MM2ID FJ456996; SV 1; circular; other DNA; STD; SYN; 8019 BP. AC FJ456996; DT 30-JAN-2009 (Rel. 99, Created) DT 30-JAN-2009 (Rel. 99, Last updated, Version 1) DE Neurospora expression vector pCCG::C-3xFLAG, complete sequence. KW . OS Neurospora expression vector pCCG::C-3xFLAG OC other sequences; artificial sequences; vectors. RN [1] RP 1-8019 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 1-8019 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..8019 FT /organism="Neurospora expression vector pCCG::C-3xFLAG" FT /mol_type="other DNA" FT /db_xref="taxon:591003" 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="ACM09996.1" FT /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI FT ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS FT PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW FT EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA FT LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS FT LIKHW" FT misc_feature complement(2345..4006) FT /note="3' flank region of Neurospora crassa his-3 gene" FT promoter complement(2345..2365) FT /note="T7" FT misc_feature 5013..5078 FT /note="3xFLAG" FT promoter 5013..5078 FT /note="Neurospora crassa ccg-1 (grg-1)" FT misc_feature complement(5237..5357) FT /note="T3 promoter and multiple cloning site from FT pBluescript KS+" FT misc_feature complement(6093..8000) FT /note="5' truncated histidine biosynthesis trifunctional FT protein; includes phosphoribosyl-AMP cyclohydrolase, FT phosphoribosyl-ATP cyclohydrolase and histindinol FT dehydrogenase" SQ Sequence 8019 BP; 1985 A; 2098 C; 2064 G; 1872 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 aagactacaa agaccatgac ggtgattata 5040 aagatcatga catcgactac aaggatgacg atgacaagta gtgatcccgg gtggcatccc 5100 tgtgacccct ccccagtgcc tctcctggcc ctggaagttg ccactccagt gcccaccagc 5160 cttgtcctaa taaaattaag ttgcatcatt ttgtctgact aggtgtcctt ctataatatt 5220 atggggtgga ggggggtggt atggagcaag gggcaagttg ggaagacaac ctgaattcga 5280 tatcaagctt atcgataccg tcgacctcga gggggggccc ggtacccagc ttttgttccc 5340 tttagtgagg gttaattagg gcgtggacgg ctaatggggt ctgaatgcta aaagacacca 5400 tttcccacac tccctctgat tttttttatg gttttttttt atcttcattt ttcgccgtct 5460 tttattcggt tgtttgggag gttcacatgc aacgtgcaag actgcaagtt ttttgagatg 5520 ggcgcttcat gcaatgcagc atcactgcag ggacattgtg acatccacgt ttcgatcgtc 5580 tgtgcttccc ccgttcatgt acgcatgatc tgaaaaaaaa aaaaaaaaaa aaggttgatg 5640 taataatgaa aataaacacc cagggacgag cacctcatct gctggaatag ggagaaaaac 5700 tcaagggccg tttgggtctc cacacttacc aacagattca tgtactgatc ttcagcctgt 5760 acagaacagt acaatgtcca aaactcgtta tcattactct atgtgcccaa aatgacccag 5820 atatacatca acccactaca tccttttgtt cttgtttttt tttttttttt ttgctacttc 5880 gaaaagaatt tccgtttagt tgctcttgct catgtgctca agacggatgc tgaccgccct 5940 tctgtgagcc tcgagctcct caaccttagc caactgcatg acagcctggc cgacgttttt 6000 gagaccctcg gcagtcaagt tggagctggt aatgtgcttg acgaacgagg cgaaattgac 6060 gccagagtac tgcttgccaa agccataggt agctatgtgt cttgacgtta gcagaagaat 6120 ctgtgaagcg gggctattta tgatatgtaa cttacgcagc gagtggttaa caccagcaga 6180 gtaatcgcca acggactcag gagtccaggc gccaatgaag acactaccgg cgttcatgac 6240 aagatcgaca gccttctcgg cctccttgat ctggaggatc aagtgctcag gagcgtactt 6300 gttgctgagc tccatggcct cctcgacggt cttgacctgc acggtgatcg agtgggcgat 6360 ggagccacgg acaatctgga