LOCUS       sequence-347895-        7363 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 1 module containing 35S-CTP-mCHERRY-cYFP_P3 F.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7363)
  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 7363)
  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: pLBT0130
FEATURES             Location/Qualifiers
     source          1..7363
                     /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"
     primer_bind     2624..2645
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2688..2777
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     polyA_signal    2818..2992
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    3037..3061
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(3217..4365)
                     /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(4701..5289)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4781..4800)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5460..6251,6252..6320))
                     /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     6083..6102
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6321..6425)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6512..7143
                     /label=oriV
                     /note="incP origin of replication"
     misc_feature    7235..7259
                     /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 cgggcagcgt gcagctcgcc gaccactacc
     2521 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagct
     2581 accagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     2641 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtac ccatacgatg
     2701 ttcctgacta tgcgggctat ccctatgacg tcccggacta tgcaggatcc tatccatatg
     2761 acgttccaga ttacgctgct taagcttctc tagctagagt cgatcgacaa gctcgagttt
     2821 ctccataata atgtgtgagt agttcccaga taagggaatt agggttccta tagggtttcg
     2881 ctcatgtgtt gagcatataa gaaaccctta gtatgtattt gtatttgtaa aatacttcta
     2941 tcaataaaat ttctaattcc taaaaccaaa atccagtact aaaatccaga tcgctttact
     3001 tgtcttctgc acgaagtggt ttaaactatc agtgtttgac aggatatatt ggcgggtaaa
     3061 cctaagagaa aagagcgttt attagaataa tcggatattt aaaagggcgt gaaaaggttt
     3121 atccgttcgt ccatttgtat gtgcatgcca accacagggt tccccagatc aggcgctggc
     3181 tgctgaaccc ccagccggaa ctgaccccac aaggccctag cgtttgcaat gcaccaggtc
     3241 atcattgacc caggcgtgtt ccaccaggcc gctgcctcgc aactcttcgc aggcttcgcc
     3301 gacctgctcg cgccacttct tcacgcgggt ggaatccgat ccgcacatga ggcggaaggt
     3361 ttccagcttg agcgggtacg gctcccggtg cgagctgaaa tagtcgaaca tccgtcgggc
     3421 cgtcggcgac agcttgcggt acttctccca tatgaatttc gtgtagtggt cgccagcaaa
     3481 cagcacgacg atttcctcgt cgatcaggac ctggcaacgg gacgttttct tgccacggtc
     3541 caggacgcgg aagcggtgca gcagcgacac cgattccagg tgcccaacgc ggtcggacgt
     3601 gaagcccatc gccgtcgcct gtaggcgcga caggcattcc tcggccttcg tgtaataccg
     3661 gccattgatc gaccagccca ggtcctggca aagctcgtag aacgtgaagg tgatcggctc
     3721 gccgataggg gtgcgcttcg cgtactccaa cacctgctgc cacaccagtt cgtcatcgtc
     3781 ggcccgcagc tcgacgccgg tgtaggtgat cttcacgtcc ttgttgacgt ggaaaatgac
     3841 cttgttttgc agcgcctcgc gcgggatttt cttgttgcgc gtggtgaaca gggcagagcg
     3901 ggccgtgtcg tttggcatcg ctcgcatcgt gtccggccac ggcgcaatat cgaacaagga
     3961 aagctgcatt tccttgatct gctgcttcgt gtgtttcagc aacgcggcct gcttggcctc
     4021 gctgacctgt tttgccaggt cctcgccggc ggtttttcgc ttcttggtcg tcatagttcc
     4081 tcgcgtgtcg atggtcatcg acttcgccaa acctgccgcc tcctgttcaa gacgacgcga
     4141 acgctccacg gcggccgatg gcgcgggcag ggcaggggga gccagttgca cgctgtcgcg
     4201 ctcgatcttg gccgtagctt gctggaccat cgagccgacg gactggaagg tttcgcgggg
     4261 cgcacgcatg acggtgcggc ttgcgatggt ttcggcatcc tcggcggaaa accccgcgtc
     4321 gatcagttct tgcctgtatg ccttccggtc aaacgtccga ttcattcacc ctccttgcgg
     4381 gattgccccg actcacgccg gggcaatgtg cccttattcc tgatttgacc cgcctggtgc
     4441 cttggtgtcc agataatcca ccttatcggc aatgaagtcg gtcccgtaga ccgtctggcc
     4501 gtccttctcg tacttggtat tccgaatctt gccctgcacg aataccagcg accccttgcc
     4561 caaatacttg ccgtgggcct cggcctgaga gccaaaacac ttgatgcgga agaagtcggt
     4621 gcgctcctgc ttgtcgccgg catcgttgcg ccacatctag gatctgccag gaaccgtaaa
     4681 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     4741 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     4801 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     4861 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     4921 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     4981 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     5041 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     5101 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     5161 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     5221 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     5281 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     5341 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     5401 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     5461 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     5521 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     5581 agtgctgcaa tgataccgcg agaaccacgc tcaccggctc cagatttatc agcaataaac
     5641 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     5701 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     5761 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     5821 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     5881 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     5941 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     6001 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     6061 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     6121 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     6181 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     6241 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     6301 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcaaggt
     6361 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     6421 ccgcgcacga attggccagc gctgccattt ttggggtgag gccgttcgcg gccgaggggc
     6481 gcagcccctg gggggatggg aggcccgcgt tagcgggccg ggagggttcg agaagggggg
     6541 gcacccccct tcggcgtgcg cggtcacgcg cacagggcgc agccctggtt aaaaacaagg
     6601 tttataaata ttggtttaaa agcaggttaa aagacaggtt agcggtggcc gaaaaacggg
     6661 cggaaaccct tgcaaatgct ggattttctg cctgtggaca gcccctcaaa tgtcaatagg
     6721 tgcgcccctc atctgtcagc actctgcccc tcaagtgtca aggatcgcgc ccctcatctg
     6781 tcagtagtcg cgcccctcaa gtgtcaatac cgcagggcac ttatccccag gcttgtccac
     6841 atcatctgtg ggaaactcgc gtaaaatcag gcgttttcgc cgatttgcga ggctggccag
     6901 ctccacgtcg ccggccgaaa tcgagcctgc ccctcatctg tcaacgccgc gccgggtgag
     6961 tcggcccctc aagtgtcaac gtccgcccct catctgtcag tgagggccaa gttttccgcg
     7021 aggtatccac aacgccggcg gccgcggtgt ctcgcacacg gcttcgacgg cgtttctggc
     7081 gcgtttgcag ggccatagac ggccgccagc ccagcggcga gggcaaccag cccggtgagc
     7141 gtcgcaaagg agatcctgat ctgactgatg ggctgcctgt atcgagtggt gattttgtgc
     7201 cgagctgccg gtcggggagc tgttggctgg ctggtggcag gatatattgt ggtgtaaaca
     7261 aattgacgct tagacaactt aataacacat tgcggacgtt tttaatgtac tggggtggat
     7321 gcagtgggcc ccactctgtg aagacaaact agaattcgag ctc
//
