yOEyOE`4ID FJ457003; SV 1; circular; other DNA; STD; SYN; 8152 BP. AC FJ457003; DT 30-JAN-2009 (Rel. 99, Created) DT 30-JAN-2009 (Rel. 99, Last updated, Version 1) DE Neurospora expression vector pCCG::C-Gly::HAT::FLAG, complete sequence. KW . OS Neurospora expression vector pCCG::C-Gly::HAT::FLAG OC other sequences; artificial sequences; vectors. RN [1] RP 1-8152 RA Honda S., Selker E.U.; RT "Tools for fungal proteomics: Multifunctional Neurospora vectors for gene RT replacement, protein expression and protein purification"; RL Genetics 0:0-0(2009). RN [2] RP 1-8152 RA Honda S., Selker E.U.; RT ; RL Submitted (11-NOV-2008) to the EMBL/GenBank/DDBJ databases. RL Institute of Molecular Biology, University of Oregon, Eugene, OR 97403, USA FH Key Location/Qualifiers FT source 1..8152 FT /organism="Neurospora expression vector FT pCCG::C-Gly::HAT::FLAG" FT /mol_type="other DNA" FT /db_xref="taxon:591135" FT promoter complement(1..17) FT /note="SP6" FT CDS 468..1328 FT /codon_start=1 FT /transl_table=11 FT /product="beta-lactamase" FT /note="ampicillin resistance" FT /protein_id="ACM10004.1" FT /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI FT ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS FT PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW FT EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA FT LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS FT LIKHW" FT promoter complement(2345..2365) FT /note="T7" FT misc_feature complement(2374..4006) FT /note="3' flank region of Neurospora crassa his-3 gene" FT promoter 4057..4985 FT /note="Neurospora crassa ccg-1 (grg-1)" FT misc_feature 5013..5042 FT /note="10xGly" FT misc_feature 5055..5110 FT /note="HAT" FT misc_feature 5112..5126 FT /note="5xGly" FT misc_feature 5127..5150 FT /note="FLAG" FT misc_feature complement(5406..5490) FT /note="T3 promoter and multiple cloning site from FT pBluescript KS+" FT misc_feature complement(6226..8133) FT /note="5' truncated histidine biosynthesis trifunctional FT protein; includes phosphoribosyl-AMP cyclohydrolase, FT phosphoribosyl-ATP cyclohydrolase and histindinol FT dehydrogenase" SQ Sequence 8152 BP; 2036 A; 2115 C; 2092 G; 1909 T; 0 other; tatagtgtca cctaaatcgt atgtgtatga tacataaggt tatgtattaa ttgtagccgc 60 gttctaacga caatatgtcc atatggtgca ctctcagtac aatctgctct gatgccgcat 120 agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc 180 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt 240 tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat 300 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg 360 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga 420 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac 480 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc 540 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca 600 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc 660 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg 720 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac 780 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca 840 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg 900 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac 960 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg 