i~5Ji~5J(ID FJ457012; SV 1; circular; other DNA; STD; SYN; 5316 BP. AC FJ457012; DT 30-JAN-2009 (Rel. 99, Created) DT 30-JAN-2009 (Rel. 99, Last updated, Version 1) DE Neurospora knock-in vector p13xMyc::hph::loxP, complete sequence. KW . OS Neurospora knock-in vector p13xMyc::hph::loxP OC other sequences; artificial sequences; vectors. RN [1] RP 1-5316 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 1-5316 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..5316 FT /organism="Neurospora knock-in vector p13xMyc::hph::loxP" FT /mol_type="other DNA" FT /db_xref="taxon:591015" 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="ACM10018.1" 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(1811..2338) FT /note="13xMyc" FT misc_feature complement(2348..2377) FT /note="10xGly" FT primer_bind complement(2434..2449) FT /note="M13 (-20) forward primer" FT primer_bind complement(2453..2469) FT /note="M13 (-40) forward primer" FT rep_origin 2914..3329 FT /note="f1" FT CDS complement(3341..4135) FT /codon_start=1 FT /transl_table=11 FT /product="Kan(R)" FT /protein_id="ACM10019.1" FT /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP FT VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS FT SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ FT GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA FT LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" FT promoter complement(4136..4273) FT /note="Kan" FT rep_origin 4521..5194 FT /note="pUC" SQ Sequence 5316 BP; 1209 A; 1480 C; 1312 G; 1315 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 gaattcacta gtgattgatt aatttttgtt caccgttcaa gtcttcctcg gagattagct 1860 tttgttcacc gttcaaatct tcttcagaaa tcaacttttg ttcaccgtcg agtccgttca 1920 agtcttcttc tgagattaat ttttgttcac cgttcaagtc ttcctcggag attagctttt 1980 gttcaccgtt caaatcttct tcagaaatca acttttgttc accgtcgagt ccgttcaagt 2040 cttcttctga gattaatttt tgttcaccgt tcaagtcttc ctcggagatt agcttttgtt 2100 caccgttcaa atcttcttca gaaatcaact tttgttcacc gtcgagtccg ttcaagtctt 2160 cttctgagat taatttttgt tcaccgttca agtcttcctc ggagattagc ttttgttcac 2220 cgttcaaatc ttcttcagaa atcaactttt gttcaccgtc gagtccgttc aagtcttctt 2280 ctgagattaa tttttgttca ccgttcaagt cttcctcgga gattagcttt tgttcaccgt 2340 taattaagcc tccgcctccg cctccgccgc ctccgcctcg agcatgcatc tagagggccc 2400 aattcgccct attcgaagtc gtattacaat tcactggccg tcgttttaca acgtcgtgac 2460 tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc 2520 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctatac 2580 gtacggcagt ttaaggttta cacctataaa agagagagcc gttatcgtct gtttgtggat 2640 gtacagagtg atattattga cacgccgggg cgacggatgg tgatccccct ggccagtgca 2700 cgtctgctgt cagataaagt ctcccgtgaa ctttacccgg tggtgcatat cggggatgaa 2760 agctggcgca tgatgaccac cgatatggcc agtgtgccgg tctccgttat cggggaagaa 2820 gtggctgatc tcagccaccg cgaaaatgac atcaaaaacg ccattaacct gatgttctgg 2880 ggaatataaa tgtcaggcct gaatggcgaa tggacgcgcc ctgtagcggc gcattaagcg 2940 cgcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc 3000 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct 3060 aaatcggggg ctccctttag ggttccgatt tagagcttta cggcacctcg accgcaaaaa 3120 acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc 3180 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact 3240 caaccctatc gcggtctatt