LOCUS       sequence-361248-        4904 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transient expression of KOC155-MXMT in plant cell.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4904)
  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 4904)
  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-KOC155-MXMT
FEATURES             Location/Qualifiers
     source          1..4904
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    566..590
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(792..816)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    860..915
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5'-leader sequence (Gallie et al., 1988)"
     promoter        complement(935..1279)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(972..992)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     primer_bind     complement(1355..1374)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1356..1374)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1384..1401)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1384..1400)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1393..1415)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1609..1628)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1728..1750
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1788..1806)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1980..1999)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1991..2806
                     /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     complement(2064..2083)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2964..3552
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3453..3472
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3695..3712
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3829..3850
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3865..3870,3871..3888,3889..3895)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3903..3919
                     /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)."
     primer_bind     3908..3930
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3927..3943
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3927..3943
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        3961..3979
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     3961..3978
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     terminator      complement(4044..4294)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
ORIGIN
        1 ccctttgttt gcaccaatcc ccaattcaat caccaaaccg ctgggaacct gagataacca
       61 atgaacactg taacaagagt gcaaaaaatg catggactcc tcggggaaga gtctgccgta
      121 gaaagagcca ggcattgcgc ttattaggca cgatcctatc ttgcgtccat tttctttctc
      181 gagtttgcgg tagaagcttg gcagcaactt gaaaaccgaa ttgaaatcat tttggaaaag
      241 atcattcaga aaaatctgaa tggtgggacg ttctaattca ttcttctctt cctggccaac
      301 tttgtcaata ctttgcacaa tgtcccgcac tgttaaaagt gtgtttggtc cagaagcgca
      361 tcccaaatcc gcaactttaa tgcacttgtt gatgttgggc aagttggccc gcaacaattc
      421 tcgtatgcat tgttcaagga aaggtttcac cttggcaaga gccagattgt aggatgcatt
      481 cttggcgtag cttgtatcgc cttcaccttc attcatatgc aggacttctt ggagctccat
      541 ccatggggag cctcctgagc ctccaaccac tttgtacaag aaagctgggt cttaggaatg
      601 agctactgca tcttctacct gctcagtaat gttgccttcg gttttcctga cgaggcgatg
      661 gccgatgtaa tggtctcctg gcatttcaag aatctctttt gccgccttgt aagtagtctt
      721 catttggcat ttgtgattgc cgcctccttc aagttttagg tacatcgtaa catcaccctt
      781 cagaaccatg aagcctgctt ttttgtacaa acttgtggag cctcctgagc ctccaagctt
      841 ggtaccgagc tcgaattctg taattgtaaa tagtaattgt aatgttgttt gttgtttgtt
      901 gttgttggta attgttgaag gatctcaaga agcctgtcct ctccaaatga aatgaacttc
      961 cttatataga ggaagggtct tgcgaaggat agtgggattg tgcgtcatcc cttacgtcag
     1021 tggagatatc acatcaatcc acttgctttg aagacgtggt tggaacgtct tctttttcca
     1081 cgatgttcct cgtgggtggg ggtccatctt tgggaccact gtcggtagag gcatcttgaa
     1141 cgatagcctt tcctttatcg caatgatggc atttgtagga gccaccttcc ttttctactg
     1201 tcctttcgat aagtgacaga tagctgggca atggaatccg aggaggtttc ccgatattac
     1261 cctttgttga aaagtctcaa tagccctcgg tcttctgagc ccgggagatc tggcctcgag
     1321 atcccgggta ccgagctcga attctgcggc cgcgccctat agtgagtcgt attacactag
     1381 ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa
     1441 tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga
     1501 tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg cgcctgtagt aatattttct
     1561 ccttacgcat ctgtgcggta tttcacaccg catatggtgc actctcagta caatctgctc
     1621 tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg
     1681 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     1741 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg
     1801 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
     1861 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     1921 tccgctcatg agacaataac cctgataaat gcttatgtct gcttacataa acagtaatac
     1981 aaggggtgtt atgagccata ttcaacggga aacgtcttgc tcaaggccgc gattaaattc
     2041 caacatggat gctgatttat atgggtataa atgggctcgc gataatgtcg ggcaatcagg
     2101 tgcgacaatc tatcgattgt atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg
     2161 caaaggtagc gttgccaatg atgttacaga tgagatggtc agactaaact ggctgacgga
     2221 atttatgcct cttccgacca tcaagcattt tatccgtact cctgatgatg catggttact
     2281 cacctctgcg atcccaggga aaacagcatt ccaggtatta gaagaatatc ctgattcagg
     2341 tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg
     2401 taattgtcct tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa
     2461 taacggtttg gttgatgcga gtgattttga tgacgagcgt aatggctggc ctgttgaaca
     2521 agtctggaaa gaaatgcata agctattgcc attctcaccg gattcagtcg tcactcatgg
     2581 tgatttctca cttgataacc ttatttttga cgaggggaaa ttaataggtt gtattgatgt
     2641 tggacgagtc ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg
     2701 tgagttttct ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga
     2761 tatgaataaa ttgcagtttc atttgatgct cgatgagttt ttctaattaa aacatatata
     2821 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     2881 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     2941 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     3001 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     3061 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     3121 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     3181 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     3241 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     3301 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     3361 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     3421 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     3481 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     3541 agcctatgga aaaacgcctg cggcgttggc ttcttccggt gctttgcttt ttgctcacat
     3601 gttctttccg gctttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     3661 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     3721 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg
     3781 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta
     3841 gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg
     3901 aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt acgccatgct
     3961 atttaggtga cactatagaa tacgcggccg caagcttgca tgcctgcagg tcgactctag
     4021 agcggatcca caattgcggc cgcatctagt aacatagatg acaccgcgcg cgataattta
     4081 tcctagtttg cgcgctatat tttgttttct atcgcgtatt aaatgtataa ttgcgggact
     4141 ctaatcataa aaacccatct cataaataac gtcatgcatt acatgttaat tattacatgc
     4201 ttaacgtaat tcaacagaaa ttatatgata atcatcgcaa gaccggcaac aggattcaat
     4261 cttaagaaac tttattgcca aatgtttgaa cgattctaga tactagttta cacgtctgac
     4321 ttctctggct ttttggcgag agaaatgata agattattat agcagccttt gcccatgtgg
     4381 agaacctttg ctgcatgctt cgcaagcctg tggaataagt caggcataat agcttctcca
     4441 aaatgacttg cgaggatggg ttcgtaaact gatctaatta atgatgccac atactctgct
     4501 ttaatttgtt catgggatct tactgggtaa tcatcatcaa tagagaaggc agcatcataa
     4561 tgggccttaa aagtctccag atataaaatt tcgcaagaac cttcctcctc aactatgcac
     4621 tttacttctt ctgctgaagg tgtaaagaat ggaatattga aactatccaa tttttcttcc
     4681 tccagaagtc cctcaacaat caagtcgttt attgccatgt caagtaagtc taggggattc
     4741 ggttcgtcga attcatctac tttacaaatg caggtaagga gcattcggcc acgtgaaaac
     4801 aactctttcg aatgaatcct tagaaatgtg gtaaaatctt tcgtaaattg atccaaatat
     4861 gccttctgga cgggcggacg acatcctttg gaagagtaaa tact
//
