LOCUS       sequence-346340-        7633 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 1 module containing 35S-HSP21-GFP_P2 F.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7633)
  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 7633)
  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: pLBT0010
FEATURES             Location/Qualifiers
     source          1..7633
                     /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        5366..5711
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     5654..5674
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             join(6709..6711,6712..7425)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(6751..6772)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(7012..7031)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7359..7380
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    join(7460..7633,1)
                     /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 ttggaggaat
     4381 tccaatccca caaaaatctg agcttaacag cacagttgct cctctcagag cagaatcggg
     4441 tattcaacac cctcatatca actactacgt tgtgtataac ggtccacatg ccggtatata
     4501 cgatgactgg ggttgtacaa aggcggcaac aaacggcgtt cccggagttg cacacaagaa
     4561 atttgccact attacagagg caagagcagc agctgacgcg tacacaacaa gtcagcaaac
     4621 agacaggttg aacttcatcc ccaaaggaga agctcaactc aagcccaaga gctttgctaa
     4681 ggccctaaca agcccaccaa agcaaaaagc ccactggctc acgctaggaa ccaaaaggcc
     4741 cagcagtgat ccagccccaa aagagatctc ctttgccccg gagattacaa tggacgattt
     4801 cctctatctt tacgatctag gaaggaagtt cgaaggtgaa ggtgacgaca ctatgttcac
     4861 cactgataat gagaaggtta gcctcttcaa tttcagaaag aatgctgacc cacagatggt
     4921 tagagaggcc tacgcagcaa gtctcatcaa gacgatctac ccgagtaaca atctccagga
     4981 gatcaaatac cttcccaaga aggttaaaga tgcagtcaaa agattcagga ctaattgcat
     5041 caagaacaca gagaaagaca tatttctcaa gatcagaagt actattccag tatggacgat
     5101 tcaaggcttg cttcataaac caaggcaagt aatagagatt ggagtctcta aaaaggtagt
     5161 tcctactgaa tctaaggcca tgcatggagt ctaagattca aatcgaggat ctaacagaac
     5221 tcgccgtcaa gactggcgaa cagttcatac agagtctttt acgactcaat gacaagaaga
     5281 aaatcttcgt caacatggtg gagcacgaca ctctggtcta ctccaaaaat gtcaaagata
     5341 cagtctcaga agatcaaagg gctattgaga cttttcaaca aaggataatt tcgggaaacc
     5401 tcctcggatt ccattgccca gctatctgtc acttcatcga aaggacagta gaaaaggaag
     5461 gtggctccta caaatgccat cattgcgata aaggaaaggc tatcattcaa gatctctctg
     5521 ccgacagtgg tcccaaagat ggacccccac ccacgaggag catcgtggaa aaagaagagg
     5581 ttccaaccac gtctacaaag caagtggatt gatgtgacat ctccactgac gtaagggatg
     5641 acgcacaatc ccactatcct tcgcaagacc cttcctctat ataaggaagt tcatttcatt
     5701 tggagaggac acgctcgagt ataagagctc atttttacaa caattaccaa caacaacaaa
     5761 caacaaacaa cattacaatt acatttacaa ttatcgatac aatggcttcc tctatgctct
     5821 cttccgctac tatggttgcc tctccggctc aggccactat ggtcgctcct ttcaacggac
     5881 ttaagtcctc cgctgccttc ccagccaccc gcaaggctaa caacgacatt acttccatca
     5941 caagcaacgg cggaagagtt aactgcatgc aggtcattta tatttcttct ttcacttttt
     6001 tattattcca tatgattttt ttcggttctt tcttcgaatc tacataaact aatatcattg
     6061 gaaaaatcga aaaaataggt gtggcctccg attggaaaga agaagtttga gactctctct
     6121 taccttcctg accttaccga ttccgaaggt gctcaagacc agagagaaaa ctccattgat
     6181 gttgttcagc aaggacaaca aaaagggaac caaggatcaa gtgtcgaaaa aagacctcaa
     6241 caacgcttaa ccatggatgt ctctcctttc ggattgttgg atcctttgtc accaatgagg
     6301 acgatgcgac aaatgttaga tactatggac aggatgttcg aggacactat gcctgtctca
     6361 ggaagaaaca gaggaggaag tggagtgtca gagattcgtg caccgtggga catcaaagag
     6421 gaagaacacg agatcaagat gcgtttcgac atgcctggac tctctaaaga ggacgtcaaa
     6481 atctctgtag aagataacgt acttgtgatc aaaggagagc agaagaagga ggacagtgat
     6541 gattcttggt ctggaagaag cgttagctca tatggaacac gacttcagct cccagacaac
     6601 tgtgagaaag acaagatcaa agctgagctc aagaacggag tcctctttat cactatccct
     6661 aagaccaaag tcgaacgcaa agtcatcgat gtccagattc aatcttcggt gagcaagggc
     6721 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
     6781 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
     6841 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccttc
     6901 agctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc
     6961 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
     7021 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     7081 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
     7141 tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac
     7201 ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag
     7261 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag
     7321 tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
     7381 accgccgccg ggatcactca cggcatggac gagctgtaca agtaagcttc tctagctaga
     7441 gtcgatcgac aagctcgagt ttctccataa taatgtgtga gtagttccca gataagggaa
     7501 ttagggttcc tatagggttt cgctcatgtg ttgagcatat aagaaaccct tagtatgtat
     7561 ttgtatttgt aaaatacttc tatcaataaa atttctaatt cctaaaacca aaatccagta
     7621 ctaaaatcca gat
//