1020 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat 1080 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg 1140 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg 1200 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc 1260 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc 1320 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt 1380 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt 1440 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt 1500 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag 1560 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca 1620 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca 1680 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg 1740 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg 1800 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct 1860 acaccgaact gagataccta cagcgtgagc attgagaaag cgccacgctt cccgaaggga 1920 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc 1980 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg 2040 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg 2100 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt 2160 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc 2220 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac 2280 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg caggttaacc tggcttatcg 2340 aaattaatac gactcactat agggagaccg gcagatccaa tgcggatgga ttcgctcact 2400 cggtgccgcg gcggtgcgaa gttcctgccg tggattgggg gattgcacag acggtggagt 2460 gcttgctaca gctaggtacc cagttgcaca ccaaacggtg cccctgtgcc cctgcgtcta 2520 ggtgtgtcct gtgagcaact gcgcccgcgc gcggcaccgg gaagctgcgg gaacctcaat 2580 ggatgattgg cgccagatcc ccggcatcac ctcagcacat gggcagcccg agaaggaatc 2640 gagggtagaa gatattccag ggtttccgct agcacaactt ccaagcgcct ctaccgctac 2700 tccacacacc cacgctgaat ggcacaacca ctgtaggcct agcggtccca gctccccgta 2760 accacgcagc cccagcgacc gaggtcatca tgacactatg acgacaatct ggcgggagac 2820 ttgggaaaat cgatgacggg cacctacctt gggaacaata gagcaagtcc agtcaacgcc 2880 ctagggaagg gggaaaaaaa aaaagagccc attctcacaa ggagcaacac tgtgctgcta 2940 tcatagtaca ctacaagcaa agtcgggatg ttgctagaat ccgtcggcta cgtcggtaga 3000 tacctcccat aggcagtaag tctaggtatc caatttatag ttccgcgtat ttcttcgtcg 3060 aacggggcac agcaaagcct aacagccgca gtgcctcgca accaacacca agcagtaatc 3120 tcatccatcc atacagatcc tcccacaata tctttacatg cgcacttgcg ctgtgttagc 3180 agtttaggtt ccactaccaa ggctgacacg agcagcactg acagcaaact gctgcttggg 3240 gccaagaaat tggcagtttg ctgagcacct acatatcagt acacatcaac cctggtttaa 3300 ttgttagcct ggtactgtta accgtcatga acgaggacgt ggaggaagaa ccacgctaac 3360 aattgaacca gggttgatgt atacctctct taccatattt gcttgcttcc acgtccctat 3420 cctcaagcga ttttgttctt gactttttgt atttcaacgc aaaggccatt caggtttctc 3480 atgttcaacc ctttggatgg cgatgccgtt cgtcccagga cctcgtgacc tttgaaagac 3540 gagaaatata tcccaaagca acttctgctt atggacaagc aactgcatgg cggcccacaa 3600 ctcatatctc gcgtaatcat ccaaacgcta ggcccgagta ctctactgtt ctcgagacac 3660 ctaccataat atgtacccaa tcagacatga caacagtcaa ccaaacatac cctgatccgg 3720 ccaaaccctg acatccctag atgatggtta ggcacacaaa tggaaaacgg gactcgttgg 3780 ttaggtggga acgcttgtat tgcagccagc caaccgactc tcctcgtccc tctcgagtcc 3840 cgttattgcc gtttgaccca gccgcccagc cgccaaacag ctgagggagc caatggtcta 3900 