~J~J,ID FJ457008; SV 1; circular; other DNA; STD; SYN; 6969 BP. AC FJ457008; DT 30-JAN-2009 (Rel. 99, Created) DT 30-JAN-2009 (Rel. 99, Last updated, Version 1) DE Neurospora expression vector pN-3xHA, complete sequence. KW . OS Neurospora expression vector pN-3xHA OC other sequences; artificial sequences; vectors. RN [1] RP 1-6969 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 1-6969 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..6969 FT /organism="Neurospora expression vector pN-3xHA" FT /mol_type="other DNA" FT /db_xref="taxon:591014" 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="ACM10010.1" 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 misc_feature 4109..4207 FT /note="3xHA" FT misc_feature 4208..4231 FT /note="8xGly" FT misc_feature complement(4849..6950) FT /note="5' truncated histidine biosynthesis trifunctional FT protein; includes phosphoribosyl-AMP cyclohydrolase, FT phosphoribosyl-ATP cyclohydrolase and histindinol FT dehydrogenase" SQ Sequence 6969 BP; 1695 A; 1825 C; 1821 G; 1628 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 tcctgcagcc caagatctat gatcttttac ccatacgatg ttcctgacta 4140 tgcgggctat ccgtatgacg tcccggacta tgcaggatcc tatccatatg acgttccaga 4200 ttacgctggc ggcggaggcg gcggaggcgg cgcgccatta attaactagt tctagagcgg 4260 ccgccaccgc ggtggagctc caattcgccc tatagtgagt cgtattaagg gcgtggacgg 4320 ctaatggggt ctgaatgcta aaagacacca tttcccacac tccctctgat tttttttatg 4380 gttttttttt atcttcattt ttcgccgtct tttattcggt tgtttgggag gttcacatgc 4440 aacgtgcaag actgcaagtt ttttgagatg ggcgcttcat gcaatgcagc atcactgcag 4500 ggacattgtg acatccacgt ttcgatcgtc tgtgcttccc ccgttcatgt acgcatgatc 4560 tgaaaaaaaa aaaaaaaaaa aaggttgatg taataatgaa aataaacacc cagggacgag 4620 cacctcatct gctggaatag ggagaaaaac tcaagggccg tttgggtctc cacacttacc 4680 aacagattca tgtactgatc ttcagcctgt acagaacagt acaatgtcca aaactcgtta 4740 tcattactct atgtgcccaa aatgacccag atatacatca acccactaca tccttttgtt 4800 cttgtttttt tttttttttt ttgctacttc gaaaagaatt tccgtttagt tgctcttgct 4860 catgtgctca agacggatgc tgaccgccct tctgtgagcc tcgagctcct caaccttagc 4920 caactgcatg acagcctggc cgacgttttt gagaccctcg gcagtcaagt tggagctggt 4980 aatgtgcttg acgaacgagg cgaaattgac gccagagtac tgcttgccaa agccataggt 5040 agctatgtgt cttgacgtta gcagaagaat ctgtgaagcg gggctattta tgatatgtaa 5100 cttacgcagc gagtggttaa caccagcaga gtaatcgcca acggactcag gagtccaggc 5160 gccaatgaag acactaccgg cgttcatgac aagatcgaca gccttctcgg cctccttgat 5220 ctggaggatc aagtgctcag gagcgtactt gttgctgagc tccatggcct cctcgacggt 5280 cttgacctgc acggtgatcg agtgggcgat ggagccacgg acaatctgga cgcgaggaag 5340 ctccgtagcc tgacggtgaa cctcgtcctc aatagcctga agatgctcct cgtcgaggtc 5400 aatagcgatc aggatgacct gactgtcaac gccgtgctca gcctgggaca ggagatccga 5460 ggcaacgaac gcggggttgg cgtccttgtc agcgatgacc agcacctcgg acgggccagc 5520 gggcatgtca ataccaacgg cagcgttggt gtcgttgctg acgaacatct tggcagcagt 5580 