LOCUS       sequence-361027-        5224 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transient expression of KON168-MXMT in plant cell.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5224)
  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 5224)
  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-KON168-MXMT
FEATURES             Location/Qualifiers
     source          1..5224
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(165..753)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(245..264)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(911..1726)
                     /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     1634..1653
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1718..1737
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     1911..1929
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1967..1989)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2089..2108
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2302..2324
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2316..2333
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2317..2333
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2343..2362
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2343..2361
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        2438..2782
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     2725..2745
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     misc_feature    2802..2857
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5'-leader sequence (Gallie et al., 1988)"
     protein_bind    2901..2925
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3436..3460)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      4636..4886
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(4951..4969)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4952..4969)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(4987..5003)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4987..5003)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5000..5022)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5011..5027
                     /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(5035..5041,5042..5059,5060..5065))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5080..5101
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(join(5218..5224,1..11))
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
       61 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
      121 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
      181 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
      241 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
      301 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
      361 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
      421 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
      481 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
      541 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
      601 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
      661 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
      721 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
      781 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
      841 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
      901 tatgttttaa ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag
      961 gattatcaat accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga
     1021 ggcagttcca taggatggca agatcctggt atcggtctgc gattccgact cgtccaacat
     1081 caatacaacc tattaatttc ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat
     1141 gagtgacgac tgaatccggt gagaatggca atagcttatg catttctttc cagacttgtt
     1201 caacaggcca gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca
     1261 ttcgtgattg cgcctgagcg agacgaaata cgcgatcgct gttaaaagga caattacaaa
     1321 caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata ttttcacctg
     1381 aatcaggata ttcttctaat acctggaatg ctgttttccc tgggatcgca gaggtgagta
     1441 accatgcatc atcaggagta cggataaaat gcttgatggt cggaagaggc ataaattccg
     1501 tcagccagtt tagtctgacc atctcatctg taacatcatt ggcaacgcta cctttgccat
     1561 gtttcagaaa caactctggc gcatcgggct tcccatacaa tcgatagatt gtcgcacctg
     1621 attgcccgac attatcgcga gcccatttat acccatataa atcagcatcc atgttggaat
     1681 ttaatcgcgg ccttgagcaa gacgtttccc gttgaatatg gctcataaca ccccttgtat
     1741 tactgtttat gtaagcagac ataagcattt atcagggtta ttgtctcatg agcggataca
     1801 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     1861 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     1921 tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc
     1981 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
     2041 agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc
     2101 agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa
     2161 aatattacta caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg
     2221 tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa
     2281 gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgagctag
     2341 tgtaatacga ctcactatag ggcgcggccg cagaattcga gctcggtacc cgggatctcg
     2401 aggccagatc tcccgggctc agaagaccga gggctattga gacttttcaa caaagggtaa
     2461 tatcgggaaa cctcctcgga ttccattgcc cagctatctg tcacttatcg aaaggacagt
     2521 agaaaaggaa ggtggctcct acaaatgcca tcattgcgat aaaggaaagg ctatcgttca
     2581 agatgcctct accgacagtg gtcccaaaga tggaccccca cccacgagga acatcgtgga
     2641 aaaagaagac gttccaacca cgtcttcaaa gcaagtggat tgatgtgata tctccactga
     2701 cgtaagggat gacgcacaat cccactatcc ttcgcaagac ccttcctcta tataaggaag
     2761 ttcatttcat ttggagagga caggcttctt gagatccttc aacaattacc aacaacaaca
     2821 aacaacaaac aacattacaa ttactattta caattacaga attcgagctc ggtaccaagc
     2881 ttggaggctc aggaggctcc acaagtttgt acaaaaaagc aggcttcatg 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 ataagaccca gctttcttgt acaaagtggt tggaggctca ggaggctccc
     3481 catggatgga gctccaagaa gtcctgcata tgaatgaagg tgaaggcgat acaagctacg
     3541 ccaagaatgc atcctacaat ctggctcttg ccaaggtgaa acctttcctt gaacaatgca
     3601 tacgagaatt gttgcgggcc aacttgccca acatcaacaa gtgcattaaa gttgcggatt
     3661 tgggatgcgc ttctggacca aacacacttt taacagtgcg ggacattgtg caaagtattg
     3721 acaaagttgg ccaggaagag aagaatgaat tagaacgtcc caccattcag atttttctga
     3781 atgatctttt ccaaaatgat ttcaattcgg ttttcaagtt gctgccaagc ttctaccgca
     3841 aactcgagaa agaaaatgga cgcaagatag gatcgtgcct aataagcgca atgcctggct
     3901 ctttctacgg cagactcttc cccgaggagt ccatgcattt tttgcactct tgttacagtg
     3961 ttcattggtt atctcaggtt cccagcggtt tggtgattga attggggatt ggtgcaaaca
     4021 aagggagtat ttactcttcc aaaggatgtc gtccgcccgt ccagaaggca tatttggatc
     4081 aatttacgaa agattttacc acatttctaa ggattcattc gaaagagttg ttttcacgtg
     4141 gccgaatgct ccttacctgc atttgtaaag tagatgaatt cgacgaaccg aatcccctag
     4201 acttacttga catggcaata aacgacttga ttgttgaggg acttctggag gaagaaaaat
     4261 tggatagttt caatattcca ttctttacac cttcagcaga agaagtaaag tgcatagttg
     4321 aggaggaagg ttcttgcgaa attttatatc tggagacttt taaggcccat tatgatgctg
     4381 ccttctctat tgatgatgat tacccagtaa gatcccatga acaaattaaa gcagagtatg
     4441 tggcatcatt aattagatca gtttacgaac ccatcctcgc aagtcatttt ggagaagcta
     4501 ttatgcctga cttattccac aggcttgcga agcatgcagc aaaggttctc cacatgggca
     4561 aaggctgcta taataatctt atcatttctc tcgccaaaaa gccagagaag tcagacgtgt
     4621 aaactagtat ctagaatcgt tcaaacattt ggcaataaag tttcttaaga ttgaatcctg
     4681 ttgccggtct tgcgatgatt atcatataat ttctgttgaa ttacgttaag catgtaataa
     4741 ttaacatgta atgcatgacg ttatttatga gatgggtttt tatgattaga gtcccgcaat
     4801 tatacattta atacgcgata gaaaacaaaa tatagcgcgc aaactaggat aaattatcgc
     4861 gcgcggtgtc atctatgtta ctagatgcgg ccgcaattgt ggatccgctc tagagtcgac
     4921 ctgcaggcat gcaagcttgc ggccgcgtat tctatagtgt cacctaaata gcatggcgta
     4981 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     5041 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     5101 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     5161 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     5221 gctc
//
