LOCUS       sequence-347894-        7675 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 1 module containing 35S-CTP-mCHERRY-nYFP_P2 F.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7675)
  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 7675)
  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: pLBT0120
FEATURES             Location/Qualifiers
     source          1..7675
                     /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             2550..2573
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2877..2896)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    3130..3304
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    3349..3373
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(3529..4677)
                     /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(5013..5601)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5093..5112)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5772..6563,6564..6632))
                     /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     6395..6414
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6633..6737)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6824..7455
                     /label=oriV
                     /note="incP origin of replication"
     misc_feature    7547..7571
                     /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 cggattacaa ggaccacgac ggggactaca
     2521 aggatcatga tttggatgct gctgccgccg actataagga tgatgacgat aaggtgagca
     2581 agggcgaaga gttgttcact ggtgtggttc ctatcctcgt tgagcttgat ggtgatgtga
     2641 acgggcacaa gttctccgtt tctggtgaag gtgagggtga tgctacttac ggaaagctca
     2701 ccctcaagtt catctgtacc actggaaagc tccctgtgcc ttggcctact ctcgttacta
     2761 ctttcggata cggactccag tgcttcgcta gataccctga tcatatgaag cagcacgact
     2821 tcttcaagag cgctatgcct gagggatacg tgcaagagag aaccattttc ttcaaggacg
     2881 acggcaacta caagaccaga gctgaggtta agttcgaagg tgacaccctc gtgaacagga
     2941 tcgagcttaa gggcatcgac ttcaaagagg acggaaacat cctcggacac aagctcgagt
     3001 acaactacaa cagccacaac gtgtacatca tggccgacaa gcagaagaac ggcatcaagg
     3061 tcaacttcaa gatcaggcac aacatcgagg actaagcttc tctagctaga gtcgatcgac
     3121 aagctcgagt ttctccataa taatgtgtga gtagttccca gataagggaa ttagggttcc
     3181 tatagggttt cgctcatgtg ttgagcatat aagaaaccct tagtatgtat ttgtatttgt
     3241 aaaatacttc tatcaataaa atttctaatt cctaaaacca aaatccagta ctaaaatcca
     3301 gatcgctact attgtcttct gcacgaagtg gtttaaacta tcagtgtttg acaggatata
     3361 ttggcgggta aacctaagag aaaagagcgt ttattagaat aatcggatat ttaaaagggc
     3421 gtgaaaaggt ttatccgttc gtccatttgt atgtgcatgc caaccacagg gttccccaga
     3481 tcaggcgctg gctgctgaac ccccagccgg aactgacccc acaaggccct agcgtttgca
     3541 atgcaccagg tcatcattga cccaggcgtg ttccaccagg ccgctgcctc gcaactcttc
     3601 gcaggcttcg ccgacctgct cgcgccactt cttcacgcgg gtggaatccg atccgcacat
     3661 gaggcggaag gtttccagct tgagcgggta cggctcccgg tgcgagctga aatagtcgaa
     3721 catccgtcgg gccgtcggcg acagcttgcg gtacttctcc catatgaatt tcgtgtagtg
     3781 gtcgccagca aacagcacga cgatttcctc gtcgatcagg acctggcaac gggacgtttt
     3841 cttgccacgg tccaggacgc ggaagcggtg cagcagcgac accgattcca ggtgcccaac
     3901 gcggtcggac gtgaagccca tcgccgtcgc ctgtaggcgc gacaggcatt cctcggcctt
     3961 cgtgtaatac cggccattga tcgaccagcc caggtcctgg caaagctcgt agaacgtgaa
     4021 ggtgatcggc tcgccgatag gggtgcgctt cgcgtactcc aacacctgct gccacaccag
     4081 ttcgtcatcg tcggcccgca gctcgacgcc ggtgtaggtg atcttcacgt ccttgttgac
     4141 gtggaaaatg accttgtttt gcagcgcctc gcgcgggatt ttcttgttgc gcgtggtgaa
     4201 cagggcagag cgggccgtgt cgtttggcat cgctcgcatc gtgtccggcc acggcgcaat
     4261 atcgaacaag gaaagctgca tttccttgat ctgctgcttc gtgtgtttca gcaacgcggc
     4321 ctgcttggcc tcgctgacct gttttgccag gtcctcgccg gcggtttttc gcttcttggt
