q[=q[=3ID FJ456999; SV 1; circular; other DNA; STD; SYN; 8112 BP. AC FJ456999; DT 30-JAN-2009 (Rel. 99, Created) DT 30-JAN-2009 (Rel. 99, Last updated, Version 1) DE Neurospora expression vector pCCG::C-HAT::FLAG, complete sequence. KW . OS Neurospora expression vector pCCG::C-HAT::FLAG OC other sequences; artificial sequences; vectors. RN [1] RP 1-8112 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 1-8112 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..8112 FT /organism="Neurospora expression vector pCCG::C-HAT::FLAG" FT /mol_type="other DNA" FT /db_xref="taxon:591006" 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="ACM09999.1" 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 5013..5078 FT /note="Neurospora crassa ccg-1 (grg-1)" FT misc_feature 5013..5063 FT /note="HAT" FT misc_feature 5070..5084 FT /note="5xGly" FT misc_feature 5085..5108 FT /note="1xFLAG" FT misc_feature complement(5366..5450) FT /note="T3 promoter and multiple cloning site from FT pBluescript KS+" FT misc_feature complement(6186..8093) FT /note="5' truncated histidine biosynthesis trifunctional FT protein; includes phosphoribosyl-AMP cyclohydrolase, FT phosphoribosyl-ATP cyclohydrolase and histindinol FT dehydrogenase" SQ Sequence 8112 BP; 2029 A; 2108 C; 2070 G; 1905 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 agaaggatca tctcatccac aatgtccaca 5040 aagaggagca cgctcatgcc cacaacaagg gcggaggcgg aggcgactac aaagacgatg 5100 acgacaagta ggcgcgccac ttctaaataa gcgaatttct tatgatttat gatttttatt 5160 attaaataag ttataaaaaa aataagtgta tacaaatttt aaagtgactc ttaggtttta 5220 aaacgaaaat tcttattctt gagtaactct ttcctgtagg tcaggttgct ttctcaggta 5280 tagtatgagg tcgctcttat tgaccacacc tctaccggca gatccgctag ggataacagg 5340 gtaatataga tcccccgggc tgcaggaatt cgatatcaag cttatcgata ccgtcgacct 5400 cgaggggggg cccggtaccc agcttttgtt ccctttagtg agggttaatt agggcgtgga 5460 cggctaatgg ggtctgaatg ctaaaagaca ccatttccca cactccctct gatttttttt 5520 atggtttttt tttatcttca tttttcgccg tcttttattc ggttgtttgg gaggttcaca 5580 tgcaacgtgc aagactgcaa gttttttgag atgggcgctt catgcaatgc agcatcactg 5640 cagggacatt gtgacatcca cgtttcgatc gtctgtgctt cccccgttca tgtacgcatg 5700 atctgaaaaa aaaaaaaaaa aaaaaggttg atgtaataat gaaaataaac acccagggac 5760 gagcacctca tctgctggaa tagggagaaa aactcaaggg ccgtttgggt ctccacactt 5820 accaacagat tcatgtactg atcttcagcc tgtacagaac agtacaatgt ccaaaactcg 5880 ttatcattac tctatgtgcc caaaatgacc cagatataca tcaacccact acatcctttt 5940 gttcttgttt tttttttttt tttttgctac ttcgaaaaga atttccgttt agttgctctt 6000 gctcatgtgc tcaagacgga tgctgaccgc ccttctgtga gcctcgagct cctcaacctt 6060 agccaactgc atgacagcct ggccgacgtt tttgagaccc tcggcagtca agttggagct 6120 ggtaatgtgc ttgacgaacg aggcgaaatt gacgccagag tactgcttgc caaagccata 6180 ggtagctatg tgtcttgacg ttagcagaag aatctgtgaa gcggggctat ttatgatatg 6240 taacttacgc agcgagtggt taacaccagc agagtaatcg ccaacggact caggagtcca 6300 ggcgccaatg aagacactac cggcgttcat gacaagatcg acagccttct cggcctcctt 6360 gatctggagg atcaagtgct caggagcgta cttgttgctg agctccatgg cctcctcgac 6420 ggtcttgacc tgcacggtga tcgagtgggc gatggagcca cggacaatct ggacgcgagg 6480 aagctccgta gcctgacggt gaacctcgtc ctcaatagcc tgaagatgct cctcgtcgag 6540 gtcaatagcg atcaggatga cctgactgtc aacgccgtgc tcagcctggg acaggagatc 6600 cgaggcaacg