gacgaactgg ttaccggggc cgagaatctt gtcgaccttg gtgatgctct cggtgccgta 5640 ggccatggca gctacggcct gggcaccgcc ggcaagcacg atggactcgg ccccaacctt 5700 gtgagcgacg tggacaatct cgggagtgat ggttccgtcg gcgcggggag gagaggcgaa 5760 cacaatcttg ttgcagccgg cgaccatggc gggaacaccc agcataaggg cagtgctggg 5820 gagaacggcg gtaccgccgg ggatgtagca gccgacggcc tcgatgggac gagagaaacg 5880 gctgcagaca acaccgggca tggtctcgac ctggaggggc ttctcctcct tctgggcggc 5940 gtggaacttg cggatgttct cgaaggacac gtcgatggca gcaatggtct cctcagggag 6000 ctgcataagc tccttgggga agggcgcctt caggacgggg ctagtaagag aggtagcctt 6060 ctcgaacttg tgagtgtacg acagaacagc cttgtcgccg ttcttgcgga cgtcctcgat 6120 gatggggaca atgatcttgt agatggcatc ggacgacttt tgcgcaggac gcttgagagc 6180 agcatcgagc tcctcggtag agaccttgga ggcgtcgaaa cgtctcatgg tgatcttctc 6240 aggggtggtc tcctgggcgg cctccttggt gacaggggcc gaagtggcag cgagagcgga 6300 cgccgcaggc ttgatgccct ccttctcagc ccacttaccc ttagcatctc cagtcctgcg 6360 cttgaccttc cagctcttgg cgtcaaggct cctttcgata tcggcaagag taacgccggc 6420 agcaacggcc ctggtaagag caaagtagaa gagatcggca gcctcaaagg cgatttcctg 6480 gggggtctga gcggtgcaga gctcctcagc ctcctccatg atcttggccc ggactagctt 6540 ctcatcggag aagagacggg cagtgtagga gccctcgggg gcagactgtt tcctcgaaat 6600 caaagtctgc tcgagcttgg gaaggccttt gagctgacca aagcagccgg actgatcgag 6660 gtggcagaaa cgacccttct gcttcacgac aaacttgaga gcatcgttat cgcagtcaag 6720 cgagatgcgg acgagctcct gagtgtctcc ggaagtagca cccttgtacc agagaccgcg 6780 cttccggctc tgatagacac cagtctgtgt cctgagggcc tcgttcacac tctcggcact 6840 gctgtagacc agacccagag cagtgtcgtg ctcatcaacg acaacggtgg ggagaagacc 6900 atcgggacgg tcagacttcc agacgctcga gaggatggtg gagatggcaa gcttcagctg 6960 ctcgagttc 6969 9pN-3xHA"Times New Roman ~JNdeI ArialR9~tj^j ~JScaI Arial9,399g ~J HpaI(2327),KpnI(2480),NheI(2669) Arial 9NV]]! ~JClaI Arial 9"*$2$2U> ~J HpaI(3320),ScaI(3640),XhoI(3652) Arial89xGwQuZbZ ~JXhoI(3833),PvuII(3881),KpnI(4044),ApaI(4050),XhoI(4055),SalI(4061),ClaI(4071),HindIII(4076),EcoRV(4084),EcoRI(4088),PstI(4098),BglII(4104),BamHI(4176),NdeI(4187),AscI(4231),PacI(4243),SpeI(4246),XbaI(4252),EagI(4259),NotI(4259),SacI(4280) Arial9")1995 ~J PstI(4500),XhoI(4901),SacI(4908) Arial%9s ~J+ScaI(5019),HpaI(5119),SacI(5262),NcoI(5263) Arial9|}~if ~JCSalI(5612),NcoI(5644),NcoI(5785),KpnI(5835),PstI(5887),Bsu36I(5993) Arial9| ~J,SacI(6192),EcoRV(6400),SacI(6504),XhoI(6611) Arial9% [ ~JGSacI(6738),Bsu36I(6813),XhoI(6927),HindIII(6950),PvuII(6958),XhoI(6962) Arial9~l{^;{^~JCɆ9 Sp6_primer"Times New Roman~J9 pGEX_3_primer"Times New Roman~JC9 AmpR_promoter"Times New Roman~JC90 Ampicillin"Times New Roman~J90ORF 2"Times New Roman~JCɆ96 pBR322_origin"Times New Roman~JC9) ;  T7_promoter"Times New Roman~JC9 T3_promoter"Times New Roman~JC9pBluescriptKS_primer"Times New Roman~J9 T7_promoter"Times New Roman~J9wORF 1"Times New Roman~JCɆ9ORF 1"Times New Roman ~Jtest"Times New RomanA9}sWtsh ~Jtest"Times New Roman=9""66%