LOCUS       sequence-382266-        3876 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transient expression of VN155 (mVenus beta-strands 8 to 11, amino 
            acids 155−239) in plant cell (Cytosol).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3876)
  AUTHORS   Ichikawa S, Kato S, Fujii Y, Ishikawa K, Numata K, Kodama Y
  TITLE     Organellar Glue: A Molecular Tool to Artificially Control 
            Chloroplast-Chloroplast Interactions.
  JOURNAL   ACS Synth Biol. 2022 Oct 21;11(10):3190-3197. doi: 
            10.1021/acssynbio.2c00367. Epub 2022 Sep 30.
  PUBMED    36178266
REFERENCE   2  (bases 1 to 3876)
  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: 
            "10.1021/acssynbio.2c00367"; journalName: "ACS Synth Biol"; date: 
            "2022-10-21- 21"; volume: "11"; issue: "10"; pages: "3190-3197"
COMMENT     Sequence Label: pGWT35S-VC155
FEATURES             Location/Qualifiers
     source          1..3876
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     405..424
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        405..423
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        500..844
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     787..807
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     misc_feature    864..919
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5'-leader sequence (Gallie et al., 1988)"
     protein_bind    963..987
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1017..1268
                     /codon_start=1
                     /product="C-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VC155
                     /translation="DKQKNGIKANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNH
                     YLSYQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYK"
     primer_bind     1205..1226
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(1281..1305)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      1350..1600
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(1665..1683)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1666..1683)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1701..1717)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1701..1717)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1714..1736)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1725..1741
                     /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(1749..1755,1756..1773,1774..1779))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1794..1815
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1932..1949)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2103..2691)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2183..2202)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2849..3664)
                     /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     3572..3591
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3656..3675
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     3849..3867
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaatattac tacaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgagct agtgtaatac gactcactat
      421 agggcgcggc cgcagaattc gagctcggta cccgggatct cgaggccaga tctcccgggc
      481 tcagaagacc gagggctatt gagacttttc aacaaagggt aatatcggga aacctcctcg
      541 gattccattg cccagctatc tgtcacttat cgaaaggaca gtagaaaagg aaggtggctc
      601 ctacaaatgc catcattgcg ataaaggaaa ggctatcgtt caagatgcct ctaccgacag
      661 tggtcccaaa gatggacccc cacccacgag gaacatcgtg gaaaaagaag acgttccaac
      721 cacgtcttca aagcaagtgg attgatgtga tatctccact gacgtaaggg atgacgcaca
      781 atcccactat ccttcgcaag acccttcctc tatataagga agttcatttc atttggagag
      841 gacaggcttc ttgagatcct tcaacaatta ccaacaacaa caaacaacaa acaacattac
      901 aattactatt tacaattaca gaattcgagc tcggtaccaa gcttggaggc tcaggaggct
      961 ccacaagttt gtacaaaaaa gcaggcttta aaggaaccaa ttcagtcgac atggtggaca
     1021 agcagaagaa cggcatcaag gccaacttca agatccgcca caacatcgag gacggcggcg
     1081 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     1141 ccgacaacca ctacctgagc taccagtccg ccctgagcaa agaccccaac gagaagcgcg
     1201 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
     1261 tgtacaagta agatatctag acccagcttt cttgtacaaa gtggttggag gctcaggagg
     1321 ctccccatgg gctagcacta gtatctagaa tcgttcaaac atttggcaat aaagtttctt
     1381 aagattgaat cctgttgccg gtcttgcgat gattatcata taatttctgt tgaattacgt
     1441 taagcatgta ataattaaca tgtaatgcat gacgttattt atgagatggg tttttatgat
     1501 tagagtcccg caattataca tttaatacgc gatagaaaac aaaatatagc gcgcaaacta
     1561 ggataaatta tcgcgcgcgg tgtcatctat gttactagat gcggccgcaa ttgtggatcc
     1621 gctctagagt cgacctgcag gcatgcaagc ttgcggccgc gtattctata gtgtcaccta
     1681 aatagcatgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     1741 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     1801 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     1861 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     1921 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     1981 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     2041 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     2101 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     2161 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     2221 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     2281 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     2341 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     2401 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     2461 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     2521 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     2581 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     2641 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     2701 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     2761 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     2821 aaatcaatct aaagtatata tgttttaatt agaaaaactc atcgagcatc aaatgaaact
     2881 gcaatttatt catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg
     2941 aaggagaaaa ctcaccgagg cagttccata ggatggcaag atcctggtat cggtctgcga
     3001 ttccgactcg tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat
     3061 caagtgagaa atcaccatga gtgacgactg aatccggtga gaatggcaat agcttatgca
     3121 tttctttcca gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat
     3181 caaccaaacc gttattcatt cgtgattgcg cctgagcgag acgaaatacg cgatcgctgt
     3241 taaaaggaca attacaaaca ggaatcgaat gcaaccggcg caggaacact gccagcgcat
     3301 caacaatatt ttcacctgaa tcaggatatt cttctaatac ctggaatgct gttttccctg
     3361 ggatcgcaga ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg
     3421 gaagaggcat aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg
     3481 caacgctacc tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaatc
     3541 gatagattgt cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat
     3601 cagcatccat gttggaattt aatcgcggcc ttgagcaaga cgtttcccgt tgaatatggc
     3661 tcataacacc ccttgtatta ctgtttatgt aagcagacat aagcatttat cagggttatt
     3721 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     3781 gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa
     3841 cctataaaaa taggcgtatc acgaggccct ttcgtc
//
