LOCUS       sequence-347623-        7564 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 1 module containing 35S-HSP21-nVENUS_P2 F.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7564)
  AUTHORS   Velay F, Soula M, Mehrez M, Belbachir C, D'Alessandro S, Laloi C, 
            Crete P, Field B
  TITLE     MoBiFC: development of a modular bimolecular fluorescence 
            complementation toolkit for the analysis of chloroplast 
            protein-protein interactions.
  JOURNAL   Plant Methods. 2022 May 26;18(1):69. doi: 
            10.1186/s13007-022-00902-1.
  PUBMED    35619173
REFERENCE   2  (bases 1 to 7564)
  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"; journalName: "Plant 
            Methods."; date: "2022-05-26"; volume: "18(1):69. doi"; pages: " 
            10.1186/s13007-022-00902-1"
COMMENT     Sequence Label: pLBT0080
FEATURES             Location/Qualifiers
     source          1..7564
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1023..1368
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     1311..1331
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             join(2381..2383,2384..2896)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     LICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIE"
     primer_bind     complement(2423..2444)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2684..2703)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    3048..3222
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    3267..3291
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(3447..4595)
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     rep_origin      complement(4931..5519)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5011..5030)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5690..6481,6482..6550))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6313..6332
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6551..6655)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6742..7373
                     /label=oriV
                     /note="incP origin of replication"
     misc_feature    7465..7489
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
ORIGIN
        1 tctgtgaaga caagcaagaa ttcaagcttg gaggaattcc aatcccacaa aaatctgagc
       61 ttaacagcac agttgctcct ctcagagcag aatcgggtat tcaacaccct catatcaact
      121 actacgttgt gtataacggt ccacatgccg gtatatacga tgactggggt tgtacaaagg
      181 cggcaacaaa cggcgttccc ggagttgcac acaagaaatt tgccactatt acagaggcaa
      241 gagcagcagc tgacgcgtac acaacaagtc agcaaacaga caggttgaac ttcatcccca
      301 aaggagaagc tcaactcaag cccaagagct ttgctaaggc cctaacaagc ccaccaaagc
      361 aaaaagccca ctggctcacg ctaggaacca aaaggcccag cagtgatcca gccccaaaag
      421 agatctcctt tgccccggag attacaatgg acgatttcct ctatctttac gatctaggaa
      481 ggaagttcga aggtgaaggt gacgacacta tgttcaccac tgataatgag aaggttagcc
      541 tcttcaattt cagaaagaat gctgacccac agatggttag agaggcctac gcagcaagtc
      601 tcatcaagac gatctacccg agtaacaatc tccaggagat caaatacctt cccaagaagg
      661 ttaaagatgc agtcaaaaga ttcaggacta attgcatcaa gaacacagag aaagacatat
      721 ttctcaagat cagaagtact attccagtat ggacgattca aggcttgctt cataaaccaa
      781 ggcaagtaat agagattgga gtctctaaaa aggtagttcc tactgaatct aaggccatgc
      841 atggagtcta agattcaaat cgaggatcta acagaactcg ccgtcaagac tggcgaacag
      901 ttcatacaga gtcttttacg actcaatgac aagaagaaaa tcttcgtcaa catggtggag
      961 cacgacactc tggtctactc caaaaatgtc aaagatacag tctcagaaga tcaaagggct
     1021 attgagactt ttcaacaaag gataatttcg ggaaacctcc tcggattcca ttgcccagct
     1081 atctgtcact tcatcgaaag gacagtagaa aaggaaggtg gctcctacaa atgccatcat
     1141 tgcgataaag gaaaggctat cattcaagat ctctctgccg acagtggtcc caaagatgga
     1201 cccccaccca cgaggagcat cgtggaaaaa gaagaggttc caaccacgtc tacaaagcaa
