LOCUS       sequence-347994-        7720 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 1 module containing 35S-CTP-mCHERRY-nVENUS_P2 F.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7720)
  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 7720)
  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: pLBT0140
FEATURES             Location/Qualifiers
     source          1..7720
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        994..1339
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     1282..1302
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             1779..2486
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1924..1942)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     CDS             join(2508..2510,2511..3023)
                     /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(2550..2571)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2811..2830)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    3175..3349
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    3394..3418
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(3574..4722)
                     /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(5058..5646)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5138..5157)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5817..6608,6609..6677))
                     /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     6440..6459
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6678..6782)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6869..7500
                     /label=oriV
                     /note="incP origin of replication"
     misc_feature    7592..7616
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
ORIGIN
        1 ggaggaattc caatcccaca aaaatctgag cttaacagca cagttgctcc tctcagagca
       61 gaatcgggta ttcaacaccc tcatatcaac tactacgttg tgtataacgg tccacatgcc
      121 ggtatatacg atgactgggg ttgtacaaag gcggcaacaa acggcgttcc cggagttgca
      181 cacaagaaat ttgccactat tacagaggca agagcagcag ctgacgcgta cacaacaagt
      241 cagcaaacag acaggttgaa cttcatcccc aaaggagaag ctcaactcaa gcccaagagc
      301 tttgctaagg ccctaacaag cccaccaaag caaaaagccc actggctcac gctaggaacc
      361 aaaaggccca gcagtgatcc agccccaaaa gagatctcct ttgccccgga gattacaatg
      421 gacgatttcc tctatcttta cgatctagga aggaagttcg aaggtgaagg tgacgacact
      481 atgttcacca ctgataatga gaaggttagc ctcttcaatt tcagaaagaa tgctgaccca
      541 cagatggtta gagaggccta cgcagcaagt ctcatcaaga cgatctaccc gagtaacaat
      601 ctccaggaga tcaaatacct tcccaagaag gttaaagatg cagtcaaaag attcaggact
      661 aattgcatca agaacacaga gaaagacata tttctcaaga tcagaagtac tattccagta
      721 tggacgattc aaggcttgct tcataaacca aggcaagtaa tagagattgg agtctctaaa
      781 aaggtagttc ctactgaatc taaggccatg catggagtct aagattcaaa tcgaggatct
      841 aacagaactc gccgtcaaga ctggcgaaca gttcatacag agtcttttac gactcaatga
      901 caagaagaaa atcttcgtca acatggtgga gcacgacact ctggtctact ccaaaaatgt
      961 caaagataca gtctcagaag atcaaagggc tattgagact tttcaacaaa ggataatttc
     1021 gggaaacctc ctcggattcc attgcccagc tatctgtcac ttcatcgaaa ggacagtaga
     1081 aaaggaaggt ggctcctaca aatgccatca ttgcgataaa ggaaaggcta tcattcaaga
     1141 tctctctgcc gacagtggtc ccaaagatgg acccccaccc acgaggagca tcgtggaaaa
     1201 agaagaggtt ccaaccacgt ctacaaagca agtggattga tgtgacatct ccactgacgt
     1261 aagggatgac gcacaatccc actatccttc gcaagaccct tcctctatat aaggaagttc
     1321 atttcatttg gagaggacac gctcgagtat aagagctcat ttttacaaca attaccaaca
     1381 acaacaaaca acaaacaaca ttacaattac atttacaatt atcgatacaa tggcttcctc
     1441 tatgctctct tccgctacta tggttgcctc tccggctcag gccactatgg tcgctccttt
     1501 caacggactt aagtcctccg ctgccttccc agccacccgc aaggctaaca acgacattac
