[[1ID FJ457007; SV 1; circular; other DNA; STD; SYN; 7793 BP. AC FJ457007; DT 30-JAN-2009 (Rel. 99, Created) DT 30-JAN-2009 (Rel. 99, Last updated, Version 1) DE Neurospora expression vector pCCG::N-FLAG::HAT, complete sequence. KW . OS Neurospora expression vector pCCG::N-FLAG::HAT OC other sequences; artificial sequences; vectors. RN [1] RP 1-7793 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 1-7793 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..7793 FT /organism="Neurospora expression vector pCCG::N-FLAG::HAT" FT /mol_type="other DNA" FT /db_xref="taxon:591010" 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="ACM10009.1" 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 4092..4934 FT /note="Neurospora crassa ccg-1 (grg-1)" FT misc_feature 4936..4962 FT /note="FLAG" FT misc_feature 4963..4971 FT /note="3xGly" FT misc_feature 4981..5040 FT /note="HAT" FT misc_feature 5041..5055 FT /note="HAT" FT misc_feature complement(5673..7774) FT /note="5' truncated histidine biosynthesis trifunctional FT protein; includes phosphoribosyl-AMP cyclohydrolase, FT phosphoribosyl-ATP cyclohydrolase and histindinol FT dehydrogenase" SQ Sequence 7793 BP; 1934 A; 2054 C; 2005 G; 1800 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 tggccctaat taaccctcac 4020 taaagggaac aaaagctggg taccgggccc cccctcgagg tcgacggtat cgataagctt 4080 gatatcgaat tcgttcaaag ccacatcact gggcacttcc attgggacag gcattgatcg 4140 gacgagaccg acttctggcc gctttcaaca gccacattat atccatgtca cggctacgcg 4200 tcggccttcg gtaacagaaa agcacacaga cagcgattgt gacatggatt cgggcaaagc 4260 attggtggtc gcaccaggtc atcctgagtg tgcagtggct gctattcaga tttcatctaa 4320 ctgcgggaga ggggttcaaa ggggcgtgac gtcacagaca acgggtgaag gacgaagatt 4380 gcctcacttc tttgctagca atttgcctgc aaagaaggcg cacatgacaa gcaaacaact 4440 gggaaaccac tattgaatac ccacaatgca aagctcggaa ggtacgtctt gattgcagtg 4500 tgtcgagtgt caaaaaagaa gcaagtgttc atgcaagcca aaattggcac ctcctccact 4560 tctccgagtg ccccacccga acctccaggc gagatggccg gaacatacca tccgcgttgg 4620 gattatgacg tatctccttc ttcttcacat gattccatcc cgttgttgct tgtttgcgag 4680 ctgtgacggg agatcgtaga tgccacttcg ggccaggcag gcagtgcagg cagccaggaa 4740 cacaagcttc caacttggtc atctcgattg ccgattaagg gaaccaaatg cctatataag 4800 actgtcctcc cacctcccca ataccattct tttcttcttc catcatcagc caacaaagca 4860 atcacatctt cactacttca aatcaacaca acactcaaac cactttcaca acccctcaca 4920 tcaaccaaag gatccatgga ctacaaagac gatgacgaca agggcggagg cttaatcggc 4980 ttgaaggatc atctcatcca caatgtccac aaagaggagc acgctcatgc ccacaacaag 5040 ggcggcggag gcggcgcgcc attaattaac tagttctaga gcggccgcca ccgcggtgga 5100 gctccaattc gccctatagt gagtcgtatt aagggcgtgg acggctaatg gggtctgaat 5160 gctaaaagac accatttccc acactccctc tgattttttt tatggttttt ttttatcttc 5220 atttttcgcc gtcttttatt cggttgtttg ggaggttcac atgcaacgtg caagactgca 5280 agttttttga gatgggcgct tcatgcaatg cagcatcact gcagggacat tgtgacatcc 5340 acgtttcgat cgtctgtgct tcccccgttc atgtacgcat gatctgaaaa aaaaaaaaaa 5400 aaaaaaggtt gatgtaataa tgaaaataaa cacccaggga cgagcacctc atctgctgga 5460 atagggagaa aaactcaagg gccgtttggg tctccacact taccaacaga ttcatgtact 5520 gatcttcagc ctgtacagaa cagtacaatg tccaaaactc gttatcatta ctctatgtgc 5580 ccaaaatgac ccagatatac atcaacccac tacatccttt tgttcttgtt tttttttttt 5640 ttttttgcta cttcgaaaag aatttccgtt tagttgctct tgctcatgtg ctcaagacgg 5700 atgctgaccg cccttctgtg agcctcgagc tcctcaacct tagccaactg catgacagcc 5760 tggccgacgt ttttgagacc ctcggcagtc aagttggagc tggtaatgtg cttgacgaac 5820 gaggcgaaat tgacgccaga gtactgcttg ccaaagccat aggtagctat gtgtcttgac 5880 gttagcagaa gaatctgtga