     4381 cgtcatagtt cctcgcgtgt cgatggtcat cgacttcgcc aaacctgccg cctcctgttc
     4441 aagacgacgc gaacgctcca cggcggccga tggcgcgggc agggcagggg gagccagttg
     4501 cacgctgtcg cgctcgatct tggccgtagc ttgctggacc atcgagccga cggactggaa
     4561 ggtttcgcgg ggcgcacgca tgacggtgcg gcttgcgatg gtttcggcat cctcggcgga
     4621 aaaccccgcg tcgatcagtt cttgcctgta tgccttccgg tcaaacgtcc gattcattca
     4681 ccctccttgc gggattgccc cgactcacgc cggggcaatg tgcccttatt cctgatttga
     4741 cccgcctggt gccttggtgt ccagataatc caccttatcg gcaatgaagt cggtcccgta
     4801 gaccgtctgg ccgtccttct cgtacttggt attccgaatc ttgccctgca cgaataccag
     4861 cgaccccttg cccaaatact tgccgtgggc ctcggcctga gagccaaaac acttgatgcg
     4921 gaagaagtcg gtgcgctcct gcttgtcgcc ggcatcgttg cgccacatct aggatctgcc
     4981 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     5041 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     5101 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     5161 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     5221 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     5281 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     5341 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     5401 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     5461 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     5521 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     5581 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     5641 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     5701 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     5761 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     5821 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     5881 ccatctggcc ccagtgctgc aatgataccg cgagaaccac gctcaccggc tccagattta
     5941 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     6001 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     6061 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     6121 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     6181 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     6241 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     6301 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     6361 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     6421 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     6481 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     6541 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     6601 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     6661 atttatcaag gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     6721 caaatagggg ttccgcgcac gaattggcca gcgctgccat ttttggggtg aggccgttcg
     6781 cggccgaggg gcgcagcccc tggggggatg ggaggcccgc gttagcgggc cgggagggtt
     6841 cgagaagggg gggcaccccc cttcggcgtg cgcggtcacg cgcacagggc gcagccctgg
     6901 ttaaaaacaa ggtttataaa tattggttta aaagcaggtt aaaagacagg ttagcggtgg
     6961 ccgaaaaacg ggcggaaacc cttgcaaatg ctggattttc tgcctgtgga cagcccctca
     7021 aatgtcaata ggtgcgcccc tcatctgtca gcactctgcc cctcaagtgt caaggatcgc
     7081 gcccctcatc tgtcagtagt cgcgcccctc aagtgtcaat accgcagggc acttatcccc
     7141 aggcttgtcc acatcatctg tgggaaactc gcgtaaaatc aggcgttttc gccgatttgc
     7201 gaggctggcc agctccacgt cgccggccga aatcgagcct gcccctcatc tgtcaacgcc
     7261 gcgccgggtg agtcggcccc tcaagtgtca acgtccgccc ctcatctgtc agtgagggcc
     7321 aagttttccg cgaggtatcc acaacgccgg cggccgcggt gtctcgcaca cggcttcgac
     7381 ggcgtttctg gcgcgtttgc agggccatag acggccgcca gcccagcggc gagggcaacc
     7441 agcccggtga gcgtcgcaaa ggagatcctg atctgactga tgggctgcct gtatcgagtg
     7501 gtgattttgt gccgagctgc cggtcgggga gctgttggct ggctggtggc aggatatatt
     7561 gtggtgtaaa caaattgacg cttagacaac ttaataacac attgcggacg tttttaatgt
     7621 actggggtgg atgcagtggg ccccactctg tgaagacaag caagaattca agctt
//
