LOCUS       sequence-347446-        6536 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 1 module containing 35S-nYFP-HSP21_P2 F.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6536)
  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 6536)
  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: pLBT0060
FEATURES             Location/Qualifiers
     source          1..6536
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    46..70
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(226..1374)
                     /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(1710..2298)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1790..1809)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2469..3260,3261..3329))
                     /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     3092..3111
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3330..3434)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3521..4152
                     /label=oriV
                     /note="incP origin of replication"
     misc_feature    4244..4268
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        4454..4799
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     4742..4762
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             5211..5234
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(5538..5557)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    join(6363..6536,1)
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
ORIGIN
        1 cgctactatt gtcttctgca cgaagtggtt taaactatca gtgtttgaca ggatatattg
       61 gcgggtaaac ctaagagaaa agagcgttta ttagaataat cggatattta aaagggcgtg
      121 aaaaggttta tccgttcgtc catttgtatg tgcatgccaa ccacagggtt ccccagatca
      181 ggcgctggct gctgaacccc cagccggaac tgaccccaca aggccctagc gtttgcaatg
      241 caccaggtca tcattgaccc aggcgtgttc caccaggccg ctgcctcgca actcttcgca
      301 ggcttcgccg acctgctcgc gccacttctt cacgcgggtg gaatccgatc cgcacatgag
      361 gcggaaggtt tccagcttga gcgggtacgg ctcccggtgc gagctgaaat agtcgaacat
      421 ccgtcgggcc gtcggcgaca gcttgcggta cttctcccat atgaatttcg tgtagtggtc
      481 gccagcaaac agcacgacga tttcctcgtc gatcaggacc tggcaacggg acgttttctt
      541 gccacggtcc aggacgcgga agcggtgcag cagcgacacc gattccaggt gcccaacgcg
      601 gtcggacgtg aagcccatcg ccgtcgcctg taggcgcgac aggcattcct cggccttcgt
      661 gtaataccgg ccattgatcg accagcccag gtcctggcaa agctcgtaga acgtgaaggt
      721 gatcggctcg ccgatagggg tgcgcttcgc gtactccaac acctgctgcc acaccagttc
      781 gtcatcgtcg gcccgcagct cgacgccggt gtaggtgatc ttcacgtcct tgttgacgtg
      841 gaaaatgacc ttgttttgca gcgcctcgcg cgggattttc ttgttgcgcg tggtgaacag
      901 ggcagagcgg gccgtgtcgt ttggcatcgc tcgcatcgtg tccggccacg gcgcaatatc
      961 gaacaaggaa agctgcattt ccttgatctg ctgcttcgtg tgtttcagca acgcggcctg
     1021 cttggcctcg ctgacctgtt ttgccaggtc ctcgccggcg gtttttcgct tcttggtcgt
     1081 catagttcct cgcgtgtcga tggtcatcga cttcgccaaa cctgccgcct cctgttcaag
     1141 acgacgcgaa cgctccacgg cggccgatgg cgcgggcagg gcagggggag ccagttgcac
     1201 gctgtcgcgc tcgatcttgg ccgtagcttg ctggaccatc gagccgacgg actggaaggt
     1261 ttcgcggggc gcacgcatga cggtgcggct tgcgatggtt tcggcatcct cggcggaaaa
     1321 ccccgcgtcg atcagttctt gcctgtatgc cttccggtca aacgtccgat tcattcaccc
     1381 tccttgcggg attgccccga ctcacgccgg ggcaatgtgc ccttattcct gatttgaccc
     1441 gcctggtgcc ttggtgtcca gataatccac cttatcggca atgaagtcgg tcccgtagac
     1501 cgtctggccg tccttctcgt acttggtatt ccgaatcttg ccctgcacga ataccagcga
     1561 ccccttgccc aaatacttgc cgtgggcctc ggcctgagag ccaaaacact tgatgcggaa
     1621 gaagtcggtg cgctcctgct tgtcgccggc atcgttgcgc cacatctagg atctgccagg
     1681 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     1741 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     1801 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     1861 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     1921 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     1981 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     2041 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     2101 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     2161 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     2221 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     2281 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     2341 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     2401 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     2461 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     2521 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     2581 tctggcccca gtgctgcaat gataccgcga gaaccacgct caccggctcc agatttatca
