DDAID EU855181; SV 1; circular; other DNA; STD; SYN; 10622 BP. AC EU855181; DT 03-AUG-2008 (Rel. 96, Created) DT 03-AUG-2008 (Rel. 96, Last updated, Version 1) DE Cloning vector pAW8_mCherry, complete sequence. KW . OS Cloning vector pAW8_mCherry OC other sequences; artificial sequences; vectors. RN [1] RP 1-10622 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 1-10622 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..10622 FT /organism="Cloning vector pAW8_mCherry" FT /mol_type="other DNA" FT /db_xref="taxon:550444" FT misc_feature 1..3873 FT /note="Cre-expression plasmid pAW8 backbone" FT misc_recomb 3874..3907 FT /note="loxP" FT CDS 3916..4626 FT /codon_start=1 FT /transl_table=11 FT /product="monomeric red fluorescent protein" FT /note="mCherry; engineered variant of GenBank Accession FT Number AF506027; N- and C-termini replaced with equivalent FT residues from EGFP" FT /protein_id="ACF75945.1" FT /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG FT TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF FT EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK FT GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA FT EGRHSTGGMDELYK" FT terminator 4635..4863 FT /note="Saccharomyces cerevisiae ADH1" FT gene 5253..6062 FT /gene="kanR" FT CDS 5253..6062 FT /codon_start=1 FT /transl_table=11 FT /gene="kanR" FT /product="kanamycin and geneticin G418 resistance protein" FT /protein_id="ACF75946.1" FT /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP FT DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA FT FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD FT FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD FT RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" FT misc_feature 6185..10466 FT /note="Cre-expression plasmid pAW8 backbone" FT misc_recomb 6310..6343 FT /note="loxM3" SQ Sequence 10622 BP; 2948 A; 2352 C; 2291 G; 3031 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 tgcttatggt gagcaagggc gaggaggata acatggccat catcaaggag 3960 ttcatgcgct tcaaggtgca catggagggc tccgtgaacg gccacgagtt cgagatcgag 4020 ggcgagggcg agggccgccc ctacgagggc acccagaccg ccaagctgaa ggtgaccaag 4080 ggtggccccc tgcccttcgc ctgggacatc ctgtcccctc agttcatgta cggctccaag 4140 gcctacgtga agcaccccgc cgacatcccc gactacttga agctgtcctt ccccgagggc 4200 ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt gacccaggac 4260 tcctccctgc aggacggcga gttcatctac aaggtgaagc tgcgcggcac caacttcccc 4320 tccgacggcc ccgtaatgca gaagaagacc atgggctggg aggcctcctc cgagcggatg 4380 taccccgagg acggcgccct gaagggcgag atcaagcaga ggctgaagct gaaggacggc 4440 ggccactacg acgctgaggt caagaccacc tacaaggcca agaagcccgt gcagctgccc 4500 ggcgcctaca acgtcaacat caagttggac atcacctccc acaacgagga ctacaccatc 4560 gtggaacagt acgaacgcgc cgagggccgc cactccaccg gcggcatgga cgagctgtac 4620 aagtaaggcg