     1261 gtggattgat gtgacatctc cactgacgta agggatgacg cacaatccca ctatccttcg
     1321 caagaccctt cctctatata aggaagttca tttcatttgg agaggacacg ctcgagtata
     1381 agagctcatt tttacaacaa ttaccaacaa caacaaacaa caaacaacat tacaattaca
     1441 tttacaatta tcgatacaat ggcttcctct atgctctctt ccgctactat ggttgcctct
     1501 ccggctcagg ccactatggt cgctcctttc aacggactta agtcctccgc tgccttccca
     1561 gccacccgca aggctaacaa cgacattact tccatcacaa gcaacggcgg aagagttaac
     1621 tgcatgcagg tcatttatat ttcttctttc acttttttat tattccatat gatttttttc
     1681 ggttctttct tcgaatctac ataaactaat atcattggaa aaatcgaaaa aataggtgtg
     1741 gcctccgatt ggaaagaaga agtttgagac tctctcttac cttcctgacc ttaccgattc
     1801 cgaaggtgct caagaccaga gagaaaactc cattgatgtt gttcagcaag gacaacaaaa
     1861 agggaaccaa ggatcaagtg tcgaaaaaag acctcaacaa cgcttaacca tggatgtctc
     1921 tcctttcgga ttgttggatc ctttgtcacc aatgaggacg atgcgacaaa tgttagatac
     1981 tatggacagg atgttcgagg acactatgcc tgtctcagga agaaacagag gaggaagtgg
     2041 agtgtcagag attcgtgcac cgtgggacat caaagaggaa gaacacgaga tcaagatgcg
     2101 tttcgacatg cctggactct ctaaagagga cgtcaaaatc tctgtagaag ataacgtact
     2161 tgtgatcaaa ggagagcaga agaaggagga cagtgatgat tcttggtctg gaagaagcgt
     2221 tagctcatat ggaacacgac ttcagctccc agacaactgt gagaaagaca agatcaaagc
     2281 tgagctcaag aacggagtcc tctttatcac tatccctaag accaaagtcg aacgcaaagt
     2341 catcgatgtc cagattcaat cttcgagatc tattgctact gtgagcaagg gcgaggagct
     2401 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
     2461 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagctgat
     2521 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgggctacgg
     2581 cctgcagtgc ttcgcccgct accccgacca catgaagcag cacgacttct tcaagtccgc
     2641 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa
     2701 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
     2761 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag
     2821 ccacaacgtc tatatcaccg ccgacaagca gaagaacggc atcaaggcca acttcaagat
     2881 ccgccacaac atcgaggacg gcggcgtgca gctcgccgac cactaccagc agaacacccc
     2941 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagctacc agtccaagct
     3001 gagcaaggat tgagcttctc tagctagagt cgatcgacaa gctcgagttt ctccataata
     3061 atgtgtgagt agttcccaga taagggaatt agggttccta tagggtttcg ctcatgtgtt
     3121 gagcatataa gaaaccctta gtatgtattt gtatttgtaa aatacttcta tcaataaaat
     3181 ttctaattcc taaaaccaaa atccagtact aaaatccaga tcgctactat tgtcttctgc
     3241 acgaagtggt ttaaactatc agtgtttgac aggatatatt ggcgggtaaa cctaagagaa
     3301 aagagcgttt attagaataa tcggatattt aaaagggcgt gaaaaggttt atccgttcgt
     3361 ccatttgtat gtgcatgcca accacagggt tccccagatc aggcgctggc tgctgaaccc
     3421 ccagccggaa ctgaccccac aaggccctag cgtttgcaat gcaccaggtc atcattgacc
     3481 caggcgtgtt ccaccaggcc gctgcctcgc aactcttcgc aggcttcgcc gacctgctcg
     3541 cgccacttct tcacgcgggt ggaatccgat ccgcacatga ggcggaaggt ttccagcttg
     3601 agcgggtacg gctcccggtg cgagctgaaa tagtcgaaca tccgtcgggc cgtcggcgac
     3661 agcttgcggt acttctccca tatgaatttc gtgtagtggt cgccagcaaa cagcacgacg
     3721 atttcctcgt cgatcaggac ctggcaacgg gacgttttct tgccacggtc caggacgcgg
     3781 aagcggtgca gcagcgacac cgattccagg tgcccaacgc ggtcggacgt gaagcccatc
     3841 gccgtcgcct gtaggcgcga caggcattcc tcggccttcg tgtaataccg gccattgatc
     3901 gaccagccca ggtcctggca aagctcgtag aacgtgaagg tgatcggctc gccgataggg
     3961 gtgcgcttcg cgtactccaa cacctgctgc cacaccagtt cgtcatcgtc ggcccgcagc
     4021 tcgacgccgg tgtaggtgat cttcacgtcc ttgttgacgt ggaaaatgac cttgttttgc
     4081 agcgcctcgc gcgggatttt cttgttgcgc gtggtgaaca gggcagagcg ggccgtgtcg
     4141 tttggcatcg ctcgcatcgt gtccggccac ggcgcaatat cgaacaagga aagctgcatt
     4201 tccttgatct gctgcttcgt gtgtttcagc aacgcggcct gcttggcctc gctgacctgt
     4261 tttgccaggt cctcgccggc ggtttttcgc ttcttggtcg tcatagttcc tcgcgtgtcg
