LOCUS       sequence-361294-        4081 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transient expression of humanized mKO1 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-hmKO1
FEATURES             Location/Qualifiers
     source          1..4081
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             32..688
                     /codon_start=1
                     /product="humanized monomeric Kusabira-Orange fluorescent 
                     protein"
                     /label=hmKO
                     /note="mammalian codon-optimized"
                     /translation="MVSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRV
                     TMAKGGPMPFAFDLVSHVFCYGHRPFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSA
                     SVSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMY
                     LKLEGGGNHKCQFKTTYKAAKKILKMPGSHYISHRLVRKTEGNITELVEDAVAHS"
     terminator      707..959
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(997..1013)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(997..1013)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1010..1032)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1021..1037
                     /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(1045..1051,1052..1069,1070..1075))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1090..1111
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1228..1245)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1399..1987)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1479..1498)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2158..2949,2950..3018))
                     /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     2781..2800
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3019..3123)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3191..3209
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3247..3269)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3369..3388
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3582..3604
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3596..3613
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3597..3613
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3673..4018
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     3961..3981
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     misc_feature    join(4038..4081,1..12)
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5'-leader sequence (Gallie et al., 1988)"
ORIGIN
        1 atttacaatt acagtcgact ctagaggatc catggtgagc gtgatcaagc ccgagatgaa
       61 gatgaggtac tacatggacg gctccgtcaa tgggcatgag ttcacaatcg agggtgaggg
      121 cacaggcaga ccttacgagg gacatcagga gatgacactg cgcgtcacaa tggccaaggg
      181 cgggccaatg cctttcgcct tcgacctggt gtcccacgtg ttctgttacg gccacagacc
      241 ttttactaaa tatccagaag agatcccaga ctatttcaag caggcctttc ctgagggcct
      301 gtcctgggag aggtccctgg agttcgagga cggcggctcc gcctccgtga gcgcccacat
      361 cagcctgagg ggcaacacct tctaccacaa gtccaagttc accggcgtga acttccccgc
      421 cgacggcccc atcatgcaga accagagcgt ggactgggag ccctccaccg agaagatcac
      481 cgccagcgac ggcgtgctga agggcgacgt gaccatgtac ctgaagctgg agggcggcgg
      541 caaccacaag tgccagttca agaccaccta caaggccgcc aagaagatcc tgaagatgcc
      601 cggcagccac tacatcagcc acaggctggt gaggaagacc gagggcaaca tcaccgagct
      661 ggtggaggac gccgtggccc actcctaaag cggccgcccg gctgcagatc gttcaaacat
      721 ttggcaataa agtttcttaa gattgaatcc tgttgccggt cttgcgatga ttatcatata
      781 atttctgttg aattacgtta agcatgtaat aattaacatg taatgcatga cgttatttat
      841 gagatgggtt tttatgatta gagtcccgca attatacatt taatacgcga tagaaaacaa
      901 aatatagcgc gcaaactagg ataaattatc gcgcgcggtg tcatctatgt tactagatcc
      961 gatgataagc tgtcaaacat gagaattcgt aatcatgtca tagctgtttc ctgtgtgaaa
     1021 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     1081 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     1141 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     1201 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     1261 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     1321 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     1381 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     1441 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     1501 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     1561 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     1621 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     1681 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     1741 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     1801 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     1861 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     1921 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     1981 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     2041 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     2101 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     2161 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     2221 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     2281 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     2341 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     2401 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     2461 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     2521 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     2581 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     2641 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     2701 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     2761 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     2821 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     2881 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     2941 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     3001 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     3061 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     3121 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     3181 aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg
     3241 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
     3301 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct
     3361 taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc
     3421 gcacagatgc gtaaggagaa aataccgcat caggcgccat tcgccattca ggctgcgcaa
     3481 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg
     3541 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa
     3601 aacgacggcc agtgccaagc ttgcatgcct gcaggctcta gaggatcccc cctcagaaga
     3661 ccagagggct attgagactt ttcaacaaag ggtaatatcg ggaaacctcc tcggattcca
     3721 ttgcccagct atctgtcact tcatcgaaag gacagtagaa aaggaaggtg gctcctacaa
     3781 atgccatcat tgcgataaag gaaaggctat cgttcaagat gcctctaccg acagtggtcc
     3841 caaagatgga cccccaccca cgaggaacat cgtggaaaaa gaagacgttc caaccacgtc
     3901 ttcaaagcaa gtggattgat gtgatatctc cactgacgta agggatgacg cacaatccca
     3961 ctatccttcg caagaccctt cctctatata aggaagttca tttcatttgg agaggacagg
     4021 cttcttgaga tccttcaaca attaccaaca acaacaaaca acaaacaaca ttacaattac
     4081 t
//
