LOCUS       sequence-356087-        9799 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 2 module containing HSP21-GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9799)
  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 9799)
  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: pLBM0010
FEATURES             Location/Qualifiers
     source          1..9799
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    36..60
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      148..858
                     /label=oriV
                     /note="incP origin of replication"
     promoter        879..983
                     /gene="bla"
                     /label=AmpR promoter
     CDS             984..1778
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      1949..2537
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2438..2457
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             2873..4021
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     misc_feature    4167..4191
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        5269..5614
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     terminator      6236..6488
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        7504..7849
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     CDS             join(8847..8849,8850..9563)
                     /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(8889..8910)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(9150..9169)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     9497..9518
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    9598..9772
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
ORIGIN
        1 gggacacgaa gtgatccgtt taaactatca gtgtttgaca ggatatattg gcgggtaaac
       61 ctaagagaaa agagcgttta ttagaataat cggatattta aaagggcgtg aaaaggttta
      121 tccgttcgtc catttgtatg tgccagccgc ctttgcgacg ctcaccgggc tggttgccct
      181 cgccgctggg ctggcggccg tctatggccc tgcaaacgcg ccagaaacgc cgtcgaagcc
      241 gtgtgcgaga caccgcggcc gccggcgttg tggatacctc gcggaaaact tggccctcac
      301 tgacagatga ggggcggacg ttgacacttg aggggccgac tcacccggcg cggcgttgac
      361 agatgagggg caggctcgat ttcggccggc gacgtggagc tggccagcct cgcaaatcgg
      421 cgaaaacgcc tgattttacg cgagtttccc acagatgatg tggacaagcc tggggataag
      481 tgccctgcgg tattgacact tgaggggcgc gactactgac agatgagggg cgcgatcctt
      541 gacacttgag gggcagagtg ctgacagatg aggggcgcac ctattgacat ttgaggggct
      601 gtccacaggc agaaaatcca gcatttgcaa gggtttccgc ccgtttttcg gccaccgcta
      661 acctgtcttt taacctgctt ttaaaccaat atttataaac cttgttttta accagggctg
      721 cgccctgtgc gcgtgaccgc gcacgccgaa ggggggtgcc cccccttctc gaaccctccc
      781 ggcccgctaa cgcgggcctc ccatcccccc aggggctgcg cccctcggcc gcgaacggcc
      841 tcaccccaaa aatggcagcg ctggccaatt cccgagtgcg cggaacccct atttgtttat
      901 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
      961 aataatattg aaaaaggaag agtatggcta aaatgagaat atcaccggaa ttgaaaaaac
     1021 tgatcgaaaa ataccgctgc gtaaaagata cggaaggaat gtctcctgct aaggtatata
     1081 agctggtggg agaaaatgaa aacctatatt taaaaatgac ggacagccgg tataaaggga
     1141 ccacctatga tgtggaacgg gaaaaggaca tgatgctatg gctggaagga aagctgcctg
     1201 ttccaaaggt cctgcacttt gaacggcatg atggctggag caatctgctc atgagtgagg
     1261 ccgatggcgt cctttgctcg gaagagtatg aagatgaaca aagccctgaa aagattatcg
     1321 agctgtatgc ggagtgcatc aggctctttc actccatcga catatcggat tgtccctata
