LOCUS Exported 3221 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION mKO1 ORF with stop codon between attL1-attL2 on Gateway entry vector. ACCESSION . VERSION . KEYWORDS pENTR-mKO1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3221) TITLE Minoru Ko Lab plasmids REFERENCE 2 (bases 1 to 3221) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3221 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 92..109 /label=L4440 /note="L4440 vector, forward primer" terminator 268..295 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" terminator 387..473 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" primer_bind 522..544 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 536..553 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 537..553 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" protein_bind 569..668 /gene="mutant version of attL" /label=attL1 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" CDS 681..1337 /codon_start=1 /product="monomeric Kusabira-Orange fluorescent protein (Karasawa et al., 2004)" /label=mKO /translation="MVSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRV TMAKGGPMPFAFDLVSHVFCYGHRPFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSA SVSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMY LKLEGGGNHKCQFKTTYKAAKKILKMPGSHYISHRLVRKTEGNITELVEDAVAHS" protein_bind complement(1346..1445) /gene="mutant version of attL" /label=attL2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(1462..1481) /label=T7 /note="T7 promoter, forward primer" promoter complement(1463..1481) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(1486..1502) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1486..1502) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(1604..1623) /label=pENTR-R /note="pENTR vectors, reverse primer" CDS 1615..2424 /codon_start=1 /gene="aph(3')-Ia" /product="aminoglycoside phosphotransferase" /label=KanR /note="confers resistance to kanamycin in bacteria or G418 (Geneticin(R)) in eukaryotes" /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" primer_bind complement(1682..1701) /label=Kan-R /note="Kanamycin resistance gene, reverse primer" rep_origin 2571..3159 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3060..3079 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga 61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga 121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca 181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc 241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta 301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc 361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa 421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg 481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa 541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac 601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa 661 agcaggctct cgagccacca atggtgagtg tgattaaacc agagatgaag atgaggtact 721 acatggacgg ctccgtcaat gggcatgagt tcacaattga aggtgaaggc acaggcagac 781 cttacgaggg acatcaagag atgacactac gcgtcacaat ggccaagggc gggccaatgc 841 ctttcgcgtt tgacttagtg tcacacgtgt tctgttacgg ccacagacct tttactaaat 901 atccagaaga gataccagac tatttcaaac aagcatttcc tgaaggcctg tcatgggaaa 961 ggtcgttgga gttcgaagat ggtgggtccg cttcagtcag tgcgcatata agccttagag 1021 gaaacacctt ctaccacaaa tccaaattta ctggggttaa ctttcctgcc gatggtccta 1081 tcatgcaaaa ccaaagtgtt gattgggagc catcaaccga gaaaattact gccagcgacg 1141 gagttctgaa gggtgatgtt acgatgtacc taaaacttga aggaggcggc aatcacaaat 1201 gccaattcaa gactacttac aaggcggcaa aaaagattct taaaatgcca ggaagccatt 1261 acatcagcca tcgcctcgtc aggaaaaccg aaggcaacat tactgagctg gtagaagatg 1321 cagtagctca ttcctaagcg gccgcaccca gctttcttgt acaaagttgg cattataaga 1381 aagcattgct tatcaatttg ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt 1441 atttgccatc cagctgatat cccctatagt gagtcgtatt acatggtcat agctgtttcc 1501 tggcagctct ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga taaaaatata 1561 tcatcatgaa caataaaact gtctgcttac ataaacagta atacaagggg tgttatgagc 1621 catattcaac gggaaacgtc gaggccgcga ttaaattcca acatggatgc tgatttatat 1681 gggtataaat gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgcttgtat 1741 gggaagcccg atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat 1801 gttacagatg agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc 1861 aagcatttta tccgtactcc tgatgatgca tggttactca ccactgcgat ccccggaaaa 1921 acagcattcc aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg 1981 gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagcgat 2041 cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt 2101 gattttgatg acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataaa 2161 cttttgccat tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt 2221 atttttgacg aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac 2281 cgataccagg atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag 2341 aaacggcttt ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat 2401 ttgatgctcg atgagttttt ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc 2461 attacgctga cttgacggga cggcgcaagc tcatgaccaa aatcccttaa cgtgagttac 2521 gcgtcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct 2581 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt 2641 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg 2701 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct 2761 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc 2821 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg 2881 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa 2941 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg 3001 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg 3061 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga 3121 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt 3181 ttacggttcc tggccttttg ctggcctttt gctcacatgt t //