cttttgattt ataagggatg ttgccgattt cggcctattg 3300 gttaaaaaat gagctgattt aacaaaaatt ttaacaaaat tcagaagaac tcgtcaagaa 3360 ggcgatagaa ggcgatgcgc tgcgaatcgg gagcggcgat accgtaaagc acgaggaagc 3420 ggtcagccca ttcgccgcca agctcttcag caatatcacg ggtagccaac gctatgtcct 3480 gatagcggtc cgccacaccc agccggccac agtcgatgaa tccagaaaag cggccatttt 3540 ccaccatgat attcggcaag caggcatcgc catgggtcac gacgagatcc tcgccgtcgg 3600 gcatgctcgc cttgagcctg gcgaacagtt cggctggcgc gagcccctga tgctcttcgt 3660 ccagatcatc ctgatcgaca agaccggctt ccatccgagt acgtgctcgc tcgatgcgat 3720 gtttcgcttg gtggtcgaat gggcaggtag ccggatcaag cgtatgcagc cgccgcattg 3780 catcagccat gatggatact ttctcggcag gagcaaggtg agatgacagg agatcctgcc 3840 ccggcacttc gcccaatagc agccagtccc ttcccgcttc agtgacaacg tcgagcacag 3900 ctgcgcaagg aacgcccgtc gtggccagcc acgatagccg cgctgcctcg tcttgcagtt 3960 cattcagggc accggacagg tcggtcttga caaaaagaac cgggcgcccc tgcgctgaca 4020 gccggaacac ggcggcatca gagcagccga ttgtctgttg tgcccagtca tagccgaata 4080 gcctctccac ccaagcggcc ggagaacctg cgtgcaatcc atcttgttca atcatgcgaa 4140 acgatcctca tcctgtctct tgatcagagc ttgatcccct gcgccatcag atccttggcg 4200 gcgagaaagc catccagttt actttgcagg gcttcccaac cttaccagag ggcgccccag 4260 ctggcaattc cggttcgctt gctgtccata aaaccgccca gtctagctat cgccatgtaa 4320 gcccactgca agctacctgc tttctctttg cgcttgcgtt ttcccttgtc cagatagccc 4380 agtagctgac attcatccgg ggtcagcacc gtttctgcgg actggctttc tacgtgaaaa 4440 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt 4500 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt 4560 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt 4620 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga 4680 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag 4740 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata 4800 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg 4860 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga 4920 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca 4980 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa 5040 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt 5100 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac 5160 ggttcctggg cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt 5220 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga 5280 ccgagcgcag cgagtcagtg agcgaggaag cggaag 5316 p13xMyc::hph::loxP"Times New Roman i~5J?PvuII(56),HindIII(277),KpnI(287),SacI(293),BamHI(295),ScaI(406) Arialt,t i~5JEagI Arial} i~5JANdeI(924),EagI(1002),NcoI(1020),PstI(1043),EcoRI(1129),EagI(1167) Arial%/ 8 8u- i~5JClaI Arialc4p>pGpGjxv i~5JFSalI(1742),ClaI(1785),HindIII(1790),EcoRV(1798),EcoRI(1802),SpeI(1808) Arial$$d, i~5J7PacI(2345),XhoI(2378),XbaI(2390),ApaI(2400),PvuII(2520) ArialV =FOO i~5JNcoI(3571),PvuII(3901) Arial }t:t i~5J!EagI(4097),BclI(4162),PvuII(4261) ArialBm8c6Y4"YFi~5JC lac_promoter"Times New Romani~5JM13_reverse_primer"Times New Romani~5JC Sp6_primer"Times New Romani~5JC,MloxP"Times New Romani~5JCɆ_`ORF 1"Times New Romani~5JCjuORF 1"Times New Romani~5JCɆloxP"Times New Romani~5JCɆU ORF 3"Times New Romani~5JC' 13xMyc"Times New Romani~5JCɆ M13_forward20_primer"Times New Romani~5J M13_pUC_fwd_primer"Times New Romani~5JC 'ORF 2"Times New Romani~5JCNEOKAN_promoter"Times New Roman i~5Jtest"Times New Roman=upe;pe8 i~5Jtest"Times New Roman; d