LOCUS       sequence-361326-        4873 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transient expression of KOC169-MXMT in plant cell.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4873)
  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 4873)
  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: pGWT35S-KOC169-MXMT
FEATURES             Location/Qualifiers
     source          1..4873
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(51..73)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     173..192
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     386..408
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     400..417
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     401..417
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     427..446
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        427..445
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        522..866
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     809..829
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     misc_feature    886..941
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5'-leader sequence (Gallie et al., 1988)"
     protein_bind    985..1009
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1169..1193)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      2369..2619
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(2684..2702)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2685..2702)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2720..2736)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2720..2736)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2733..2755)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2744..2760
                     /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(2768..2774,2775..2792,2793..2798))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2813..2834
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2951..2968)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3122..3710)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3202..3221)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3868..4683)
                     /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="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTSAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     4591..4610
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4675..4694
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     join(4868..4873,1..13)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
       61 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
      121 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca
      181 gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg
      241 agaaaatatt actacaggcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga
      301 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga
      361 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgag
      421 ctagtgtaat acgactcact atagggcgcg gccgcagaat tcgagctcgg tacccgggat
      481 ctcgaggcca gatctcccgg gctcagaaga ccgagggcta ttgagacttt tcaacaaagg
      541 gtaatatcgg gaaacctcct cggattccat tgcccagcta tctgtcactt atcgaaagga
      601 cagtagaaaa ggaaggtggc tcctacaaat gccatcattg cgataaagga aaggctatcg
      661 ttcaagatgc ctctaccgac agtggtccca aagatggacc cccacccacg aggaacatcg
      721 tggaaaaaga agacgttcca accacgtctt caaagcaagt ggattgatgt gatatctcca
      781 ctgacgtaag ggatgacgca caatcccact atccttcgca agacccttcc tctatataag
      841 gaagttcatt tcatttggag aggacaggct tcttgagatc cttcaacaat taccaacaac
      901 aacaaacaac aaacaacatt acaattacta tttacaatta cagaattcga gctcggtacc
      961 aagcttggag gctcaggagg ctccacaagt ttgtacaaaa aagcaggctt catgggcggc
     1021 aatcacaaat gccaaatgaa gactacttac aaggcggcaa aagagattct tgaaatgcca
     1081 ggagaccatt acatcggcca tcgcctcgtc aggaaaaccg aaggcaacat tactgagcag
     1141 gtagaagatg cagtagctca ttcctaagac ccagctttct tgtacaaagt ggttggaggc
     1201 tcaggaggct ccccatggat ggagctccaa gaagtcctgc atatgaatga aggtgaaggc
     1261 gatacaagct acgccaagaa tgcatcctac aatctggctc ttgccaaggt gaaacctttc
     1321 cttgaacaat gcatacgaga attgttgcgg gccaacttgc ccaacatcaa caagtgcatt
     1381 aaagttgcgg atttgggatg cgcttctgga ccaaacacac ttttaacagt gcgggacatt
     1441 gtgcaaagta ttgacaaagt tggccaggaa gagaagaatg aattagaacg tcccaccatt
     1501 cagatttttc tgaatgatct tttccaaaat gatttcaatt cggttttcaa gttgctgcca
     1561 agcttctacc gcaaactcga gaaagaaaat ggacgcaaga taggatcgtg cctaataagc
     1621 gcaatgcctg gctctttcta cggcagactc ttccccgagg agtccatgca ttttttgcac
     1681 tcttgttaca gtgttcattg gttatctcag gttcccagcg gtttggtgat tgaattgggg
     1741 attggtgcaa acaaagggag tatttactct tccaaaggat gtcgtccgcc cgtccagaag
     1801 gcatatttgg atcaatttac gaaagatttt accacatttc taaggattca ttcgaaagag
     1861 ttgttttcac gtggccgaat gctccttacc tgcatttgta aagtagatga attcgacgaa
     1921 ccgaatcccc tagacttact tgacatggca ataaacgact tgattgttga gggacttctg
     1981 gaggaagaaa aattggatag tttcaatatt ccattcttta caccttcagc agaagaagta
     2041 aagtgcatag ttgaggagga aggttcttgc gaaattttat atctggagac ttttaaggcc
     2101 cattatgatg ctgccttctc tattgatgat gattacccag taagatccca tgaacaaatt
     2161 aaagcagagt atgtggcatc attaattaga tcagtttacg aacccatcct cgcaagtcat
     2221 tttggagaag ctattatgcc tgacttattc cacaggcttg cgaagcatgc agcaaaggtt
     2281 ctccacatgg gcaaaggctg ctataataat cttatcattt ctctcgccaa aaagccagag
     2341 aagtcagacg tgtaaactag tatctagaat cgttcaaaca tttggcaata aagtttctta
     2401 agattgaatc ctgttgccgg tcttgcgatg attatcatat aatttctgtt gaattacgtt
     2461 aagcatgtaa taattaacat gtaatgcatg acgttattta tgagatgggt ttttatgatt
     2521 agagtcccgc aattatacat ttaatacgcg atagaaaaca aaatatagcg cgcaaactag
     2581 gataaattat cgcgcgcggt gtcatctatg ttactagatg cggccgcaat tgtggatccg
     2641 ctctagagtc gacctgcagg catgcaagct tgcggccgcg tattctatag tgtcacctaa
     2701 atagcatggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     2761 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     2821 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     2881 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     2941 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     3001 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     3061 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     3121 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     3181 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     3241 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     3301 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     3361 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     3421 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     3481 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     3541 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     3601 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     3661 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     3721 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     3781 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     3841 aatcaatcta aagtatatat gttttaatta gaaaaactca tcgagcatca aatgaaactg
     3901 caatttattc atatcaggat tatcaatacc atatttttga aaaagccgtt tctgtaatga
     3961 aggagaaaac tcaccgaggc agttccatag gatggcaaga tcctggtatc ggtctgcgat
     4021 tccgactcgt ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa taaggttatc
     4081 aagtgagaaa tcaccatgag tgacgactga atccggtgag aatggcaata gcttatgcat
     4141 ttctttccag acttgttcaa caggccagcc attacgctcg tcatcaaaat cactcgcatc
     4201 aaccaaaccg ttattcattc gtgattgcgc ctgagcgaga cgaaatacgc gatcgctgtt
     4261 aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc aggaacactg ccagcgcatc
     4321 aacaatattt tcacctgaat caggatattc ttctaatacc tggaatgctg ttttccctgg
     4381 gatcgcagag gtgagtaacc atgcatcatc aggagtacgg ataaaatgct tgatggtcgg
     4441 aagaggcata aattccgtca gccagtttag tctgaccatc tcatctgtaa catcattggc
     4501 aacgctacct ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc catacaatcg
     4561 atagattgtc gcacctgatt gcccgacatt atcgcgagcc catttatacc catataaatc
     4621 agcatccatg ttggaattta atcgcggcct tgagcaagac gtttcccgtt gaatatggct
     4681 cataacaccc cttgtattac tgtttatgta agcagacata agcatttatc agggttattg
     4741 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     4801 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     4861 ctataaaaat agg
//