cgccacttct aaataagcga atttcttatg atttatgatt tttattatta 4680 aataagttat aaaaaaaata agtgtataca aattttaaag tgactcttag gttttaaaac 4740 gaaaattctt attcttgagt aactctttcc tgtaggtcag gttgctttct caggtatagt 4800 atgaggtcgc tcttattgac cacacctcta ccggcagatc cgctagggat aacagggtaa 4860 tatagatctg tttagcttgc ctcgtccccg ccgggtcacc cggccagcga catggaggcc 4920 cagaataccc tccttgacag tcttgacgtg cgcagctcag gggcatgatg tgactgtcgc 4980 ccgtacattt agcccataca tccccatgta taatcatttg catccataca ttttgatggc 5040 cgcacggcgc gaagcaaaaa ttacggctcc tcgctgcaga cctgcgagca gggaaacgct 5100 cccctcacag acgcgttgaa ttgtccccac gccgcgcccc tgtagagaaa tataaaaggt 5160 taggatttgc cactgaggtt cttctttcat atacttcctt ttaaaatctt gctaggatac 5220 agttctcaca tcacatccga acataaacaa ccatgggtaa ggaaaagact cacgtttcga 5280 ggccgcgatt aaattccaac atggatgctg atttatatgg gtataaatgg gctcgcgata 5340 atgtcgggca atcaggtgcg acaatctatc gattgtatgg gaagcccgat gcgccagagt 5400 tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt tacagatgag atggtcagac 5460 taaactggct gacggaattt atgcctcttc cgaccatcaa gcattttatc cgtactcctg 5520 atgatgcatg gttactcacc actgcgatcc ccggcaaaac agcattccag gtattagaag 5580 aatatcctga ttcaggtgaa aatattgttg atgcgctggc agtgttcctg cgccggttgc 5640 attcgattcc tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg 5700 cgcaatcacg aatgaataac ggtttggttg atgcgagtga ttttgatgac gagcgtaatg 5760 gctggcctgt tgaacaagtc tggaaagaaa tgcataagct tttgccattc tcaccggatt 5820 cagtcgtcac tcatggtgat ttctcacttg ataaccttat ttttgacgag gggaaattaa 5880 taggttgtat tgatgttgga cgagtcggaa tcgcagaccg ataccaggat cttgccatcc 5940 tatggaactg cctcggtgag ttttctcctt cattacagaa acggcttttt caaaaatatg 6000 gtattgataa tcctgatatg aataaattgc agtttcattt gatgctcgat gagtttttct 6060 aatcagtact gacaataaaa agattcttgt tttcaagaac ttgtcatttg tatagttttt 6120 ttatattgta gttgttctat tttaatcaaa tgttagcgtg atttatattt tttttcgcct 6180 cgacatcatc tgcccagatg cgaagttaag tgcgcagaaa gtaatatcat gcgtcaatcg 6240 tatgtgaatg ctggtcgcta tactgctgtc gattcgatac taacgccgcc atccagtcga 6300 cgaactagta taacttcgta tatggtatta tatacgaagt tatgcggccg cgcgaattca 6360 ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc 6420 cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc 6480 ccttcccaac agttgcgcag cctgaatggc gaatggcgcc tgatgcggta ttttctcctt 6540 acgcatctgt gcggtatttc acaccgcata ccatcgtccg gatcaatgaa gtagatgagt 6600 ttactacctg tatatacttt tttttttgtc aaagtcactt ttatggcatt tccatgttga 6660 aggattagcc acagcactaa attttcagcg tatgattgct tttaaatatt taattttcat 6720 cgttttttaa ttaatattca aaacgattta atgccttcta catctgaagt caagctcata 6780 gactcgggtt gggtagctaa aactacatct aacacgaagg ggtatagcct tcttaaagta 6840 tactcatgta taaacgtttt gggttcatat ttttgggcca cgtggtttat aaaaattctt 6900 aactacacca ctcggtatcc cgcacccgtc tacgtttcta cgatttcgag ctaatattga 6960 gtaaagtgat tagcaaaaaa aataaaagta ctcgttgtcg gagatcaaga