aacgcggggt tggcgtcctt gtcagcgatg accagcacct cggacgggcc 6660 agcgggcatg tcaataccaa cggcagcgtt ggtgtcgttg ctgacgaaca tcttggcagc 6720 agtgacgaac tggttaccgg ggccgagaat cttgtcgacc ttggtgatgc tctcggtgcc 6780 gtaggccatg gcagctacgg cctgggcacc gccggcaagc acgatggact cggccccaac 6840 cttgtgagcg acgtggacaa tctcgggagt gatggttccg tcggcgcggg gaggagaggc 6900 gaacacaatc ttgttgcagc cggcgaccat ggcgggaaca cccagcataa gggcagtgct 6960 ggggagaacg gcggtaccgc cggggatgta gcagccgacg gcctcgatgg gacgagagaa 7020 acggctgcag acaacaccgg gcatggtctc gacctggagg ggcttctcct ccttctgggc 7080 ggcgtggaac ttgcggatgt tctcgaagga cacgtcgatg gcagcaatgg tctcctcagg 7140 gagctgcata agctccttgg ggaagggcgc cttcaggacg gggctagtaa gagaggtagc 7200 cttctcgaac ttgtgagtgt acgacagaac agccttgtcg ccgttcttgc ggacgtcctc 7260 gatgatgggg acaatgatct tgtagatggc atcggacgac ttttgcgcag gacgcttgag 7320 agcagcatcg agctcctcgg tagagacctt ggaggcgtcg aaacgtctca tggtgatctt 7380 ctcaggggtg gtctcctggg cggcctcctt ggtgacaggg gccgaagtgg cagcgagagc 7440 ggacgccgca ggcttgatgc cctccttctc agcccactta cccttagcat ctccagtcct 7500 gcgcttgacc ttccagctct tggcgtcaag gctcctttcg atatcggcaa gagtaacgcc 7560 ggcagcaacg gccctggtaa gagcaaagta gaagagatcg gcagcctcaa aggcgatttc 7620 ctggggggtc tgagcggtgc agagctcctc agcctcctcc atgatcttgg cccggactag 7680 cttctcatcg gagaagagac gggcagtgta ggagccctcg ggggcagact gtttcctcga 7740 aatcaaagtc tgctcgagct tgggaaggcc tttgagctga ccaaagcagc cggactgatc 7800 gaggtggcag aaacgaccct tctgcttcac gacaaacttg agagcatcgt tatcgcagtc 7860 aagcgagatg cggacgagct cctgagtgtc tccggaagta gcacccttgt accagagacc 7920 gcgcttccgg ctctgataga caccagtctg tgtcctgagg gcctcgttca cactctcggc 7980 actgctgtag accagaccca gagcagtgtc gtgctcatca acgacaacgg tggggagaag 8040 accatcggga cggtcagact tccagacgct cgagaggatg gtggagatgg caagcttcag 8100 ctgctcgagt tc 8112 pCCG::C-HAT::FLAG"Times New Roman q[=NdeI ArialR qg\P\ q[=ScaI Arial {jd2p q[=+HpaI(2327),KpnI(2480),NheI(2669),ClaI(2831) Arial MV__ q[=+HpaI(3320),ScaI(3640),XhoI(3652),XhoI(3833) Arial809BB q[=NPvuII(3881),SacI(4040),EagI(4053),NotI(4053),ClaI(4111),MluI(4248),SexAI(4325) Arialy<yFyOLO q[=#NheI(4446),HindIII(4796),XbaI(4982) Arial"**N q[=SpeI(4988),BamHI(4994),XmaI(4999),SmaI(5001),PacI(5009),AscI(5113),XmaI(5355),SmaI(5357),PstI(5365),EcoRI(5367),EcoRV(5375),HindIII(5379),ClaI(5386),SalI(5394),XhoI(5400),ApaI(5413),KpnI(5419) ArialAS q[=+PstI(5643),XhoI(6044),SacI(6051),ScaI(6162) Arial__{_EG{w q[=6HpaI(6262),SacI(6405),NcoI(6406),SalI(6755),NcoI(6787) ArialK7 q[=8NcoI(6928),KpnI(6978),PstI(7030),Bsu36I(7136),SacI(7335) Arialvwv q[=DEcoRV(7543),SacI(7647),XhoI(7754),SacI(7881),Bsu36I(7956),XhoI(8070) ArialJ5y2p+e+e q[=$HindIII(8093),PvuII(8101),XhoI(8105) Arialpqp_|L|(Lq[=C Sp6_primer"Times New Romanq[=C pGEX_3_primer"Times New Romanq[=C AmpR_promoter"Times New Romanq[=C0 Ampicillin"Times New Romanq[=0ORF 2"Times New Romanq[=6 pBR322_origin"Times New Romanq[=C) ;  T7_promoter"Times New Romanq[=CɆ T7_promoter"Times New Romanq[=CɆupBluescriptSK_primer"Times New Romanq[=C HAT_primer"Times New Romanq[=CBZpHybLex_rev_primer"Times New Romanq[= pBluescriptKS_primer"Times New Romanq[=CɆ7J T3_promoter"Times New Romanq[=CeORF 1"Times New Romanq[=CɆ~ORF 1"Times New Roman q[=test"Times New RomanAwn_CD_h q[=test"Times New Roman_