     1381 cgaatagctt agacagccgc ttagccgaat tggattactt actgaataac gatctggccg
     1441 atgtggattg cgaaaactgg gaagaggaca ctccatttaa agatccgcgc gagctgtatg
     1501 attttttaaa gacggaaaag cccgaagagg aacttgtctt ttcccacggc gacctgggag
     1561 acagcaacat ctttgtgaaa gatggcaaag taagtggctt tattgatctt gggagaagcg
     1621 gcagggcgga caagtggtat gacattgcct tctgcgtccg gtcgctcagg gaggatatcg
     1681 gggaagaaca gtatgtcgag ctattttttg acttactggg gatcaagcct gattgggaga
     1741 aaataaaata ttatatttta ctggatgaat tgttttagct gtcagaccaa gtttactcat
     1801 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     1861 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     1921 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     1981 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     2041 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     2101 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     2161 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     2221 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     2281 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     2341 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     2401 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     2461 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     2521 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gcagatccta
     2581 gatgtggcgc aacgatgccg gcgacaagca ggagcgcacc gacttcttcc gcatcaagtg
     2641 ttttggctct caggccgagg cccacggcaa gtatttgggc aaggggtcgc tggtattcgt
     2701 gcagggcaag attcggaata ccaagtacga gaaggacggc cagacggtct acgggaccga
     2761 cttcattgcc gataaggtgg attatctgga caccaaggca ccaggcgggt caaatcagga
     2821 ataagggcac attgccccgg cgtgagtcgg ggcaatcccg caaggagggt gaatgaatcg
     2881 gacgtttgac cggaaggcat acaggcaaga actgatcgac gcggggtttt ccgccgagga
     2941 tgccgaaacc atcgcaagcc gcaccgtcat gcgtgcgccc cgcgaaacct tccagtccgt
     3001 cggctcgatg gtccagcaag ctacggccaa gatcgagcgc gacagcgtgc aactggctcc
     3061 ccctgccctg cccgcgccat cggccgccgt ggagcgttcg cgtcgtcttg aacaggaggc
     3121 ggcaggtttg gcgaagtcga tgaccatcga cacgcgagga actatgacga ccaagaagcg
     3181 aaaaaccgcc ggcgaggacc tggcaaaaca ggtcagcgag gccaagcagg ccgcgttgct
     3241 gaaacacacg aagcagcaga tcaaggaaat gcagctttcc ttgttcgata ttgcgccgtg
     3301 gccggacacg atgcgagcga tgccaaacga cacggcccgc tctgccctgt tcaccacgcg
     3361 caacaagaaa atcccgcgcg aggcgctgca aaacaaggtc attttccacg tcaacaagga
     3421 cgtgaagatc acctacaccg gcgtcgagct gcgggccgac gatgacgaac tggtgtggca
     3481 gcaggtgttg gagtacgcga agcgcacccc tatcggcgag ccgatcacct tcacgttcta
     3541 cgagctttgc caggacctgg gctggtcgat caatggccgg tattacacga aggccgagga
     3601 atgcctgtcg cgcctacagg cgacggcgat gggcttcacg tccgaccgcg ttgggcacct
     3661 ggaatcggtg tcgctgctgc accgcttccg cgtcctggac cgtggcaaga aaacgtcccg
     3721 ttgccaggtc ctgatcgacg aggaaatcgt cgtgctgttt gctggcgacc actacacgaa
     3781 attcatatgg gagaagtacc gcaagctgtc gccgacggcc cgacggatgt tcgactattt
     3841 cagctcgcac cgggagccgt acccgctcaa gctggaaacc ttccgcctca tgtgcggatc
     3901 ggattccacc cgcgtgaaga agtggcgcga gcaggtcggc gaagcctgcg aagagttgcg
     3961 aggcagcggc ctggtggaac acgcctgggt caatgatgac ctggtgcatt gcaaacgcta
     4021 gggccttgtg gggtcagttc cggctggggg ttcagcagcc cctgctcgga tctgttggac
     4081 cggacagtag tcatggttga tgggctgcct gtatcgagtg gtgattttgt gccgagctgc
     4141 cggtcgggga gctgttggct ggctggtggc aggatatatt gtggtgtaaa caaattgacg
     4201 cttagacaac ttaataacac attgcggacg tttttaatgt actggggttg aacactctgt
