LOCUS       sequence-363984-        4759 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transient expression of KOC207-MXMT in plant cell.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4759)
  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 4759)
  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-KOC207-MXMT
FEATURES             Location/Qualifiers
     source          1..4759
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     66..84
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(122..144)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     244..263
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     457..479
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     471..488
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     472..488
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     498..517
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        498..516
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        593..937
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     880..900
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     misc_feature    957..1012
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5'-leader sequence (Gallie et al., 1988)"
     protein_bind    1056..1080
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1126..1150)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      2326..2576
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(2641..2659)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2642..2659)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2677..2693)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2677..2693)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2690..2712)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2701..2717
                     /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(2725..2731,2732..2749,2750..2755))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2770..2791
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2908..2925)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3079..3667)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3159..3178)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3825..4640)
                     /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     4548..4567
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4632..4651
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
       61 ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa
      121 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
      181 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact
      241 atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca
      301 gatgcgtaag gagaaaatat tactacaggc gccattcgcc attcaggctg cgcaactgtt
      361 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg
      421 ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga
      481 cggccagtga gctagtgtaa tacgactcac tatagggcgc ggccgcagaa ttcgagctcg
      541 gtacccggga tctcgaggcc agatctcccg ggctcagaag accgagggct attgagactt
      601 ttcaacaaag ggtaatatcg ggaaacctcc tcggattcca ttgcccagct atctgtcact
      661 tatcgaaagg acagtagaaa aggaaggtgg ctcctacaaa tgccatcatt gcgataaagg
      721 aaaggctatc gttcaagatg cctctaccga cagtggtccc aaagatggac ccccacccac
      781 gaggaacatc gtggaaaaag aagacgttcc aaccacgtct tcaaagcaag tggattgatg
      841 tgatatctcc actgacgtaa gggatgacgc acaatcccac tatccttcgc aagacccttc
      901 ctctatataa ggaagttcat ttcatttgga gaggacaggc ttcttgagat ccttcaacaa
      961 ttaccaacaa caacaaacaa caaacaacat tacaattact atttacaatt acagaattcg
     1021 agctcggtac caagcttgga ggctcaggag gctccacaag tttgtacaaa aaagcaggct
     1081 tcatgattac tgagcaggta gaagatgcag tagctcattc ctaagaccca gctttcttgt
     1141 acaaagtggt tggaggctca ggaggctccc catggatgga gctccaagaa gtcctgcata
     1201 tgaatgaagg tgaaggcgat acaagctacg ccaagaatgc atcctacaat ctggctcttg
     1261 ccaaggtgaa acctttcctt gaacaatgca tacgagaatt gttgcgggcc aacttgccca
     1321 acatcaacaa gtgcattaaa gttgcggatt tgggatgcgc ttctggacca aacacacttt
     1381 taacagtgcg ggacattgtg caaagtattg acaaagttgg ccaggaagag aagaatgaat
     1441 tagaacgtcc caccattcag atttttctga atgatctttt ccaaaatgat ttcaattcgg
     1501 ttttcaagtt gctgccaagc ttctaccgca aactcgagaa agaaaatgga cgcaagatag
     1561 gatcgtgcct aataagcgca atgcctggct ctttctacgg cagactcttc cccgaggagt
     1621 ccatgcattt tttgcactct tgttacagtg ttcattggtt atctcaggtt cccagcggtt
     1681 tggtgattga attggggatt ggtgcaaaca aagggagtat ttactcttcc aaaggatgtc
     1741 gtccgcccgt ccagaaggca tatttggatc aatttacgaa agattttacc acatttctaa
     1801 ggattcattc gaaagagttg ttttcacgtg gccgaatgct ccttacctgc atttgtaaag
     1861 tagatgaatt cgacgaaccg aatcccctag acttacttga catggcaata aacgacttga
     1921 ttgttgaggg acttctggag gaagaaaaat tggatagttt caatattcca ttctttacac
     1981 cttcagcaga agaagtaaag tgcatagttg aggaggaagg ttcttgcgaa attttatatc
     2041 tggagacttt taaggcccat tatgatgctg ccttctctat tgatgatgat tacccagtaa
     2101 gatcccatga acaaattaaa gcagagtatg tggcatcatt aattagatca gtttacgaac
     2161 ccatcctcgc aagtcatttt ggagaagcta ttatgcctga cttattccac aggcttgcga
     2221 agcatgcagc aaaggttctc cacatgggca aaggctgcta taataatctt atcatttctc
     2281 tcgccaaaaa gccagagaag tcagacgtgt aaactagtat ctagaatcgt tcaaacattt
     2341 ggcaataaag tttcttaaga ttgaatcctg ttgccggtct tgcgatgatt atcatataat
     2401 ttctgttgaa ttacgttaag catgtaataa ttaacatgta atgcatgacg ttatttatga
     2461 gatgggtttt tatgattaga gtcccgcaat tatacattta atacgcgata gaaaacaaaa
     2521 tatagcgcgc aaactaggat aaattatcgc gcgcggtgtc atctatgtta ctagatgcgg
     2581 ccgcaattgt ggatccgctc tagagtcgac ctgcaggcat gcaagcttgc ggccgcgtat
     2641 tctatagtgt cacctaaata gcatggcgta atcatggtca tagctgtttc ctgtgtgaaa
     2701 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     2761 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     2821 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     2881 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     2941 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     3001 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     3061 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     3121 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     3181 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     3241 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     3301 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     3361 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     3421 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     3481 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     3541 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     3601 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     3661 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     3721 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     3781 ttaaaaatga agttttaaat caatctaaag tatatatgtt ttaattagaa aaactcatcg
     3841 agcatcaaat gaaactgcaa tttattcata tcaggattat caataccata tttttgaaaa
     3901 agccgtttct gtaatgaagg agaaaactca ccgaggcagt tccataggat ggcaagatcc
     3961 tggtatcggt ctgcgattcc gactcgtcca acatcaatac aacctattaa tttcccctcg
     4021 tcaaaaataa ggttatcaag tgagaaatca ccatgagtga cgactgaatc cggtgagaat
     4081 ggcaatagct tatgcatttc tttccagact tgttcaacag gccagccatt acgctcgtca
     4141 tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg agcgagacga
     4201 aatacgcgat cgctgttaaa aggacaatta caaacaggaa tcgaatgcaa ccggcgcagg
     4261 aacactgcca gcgcatcaac aatattttca cctgaatcag gatattcttc taatacctgg
     4321 aatgctgttt tccctgggat cgcagaggtg agtaaccatg catcatcagg agtacggata
     4381 aaatgcttga tggtcggaag aggcataaat tccgtcagcc agtttagtct gaccatctca
     4441 tctgtaacat cattggcaac gctacctttg ccatgtttca gaaacaactc tggcgcatcg
     4501 ggcttcccat acaatcgata gattgtcgca cctgattgcc cgacattatc gcgagcccat
     4561 ttatacccat ataaatcagc atccatgttg gaatttaatc gcggccttga gcaagacgtt
     4621 tcccgttgaa tatggctcat aacacccctt gtattactgt ttatgtaagc agacataagc
     4681 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     4741 caaatagggg ttccgcgca
//
