LOCUS       sequence-361250-        5182 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transient expression of KON154-MXMT in plant cell.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5182)
  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 5182)
  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-KON154-MXMT
FEATURES             Location/Qualifiers
     source          1..5182
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(30..48)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(31..48)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(66..82)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(66..82)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(79..101)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    90..106
                     /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(114..120,121..138,139..144))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    159..180
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(297..314)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(468..1056)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(548..567)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1214..2029)
                     /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     1937..1956
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2021..2040
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     2214..2232
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2270..2292)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2392..2411
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2605..2627
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2619..2636
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2620..2636
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2646..2665
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2646..2664
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        2741..3085
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     3028..3048
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     misc_feature    3105..3160
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5'-leader sequence (Gallie et al., 1988)"
     protein_bind    3204..3228
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3697..3721)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      4897..5147
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
ORIGIN
        1 tgcaggcatg caagcttgcg gccgcgtatt ctatagtgtc acctaaatag catggcgtaa
       61 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
      121 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
      181 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
      241 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
      301 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
      361 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
      421 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
      481 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
      541 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
      601 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
      661 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
      721 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
      781 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
      841 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
      901 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
      961 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     1021 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     1081 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     1141 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     1201 atatatgttt taattagaaa aactcatcga gcatcaaatg aaactgcaat ttattcatat
     1261 caggattatc aataccatat ttttgaaaaa gccgtttctg taatgaagga gaaaactcac
     1321 cgaggcagtt ccataggatg gcaagatcct ggtatcggtc tgcgattccg actcgtccaa
     1381 catcaataca acctattaat ttcccctcgt caaaaataag gttatcaagt gagaaatcac
     1441 catgagtgac gactgaatcc ggtgagaatg gcaatagctt atgcatttct ttccagactt
     1501 gttcaacagg ccagccatta cgctcgtcat caaaatcact cgcatcaacc aaaccgttat
     1561 tcattcgtga ttgcgcctga gcgagacgaa atacgcgatc gctgttaaaa ggacaattac
     1621 aaacaggaat cgaatgcaac cggcgcagga acactgccag cgcatcaaca atattttcac
