LOCUS       sequence-274734-        3221 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mKO1 ORF with stop codon between attL1-attL2 on Gateway entry 
            vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
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 Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pENTR-mKO1
FEATURES             Location/Qualifiers
     source          1..3221
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(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
//
