LOCUS       sequence-361317-        4081 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transient expression of mKO2 in plant cell.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4081)
  AUTHORS   Fujii Y, Yoshimura A, Kodama Y
  TITLE     A novel orange-colored bimolecular fluorescence complementation 
            (BiFC) assay using monomeric Kusabira-Orange protein.
  JOURNAL   Biotechniques. 2018 Apr;64(4):153-161. doi: 10.2144/btn-2017-0121.
  PUBMED    29661017
REFERENCE   2  (bases 1 to 4081)
  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"; doi: "10"; journalName: 
            "Biotechniques"; date: "2018-04"; volume: "64"; issue: "4"; pages: 
            "153-161"
COMMENT     Sequence Label: p35S-mKO2
FEATURES             Location/Qualifiers
     source          1..4081
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(43..60)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(214..802)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(294..313)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(973..1764,1765..1833))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1596..1615
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1834..1938)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2006..2024
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2062..2084)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2184..2203
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2397..2419
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2411..2428
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2412..2428
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2488..2833
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     2776..2796
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     misc_feature    2853..2908
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5'-leader sequence (Gallie et al., 1988)"
     CDS             2928..3584
                     /codon_start=1
                     /product="monomeric Kusabira-Orange 2 fluorescent protein"
                     /label=mKO2
                     /translation="MVSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRV
                     TMAEGGPMPFAFDLVSHVFCYGHRVFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSA
                     SVSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMY
                     LKLEGGGNHKCQMKTTYKAAKEILEMPGDHYIGHRLVRKTEGNITEQVEDAVAHS"
     terminator      3603..3855
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(3893..3909)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3893..3909)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3906..3928)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3917..3933
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3941..3947,3948..3965,3966..3971))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3986..4007
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
       61 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
      121 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
      181 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
      241 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
      301 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
      361 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
      421 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
      481 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
      541 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
      601 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
      661 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
      721 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
      781 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
      841 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
      901 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
      961 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     1021 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     1081 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     1141 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     1201 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     1261 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     1321 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     1381 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     1441 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     1501 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     1561 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     1621 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     1681 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     1741 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     1801 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     1861 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     1921 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
     1981 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg
     2041 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     2101 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     2161 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat
     2221 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc
     2281 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
     2341 gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca
     2401 gtcacgacgt tgtaaaacga cggccagtgc caagcttgca tgcctgcagg ctctagagga
     2461 tcccccctca gaagaccaga gggctattga gacttttcaa caaagggtaa tatcgggaaa
     2521 cctcctcgga ttccattgcc cagctatctg tcacttcatc gaaaggacag tagaaaagga
     2581 aggtggctcc tacaaatgcc atcattgcga taaaggaaag gctatcgttc aagatgcctc
     2641 taccgacagt ggtcccaaag atggaccccc acccacgagg aacatcgtgg aaaaagaaga
     2701 cgttccaacc acgtcttcaa agcaagtgga ttgatgtgat atctccactg acgtaaggga
     2761 tgacgcacaa tcccactatc cttcgcaaga cccttcctct atataaggaa gttcatttca
     2821 tttggagagg acaggcttct tgagatcctt caacaattac caacaacaac aaacaacaaa
     2881 caacattaca attactattt acaattacag tcgactctag aggatccatg gtgagtgtga
     2941 ttaaaccaga gatgaagatg aggtactaca tggacggctc cgtcaatggg catgagttca
     3001 caattgaagg tgaaggcaca ggcagacctt acgagggaca tcaagagatg acactacgcg
     3061 tcacaatggc cgagggcggg ccaatgcctt tcgcgtttga cttagtgtca cacgtgttct
     3121 gttacggcca cagagtattt actaaatatc cagaagagat accagactat ttcaaacaag
     3181 catttcctga aggcctgtca tgggaaaggt cgttggagtt cgaagatggt gggtccgctt
     3241 cagtcagtgc gcatataagc cttagaggaa acaccttcta ccacaaatcc aaatttactg
     3301 gggttaactt tcctgccgat ggtcctatca tgcaaaacca aagtgttgat tgggagccat
     3361 caaccgagaa aattactgcc agcgacggag ttctgaaggg tgatgttacg atgtacctaa
     3421 aacttgaagg aggcggcaat cacaaatgcc aaatgaagac tacttacaag gcggcaaaag
     3481 agattcttga aatgccagga gaccattaca tcggccatcg cctcgtcagg aaaaccgaag
     3541 gcaacattac tgagcaggta gaagatgcag tagctcattc ctaaagcggc cgcccggctg
     3601 cagatcgttc aaacatttgg caataaagtt tcttaagatt gaatcctgtt gccggtcttg
     3661 cgatgattat catataattt ctgttgaatt acgttaagca tgtaataatt aacatgtaat
     3721 gcatgacgtt atttatgaga tgggttttta tgattagagt cccgcaatta tacatttaat
     3781 acgcgataga aaacaaaata tagcgcgcaa actaggataa attatcgcgc gcggtgtcat
     3841 ctatgttact agatccgatg ataagctgtc aaacatgaga attcgtaatc atgtcatagc
     3901 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     3961 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     4021 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     4081 g
//
