e[1e[13ID FJ457001; SV 1; circular; other DNA; STD; SYN; 8049 BP. AC FJ457001; DT 30-JAN-2009 (Rel. 99, Created) DT 30-JAN-2009 (Rel. 99, Last updated, Version 1) DE Neurospora expression vector pCCG::C-Gly::3xFLAG, complete sequence. KW . OS Neurospora expression vector pCCG::C-Gly::3xFLAG OC other sequences; artificial sequences; vectors. RN [1] RP 1-8049 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 1-8049 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..8049 FT /organism="Neurospora expression vector FT pCCG::C-Gly::3xFLAG" FT /mol_type="other DNA" FT /db_xref="taxon:591004" 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="ACM10001.1" 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 5043..5108 FT /note="3xFLAG" FT misc_feature complement(5303..5387) FT /note="T3 promoter and multiple cloning site from FT pBluescript KS+" FT misc_feature complement(6123..8030) FT /note="5' truncated histidine biosynthesis trifunctional FT protein; includes phosphoribosyl-AMP cyclohydrolase, FT phosphoribosyl-ATP cyclohydrolase and histindinol FT dehydrogenase" SQ Sequence 8049 BP; 1988 A; 2104 C; 2085 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 agggcggagg cggcggaggc ggaggcggag 5040 gcgactacaa agaccatgac ggtgattata aagatcatga catcgactac aaggatgacg 5100 atgacaagta gtgatcccgg gtggcatccc tgtgacccct ccccagtgcc tctcctggcc 5160 ctggaagttg ccactccagt gcccaccagc cttgtcctaa taaaattaag ttgcatcatt 5220 ttgtctgact aggtgtcctt ctataatatt atggggtgga ggggggtggt atggagcaag 5280 gggcaagttg ggaagacaac ctgaattcga tatcaagctt atcgataccg tcgacctcga 5340 gggggggccc ggtacccagc ttttgttccc tttagtgagg gttaattagg gcgtggacgg 5400 ctaatggggt ctgaatgcta aaagacacca tttcccacac tccctctgat tttttttatg 5460 gttttttttt atcttcattt ttcgccgtct tttattcggt tgtttgggag gttcacatgc 5520 aacgtgcaag actgcaagtt ttttgagatg ggcgcttcat gcaatgcagc atcactgcag 5580 ggacattgtg acatccacgt ttcgatcgtc tgtgcttccc ccgttcatgt acgcatgatc 5640 tgaaaaaaaa aaaaaaaaaa aaggttgatg taataatgaa aataaacacc cagggacgag 5700 cacctcatct gctggaatag ggagaaaaac tcaagggccg tttgggtctc cacacttacc 5760 aacagattca tgtactgatc ttcagcctgt acagaacagt acaatgtcca aaactcgtta 5820 tcattactct atgtgcccaa aatgacccag atatacatca acccactaca tccttttgtt 5880 cttgtttttt tttttttttt ttgctacttc gaaaagaatt tccgtttagt tgctcttgct 5940 catgtgctca agacggatgc tgaccgccct tctgtgagcc tcgagctcct caaccttagc 6000 caactgcatg acagcctggc cgacgttttt gagaccctcg gcagtcaagt tggagctggt 6060 aatgtgcttg acgaacgagg cgaaattgac gccagagtac tgcttgccaa agccataggt 6120 agctatgtgt cttgacgtta gcagaagaat ctgtgaagcg gggctattta tgatatgtaa 6180 cttacgcagc gagtggttaa caccagcaga gtaatcgcca acggactcag gagtccaggc 6240 gccaatgaag acactaccgg cgttcatgac aagatcgaca gccttctcgg cctccttgat 6300 ctggaggatc aagtgctcag gagcgtactt gttgctgagc tccatggcct cctcgacggt 6360 cttgacctgc acggtgatcg agtgggcgat ggagccacgg acaatctgga cgcgaggaag 6420 ctccgtagcc tgacggtgaa cctcgtcctc aatagcctga agatgctcct cgtcgaggtc 6480 aatagcgatc aggatgacct gactgtcaac gccgtgctca gcctgggaca ggagatccga 6540 ggcaacgaac gcggggttgg cgtccttgtc agcgatgacc agcacctcgg acgggccagc 6600 gggcatgtca ataccaacgg