     2641 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     2701 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     2761 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     2821 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     2881 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     2941 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     3001 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     3061 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     3121 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     3181 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     3241 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     3301 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     3361 tatcaaggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     3421 ataggggttc cgcgcacgaa ttggccagcg ctgccatttt tggggtgagg ccgttcgcgg
     3481 ccgaggggcg cagcccctgg ggggatggga ggcccgcgtt agcgggccgg gagggttcga
     3541 gaaggggggg cacccccctt cggcgtgcgc ggtcacgcgc acagggcgca gccctggtta
     3601 aaaacaaggt ttataaatat tggtttaaaa gcaggttaaa agacaggtta gcggtggccg
     3661 aaaaacgggc ggaaaccctt gcaaatgctg gattttctgc ctgtggacag cccctcaaat
     3721 gtcaataggt gcgcccctca tctgtcagca ctctgcccct caagtgtcaa ggatcgcgcc
     3781 cctcatctgt cagtagtcgc gcccctcaag tgtcaatacc gcagggcact tatccccagg
     3841 cttgtccaca tcatctgtgg gaaactcgcg taaaatcagg cgttttcgcc gatttgcgag
     3901 gctggccagc tccacgtcgc cggccgaaat cgagcctgcc cctcatctgt caacgccgcg
     3961 ccgggtgagt cggcccctca agtgtcaacg tccgcccctc atctgtcagt gagggccaag
     4021 ttttccgcga ggtatccaca acgccggcgg ccgcggtgtc tcgcacacgg cttcgacggc
     4081 gtttctggcg cgtttgcagg gccatagacg gccgccagcc cagcggcgag ggcaaccagc
     4141 ccggtgagcg tcgcaaagga gatcctgatc tgactgatgg gctgcctgta tcgagtggtg
     4201 attttgtgcc gagctgccgg tcggggagct gttggctggc tggtggcagg atatattgtg
     4261 gtgtaaacaa attgacgctt agacaactta ataacacatt gcggacgttt ttaatgtact
     4321 ggggtggatg cagtgggccc cactctgtga agacaagcaa gaattcaagc ttggaggtca
     4381 acatggtgga gcacgacact ctggtctact ccaaaaatgt caaagataca gtctcagaag
     4441 atcaaagggc tattgagact tttcaacaaa ggataatttc gggaaacctc ctcggattcc
     4501 attgcccagc tatctgtcac ttcatcgaaa ggacagtaga aaaggaaggt ggctcctaca
     4561 aatgccatca ttgcgataaa ggaaaggcta tcattcaaga tctctctgcc gacagtggtc
     4621 ccaaagatgg acccccaccc acgaggagca tcgtggaaaa agaagaggtt ccaaccacgt
     4681 ctacaaagca agtggattga tgtgacatct ccactgacgt aagggatgac gcacaatccc
     4741 actatccttc gcaagaccct tcctctatat aaggaagttc atttcatttg gagaggacac
     4801 gcccatggct tcctctatgc tctcttccgc tactatggtt gcctctccgg ctcaggccac
     4861 tatggtcgct cctttcaacg gacttaagtc ctccgctgcc ttcccagcca cccgcaaggc
     4921 taacaacgac attacttcca tcacaagcaa cggcggaaga gttaactgca tgcaggtcat
     4981 ttatatttct tctttcactt ttttattatt ccatatgatt tttttcggtt ctttcttcga
     5041 atctacataa actaatatca ttggaaaaat cgaaaaaata ggtgtggcct ccgattggaa
     5101 agaagaagtt tgagactctc tcttaccttc ctgaccttac cgattccgaa atggattaca
     5161 aggaccacga cggggactac aaggatcatg atttggatgc tgctgccgcc gactataagg
     5221 atgatgacga taaggtgagc aagggcgaag agttgttcac tggtgtggtt cctatcctcg
     5281 ttgagcttga tggtgatgtg aacgggcaca agttctccgt ttctggtgaa ggtgagggtg
     5341 atgctactta cggaaagctc accctcaagt tcatctgtac cactggaaag ctccctgtgc
     5401 cttggcctac tctcgttact actttcggat acggactcca gtgcttcgct agataccctg
     5461 atcatatgaa gcagcacgac ttcttcaaga gcgctatgcc tgagggatac gtgcaagaga
     5521 gaaccatttt cttcaaggac gacggcaact acaagaccag agctgaggtt aagttcgaag
     5581 gtgacaccct cgtgaacagg atcgagctta agggcatcga cttcaaagag gacggaaaca
     5641 tcctcggaca caagctcgag tacaactaca acagccacaa cgtgtacatc atggccgaca
     5701 agcagaagaa cggcatcaag gtcaacttca agatcaggca caacatcgag gacgaaggtg
     5761 ctcaagacca gagagaaaac tccattgatg ttgttcagca aggacaacaa aaagggaacc
     5821 aaggatcaag tgtcgaaaaa agacctcaac aacgcttaac catggatgtc tctcctttcg
     5881 gattgttgga tcctttgtca ccaatgagga cgatgcgaca aatgttagat actatggaca
     5941 ggatgttcga ggacactatg cctgtctcag gaagaaacag aggaggaagt ggagtgtcag
     6001 agattcgtgc accgtgggac atcaaagagg aagaacacga gatcaagatg cgtttcgaca
     6061 tgcctggact ctctaaagag gacgtcaaaa tctctgtaga agataacgta cttgtgatca
     6121 aaggagagca gaagaaggag gacagtgatg attcttggtc tggaagaagc gttagctcat
     6181 atggaacacg acttcagctc ccagacaact gtgagaaaga caagatcaaa gctgagctca
     6241 agaacggagt cctctttatc actatcccta agaccaaagt cgaacgcaaa gtcatcgatg
     6301 tccagattca atcttcgtga ataattgagc ttctctagct agagtcgatc gacaagctcg
     6361 agtttctcca taataatgtg tgagtagttc ccagataagg gaattagggt tcctataggg
     6421 tttcgctcat gtgttgagca tataagaaac ccttagtatg tatttgtatt tgtaaaatac
     6481 ttctatcaat aaaatttcta attcctaaaa ccaaaatcca gtactaaaat ccagat
//