     4321 atggtcatcg acttcgccaa acctgccgcc tcctgttcaa gacgacgcga acgctccacg
     4381 gcggccgatg gcgcgggcag ggcaggggga gccagttgca cgctgtcgcg ctcgatcttg
     4441 gccgtagctt gctggaccat cgagccgacg gactggaagg tttcgcgggg cgcacgcatg
     4501 acggtgcggc ttgcgatggt ttcggcatcc tcggcggaaa accccgcgtc gatcagttct
     4561 tgcctgtatg ccttccggtc aaacgtccga ttcattcacc ctccttgcgg gattgccccg
     4621 actcacgccg gggcaatgtg cccttattcc tgatttgacc cgcctggtgc cttggtgtcc
     4681 agataatcca ccttatcggc aatgaagtcg gtcccgtaga ccgtctggcc gtccttctcg
     4741 tacttggtat tccgaatctt gccctgcacg aataccagcg accccttgcc caaatacttg
     4801 ccgtgggcct cggcctgaga gccaaaacac ttgatgcgga agaagtcggt gcgctcctgc
     4861 ttgtcgccgg catcgttgcg ccacatctag gatctgccag gaaccgtaaa aaggccgcgt
     4921 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     4981 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     5041 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     5101 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     5161 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     5221 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     5281 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     5341 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     5401 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     5461 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     5521 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     5581 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     5641 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     5701 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     5761 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     5821 tgataccgcg agaaccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     5881 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     5941 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     6001 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     6061 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     6121 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     6181 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     6241 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     6301 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     6361 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     6421 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     6481 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     6541 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcaaggt tattgtctca
     6601 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacga
     6661 attggccagc gctgccattt ttggggtgag gccgttcgcg gccgaggggc gcagcccctg
     6721 gggggatggg aggcccgcgt tagcgggccg ggagggttcg agaagggggg gcacccccct
     6781 tcggcgtgcg cggtcacgcg cacagggcgc agccctggtt aaaaacaagg tttataaata
     6841 ttggtttaaa agcaggttaa aagacaggtt agcggtggcc gaaaaacggg cggaaaccct
     6901 tgcaaatgct ggattttctg cctgtggaca gcccctcaaa tgtcaatagg tgcgcccctc
     6961 atctgtcagc actctgcccc tcaagtgtca aggatcgcgc ccctcatctg tcagtagtcg
     7021 cgcccctcaa gtgtcaatac cgcagggcac ttatccccag gcttgtccac atcatctgtg
     7081 ggaaactcgc gtaaaatcag gcgttttcgc cgatttgcga ggctggccag ctccacgtcg
     7141 ccggccgaaa tcgagcctgc ccctcatctg tcaacgccgc gccgggtgag tcggcccctc
     7201 aagtgtcaac gtccgcccct catctgtcag tgagggccaa gttttccgcg aggtatccac
     7261 aacgccggcg gccgcggtgt ctcgcacacg gcttcgacgg cgtttctggc gcgtttgcag
     7321 ggccatagac ggccgccagc ccagcggcga gggcaaccag cccggtgagc gtcgcaaagg
     7381 agatcctgat ctgactgatg ggctgcctgt atcgagtggt gattttgtgc cgagctgccg
     7441 gtcggggagc tgttggctgg ctggtggcag gatatattgt ggtgtaaaca aattgacgct
     7501 tagacaactt aataacacat tgcggacgtt tttaatgtac tggggtggat gcagtgggcc
     7561 ccac
//
