LOCUS       sequence-382264-        4009 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transient expression of AtOEP7-VC155 (mVenus beta-strands 8 to 11, 
            amino acids 155−239) in plant cell (Chloroplast outer envelope 
            membrane).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4009)
  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 4009)
  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)-VC155
FEATURES             Location/Qualifiers
     source          1..4009
                     /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             1158..1409
                     /codon_start=1
                     /product="C-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VC155
                     /translation="DKQKNGIKANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNH
                     YLSYQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYK"
     primer_bind     1346..1367
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(1414..1438)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      1483..1733
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(1798..1816)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1799..1816)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1834..1850)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1834..1850)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1847..1869)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1858..1874
                     /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(1882..1888,1889..1906,1907..1912))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1927..1948
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2065..2082)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2236..2824)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2316..2335)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2982..3797)
                     /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     3705..3724
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3789..3808
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     3982..4000
                     /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 gcggtagcgg cggtgtggac aagcagaaga acggcatcaa ggccaacttc aagatccgcc
     1201 acaacatcga ggacggcggc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
     1261 gcgacggccc cgtgctgctg cccgacaacc actacctgag ctaccagtcc aagctgagca
     1321 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
     1381 tcactctcgg catggacgag ctgtacaagt gagacccagc tttcttgtac aaagtggttg
     1441 gaggctcagg aggctcccca tgggctagca ctagtatcta gaatcgttca aacatttggc
     1501 aataaagttt cttaagattg aatcctgttg ccggtcttgc gatgattatc atataatttc
     1561 tgttgaatta cgttaagcat gtaataatta acatgtaatg catgacgtta tttatgagat
     1621 gggtttttat gattagagtc ccgcaattat acatttaata cgcgatagaa aacaaaatat
     1681 agcgcgcaaa ctaggataaa ttatcgcgcg cggtgtcatc tatgttacta gatgcggccg
     1741 caattgtgga tccgctctag agtcgacctg caggcatgca agcttgcggc cgcgtattct
     1801 atagtgtcac ctaaatagca tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     1861 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     1921 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     1981 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     2041 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     2101 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     2161 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     2221 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     2281 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     2341 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     2401 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     2461 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     2521 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     2581 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     2641 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     2701 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     2761 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     2821 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     2881 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     2941 aaaatgaagt tttaaatcaa tctaaagtat atatgtttta attagaaaaa ctcatcgagc
     3001 atcaaatgaa actgcaattt attcatatca ggattatcaa taccatattt ttgaaaaagc
     3061 cgtttctgta atgaaggaga aaactcaccg aggcagttcc ataggatggc aagatcctgg
     3121 tatcggtctg cgattccgac tcgtccaaca tcaatacaac ctattaattt cccctcgtca
     3181 aaaataaggt tatcaagtga gaaatcacca tgagtgacga ctgaatccgg tgagaatggc
     3241 aatagcttat gcatttcttt ccagacttgt tcaacaggcc agccattacg ctcgtcatca
     3301 aaatcactcg catcaaccaa accgttattc attcgtgatt gcgcctgagc gagacgaaat
     3361 acgcgatcgc tgttaaaagg acaattacaa acaggaatcg aatgcaaccg gcgcaggaac
     3421 actgccagcg catcaacaat attttcacct gaatcaggat attcttctaa tacctggaat
     3481 gctgttttcc ctgggatcgc agaggtgagt aaccatgcat catcaggagt acggataaaa
     3541 tgcttgatgg tcggaagagg cataaattcc gtcagccagt ttagtctgac catctcatct
     3601 gtaacatcat tggcaacgct acctttgcca tgtttcagaa acaactctgg cgcatcgggc
     3661 ttcccataca atcgatagat tgtcgcacct gattgcccga cattatcgcg agcccattta
     3721 tacccatata aatcagcatc catgttggaa tttaatcgcg gccttgagca agacgtttcc
     3781 cgttgaatat ggctcataac accccttgta ttactgttta tgtaagcaga cataagcatt
     3841 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     3901 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt
     3961 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtc
//