     1681 ctgaatcagg atattcttct aatacctgga atgctgtttt ccctgggatc gcagaggtga
     1741 gtaaccatgc atcatcagga gtacggataa aatgcttgat ggtcggaaga ggcataaatt
     1801 ccgtcagcca gtttagtctg accatctcat ctgtaacatc attggcaacg ctacctttgc
     1861 catgtttcag aaacaactct ggcgcatcgg gcttcccata caatcgatag attgtcgcac
     1921 ctgattgccc gacattatcg cgagcccatt tatacccata taaatcagca tccatgttgg
     1981 aatttaatcg cggccttgag caagacgttt cccgttgaat atggctcata acaccccttg
     2041 tattactgtt tatgtaagca gacataagca tttatcaggg ttattgtctc atgagcggat
     2101 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     2161 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     2221 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     2281 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     2341 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag
     2401 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga
     2461 gaaaatatta ctacaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat
     2521 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat
     2581 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgagc
     2641 tagtgtaata cgactcacta tagggcgcgg ccgcagaatt cgagctcggt acccgggatc
     2701 tcgaggccag atctcccggg ctcagaagac cgagggctat tgagactttt caacaaaggg
     2761 taatatcggg aaacctcctc ggattccatt gcccagctat ctgtcactta tcgaaaggac
     2821 agtagaaaag gaaggtggct cctacaaatg ccatcattgc gataaaggaa aggctatcgt
     2881 tcaagatgcc tctaccgaca gtggtcccaa agatggaccc ccacccacga ggaacatcgt
     2941 ggaaaaagaa gacgttccaa ccacgtcttc aaagcaagtg gattgatgtg atatctccac
     3001 tgacgtaagg gatgacgcac aatcccacta tccttcgcaa gacccttcct ctatataagg
     3061 aagttcattt catttggaga ggacaggctt cttgagatcc ttcaacaatt accaacaaca
     3121 acaaacaaca aacaacatta caattactat ttacaattac agaattcgag ctcggtacca
     3181 agcttggagg ctcaggaggc tccacaagtt tgtacaaaaa agcaggcttc atggtgagtg
     3241 tgattaaacc agagatgaag atgaggtact acatggacgg ctccgtcaat gggcatgagt
     3301 tcacaattga aggtgaaggc acaggcagac cttacgaggg acatcaagag atgacactac
     3361 gcgtcacaat ggccgagggc gggccaatgc ctttcgcgtt tgacttagtg tcacacgtgt
     3421 tctgttacgg ccacagagta tttactaaat atccagaaga gataccagac tatttcaaac
     3481 aagcatttcc tgaaggcctg tcatgggaaa ggtcgttgga gttcgaagat ggtgggtccg
     3541 cttcagtcag tgcgcatata agccttagag gaaacacctt ctaccacaaa tccaaattta
     3601 ctggggttaa ctttcctgcc gatggtccta tcatgcaaaa ccaaagtgtt gattgggagc
     3661 catcaaccga gaaaattact gccagcgacg gataagaccc agctttcttg tacaaagtgg
     3721 ttggaggctc aggaggctcc ccatggatgg agctccaaga agtcctgcat atgaatgaag
     3781 gtgaaggcga tacaagctac gccaagaatg catcctacaa tctggctctt gccaaggtga
     3841 aacctttcct tgaacaatgc atacgagaat tgttgcgggc caacttgccc aacatcaaca
     3901 agtgcattaa agttgcggat ttgggatgcg cttctggacc aaacacactt ttaacagtgc
     3961 gggacattgt gcaaagtatt gacaaagttg gccaggaaga gaagaatgaa ttagaacgtc
     4021 ccaccattca gatttttctg aatgatcttt tccaaaatga tttcaattcg gttttcaagt
     4081 tgctgccaag cttctaccgc aaactcgaga aagaaaatgg acgcaagata ggatcgtgcc
     4141 taataagcgc aatgcctggc tctttctacg gcagactctt ccccgaggag tccatgcatt
     4201 ttttgcactc ttgttacagt gttcattggt tatctcaggt tcccagcggt ttggtgattg
     4261 aattggggat tggtgcaaac aaagggagta tttactcttc caaaggatgt cgtccgcccg
     4321 tccagaaggc atatttggat caatttacga aagattttac cacatttcta aggattcatt
     4381 cgaaagagtt gttttcacgt ggccgaatgc tccttacctg catttgtaaa gtagatgaat
     4441 tcgacgaacc gaatccccta gacttacttg acatggcaat aaacgacttg attgttgagg
     4501 gacttctgga ggaagaaaaa ttggatagtt tcaatattcc attctttaca ccttcagcag
     4561 aagaagtaaa gtgcatagtt gaggaggaag gttcttgcga aattttatat ctggagactt
     4621 ttaaggccca ttatgatgct gccttctcta ttgatgatga ttacccagta agatcccatg
     4681 aacaaattaa agcagagtat gtggcatcat taattagatc agtttacgaa cccatcctcg
     4741 caagtcattt tggagaagct attatgcctg acttattcca caggcttgcg aagcatgcag
     4801 caaaggttct ccacatgggc aaaggctgct ataataatct tatcatttct ctcgccaaaa
     4861 agccagagaa gtcagacgtg taaactagta tctagaatcg ttcaaacatt tggcaataaa
     4921 gtttcttaag attgaatcct gttgccggtc ttgcgatgat tatcatataa tttctgttga
     4981 attacgttaa gcatgtaata attaacatgt aatgcatgac gttatttatg agatgggttt
     5041 ttatgattag agtcccgcaa ttatacattt aatacgcgat agaaaacaaa atatagcgcg
     5101 caaactagga taaattatcg cgcgcggtgt catctatgtt actagatgcg gccgcaattg
     5161 tggatccgct ctagagtcga cc
//