atacaccgtc caacggcccc agtgatgttt caaacttgat tgacagcgaa cgaaacccct 3960 gaaacccgtt atagatatgt atagatttga cggggtagct tggcccttaa tacgactcac 4020 tatagggcga attggagctc caccgcggtg gcggccgcta gaaggagcag tccatctgcg 4080 tgaatcacga gagaatcagc tactttgaat cgatggatgc agctaccaga agtcactcag 4140 ttcgttcaaa gccacatcac tgggcacttc cattgggaca ggcattgatc ggacgagacc 4200 gacttctggc cgctttcaac agccacatta tatccatgtc acggctacgc gtcggccttc 4260 ggtaacagaa aagcacacag acagcgattg tgacatggat tcgggcaaag cattggtggt 4320 cgcaccaggt catcctgagt gtgcagtggc tgctattcag atttcatcta actgcgggag 4380 aggggttcaa aggggcgtga cgtcacagac aacgggtgaa ggacgaagat tgcctcactt 4440 ctttgctagc aatttgcctg caaagaaggc gcacatgaca agcaaacaac tgggaaacca 4500 ctattgaata cccacaatgc aaagctcgga aggtacgtct tgattgcagt gtgtcgagtg 4560 tcaaaaaaga agcaagtgtt catgcaagcc aaaattggca cctcctccac ttctccgagt 4620 gccccacccg aacctccagg cgagatggcc ggaacatacc atccgcgttg ggattatgac 4680 gtatctcctt cttcttcaca tgattccatc ccgttgttgc ttgtttgcga gctgtgacgg 4740 gagatcgtag atgccacttc gggccaggca ggcagtgcag gcagccagga acacaagctt 4800 ccaacttggt catctcgatt gccgattaag ggaaccaaat gcctatataa gactgtcctc 4860 ccacctcccc aataccattc ttttcttctt ccatcatcag ccaacaaagc aatcacatct 4920 tcactacttc aaatcaacac aacactcaaa ccactttcac aacccctcac atcaaccaaa 4980 tctagaacta gtggatcccc gggttaatta agggcggagg cggcggaggc ggaggcggag 5040 gcttaatcgg cttgaaggat catctcatcc acaatgtcca caaagaggag cacgcacatg 5100 cccacaacaa gggcggaggc ggaggcgact acaaagacga tgacgacaag taggcgccac 5160 ttctaaataa gcgaatttct tatgatttat gatttttatt attaaataag ttataaaaaa 5220 aataagtgta tacaaatttt aaagtgactc ttaggtttta aaacgaaaat tcttattctt 5280 gagtaactct ttcctgtagg tcaggttgct ttctcaggta tagtatgagg tcgctcttat 5340 tgaccacacc tctaccggca gatccgctag ggataacagg gtaatataga tcccccgggc 5400 tgcaggaatt cgatatcaag cttatcgata ccgtcgacct cgaggggggg cccggtaccc 5460 agcttttgtt ccctttagtg agggttaatt agggcgtgga cggctaatgg ggtctgaatg 5520 ctaaaagaca ccatttccca cactccctct gatttttttt atggtttttt tttatcttca 5580 tttttcgccg tcttttattc ggttgtttgg gaggttcaca tgcaacgtgc aagactgcaa 5640 gttttttgag atgggcgctt catgcaatgc agcatcactg cagggacatt gtgacatcca 5700 cgtttcgatc gtctgtgctt cccccgttca tgtacgcatg atctgaaaaa aaaaaaaaaa 5760 aaaaaggttg atgtaataat gaaaataaac acccagggac gagcacctca tctgctggaa 5820 tagggagaaa aactcaaggg ccgtttgggt ctccacactt accaacagat tcatgtactg 5880 atcttcagcc tgtacagaac agtacaatgt ccaaaactcg ttatcattac tctatgtgcc 5940 caaaatgacc cagatataca tcaacccact acatcctttt gttcttgttt tttttttttt 6000 tttttgctac ttcgaaaaga atttccgttt agttgctctt gctcatgtgc tcaagacgga 6060 tgctgaccgc ccttctgtga gcctcgagct cctcaacctt agccaactgc atgacagcct 6120 ggccgacgtt tttgagaccc tcggcagtca agttggagct ggtaatgtgc ttgacgaacg 6180 aggcgaaatt gacgccagag tactgcttgc caaagccata ggtagctatg tgtcttgacg 6240 ttagcagaag aatctgtgaa gcggggctat ttatgatatg taacttacgc agcgagtggt 6300 taacaccagc agagtaatcg ccaacggact caggagtcca ggcgccaatg aagacactac 6360 cggcgttcat gacaagatcg acagccttct cggcctcctt gatctggagg atcaagtgct 6420 caggagcgta cttgttgctg agctccatgg cctcctcgac ggtcttgacc tgcacggtga 6480 tcgagtgggc gatggagcca cggacaatct ggacgcgagg aagctccgta gcctgacggt 6540 gaacctcgtc ctcaatagcc tgaagatgct cctcgtcgag gtcaatagcg atcaggatga 6600 cctgactgtc aacgccgtgc