cagcgttggt gtcgttgctg acgaacatct tggcagcagt 6660 gacgaactgg ttaccggggc cgagaatctt gtcgaccttg gtgatgctct cggtgccgta 6720 ggccatggca gctacggcct gggcaccgcc ggcaagcacg atggactcgg ccccaacctt 6780 gtgagcgacg tggacaatct cgggagtgat ggttccgtcg gcgcggggag gagaggcgaa 6840 cacaatcttg ttgcagccgg cgaccatggc gggaacaccc agcataaggg cagtgctggg 6900 gagaacggcg gtaccgccgg ggatgtagca gccgacggcc tcgatgggac gagagaaacg 6960 gctgcagaca acaccgggca tggtctcgac ctggaggggc ttctcctcct tctgggcggc 7020 gtggaacttg cggatgttct cgaaggacac gtcgatggca gcaatggtct cctcagggag 7080 ctgcataagc tccttgggga agggcgcctt caggacgggg ctagtaagag aggtagcctt 7140 ctcgaacttg tgagtgtacg acagaacagc cttgtcgccg ttcttgcgga cgtcctcgat 7200 gatggggaca atgatcttgt agatggcatc ggacgacttt tgcgcaggac gcttgagagc 7260 agcatcgagc tcctcggtag agaccttgga ggcgtcgaaa cgtctcatgg tgatcttctc 7320 aggggtggtc tcctgggcgg cctccttggt gacaggggcc gaagtggcag cgagagcgga 7380 cgccgcaggc ttgatgccct ccttctcagc ccacttaccc ttagcatctc cagtcctgcg 7440 cttgaccttc cagctcttgg cgtcaaggct cctttcgata tcggcaagag taacgccggc 7500 agcaacggcc ctggtaagag caaagtagaa gagatcggca gcctcaaagg cgatttcctg 7560 gggggtctga gcggtgcaga gctcctcagc ctcctccatg atcttggccc ggactagctt 7620 ctcatcggag aagagacggg cagtgtagga gccctcgggg gcagactgtt tcctcgaaat 7680 caaagtctgc tcgagcttgg gaaggccttt gagctgacca aagcagccgg actgatcgag 7740 gtggcagaaa cgacccttct gcttcacgac aaacttgaga gcatcgttat cgcagtcaag 7800 cgagatgcgg acgagctcct gagtgtctcc ggaagtagca cccttgtacc agagaccgcg 7860 cttccggctc tgatagacac cagtctgtgt cctgagggcc tcgttcacac tctcggcact 7920 gctgtagacc agacccagag cagtgtcgtg ctcatcaacg acaacggtgg ggagaagacc 7980 atcgggacgg tcagacttcc agacgctcga gaggatggtg gagatggcaa gcttcagctg 8040 ctcgagttc 8049 qpCCG::C-Gly::3xFLAG"Times New Roman e[1NdeI ArialRqe[QlEQ e[1ScaI Arialq{o.{ e[1+HpaI(2327),KpnI(2480),NheI(2669),ClaI(2831) Arial qMV__ e[1+HpaI(3320),ScaI(3640),XhoI(3652),XhoI(3833) Arial8q%/88x e[1NPvuII(3881),SacI(4040),EagI(4053),NotI(4053),ClaI(4111),MluI(4248),SexAI(4325) Arialqp0p:oCBC e[1#NheI(4446),HindIII(4796),XbaI(4982) Arialq = e[1ͪSpeI(4988),BamHI(4994),XmaI(4999),SmaI(5001),PacI(5009),XmaI(5117),SmaI(5119),EcoRI(5304),EcoRV(5312),HindIII(5316),ClaI(5323),SalI(5331),XhoI(5337),ApaI(5350),KpnI(5356) ArialqM8 e[1+PstI(5580),XhoI(5981),SacI(5988),ScaI(6099) Arial]qWW{WE?{o e[16HpaI(6199),SacI(6342),NcoI(6343),SalI(6692),NcoI(6724) ArialqJ0 e[18NcoI(6865),KpnI(6915),PstI(6967),Bsu36I(7073),SacI(7272) Arial7qwj e[1DEcoRV(7480),SacI(7584),XhoI(7691),SacI(7818),Bsu36I(7893),XhoI(8007) Arial q1o.e'\'\ e[1$HindIII(8030),PvuII(8038),XhoI(8042) ArialfqpepSfD' fDe[1Cq Sp6_primer"Times New Romane[1Cq pGEX_3_primer"Times New Romane[1Cq AmpR_promoter"Times New Romane[1Cq0 Ampicillin"Times New Romane[1CɆq0ORF 2"Times New Romane[1q6 pBR322_origin"Times New Romane[1Cq) ;  T7_promoter"Times New Romane[1CɆq T7_promoter"Times New Romane[1CɆqupBluescriptSK_primer"Times New Romane[1CqEK"Times New Romane[1qpBluescriptKS_primer"Times New Romane[1Cq  T3_promoter"Times New Romane[1Cq&ORF 1"Times New Romane[1CɆq?ORF 1"Times New Roman e[1test"Times New RomanAqkbZ>DZh e[1test"Times New Roman@q