agcggggcta tttatgatat gtaacttacg cagcgagtgg 5940 ttaacaccag cagagtaatc gccaacggac tcaggagtcc aggcgccaat gaagacacta 6000 ccggcgttca tgacaagatc gacagccttc tcggcctcct tgatctggag gatcaagtgc 6060 tcaggagcgt acttgttgct gagctccatg gcctcctcga cggtcttgac ctgcacggtg 6120 atcgagtggg cgatggagcc acggacaatc tggacgcgag gaagctccgt agcctgacgg 6180 tgaacctcgt cctcaatagc ctgaagatgc tcctcgtcga ggtcaatagc gatcaggatg 6240 acctgactgt caacgccgtg ctcagcctgg gacaggagat ccgaggcaac gaacgcgggg 6300 ttggcgtcct tgtcagcgat gaccagcacc tcggacgggc cagcgggcat gtcaatacca 6360 acggcagcgt tggtgtcgtt gctgacgaac atcttggcag cagtgacgaa ctggttaccg 6420 gggccgagaa tcttgtcgac cttggtgatg ctctcggtgc cgtaggccat ggcagctacg 6480 gcctgggcac cgccggcaag cacgatggac tcggccccaa ccttgtgagc gacgtggaca 6540 atctcgggag tgatggttcc gtcggcgcgg ggaggagagg cgaacacaat cttgttgcag 6600 ccggcgacca tggcgggaac acccagcata agggcagtgc tggggagaac ggcggtaccg 6660 ccggggatgt agcagccgac ggcctcgatg ggacgagaga aacggctgca gacaacaccg 6720 ggcatggtct cgacctggag gggcttctcc tccttctggg cggcgtggaa cttgcggatg 6780 ttctcgaagg acacgtcgat ggcagcaatg gtctcctcag ggagctgcat aagctccttg 6840 gggaagggcg ccttcaggac ggggctagta agagaggtag ccttctcgaa cttgtgagtg 6900 tacgacagaa cagccttgtc gccgttcttg cggacgtcct cgatgatggg gacaatgatc 6960 ttgtagatgg catcggacga cttttgcgca ggacgcttga gagcagcatc gagctcctcg 7020 gtagagacct tggaggcgtc gaaacgtctc atggtgatct tctcaggggt ggtctcctgg 7080 gcggcctcct tggtgacagg ggccgaagtg gcagcgagag cggacgccgc aggcttgatg 7140 ccctccttct cagcccactt acccttagca tctccagtcc tgcgcttgac cttccagctc 7200 ttggcgtcaa ggctcctttc gatatcggca agagtaacgc cggcagcaac ggccctggta 7260 agagcaaagt agaagagatc ggcagcctca aaggcgattt cctggggggt ctgagcggtg 7320 cagagctcct cagcctcctc catgatcttg gcccggacta gcttctcatc ggagaagaga 7380 cgggcagtgt aggagccctc gggggcagac tgtttcctcg aaatcaaagt ctgctcgagc 7440 ttgggaaggc ctttgagctg accaaagcag ccggactgat cgaggtggca gaaacgaccc 7500 ttctgcttca cgacaaactt gagagcatcg ttatcgcagt caagcgagat gcggacgagc 7560 tcctgagtgt ctccggaagt agcacccttg taccagagac cgcgcttccg gctctgatag 7620 acaccagtct gtgtcctgag ggcctcgttc acactctcgg cactgctgta gaccagaccc 7680 agagcagtgt cgtgctcatc aacgacaacg gtggggagaa gaccatcggg acggtcagac 7740 ttccagacgc tcgagaggat ggtggagatg gcaagcttca gctgctcgag ttc 7793 qpCCG::N-FLAG::HAT"Times New Roman [NdeI ArialRq{o{ [ScaI Arialq%+00^ [+HpaI(2327),KpnI(2480),NheI(2669),ClaI(2831) Arial qt} [+HpaI(3320),ScaI(3640),XhoI(3652),XhoI(3833) Arial8qU_hh [͊PvuII(3881),KpnI(4044),ApaI(4050),XhoI(4055),SalI(4061),ClaI(4071),HindIII(4076),EcoRV(4084),EcoRI(4088),MluI(4197),SexAI(4274),NheI(4395) Arial*qXji)i [qHindIII(4745),BamHI(4931),NcoI(4935),AscI(5055),PacI(5067),SpeI(5070),XbaI(5076),EagI(5083),NotI(5083),SacI(5104) Arialq [+PstI(5324),XhoI(5725),SacI(5732),ScaI(5843) Arial]qsw [+HpaI(5943),SacI(6086),NcoI(6087),SalI(6436) ArialqIGEu-] [8NcoI(6468),NcoI(6609),KpnI(6659),PstI(6711),Bsu36I(6817) Arialq [,SacI(7016),EcoRV(7224),SacI(7328),XhoI(7435) Arial8q950f0 [GSacI(7562),Bsu36I(7637),XhoI(7751),HindIII(7774),PvuII(7782),XhoI(7786) ArialGq}nG&n[q Sp6_primer"Times New Roman[Cq pGEX_3_primer"Times New Roman[Cq AmpR_promoter"Times New Roman[Cq0 Ampicillin"Times New Roman[Cq0ORF 1"Times New Roman[q6 pBR322_origin"Times New Roman[CɆq) ;  T7_promoter"Times New Roman[Cq T3_promoter"Times New Roman[CɆqpBluescriptKS_primer"Times New Roman[Cq HAT_primer"Times New Roman[Cq  T7_promoter"Times New Roman[CɆq&ORF 1"Times New Roman[Cq?ORF 2"Times New Roman [test"Times New RomanAqxuh [test"Times New RomanqYI< G< [test"Times New Roman1Sq^ :