PQPQ'ID FJ457010; SV 1; circular; other DNA; STD; SYN; 5147 BP. AC FJ457010; DT 30-JAN-2009 (Rel. 99, Created) DT 30-JAN-2009 (Rel. 99, Last updated, Version 1) DE Neurospora knock-in vector p3xHA::hph::loxP, complete sequence. KW . OS Neurospora knock-in vector p3xHA::hph::loxP OC other sequences; artificial sequences; vectors. RN [1] RP 1-5147 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 1-5147 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..5147 FT /organism="Neurospora knock-in vector p3xHA::hph::loxP" FT /mol_type="other DNA" FT /db_xref="taxon:591136" 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="ACM10013.1" 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(2065..2169) FT /note="3xHA" FT misc_feature complement(2179..2208) FT /note="10xGly" FT primer_bind complement(2265..2280) FT /note="M13 (-20) forward primer" FT primer_bind complement(2284..2300) FT /note="M13 (-40) forward primer" FT rep_origin 2745..3160 FT /note="f1" FT CDS complement(3172..3966) FT /codon_start=1 FT /transl_table=11 FT /product="Kan(R)" FT /protein_id="ACM10014.1" FT /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP FT VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS FT SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ FT GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA FT LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" FT promoter complement(3967..4104) FT /note="Kan" FT rep_origin 4352..5025 FT /note="pUC" SQ Sequence 5147 BP; 1217 A; 1426 C; 1304 G; 1200 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 gaattcctgc agcccggggg atctatatta ccctgttatc cctagcggat ctgccggtag 1860 aggtgtggtc aataagagcg acctcatact atacctgaga aagcaacctg acctacagga 1920 aagagttact caagaataag aattttcgtt ttaaaaccta agagtcactt taaaatttgt 1980 atacacttat tttttttata acttatttaa taataaaaat cataaatcat aagaaattcg 2040 cttatttaga agtggcgcgc ctcagcactg agcagcgtaa tctggaacgt catatggata 2100 ggatcctgca tagtccggga cgtcatacgg atagcccgca tagtcaggaa catcgtatgg 2160 gtaaaagatg ttaattaagc ctccgcctcc gcctccgccg cctccgcctc gagcatgcat 2220 ctagagggcc caattcgccc tattcgaagt cgtattacaa ttcactggcc gtcgttttac 2280 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc 2340 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc 2400 gcagcctata cgtacggcag tttaaggttt acacctataa aagagagagc cgttatcgtc 2460 tgtttgtgga tgtacagagt gatattattg acacgccggg gcgacggatg gtgatccccc 2520 tggccagtgc acgtctgctg tcagataaag tctcccgtga actttacccg gtggtgcata 2580 tcggggatga aagctggcgc atgatgacca ccgatatggc cagtgtgccg gtctccgtta 2640 tcggggaaga agtggctgat ctcagccacc gcgaaaatga catcaaaaac gccattaacc 2700 tgatgttctg gggaatataa atgtcaggcc tgaatggcga atggacgcgc cctgtagcgg 2760 cgcattaagc gcgcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc 2820 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc 2880 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagagcttt acggcacctc 2940 gaccgcaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg 3000 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact 3060 ggaacaacac tcaaccctat cgcggtctat tcttttgatt tataagggat gttgccgatt 3120 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat tttaacaaaa ttcagaagaa 3180 ctcgtcaaga aggcgataga aggcgatgcg ctgcgaatcg ggagcggcga taccgtaaag 3240 cacgaggaag cggtcagccc attcgccgcc aagctcttca gcaatatcac gggtagccaa 3300 cgctatgtcc tgatagcggt ccgccacacc cagccggcca cagtcgatga atccagaaaa 3360 gcggccattt tccaccatga tattcggcaa gcaggcatcg ccatgggtca cgacgagatc 3420 ctcgccgtcg ggcatgctcg ccttgagcct ggcgaacagt tcggctggcg cgagcccctg 3480 atgctcttcg tccagatcat cctgatcgac aagaccggct tccatccgag tacgtgctcg 3540 ctcgatgcga tgtttcgctt ggtggtcgaa tgggcaggta gccggatcaa gcgtatgcag 3600 ccgccgcatt gcatcagcca tgatggatac tttctcggca ggagcaaggt gagatgacag 3660 gagatcctgc cccggcactt cgcccaatag cagccagtcc cttcccgctt cagtgacaac 3720 gtcgagcaca gctgcgcaag gaacgcccgt cgtggccagc cacgatagcc gcgctgcctc 3780 gtcttgcagt tcattcaggg caccggacag gtcggtcttg acaaaaagaa ccgggcgccc 3840 ctgcgctgac agccggaaca cggcggcatc agagcagccg attgtctgtt gtgcccagtc 3900 atagccgaat agcctctcca cccaagcggc cggagaacct gcgtgcaatc catcttgttc 3960 aatcatgcga aacgatcctc atcctgtctc ttgatcagag cttgatcccc tgcgccatca 4020 gatccttggc ggcgagaaag ccatccagtt tactttgcag ggcttcccaa ccttaccaga 4080 gggcgcccca gctggcaatt ccggttcgct tgctgtccat aaaaccgccc agtctagcta 4140 tcgccatgta agcccactgc aagctacctg ctttctcttt gcgcttgcgt tttcccttgt 4200 ccagatagcc cagtagctga cattcatccg gggtcagcac cgtttctgcg gactggcttt 4260 ctacgtgaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta 4320 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg 4380 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc 4440 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag 4500 cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa 4560 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc 4620 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc 4680 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta 4740 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag 4800 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct 4860 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga 4920 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc 4980 ggccttttta cggttcctgg gcttttgctg gccttttgct cacatgttct ttcctgcgtt 5040 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg 5100 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaag 5147 p3xHA::hph::loxP"Times New Roman PQ5PvuII(56),HindIII(277),KpnI(287),SacI(293),BamHI(295) Arial^UBB PQScaI(406),EagI(432) ArialkTMAY PQANdeI(924),EagI(1002),NcoI(1020),PstI(1043),EcoRI(1129),EagI(1167) Arial!!^ PQClaI ArialcO$P-Q-K^W PQgSalI(1742),ClaI(1785),HindIII(1790),EcoRV(1798),EcoRI(1802),PstI(1812),XmaI(1814),SmaI(1816),AscI(2056) Arial @5 PQNNdeI(2093),BamHI(2102),PacI(2176),XhoI(2209),XbaI(2221),ApaI(2231),PvuII(2351) Arialv` PQNcoI(3402),PvuII(3732) ArialJ ulc)c PQ!EagI(3928),BclI(3993),PvuII(4092) ArialRH>>!PQC lac_promoter"Times New RomanPQM13_reverse_primer"Times New RomanPQC Sp6_primer"Times New RomanPQC,MloxP"Times New RomanPQ_`ORF 1"Times New RomanPQjuORF 1"Times New RomanPQCɆloxP"Times New RomanPQCpHybLex_rev_primer"Times New RomanPQCɆM13_forward20_primer"Times New RomanPQM13_pUC_fwd_primer"Times New RomanPQCd ~ORF 2"Times New RomanPQCNEOKAN_promoter"Times New Roman PQtest"Times New RomaniVtv_m_8 PQtest"Times New Roman