     1561 ttccatcaca agcaacggcg gaagagttaa ctgcatgcag gtcatttata tttcttcttt
     1621 cactttttta ttattccata tgattttttt cggttctttc ttcgaatcta cataaactaa
     1681 tatcattgga aaaatcgaaa aaataggtgt ggcctccgat tggaaagaag aagtttgaga
     1741 ctctctctta ccttcctgac cttaccgatt ccgaaggtat ggtgagcaag ggcgaggagg
     1801 ataacatggc catcatcaag gagttcatgc gcttcaaggt gcacatggag ggctccgtga
     1861 acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga
     1921 ccgccaagct gaaggtgacc aagggtggcc ccctgccctt cgcctgggac atcctgtccc
     1981 ctcagttcat gtacggctcc aaggcctacg tgaagcaccc cgccgacatc cccgactact
     2041 tgaagctgtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg
     2101 gcgtggtgac cgtgacccag gactcctccc tgcaggacgg cgagttcatc tacaaggtga
     2161 agctgcgcgg caccaacttc ccctccgacg gccccgtaat gcagaagaaa accatgggct
     2221 gggaggcctc ctccgagcgg atgtaccccg aggacggcgc cctgaagggc gagatcaagc
     2281 agaggctgaa gctgaaggac ggcggccact acgacgctga ggtcaagacc acctacaagg
     2341 ccaagaagcc cgtgcagctg cccggcgcct acaacgtcaa catcaagttg gacatcacct
     2401 cccacaacga ggactacacc atcgtggaac agtacgaacg cgccgagggc cgccactcca
     2461 ccggcggcat ggacgagctg tacaagggtt cgagatctat tgctactgtg agcaagggcg
     2521 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
     2581 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
     2641 agctgatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctgg
     2701 gctacggcct gcagtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca
     2761 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
     2821 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     2881 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     2941 acaacagcca caacgtctat atcaccgccg acaagcagaa gaacggcatc aaggccaact
     3001 tcaagatccg ccacaacatc gaggacggcg gcgtgcagct cgccgaccac taccagcaga
     3061 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agctaccagt
     3121 ccaagctgag caaggattga gcttctctag ctagagtcga tcgacaagct cgagtttctc
     3181 cataataatg tgtgagtagt tcccagataa gggaattagg gttcctatag ggtttcgctc
     3241 atgtgttgag catataagaa acccttagta tgtatttgta tttgtaaaat acttctatca
     3301 ataaaatttc taattcctaa aaccaaaatc cagtactaaa atccagatcg ctactattgt
     3361 cttctgcacg aagtggttta aactatcagt gtttgacagg atatattggc gggtaaacct
     3421 aagagaaaag agcgtttatt agaataatcg gatatttaaa agggcgtgaa aaggtttatc
     3481 cgttcgtcca tttgtatgtg catgccaacc acagggttcc ccagatcagg cgctggctgc
     3541 tgaaccccca gccggaactg accccacaag gccctagcgt ttgcaatgca ccaggtcatc
     3601 attgacccag gcgtgttcca ccaggccgct gcctcgcaac tcttcgcagg cttcgccgac
     3661 ctgctcgcgc cacttcttca cgcgggtgga atccgatccg cacatgaggc ggaaggtttc
     3721 cagcttgagc gggtacggct cccggtgcga gctgaaatag tcgaacatcc gtcgggccgt
     3781 cggcgacagc ttgcggtact tctcccatat gaatttcgtg tagtggtcgc cagcaaacag
     3841 cacgacgatt tcctcgtcga tcaggacctg gcaacgggac gttttcttgc cacggtccag
     3901 gacgcggaag cggtgcagca gcgacaccga ttccaggtgc ccaacgcggt cggacgtgaa
     3961 gcccatcgcc gtcgcctgta ggcgcgacag gcattcctcg gccttcgtgt aataccggcc
     4021 attgatcgac cagcccaggt cctggcaaag ctcgtagaac gtgaaggtga tcggctcgcc
     4081 gataggggtg cgcttcgcgt actccaacac ctgctgccac accagttcgt catcgtcggc
     4141 ccgcagctcg acgccggtgt aggtgatctt cacgtccttg ttgacgtgga aaatgacctt
     4201 gttttgcagc gcctcgcgcg ggattttctt gttgcgcgtg gtgaacaggg cagagcgggc
     4261 cgtgtcgttt ggcatcgctc gcatcgtgtc cggccacggc gcaatatcga acaaggaaag
     4321 ctgcatttcc ttgatctgct gcttcgtgtg tttcagcaac gcggcctgct tggcctcgct
     4381 gacctgtttt gccaggtcct cgccggcggt ttttcgcttc ttggtcgtca tagttcctcg
     4441 cgtgtcgatg gtcatcgact tcgccaaacc tgccgcctcc tgttcaagac gacgcgaacg
