LOCUS       sequence-382260-        4219 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transient expression of AtOEP7-VN154 (mVenus beta-strands 1 to 7, 
            amino acids 1−154) in plant cell (Chloroplast outer envelope 
            membrane).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4219)
  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 4219)
  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-AtOEP7(1-50aa)-VN154
FEATURES             Location/Qualifiers
     source          1..4219
                     /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             join(1155..1157,1158..1160,1161..1619)
                     /codon_start=1
                     /product="improved N-terminal fragment of mVenus for use in
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN155(I152L)
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITA"
     primer_bind     complement(1200..1221)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1461..1480)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     protein_bind    complement(1624..1648)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      1693..1943
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(2008..2026)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2009..2026)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2044..2060)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2044..2060)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2057..2079)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2068..2084
                     /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(2092..2098,2099..2116,2117..2122))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2137..2158
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2275..2292)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2446..3034)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2526..2545)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3192..4007)
                     /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     3915..3934
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3999..4018
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     4192..4210
                     /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 gcaggctcca tgggaaaaac ttcgggagcg aaacaggcga
     1021 ctgtggtggt cgcagcgatg gcgttaggat ggttagccat agagatcgct ttcaagcctt
     1081 tcctcgataa attccgctcc tcaatcgaca aatctgaccc aaccaaagac cccgatgacg
     1141 gtggctctgg aggtatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
     1201 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg
     1261 gcgatgccac ctacggcaag ctgaccctga agctgatctg caccaccggc aagctgcccg
     1321 tgccctggcc caccctcgtg accaccctgg gctacggcct gcagtgcttc gcccgctacc
     1381 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
     1441 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
     1501 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
     1561 acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat atcaccgcct
     1621 gagacccagc tttcttgtac aaagtggttg gaggctcagg aggctcccca tgggctagca
     1681 ctagtatcta gaatcgttca aacatttggc aataaagttt cttaagattg aatcctgttg
     1741 ccggtcttgc gatgattatc atataatttc tgttgaatta cgttaagcat gtaataatta
     1801 acatgtaatg catgacgtta tttatgagat gggtttttat gattagagtc ccgcaattat
     1861 acatttaata cgcgatagaa aacaaaatat agcgcgcaaa ctaggataaa ttatcgcgcg
     1921 cggtgtcatc tatgttacta gatgcggccg caattgtgga tccgctctag agtcgacctg
     1981 caggcatgca agcttgcggc cgcgtattct atagtgtcac ctaaatagca tggcgtaatc
     2041 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     2101 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     2161 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     2221 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     2281 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     2341 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     2401 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     2461 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     2521 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     2581 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     2641 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     2701 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     2761 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     2821 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     2881 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     2941 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     3001 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     3061 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     3121 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     3181 atatgtttta attagaaaaa ctcatcgagc atcaaatgaa actgcaattt attcatatca
     3241 ggattatcaa taccatattt ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg
     3301 aggcagttcc ataggatggc aagatcctgg tatcggtctg cgattccgac tcgtccaaca
     3361 tcaatacaac ctattaattt cccctcgtca aaaataaggt tatcaagtga gaaatcacca
     3421 tgagtgacga ctgaatccgg tgagaatggc aatagcttat gcatttcttt ccagacttgt
     3481 tcaacaggcc agccattacg ctcgtcatca aaatcactcg catcaaccaa accgttattc
     3541 attcgtgatt gcgcctgagc gagacgaaat acgcgatcgc tgttaaaagg acaattacaa
     3601 acaggaatcg aatgcaaccg gcgcaggaac actgccagcg catcaacaat attttcacct
     3661 gaatcaggat attcttctaa tacctggaat gctgttttcc ctgggatcgc agaggtgagt
     3721 aaccatgcat catcaggagt acggataaaa tgcttgatgg tcggaagagg cataaattcc
     3781 gtcagccagt ttagtctgac catctcatct gtaacatcat tggcaacgct acctttgcca
     3841 tgtttcagaa acaactctgg cgcatcgggc ttcccataca atcgatagat tgtcgcacct
     3901 gattgcccga cattatcgcg agcccattta tacccatata aatcagcatc catgttggaa
     3961 tttaatcgcg gccttgagca agacgtttcc cgttgaatat ggctcataac accccttgta
     4021 ttactgttta tgtaagcaga cataagcatt tatcagggtt attgtctcat gagcggatac
     4081 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     4141 gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt
     4201 atcacgaggc cctttcgtc
//