atttttctat 7020 tatctcatct aaaccacttt ctaaaagcga aaaacaaaat cgtaatatgc agcttgaatg 7080 ggcttccata gtttgaaaga aaaaccctag cagtactggc aagggagaca ttccttttac 7140 ctggctaatc gccatcttcc agcaggcgca ccattgcccc tgtttcacta tccaggttac 7200 ggatatagtt catgacaata tttacattgg tccagccacc agcttgcatg atctccggta 7260 ttgaaactcc agcgcgggcc atatctcgcg cggctccgac acgggcactg tgtccagacc 7320 aggccaggta tctctgacca gagtcatcct tagcgccgta aatcaatcga tgagttgctt 7380 caaaaatccc ttccagggcg cgagttgata gctggctggt ggcagatggc gcggcaacac 7440 cattttttct gacccggcaa aacaggtagt tattcggatc atcagctaca ccagagacgg 7500 aaatccatcg ctcgaccagt ttagttaccc ccaggctaag tgccttctct acacctgcgg 7560 tgctaaccag cgttttcgtt ctgccaatat ggattaacat tctcccaccg tcagtacgtg 7620 agatatcttt aaccctgatc ctggcaattt cggctatacg taacagggtg ttataagcaa 7680 tccccagaaa tgccagatta cgtatatcct ggcagcgatc gctattttcc atgagtgaac 7740 gaacctggtc gaaatcagtg cgttcgaacg ctagagcctg ttttgcacgt tcaccggcat 7800 caacgttttc ttttcggatc cgccgcataa ccagtgaaac agcattgctg tcacttggtc 7860 gtggcagccc ggaccgacga tgaagcatgt ttagctggcc caaatgttgc tggatagttt 7920 ttactgccag accgcgcgcc tgaagatata gaagataatc gcgaacatct tcaggttctg 7980 cgggaaacca tttccggtta ttcaacttgc accatgccgc ccacgaccgg caaacggaca 8040 gaagcatttt ccaggtatgc tcagaaaacg cctggcgatc cctgaacatg tccatcaggt 8100 tcttgcgaac ctcatcactc gttgcatcga ccggtaactt ttagattaac atttgtttaa 8160 atatgaaatt aggcaatgta acatactgca ggcaaatttt ggtgtacggt aagtaaattg 8220 gacatatgat ttaacaaagc gactataagt cagaaagtga gaatgagatt gaaataatta 8280 attcaactta ttcaattgat gatatgccag gattcctctt cctttatctt caatgtttcg 8340 attaacaggt ccattatccg gtttttagtt tcgctgcaca ttgccgaatg acaatcggat 8400 tcctttaagc tttgcttttg ccctgttttc tctccccagt aatcgttggg tgactgaacc 8460 atccagtagc cgagagttac gtttttcctg atttaataaa tatccatatc tatttgtctc 8520 gtggtgtttt acagcaacta accgaaatta ctattttgca tcatccaacc ataaagagac 8580 aacaataatt ctagaaaaca tataacaagt gtttttttct atttacgtct catacgcaaa 8640 gtccacttta ccatgttttg gaaacttgta cccaaccatt catttttcga ttttgtaatg 8700 aggctctttt cttctaacaa tcattgaggt cattatcatg tgtcctcacc tattgtttaa 8760 attgaggctt gcgctattcc caaaattgtg ggtattattt tcatatatca gtatctgcaa 8820 tttcggttca actccggaag ctcggtaccc aactgtggga atactcaggt atcgtaagat 8880 gcaagagttc gaatctctta gcaaccatta ttttttttct ctcaacataa cgagaacaca 8940 caggggcgct atcgcacaga atcaaattcg atgactggaa attttttgtt aatttcagag 9000 gtcgcctgac gcatatacct ttttcaactg aaaaattggg agaaaaagga aaggtgagag 9060 cgccggaacc ggcttttcat atagaataga gaagcgttca tgactaaatg cttgcatcac 9120 aatacttgaa gttgacaata ttatttaagg acctattgtt ttttccaata ggtggttagc 9180 aatcgtctta ctttctaact tttcttacct tttacatttc agcaatatat atatatatat 9240 atatatatat ttcaaggata taccattgta atgtctgccc ctaagaagat cgtcgttttg 9300 ccaggtgacc acgttggtca agaaatcaca gccgaagcca ttaaggttct taaagctatt 9360 tctgatgttc gttccaatgt caagttcgat