     4501 ctccacggcg gccgatggcg cgggcagggc agggggagcc agttgcacgc tgtcgcgctc
     4561 gatcttggcc gtagcttgct ggaccatcga gccgacggac tggaaggttt cgcggggcgc
     4621 acgcatgacg gtgcggcttg cgatggtttc ggcatcctcg gcggaaaacc ccgcgtcgat
     4681 cagttcttgc ctgtatgcct tccggtcaaa cgtccgattc attcaccctc cttgcgggat
     4741 tgccccgact cacgccgggg caatgtgccc ttattcctga tttgacccgc ctggtgcctt
     4801 ggtgtccaga taatccacct tatcggcaat gaagtcggtc ccgtagaccg tctggccgtc
     4861 cttctcgtac ttggtattcc gaatcttgcc ctgcacgaat accagcgacc ccttgcccaa
     4921 atacttgccg tgggcctcgg cctgagagcc aaaacacttg atgcggaaga agtcggtgcg
     4981 ctcctgcttg tcgccggcat cgttgcgcca catctaggat ctgccaggaa ccgtaaaaag
     5041 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     5101 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     5161 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     5221 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     5281 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     5341 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     5401 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     5461 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     5521 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     5581 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     5641 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     5701 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     5761 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     5821 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     5881 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     5941 gctgcaatga taccgcgaga accacgctca ccggctccag atttatcagc aataaaccag
     6001 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     6061 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     6121 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     6181 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     6241 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     6301 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     6361 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     6421 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     6481 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     6541 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     6601 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     6661 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcaaggttat
     6721 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     6781 cgcacgaatt ggccagcgct gccatttttg gggtgaggcc gttcgcggcc gaggggcgca
     6841 gcccctgggg ggatgggagg cccgcgttag cgggccggga gggttcgaga agggggggca
     6901 ccccccttcg gcgtgcgcgg tcacgcgcac agggcgcagc cctggttaaa aacaaggttt
     6961 ataaatattg gtttaaaagc aggttaaaag acaggttagc ggtggccgaa aaacgggcgg
     7021 aaacccttgc aaatgctgga ttttctgcct gtggacagcc cctcaaatgt caataggtgc
     7081 gcccctcatc tgtcagcact ctgcccctca agtgtcaagg atcgcgcccc tcatctgtca
     7141 gtagtcgcgc ccctcaagtg tcaataccgc agggcactta tccccaggct tgtccacatc
     7201 atctgtggga aactcgcgta aaatcaggcg ttttcgccga tttgcgaggc tggccagctc
     7261 cacgtcgccg gccgaaatcg agcctgcccc tcatctgtca acgccgcgcc gggtgagtcg
     7321 gcccctcaag tgtcaacgtc cgcccctcat ctgtcagtga gggccaagtt ttccgcgagg
     7381 tatccacaac gccggcggcc gcggtgtctc gcacacggct tcgacggcgt ttctggcgcg
     7441 tttgcagggc catagacggc cgccagccca gcggcgaggg caaccagccc ggtgagcgtc
     7501 gcaaaggaga tcctgatctg actgatgggc tgcctgtatc gagtggtgat tttgtgccga
     7561 gctgccggtc ggggagctgt tggctggctg gtggcaggat atattgtggt gtaaacaaat
     7621 tgacgcttag acaacttaat aacacattgc ggacgttttt aatgtactgg ggtggatgca
     7681 gtgggcccca ctctgtgaag acaagcaaga attcaagctt
//
