LOCUS       sequence-363985-        3514 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Destination vector for transient expression in plant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3514)
  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 3514)
  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
FEATURES             Location/Qualifiers
     source          1..3514
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(30..47)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(201..789)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(281..300)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(947..1762)
                     /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     1670..1689
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1754..1773
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     1947..1965
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2003..2025)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2125..2144
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2338..2360
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2352..2369
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2353..2369
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2379..2398
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2379..2397
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        2474..2818
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     2761..2781
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     misc_feature    2838..2893
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5'-leader sequence (Gallie et al., 1988)"
     terminator      2962..3212
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(3277..3295)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3278..3295)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3313..3329)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3313..3329)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3326..3348)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3337..3353
                     /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(3361..3367,3368..3385,3386..3391))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3406..3427
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
       61 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
      121 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
      181 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
      241 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
      301 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
      361 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
      421 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
      481 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
      541 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
      601 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
      661 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
      721 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
      781 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
      841 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
      901 aattaaaaat gaagttttaa atcaatctaa agtatatatg ttttaattag aaaaactcat
      961 cgagcatcaa atgaaactgc aatttattca tatcaggatt atcaatacca tatttttgaa
     1021 aaagccgttt ctgtaatgaa ggagaaaact caccgaggca gttccatagg atggcaagat
     1081 cctggtatcg gtctgcgatt ccgactcgtc caacatcaat acaacctatt aatttcccct
     1141 cgtcaaaaat aaggttatca agtgagaaat caccatgagt gacgactgaa tccggtgaga
     1201 atggcaatag cttatgcatt tctttccaga cttgttcaac aggccagcca ttacgctcgt
     1261 catcaaaatc actcgcatca accaaaccgt tattcattcg tgattgcgcc tgagcgagac
     1321 gaaatacgcg atcgctgtta aaaggacaat tacaaacagg aatcgaatgc aaccggcgca
     1381 ggaacactgc cagcgcatca acaatatttt cacctgaatc aggatattct tctaatacct
     1441 ggaatgctgt tttccctggg atcgcagagg tgagtaacca tgcatcatca ggagtacgga
     1501 taaaatgctt gatggtcgga agaggcataa attccgtcag ccagtttagt ctgaccatct
     1561 catctgtaac atcattggca acgctacctt tgccatgttt cagaaacaac tctggcgcat
     1621 cgggcttccc atacaatcga tagattgtcg cacctgattg cccgacatta tcgcgagccc
     1681 atttataccc atataaatca gcatccatgt tggaatttaa tcgcggcctt gagcaagacg
     1741 tttcccgttg aatatggctc ataacacccc ttgtattact gtttatgtaa gcagacataa
     1801 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     1861 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     1921 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc
     1981 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
     2041 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     2101 ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
     2161 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ttactacagg cgccattcgc
     2221 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
     2281 agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc
     2341 agtcacgacg ttgtaaaacg acggccagtg agctagtgta atacgactca ctatagggcg
     2401 cggccgcaga attcgagctc ggtacccggg atctcgaggc cagatctccc gggctcagaa
     2461 gaccgagggc tattgagact tttcaacaaa gggtaatatc gggaaacctc ctcggattcc
     2521 attgcccagc tatctgtcac ttatcgaaag gacagtagaa aaggaaggtg gctcctacaa
     2581 atgccatcat tgcgataaag gaaaggctat cgttcaagat gcctctaccg acagtggtcc
     2641 caaagatgga cccccaccca cgaggaacat cgtggaaaaa gaagacgttc caaccacgtc
     2701 ttcaaagcaa gtggattgat gtgatatctc cactgacgta agggatgacg cacaatccca
     2761 ctatccttcg caagaccctt cctctatata aggaagttca tttcatttgg agaggacagg
     2821 cttcttgaga tccttcaaca attaccaaca acaacaaaca acaaacaaca ttacaattac
     2881 tatttacaat tacagaattc gagctcggta ccaagcttgg aggctcagga ggctccccat
     2941 gggctagcac tagtatctag aatcgttcaa acatttggca ataaagtttc ttaagattga
     3001 atcctgttgc cggtcttgcg atgattatca tataatttct gttgaattac gttaagcatg
     3061 taataattaa catgtaatgc atgacgttat ttatgagatg ggtttttatg attagagtcc
     3121 cgcaattata catttaatac gcgatagaaa acaaaatata gcgcgcaaac taggataaat
     3181 tatcgcgcgc ggtgtcatct atgttactag atgcggccgc aattgtggat ccgctctaga
     3241 gtcgacctgc aggcatgcaa gcttgcggcc gcgtattcta tagtgtcacc taaatagcat
     3301 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     3361 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     3421 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     3481 gcattaatga atcggccaac gcgcggggag aggc
//