ttcgaaaatc atttaattgg tggtgctgct 9420 atcgatgcta caggtgttcc acttccagat gaggcgctgg aagcctccaa gaaggctgat 9480 gccgttttgt taggtgctgt gggtggtcct aaatggggta ccggtagtgt tagacctgaa 9540 caaggtttac taaaaatccg taaagaactt caattgtacg ccaacttaag accatgtaac 9600 tttgcatccg actctctttt agacttatct ccaatcaagc cacaatttgc taaaggtact 9660 gacttcgttg ttgttagaga attagtggga ggtatttact ttggtaagag aaaggaagac 9720 gatggtgatg gtgtcgcttg ggatagtgaa caatacaccg ttccagaagt gcaaagaatc 9780 acaagaatgg ccgctttcat ggccctacaa catgagccac cattgcctat ttggtccttg 9840 gataaagcta atgttttggc ctcttcaaga ttatggagaa aaactgtgga ggaaaccatc 9900 aagaacgaat ttcctacatt gaaagttcaa catcaattga ttgattctgc cgccatgatc 9960 ctagttaaga acccaaccca cctaaatggt attataatca ccagcaacat gtttggtgat 10020 atcatctccg atgaagcctc cgttatccca ggctccttgg gtttgttgcc atctgcgtcc 10080 ttggcctctt tgccagacaa gaacaccgca tttggtttgt acgaaccatg ccatggttcc 10140 gctccagatt tgccaaagaa taaggtcaac cctatcgcca ctatcttgtc tgctgcaatg 10200 atgttgaaat tgtcattgaa cttgcctgaa gaaggtaaag ccattgaaga tgcagttaaa 10260 aaggttttgg atgcaggtat cagaactggt gatttaggtg gttccaacag taccaccgaa 10320 gtcggtgatg ctgtcgccga agaagttaag aaaatccttg cttaaaaaga ttctcttttt 10380 ttgtgatatt tgtacataaa ctttataaat gaaattcata atagaaacga cacgaaatta 10440 caaaatggaa tatgttcata gggtagacga aactatatac gcaatctaca tacatttatc 10500 aagaaggaga aaaaggagga tgtaaaggaa tacaggtaag caaattgata ctaatggctc 10560 aacgtgataa ggaaaaagaa ttgcacttta acattaatat tgacaaggag gagggcatca 10620 ca 10622 ~) pAW8_mCherry"Times New Roman DMluI(392),XbaI(798),ScaI(1014) Arial~)] T$K 7K D+ScaI(1111),NdeI(1192),XbaI(1214),ClaI(1429) Arial~)GyNrTjT:j DScaI ArialP~) DEPvuII(3677),HindIII(3856),EagI(3868),NotI(3868),PstI(4272),NcoI(4350) Arial~)aipp D8PvuII(4495),AscI(4629),BglII(4865),PstI(5079),MluI(5112) Arial~)Z D.NcoI(5252),NruI(5336),ClaI(5370),HindIII(5797) Arial~)$$l DYScaI(6068),SalI(6297),SpeI(6305),NotI(6346),EagI(6346),EcoRI(6355),PvuII(6447),PacI(6732) Arial ~)0*$$V D,ScaI(6990),ScaI(7115),ClaI(7368),EcoRV(7625) Arial~) DFSexAI(7744),BamHI(7817),NruI(7962),PstI(8191),NdeI(8225),HindIII(8408) Arial~)...z R DXbaI(8591),KpnI(8849) Arial!~) D"ClaI(9423),KpnI(9522),EcoRV(10021) Arial2%~)&t ld@d DNcoI Arial'~)pNmDj:N.:DC~)"8 pGEX_3_primer"Times New RomanDCɆ~) AmpR_promoter"Times New RomanDC~)  Ampicillin"Times New RomanD~)y ORF 3"Times New RomanD~)   pBR322_ori"Times New RomanD~) lac_promoter"Times New RomanDC~)M13_pUC_rev_primer"Times New RomanDCɆ~)M13_reverse_primer"Times New RomanDCɆ~)ORF 1"Times New RomanDC~)"CloxP"Times New RomanD~)^vpHybLex_rev_primer"Times New RomanDCɆ~)kanR"Times New RomanDC~)M13_forward20_primer"Times New RomanD~)M13_pUC_fwd_primer"Times New RomanDCɆ~)ORF 1"Times New RomanDC~)7$}(ORF 2"Times New Roman Dtest"Times New Roman~){cKK Dtest"Times New Roman~)><.Us.! Dtest"Times New Roman ~))/;;