LOCUS       sequence-347997-        7228 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 1 module containing 35S-CTP-mCHERRY-cVENUS_P3 F.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7228)
  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 7228)
  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: pLBT0150
FEATURES             Location/Qualifiers
     source          1..7228
                     /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             2493..2558
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     2582..2603
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2683..2857
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    2902..2926
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(3082..4230)
                     /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(4566..5154)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4646..4665)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5325..6116,6117..6185))
                     /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     5948..5967
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6186..6290)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6377..7008
                     /label=oriV
                     /note="incP origin of replication"
     misc_feature    7100..7124
                     /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 cggactacaa agaccatgac ggtgattata
     2521 aagatcatga catcgattac aaggatgacg atgacaagga ccccaacgag aagcgcgatc
     2581 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
     2641 acaagtaagc ttctctagct agagtcgatc gacaagctcg agtttctcca taataatgtg
     2701 tgagtagttc ccagataagg gaattagggt tcctataggg tttcgctcat gtgttgagca
     2761 tataagaaac ccttagtatg tatttgtatt tgtaaaatac ttctatcaat aaaatttcta
     2821 attcctaaaa ccaaaatcca gtactaaaat ccagatcgct ttacttgtct tctgcacgaa
     2881 gtggtttaaa ctatcagtgt ttgacaggat atattggcgg gtaaacctaa gagaaaagag
     2941 cgtttattag aataatcgga tatttaaaag ggcgtgaaaa ggtttatccg ttcgtccatt
     3001 tgtatgtgca tgccaaccac agggttcccc agatcaggcg ctggctgctg aacccccagc
     3061 cggaactgac cccacaaggc cctagcgttt gcaatgcacc aggtcatcat tgacccaggc
     3121 gtgttccacc aggccgctgc ctcgcaactc ttcgcaggct tcgccgacct gctcgcgcca
     3181 cttcttcacg cgggtggaat ccgatccgca catgaggcgg aaggtttcca gcttgagcgg
     3241 gtacggctcc cggtgcgagc tgaaatagtc gaacatccgt cgggccgtcg gcgacagctt
     3301 gcggtacttc tcccatatga atttcgtgta gtggtcgcca gcaaacagca cgacgatttc
     3361 ctcgtcgatc aggacctggc aacgggacgt tttcttgcca cggtccagga cgcggaagcg
     3421 gtgcagcagc gacaccgatt ccaggtgccc aacgcggtcg gacgtgaagc ccatcgccgt
     3481 cgcctgtagg cgcgacaggc attcctcggc cttcgtgtaa taccggccat tgatcgacca
     3541 gcccaggtcc tggcaaagct cgtagaacgt gaaggtgatc ggctcgccga taggggtgcg
     3601 cttcgcgtac tccaacacct gctgccacac cagttcgtca tcgtcggccc gcagctcgac
     3661 gccggtgtag gtgatcttca cgtccttgtt gacgtggaaa atgaccttgt tttgcagcgc
     3721 ctcgcgcggg attttcttgt tgcgcgtggt gaacagggca gagcgggccg tgtcgtttgg
     3781 catcgctcgc atcgtgtccg gccacggcgc aatatcgaac aaggaaagct gcatttcctt
     3841 gatctgctgc ttcgtgtgtt tcagcaacgc ggcctgcttg gcctcgctga cctgttttgc
     3901 caggtcctcg ccggcggttt ttcgcttctt ggtcgtcata gttcctcgcg tgtcgatggt
     3961 catcgacttc gccaaacctg ccgcctcctg ttcaagacga cgcgaacgct ccacggcggc
     4021 cgatggcgcg ggcagggcag ggggagccag ttgcacgctg tcgcgctcga tcttggccgt
     4081 agcttgctgg accatcgagc cgacggactg gaaggtttcg cggggcgcac gcatgacggt
     4141 gcggcttgcg atggtttcgg catcctcggc ggaaaacccc gcgtcgatca gttcttgcct
     4201 gtatgccttc cggtcaaacg tccgattcat tcaccctcct tgcgggattg ccccgactca
     4261 cgccggggca atgtgccctt attcctgatt tgacccgcct ggtgccttgg tgtccagata
     4321 atccacctta tcggcaatga agtcggtccc gtagaccgtc tggccgtcct tctcgtactt
     4381 ggtattccga atcttgccct gcacgaatac cagcgacccc ttgcccaaat acttgccgtg
     4441 ggcctcggcc tgagagccaa aacacttgat gcggaagaag tcggtgcgct cctgcttgtc
     4501 gccggcatcg ttgcgccaca tctaggatct gccaggaacc gtaaaaaggc cgcgttgctg
     4561 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     4621 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     4681 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     4741 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     4801 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     4861 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     4921 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     4981 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     5041 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     5101 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     5161 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     5221 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     5281 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     5341 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     5401 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     5461 ccgcgagaac cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     5521 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     5581 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     5641 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     5701 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     5761 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     5821 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     5881 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     5941 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     6001 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     6061 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     6121 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     6181 ctcatactct tcctttttca atattattga agcatttatc aaggttattg tctcatgagc
     6241 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacgaattgg
     6301 ccagcgctgc catttttggg gtgaggccgt tcgcggccga ggggcgcagc ccctgggggg
     6361 atgggaggcc cgcgttagcg ggccgggagg gttcgagaag ggggggcacc ccccttcggc
     6421 gtgcgcggtc acgcgcacag ggcgcagccc tggttaaaaa caaggtttat aaatattggt
     6481 ttaaaagcag gttaaaagac aggttagcgg tggccgaaaa acgggcggaa acccttgcaa
     6541 atgctggatt ttctgcctgt ggacagcccc tcaaatgtca ataggtgcgc ccctcatctg
     6601 tcagcactct gcccctcaag tgtcaaggat cgcgcccctc atctgtcagt agtcgcgccc
     6661 ctcaagtgtc aataccgcag ggcacttatc cccaggcttg tccacatcat ctgtgggaaa
     6721 ctcgcgtaaa atcaggcgtt ttcgccgatt tgcgaggctg gccagctcca cgtcgccggc
     6781 cgaaatcgag cctgcccctc atctgtcaac gccgcgccgg gtgagtcggc ccctcaagtg
     6841 tcaacgtccg cccctcatct gtcagtgagg gccaagtttt ccgcgaggta tccacaacgc
     6901 cggcggccgc ggtgtctcgc acacggcttc gacggcgttt ctggcgcgtt tgcagggcca
     6961 tagacggccg ccagcccagc ggcgagggca accagcccgg tgagcgtcgc aaaggagatc
     7021 ctgatctgac tgatgggctg cctgtatcga gtggtgattt tgtgccgagc tgccggtcgg
     7081 ggagctgttg gctggctggt ggcaggatat attgtggtgt aaacaaattg acgcttagac
     7141 aacttaataa cacattgcgg acgtttttaa tgtactgggg tggatgcagt gggccccact
     7201 ctgtgaagac aaactagaat tcgagctc
//
