`O,`O,L'ID FJ457009; SV 1; circular; other DNA; STD; SYN; 5044 BP. AC FJ457009; DT 30-JAN-2009 (Rel. 99, Created) DT 30-JAN-2009 (Rel. 99, Last updated, Version 1) DE Neurospora knock-in vector p3xFLAG::hph::loxP, complete sequence. KW . OS Neurospora knock-in vector p3xFLAG::hph::loxP OC other sequences; artificial sequences; vectors. RN [1] RP 1-5044 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 1-5044 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..5044 FT /organism="Neurospora knock-in vector p3xFLAG::hph::loxP" FT /mol_type="other DNA" FT /db_xref="taxon:591016" FT promoter 95..216 FT /note="lac" FT primer_bind 205..221 FT /note="M13 reverse primer" FT primer_bind 239..256 FT /note="SP6 primer" FT misc_binding complement(300..333) FT /bound_moiety="LoxP" FT CDS complement(351..1376) FT /codon_start=1 FT /transl_table=11 FT /product="hph" FT /note="hygromycin resistance" FT /protein_id="ACM10011.1" FT /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG FT YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP FT ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ FT TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF FT GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG FT NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK FT E" FT promoter complement(1382..1746) FT /note="trpC" FT misc_binding complement(1751..1784) FT /bound_moiety="LoxP" FT misc_feature complement(2001..2066) FT /note="3xFLAG" FT misc_feature complement(2076..2105) FT /note="10xGly" FT primer_bind complement(2162..2177) FT /note="M13 (-20) forward primer" FT primer_bind complement(2181..2197) FT /note="M13 (-40) forward primer" FT rep_origin 2642..3057 FT /note="f1" FT CDS complement(3069..3863) FT /codon_start=1 FT /transl_table=11 FT /product="Kan(R)" FT /protein_id="ACM10012.1" FT /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP FT VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS FT SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ FT GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA FT LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" FT promoter complement(3864..4001) FT /note="Kan" FT rep_origin 4249..4922 FT /note="pUC" SQ Sequence 5044 BP; 1163 A; 1424 C; 1289 G; 1168 T; 0 other; agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc 60 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc 120 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa 180 ttgtgagcga ataacaattt cacacaggaa acagctatga ccatgattac gccaagctat 240 ttaggtgaca ctatagaata ctcaagctat gcatcaagct tggtaccgag ctcggatcca 300 taacttcgta tagcatacat tatacgaagt tatcccggtc ggcatctact ctattccttt 360 gccctcggac gagtgctggg gcgtcggttt ccactatcgg cgagtacttc tacacagcca 420 tcggtccaga cggccgcgct tctgcgggcg atttgtgtac gcccgacagt cccggctccg 480 gatcggacga ttgcgtcgca tcgaccctgc gcccaagctg catcatcgaa attgccgtca 540 accaagctct gatagagttg gtcaagacca atgcggagca tatacgcccg gagccgcggc 600 gatcctgcaa gctccggatg cctccgctcg aagtagcgcg tctgctgctc catacaagcc 660 aaccacggcc tccagaagaa gatgttggcg acctcgtatt gggaatcccc gaacatcgcc 720 tcgctccagt caatgaccgc tgttatgcgg ccattgtccg tcaggacatt gttggagccg 780 aaatccgcgt gcacgaggtg ccggacttcg gggcagtcct cggcccaaag catcagctca 840 tcgagagcct gcgcgacgga cgcactgacg gtgtcgtcca tcacagtttg ccagtgatac 900 acatggggat cagcaatcgc gcatatgaaa tcacgccatg tagtgtattg accgattcct 960 tgcggtccga atgggccgaa cccgctcgtc tggctaagat cggccgcagc gatcgcatcc 1020 atggcctccg cgaccggctg cagaacagcg ggcagttcgg tttcaggcag gtcttgcaac 1080 gtgacaccct gtgcacggcg ggagatgcaa taggtcaggc tctcgctgaa ttccccaatg 1140 tcaagcactt ccggaatcgg gagcgcggcc gatgcaaagt gccgataaac ataacgatct 1200 ttgtagaaac catcggcgca gctatttacc cgcaggacat atccacgccc tcctacatcg 1260 aagctgaaag cacgagattc ttcgccctcc gagagctgca tcaggtcgga gacgctgtcg 1320 aacttttcga tcagaaactt ctcgacagac gtcgcggtga gttcaggctt tttcatatcg 1380 atgcttgggt agaataggta agtcagattg aatctgaaat aaagggagga agggcgaact 1440 taagaaggta tgaccgggtc gttcacttac cttgcttgac aaacgcacca agttatcgtg 1500 caccaagcag cagatgataa taatgtcctc gttcctgtct gctaataaga gtcacacttc 1560 gagcgccgcc gctactgcta caagtggggc tgatctgacc agttgcctaa atgaaccatc 1620 ttgtcaaacg acacaaattt tgtgctcacc gcctggacga ctaaaccaaa ataggcattc 1680 attgttgacc tccactagct ccagccaagc ccaaaaagtg ctccttcaat atcatcttct 1740 gtcgactaga ataacttcgt atagcataca ttatacgaag ttatcgataa gcttgatatc 1800 gaattcaggt tgtcttccca acttgcccct tgctccatac cacccccctc caccccataa 1860 tattatagaa ggacacctag tcagacaaaa tgatgcaact