     4261 gccgaattcg gatccggagg aattccaatc ccacaaaaat ctgagcttaa cagcacagtt
     4321 gctcctctca gagcagaatc gggtattcaa caccctcata tcaactacta cgttgtgtat
     4381 aacggtccac atgccggtat atacgatgac tggggttgta caaaggcggc aacaaacggc
     4441 gttcccggag ttgcacacaa gaaatttgcc actattacag aggcaagagc agcagctgac
     4501 gcgtacacaa caagtcagca aacagacagg ttgaacttca tccccaaagg agaagctcaa
     4561 ctcaagccca agagctttgc taaggcccta acaagcccac caaagcaaaa agcccactgg
     4621 ctcacgctag gaaccaaaag gcccagcagt gatccagccc caaaagagat ctcctttgcc
     4681 ccggagatta caatggacga tttcctctat ctttacgatc taggaaggaa gttcgaaggt
     4741 gaaggtgacg acactatgtt caccactgat aatgagaagg ttagcctctt caatttcaga
     4801 aagaatgctg acccacagat ggttagagag gcctacgcag caagtctcat caagacgatc
     4861 tacccgagta acaatctcca ggagatcaaa taccttccca agaaggttaa agatgcagtc
     4921 aaaagattca ggactaattg catcaagaac acagagaaag acatatttct caagatcaga
     4981 agtactattc cagtatggac gattcaaggc ttgcttcata aaccaaggca agtaatagag
     5041 attggagtct ctaaaaaggt agttcctact gaatctaagg ccatgcatgg agtctaagat
     5101 tcaaatcgag gatctaacag aactcgccgt caagactggc gaacagttca tacagagtct
     5161 tttacgactc aatgacaaga agaaaatctt cgtcaacatg gtggagcacg acactctggt
     5221 ctactccaaa aatgtcaaag atacagtctc agaagatcaa agggctattg agacttttca
     5281 acaaaggata atttcgggaa acctcctcgg attccattgc ccagctatct gtcacttcat
     5341 cgaaaggaca gtagaaaagg aaggtggctc ctacaaatgc catcattgcg ataaaggaaa
     5401 ggctatcatt caagatctct ctgccgacag tggtcccaaa gatggacccc cacccacgag
     5461 gagcatcgtg gaaaaagaag aggttccaac cacgtctaca aagcaagtgg attgatgtga
     5521 catctccact gacgtaaggg atgacgcaca atcccactat ccttcgcaag acccttcctc
     5581 tatataagga agttcatttc atttggagag gacacgctcg agtataagag ctcattttta
     5641 caacaattac caacaacaac aaacaacaaa caacattaca attacattta caattatcga
     5701 tacaatggaa cgagctatac aaggaaacga tgctagggaa caagcttatg gtgaacgttg
     5761 gaatggagga tcaggaagtt ccacttctcc cttcaaactt cctgacgaaa gtccgagttg
     5821 gactgagtgg cggctacata acgatgagac tatttcgaat caagataatc cccttggttt
     5881 caaggaaagc tggggtttcg ggaaagttgt atttaagaga tatctcagat acgacgggac
     5941 ggaaacttca ctgcacagag tccttggatc ttggacggga gattcggtta actatgcagc
     6001 atctcgattt ctcggtttcg accagatcgg atgtacctat agtattcggt ttcgaggagt
     6061 tagtgtcacc atttctggag ggtcgcgaac tcttcagcat ctcagtgaaa tggcaattcg
     6121 gtctaagcaa gaactgctac agcttacccc agtcaaagtg gaaagtgatg tatcaagagg
     6181 atgccctgaa ggtgttgaaa ccttcgaaga agaaagcgag taagcttgtc aagcagatcg
     6241 ttcaaacatt tggcaataaa gtttcttaag attgaatcct gttgccggtc ttgcgatgat
     6301 tatcatataa tttctgttga attacgttaa gcatgtaata attaacatgt aatgcatgac
     6361 gttatttatg agatgggttt ttatgattag agtcccgcaa ttatacattt aatacgcgat
     6421 agaaaacaaa atatagcgcg caaactagga taaattatcg cgcgcggtgt catctatgtt
     6481 actagatcga cgctgcaaga attcaagctt ggaggaattc caatcccaca aaaatctgag
     6541 cttaacagca cagttgctcc tctcagagca gaatcgggta ttcaacaccc tcatatcaac
     6601 tactacgttg tgtataacgg tccacatgcc ggtatatacg atgactgggg ttgtacaaag
     6661 gcggcaacaa acggcgttcc cggagttgca cacaagaaat ttgccactat tacagaggca
     6721 agagcagcag ctgacgcgta cacaacaagt cagcaaacag acaggttgaa cttcatcccc
     6781 aaaggagaag ctcaactcaa gcccaagagc tttgctaagg ccctaacaag cccaccaaag
     6841 caaaaagccc actggctcac gctaggaacc aaaaggccca gcagtgatcc agccccaaaa
     6901 gagatctcct ttgccccgga gattacaatg gacgatttcc tctatcttta cgatctagga