cgcgaggaag ctccgtagcc tgacggtgaa cctcgtcctc 6420 aatagcctga agatgctcct cgtcgaggtc aatagcgatc aggatgacct gactgtcaac 6480 gccgtgctca gcctgggaca ggagatccga ggcaacgaac gcggggttgg cgtccttgtc 6540 agcgatgacc agcacctcgg acgggccagc gggcatgtca ataccaacgg cagcgttggt 6600 gtcgttgctg acgaacatct tggcagcagt gacgaactgg ttaccggggc cgagaatctt 6660 gtcgaccttg gtgatgctct cggtgccgta ggccatggca gctacggcct gggcaccgcc 6720 ggcaagcacg atggactcgg ccccaacctt gtgagcgacg tggacaatct cgggagtgat 6780 ggttccgtcg gcgcggggag gagaggcgaa cacaatcttg ttgcagccgg cgaccatggc 6840 gggaacaccc agcataaggg cagtgctggg gagaacggcg gtaccgccgg ggatgtagca 6900 gccgacggcc tcgatgggac gagagaaacg gctgcagaca acaccgggca tggtctcgac 6960 ctggaggggc ttctcctcct tctgggcggc gtggaacttg cggatgttct cgaaggacac 7020 gtcgatggca gcaatggtct cctcagggag ctgcataagc tccttgggga agggcgcctt 7080 caggacgggg ctagtaagag aggtagcctt ctcgaacttg tgagtgtacg acagaacagc 7140 cttgtcgccg ttcttgcgga cgtcctcgat gatggggaca atgatcttgt agatggcatc 7200 ggacgacttt tgcgcaggac gcttgagagc agcatcgagc tcctcggtag agaccttgga 7260 ggcgtcgaaa cgtctcatgg tgatcttctc aggggtggtc tcctgggcgg cctccttggt 7320 gacaggggcc gaagtggcag cgagagcgga cgccgcaggc ttgatgccct ccttctcagc 7380 ccacttaccc ttagcatctc cagtcctgcg cttgaccttc cagctcttgg cgtcaaggct 7440 cctttcgata tcggcaagag taacgccggc agcaacggcc ctggtaagag caaagtagaa 7500 gagatcggca gcctcaaagg cgatttcctg gggggtctga gcggtgcaga gctcctcagc 7560 ctcctccatg atcttggccc ggactagctt ctcatcggag aagagacggg cagtgtagga 7620 gccctcgggg gcagactgtt tcctcgaaat caaagtctgc tcgagcttgg gaaggccttt 7680 gagctgacca aagcagccgg actgatcgag gtggcagaaa cgacccttct gcttcacgac 7740 aaacttgaga gcatcgttat cgcagtcaag cgagatgcgg acgagctcct gagtgtctcc 7800 ggaagtagca cccttgtacc agagaccgcg cttccggctc tgatagacac cagtctgtgt 7860 cctgagggcc tcgttcacac tctcggcact gctgtagacc agacccagag cagtgtcgtg 7920 ctcatcaacg acaacggtgg ggagaagacc atcgggacgg tcagacttcc agacgctcga 7980 gaggatggtg gagatggcaa gcttcagctg ctcgagttc 8019 SpCCG::C-3xFLAG"Times New Roman MNdeI ArialRSwmam MScaI ArialSE9EE{s M+HpaI(2327),KpnI(2480),NheI(2669),ClaI(2831) Arial S M+HpaI(3320),ScaI(3640),XhoI(3652),XhoI(3833) Arial8SB L UU' MNPvuII(3881),SacI(4040),EagI(4053),NotI(4053),ClaI(4111),MluI(4248),SexAI(4325) ArialSLV__ MNheI(4446),HindIII(4796) Arial^Sz@vIrReR M͵XbaI(4982),SpeI(4988),BamHI(4994),XmaI(4999),SmaI(5001),PacI(5009),XmaI(5087),SmaI(5089),EcoRI(5274),EcoRV(5282),HindIII(5286),ClaI(5293),SalI(5301),XhoI(5307),ApaI(5320),KpnI(5326) Arial ZS     M+PstI(5550),XhoI(5951),SacI(5958),ScaI(6069) Arial?Ssss[ M6HpaI(6169),SacI(6312),NcoI(6313),SalI(6662),NcoI(6694) ArialS31.L M8NcoI(6835),KpnI(6885),PstI(6937),Bsu36I(7043),SacI(7242) ArialS MDEcoRV(7450),SacI(7554),XhoI(7661),SacI(7788),Bsu36I(7863),XhoI(7977) ArialS~qw1/qw M$HindIII(8000),PvuII(8008),XhoI(8012) ArialHSulpHlMCS Sp6_primer"Times New RomanMCS pGEX_3_primer"Times New RomanMCS AmpR_promoter"Times New RomanMCS0 Ampicillin"Times New RomanMCɆS0ORF 2"Times New RomanMS6 pBR322_origin"Times New RomanMCS) ;  T7_promoter"Times New RomanMCɆS T7_promoter"Times New RomanMCɆSupBluescriptSK_primer"Times New RomanMCSEK"Times New RomanMSpBluescriptKS_primer"Times New RomanMCS T3_promoter"Times New RomanMCSORF 1"Times New RomanMCɆS!ORF 1"Times New Roman Mtest"Times New RomanASmh Mtest"Times New Roman1S//