taattttatt aggacaaggc 1920 tggtgggcac tggagtggca acttccaggg ccaggagagg cactggggag gggtcacagg 1980 gatgccaccc gggatcacta cttgtcatcg tcatccttgt agtcgatgtc atgatcttta 2040 taatcaccgt catggtcttt gtagtcttta attaagcctc cgcctccgcc tccgccgcct 2100 ccgcctcgag catgcatcta gagggcccaa ttcgccctat tcgaagtcgt attacaattc 2160 actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg 2220 ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg 2280 cccttcccaa cagttgcgca gcctatacgt acggcagttt aaggtttaca cctataaaag 2340 agagagccgt tatcgtctgt ttgtggatgt acagagtgat attattgaca cgccggggcg 2400 acggatggtg atccccctgg ccagtgcacg tctgctgtca gataaagtct cccgtgaact 2460 ttacccggtg gtgcatatcg gggatgaaag ctggcgcatg atgaccaccg atatggccag 2520 tgtgccggtc tccgttatcg gggaagaagt ggctgatctc agccaccgcg aaaatgacat 2580 caaaaacgcc attaacctga tgttctgggg aatataaatg tcaggcctga atggcgaatg 2640 gacgcgccct gtagcggcgc attaagcgcg cgggtgtggt ggttacgcgc agcgtgaccg 2700 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca 2760 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta 2820 gagctttacg gcacctcgac cgcaaaaaac ttgatttggg tgatggttca cgtagtgggc 2880 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg 2940 gactcttgtt ccaaactgga acaacactca accctatcgc ggtctattct tttgatttat 3000 aagggatgtt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaatttt 3060 aacaaaattc agaagaactc gtcaagaagg cgatagaagg cgatgcgctg cgaatcggga 3120 gcggcgatac cgtaaagcac gaggaagcgg tcagcccatt cgccgccaag ctcttcagca 3180 atatcacggg tagccaacgc tatgtcctga tagcggtccg ccacacccag ccggccacag 3240 tcgatgaatc cagaaaagcg gccattttcc accatgatat tcggcaagca ggcatcgcca 3300 tgggtcacga cgagatcctc gccgtcgggc atgctcgcct tgagcctggc gaacagttcg 3360 gctggcgcga gcccctgatg ctcttcgtcc agatcatcct gatcgacaag accggcttcc 3420 atccgagtac gtgctcgctc gatgcgatgt ttcgcttggt ggtcgaatgg gcaggtagcc 3480 ggatcaagcg tatgcagccg ccgcattgca tcagccatga tggatacttt ctcggcagga 3540 gcaaggtgag atgacaggag atcctgcccc ggcacttcgc ccaatagcag ccagtccctt 3600 cccgcttcag tgacaacgtc gagcacagct gcgcaaggaa cgcccgtcgt ggccagccac 3660 gatagccgcg ctgcctcgtc ttgcagttca ttcagggcac cggacaggtc ggtcttgaca 3720 aaaagaaccg ggcgcccctg cgctgacagc cggaacacgg cggcatcaga gcagccgatt 3780 gtctgttgtg cccagtcata gccgaatagc ctctccaccc aagcggccgg agaacctgcg 3840 tgcaatccat cttgttcaat catgcgaaac gatcctcatc ctgtctcttg atcagagctt 3900 gatcccctgc gccatcagat ccttggcggc gagaaagcca tccagtttac tttgcagggc 3960 ttcccaacct taccagaggg cgccccagct ggcaattccg gttcgcttgc tgtccataaa 4020 accgcccagt ctagctatcg ccatgtaagc ccactgcaag ctacctgctt tctctttgcg 4080 cttgcgtttt cccttgtcca gatagcccag tagctgacat tcatccgggg tcagcaccgt 4140 ttctgcggac tggctttcta cgtgaaaagg atctaggtga agatcctttt tgataatctc 4200 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag 4260 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa 4320 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg 4380 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag 4440 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg 4500 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga 4560 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc 4620 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc 4680 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga 4740 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt 4800 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg 4860 aaaaacgcca gcaacgcggc ctttttacgg ttcctgggct tttgctggcc ttttgctcac 4920 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga 4980 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg 5040 gaag 5044 p3xFLAG::hph::loxP"Times New Roman `O,5PvuII(56),HindIII(277),KpnI(287),SacI(293),BamHI(295) ArialmdOO `O,ScaI(406),EagI(432) ArialzcYM*e `O,ANdeI(924),EagI(1002),NcoI(1020),PstI(1043),EcoRI(1129),EagI(1167) ArialCMVV `O,ClaI ArialcLfVg_h_bn `O,\SalI(1742),ClaI(1785),HindIII(1790),EcoRV(1798),EcoRI(1802),XmaI(1989),SmaI(1991),PacI(2073) Arial&--m? `O,,XhoI(2106),XbaI(2118),ApaI(2128),PvuII(2248) ArialF&77g `O,NcoI(3299),PvuII(3629) Arial a `O,!EagI(3825),BclI(3890),PvuII(3989) Arial2)z(p&6p8`O,C lac_promoter"Times New Roman`O,M13_reverse_primer"Times New Roman`O,C Sp6_primer"Times New Roman`O,C,MloxP"Times New Roman`O,C_`ORF 2"Times New Roman`O,CjuORF 1"Times New Roman`O,CɆloxP"Times New Roman`O,CEK"Times New Roman`O,CɆqM13_forward20_primer"Times New Roman`O,zM13_pUC_fwd_primer"Times New Roman`O,C ORF 1"Times New Roman`O,CpNEOKAN_promoter"Times New Roman `O,test"Times New Romanf><'Q8 `O,test"Times New Roman0!2B _