tcagcctggg acaggagatc cgaggcaacg aacgcggggt 6660 tggcgtcctt gtcagcgatg accagcacct cggacgggcc agcgggcatg tcaataccaa 6720 cggcagcgtt ggtgtcgttg ctgacgaaca tcttggcagc agtgacgaac tggttaccgg 6780 ggccgagaat cttgtcgacc ttggtgatgc tctcggtgcc gtaggccatg gcagctacgg 6840 cctgggcacc gccggcaagc acgatggact cggccccaac cttgtgagcg acgtggacaa 6900 tctcgggagt gatggttccg tcggcgcggg gaggagaggc gaacacaatc ttgttgcagc 6960 cggcgaccat ggcgggaaca cccagcataa gggcagtgct ggggagaacg gcggtaccgc 7020 cggggatgta gcagccgacg gcctcgatgg gacgagagaa acggctgcag acaacaccgg 7080 gcatggtctc gacctggagg ggcttctcct ccttctgggc ggcgtggaac ttgcggatgt 7140 tctcgaagga cacgtcgatg gcagcaatgg tctcctcagg gagctgcata agctccttgg 7200 ggaagggcgc cttcaggacg gggctagtaa gagaggtagc cttctcgaac ttgtgagtgt 7260 acgacagaac agccttgtcg ccgttcttgc ggacgtcctc gatgatgggg acaatgatct 7320 tgtagatggc atcggacgac ttttgcgcag gacgcttgag agcagcatcg agctcctcgg 7380 tagagacctt ggaggcgtcg aaacgtctca tggtgatctt ctcaggggtg gtctcctggg 7440 cggcctcctt ggtgacaggg gccgaagtgg cagcgagagc ggacgccgca ggcttgatgc 7500 cctccttctc agcccactta cccttagcat ctccagtcct gcgcttgacc ttccagctct 7560 tggcgtcaag gctcctttcg atatcggcaa gagtaacgcc ggcagcaacg gccctggtaa 7620 gagcaaagta gaagagatcg gcagcctcaa aggcgatttc ctggggggtc tgagcggtgc 7680 agagctcctc agcctcctcc atgatcttgg cccggactag cttctcatcg gagaagagac 7740 gggcagtgta ggagccctcg ggggcagact gtttcctcga aatcaaagtc tgctcgagct 7800 tgggaaggcc tttgagctga ccaaagcagc cggactgatc gaggtggcag aaacgaccct 7860 tctgcttcac gacaaacttg agagcatcgt tatcgcagtc aagcgagatg cggacgagct 7920 cctgagtgtc tccggaagta gcacccttgt accagagacc gcgcttccgg ctctgataga 7980 caccagtctg tgtcctgagg gcctcgttca cactctcggc actgctgtag accagaccca 8040 gagcagtgtc gtgctcatca acgacaacgg tggggagaag accatcggga cggtcagact 8100 tccagacgct cgagaggatg gtggagatgg caagcttcag ctgctcgagt tc 8152 pCCG::C-Gly::HAT::FLAG"Times New Roman yOENdeI ArialRuyuoeYe yOEScaI Arial" yOE+HpaI(2327),KpnI(2480),NheI(2669),ClaI(2831) Arial BKTT yOE+HpaI(3320),ScaI(3640),XhoI(3652),XhoI(3833) Arial87@II yOENPvuII(3881),SacI(4040),EagI(4053),NotI(4053),ClaI(4111),MluI(4248),SexAI(4325) ArialqDqNqWDW yOE.NheI(4446),HindIII(4796),XbaI(4982),SpeI(4988) Arial$,44d yOEͪBamHI(4994),XmaI(4999),SmaI(5001),PacI(5009),XmaI(5395),SmaI(5397),PstI(5405),EcoRI(5407),EcoRV(5415),HindIII(5419),ClaI(5426),SalI(5434),XhoI(5440),ApaI(5453),KpnI(5459) ArialHC yOE+PstI(5683),XhoI(6084),SacI(6091),ScaI(6202) Arialgygog9Oo yOE6HpaI(6302),SacI(6445),NcoI(6446),SalI(6795),NcoI(6827) Arial.'$v!?v? yOE8NcoI(6968),KpnI(7018),PstI(7070),Bsu36I(7176),SacI(7375) Arialn{ yOEDEcoRV(7583),SacI(7687),XhoI(7794),SacI(7921),Bsu36I(7996),XhoI(8110) Arialr)&xm%m yOE$HindIII(8133),PvuII(8141),XhoI(8145) ArialdydgvQv-QyOEC Sp6_primer"Times New RomanyOE pGEX_3_primer"Times New RomanyOEC AmpR_promoter"Times New RomanyOEC0 Ampicillin"Times New RomanyOE0ORF 2"Times New RomanyOECɆ6 pBR322_origin"Times New RomanyOEC) ;  T7_promoter"Times New RomanyOEC T7_promoter"Times New RomanyOECupBluescriptSK_primer"Times New RomanyOEjpHybLex_rev_primer"Times New RomanyOECɆ3CpBluescriptKS_primer"Times New RomanyOEC_r T3_promoter"Times New RomanyOECORF 1"Times New RomanyOEC(ORF 1"Times New Roman yOEtest"Times New RomanAvkO;kh yOEtest"Times New Romans