍÍÍÍÆP’†ÉÍÍÍÍ<   Ž pBR322_oriõÿÿÿ"Times New RomanèÍÍÍÍÆP’†ÉÍÍÍÍ< ´ÑŽ lac_promoterõÿÿÿ"Times New RomanèÍÍÍÍÆP’†CÍÍÍÍ< ßõŽM13_pUC_rev_primerõÿÿÿ"Times New RomanèÍÍÍÍÆP’CɆÍÍÍÍ< ôŽM13_reverse_primerõÿÿÿ"Times New RomanèÍÍÍÍÆP’ÉCÍÍÍÍ< "CŽloxPõÿÿÿ"Times New RomanèÍÍÍÍÆP’†CÍÍÍÍ< –¦ŒM13_forward20_primerõÿÿÿ"Times New RomanèÍÍÍÍÆP’CɆÍÍÍÍ< ŸµŒM13_pUC_fwd_primerõÿÿÿ"Times New RomanèÍÍÍÍÆP’ÉCÍÍÍÍ< §]ŒORF 1õÿÿÿ"Times New RomanèÍÍÍÍÆP’†ÉÍÍÍÍ< õ;ŽLEU2õÿÿÿ"Times New Romanè ÍÍÍÍÆP’ÍÍÍÍtestõÿÿÿ"Times New Roman·+ < xÿÿÿ‰ömüGó³×Gc ÍÍÍÍÆP’ÍÍÍÍtestõÿÿÿ"Times New Roman42a< 9ÿÿÿ×æÔð œÔð€