     6961 aggaagttcg aaggtgaagg tgacgacact atgttcacca ctgataatga gaaggttagc
     7021 ctcttcaatt tcagaaagaa tgctgaccca cagatggtta gagaggccta cgcagcaagt
     7081 ctcatcaaga cgatctaccc gagtaacaat ctccaggaga tcaaatacct tcccaagaag
     7141 gttaaagatg cagtcaaaag attcaggact aattgcatca agaacacaga gaaagacata
     7201 tttctcaaga tcagaagtac tattccagta tggacgattc aaggcttgct tcataaacca
     7261 aggcaagtaa tagagattgg agtctctaaa aaggtagttc ctactgaatc taaggccatg
     7321 catggagtct aagattcaaa tcgaggatct aacagaactc gccgtcaaga ctggcgaaca
     7381 gttcatacag agtcttttac gactcaatga caagaagaaa atcttcgtca acatggtgga
     7441 gcacgacact ctggtctact ccaaaaatgt caaagataca gtctcagaag atcaaagggc
     7501 tattgagact tttcaacaaa ggataatttc gggaaacctc ctcggattcc attgcccagc
     7561 tatctgtcac ttcatcgaaa ggacagtaga aaaggaaggt ggctcctaca aatgccatca
     7621 ttgcgataaa ggaaaggcta tcattcaaga tctctctgcc gacagtggtc ccaaagatgg
     7681 acccccaccc acgaggagca tcgtggaaaa agaagaggtt ccaaccacgt ctacaaagca
     7741 agtggattga tgtgacatct ccactgacgt aagggatgac gcacaatccc actatccttc
     7801 gcaagaccct tcctctatat aaggaagttc atttcatttg gagaggacac gctcgagtat
     7861 aagagctcat ttttacaaca attaccaaca acaacaaaca acaaacaaca ttacaattac
     7921 atttacaatt atcgatacaa tggcttcctc tatgctctct tccgctacta tggttgcctc
     7981 tccggctcag gccactatgg tcgctccttt caacggactt aagtcctccg ctgccttccc
     8041 agccacccgc aaggctaaca acgacattac ttccatcaca agcaacggcg gaagagttaa
     8101 ctgcatgcag gtcatttata tttcttcttt cactttttta ttattccata tgattttttt
     8161 cggttctttc ttcgaatcta cataaactaa tatcattgga aaaatcgaaa aaataggtgt
     8221 ggcctccgat tggaaagaag aagtttgaga ctctctctta ccttcctgac cttaccgatt
     8281 ccgaaggtgc tcaagaccag agagaaaact ccattgatgt tgttcagcaa ggacaacaaa
     8341 aagggaacca aggatcaagt gtcgaaaaaa gacctcaaca acgcttaacc atggatgtct
     8401 ctcctttcgg attgttggat cctttgtcac caatgaggac gatgcgacaa atgttagata
     8461 ctatggacag gatgttcgag gacactatgc ctgtctcagg aagaaacaga ggaggaagtg
     8521 gagtgtcaga gattcgtgca ccgtgggaca tcaaagagga agaacacgag atcaagatgc
     8581 gtttcgacat gcctggactc tctaaagagg acgtcaaaat ctctgtagaa gataacgtac
     8641 ttgtgatcaa aggagagcag aagaaggagg acagtgatga ttcttggtct ggaagaagcg
     8701 ttagctcata tggaacacga cttcagctcc cagacaactg tgagaaagac aagatcaaag
     8761 ctgagctcaa gaacggagtc ctctttatca ctatccctaa gaccaaagtc gaacgcaaag
     8821 tcatcgatgt ccagattcaa tcttcggtga gcaagggcga ggagctgttc accggggtgg
     8881 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg
     8941 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca
     9001 agctgcccgt gccctggccc accctcgtga ccaccttcag ctacggcgtg cagtgcttca
     9061 gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
     9121 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
     9181 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
     9241 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata
     9301 tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg
     9361 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     9421 ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca
     9481 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactcacg
     9541 gcatggacga gctgtacaag taagcttctc tagctagagt cgatcgacaa gctcgagttt
     9601 ctccataata atgtgtgagt agttcccaga taagggaatt agggttccta tagggtttcg
     9661 ctcatgtgtt gagcatataa gaaaccctta gtatgtattt gtatttgtaa aatacttcta
     9721 tcaataaaat ttctaattcc taaaaccaaa atccagtact aaaatccaga tcgctactag
     9781 aggatgcaca tgtgaccga
//
