LOCUS       sequence-347875-        7696 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 1 module containing 35S-PTAC5-cYFP_P3 F.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7696)
  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 7696)
  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: pLBT0100
FEATURES             Location/Qualifiers
     source          1..7696
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1023..1368
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     1311..1331
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     primer_bind     2986..3007
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3050..3139
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     polyA_signal    3180..3354
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    3399..3423
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(3579..4727)
                     /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(5063..5651)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5143..5162)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5822..6613,6614..6682))
                     /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     6445..6464
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6683..6787)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6874..7505
                     /label=oriV
                     /note="incP origin of replication"
     misc_feature    7597..7621
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
ORIGIN
        1 tctgtgaaga caaactagaa ttcgagctcg gaggaattcc aatcccacaa aaatctgagc
       61 ttaacagcac agttgctcct ctcagagcag aatcgggtat tcaacaccct catatcaact
      121 actacgttgt gtataacggt ccacatgccg gtatatacga tgactggggt tgtacaaagg
      181 cggcaacaaa cggcgttccc ggagttgcac acaagaaatt tgccactatt acagaggcaa
      241 gagcagcagc tgacgcgtac acaacaagtc agcaaacaga caggttgaac ttcatcccca
      301 aaggagaagc tcaactcaag cccaagagct ttgctaaggc cctaacaagc ccaccaaagc
      361 aaaaagccca ctggctcacg ctaggaacca aaaggcccag cagtgatcca gccccaaaag
      421 agatctcctt tgccccggag attacaatgg acgatttcct ctatctttac gatctaggaa
      481 ggaagttcga aggtgaaggt gacgacacta tgttcaccac tgataatgag aaggttagcc
      541 tcttcaattt cagaaagaat gctgacccac agatggttag agaggcctac gcagcaagtc
      601 tcatcaagac gatctacccg agtaacaatc tccaggagat caaatacctt cccaagaagg
      661 ttaaagatgc agtcaaaaga ttcaggacta attgcatcaa gaacacagag aaagacatat
      721 ttctcaagat cagaagtact attccagtat ggacgattca aggcttgctt cataaaccaa
      781 ggcaagtaat agagattgga gtctctaaaa aggtagttcc tactgaatct aaggccatgc
      841 atggagtcta agattcaaat cgaggatcta acagaactcg ccgtcaagac tggcgaacag
      901 ttcatacaga gtcttttacg actcaatgac aagaagaaaa tcttcgtcaa catggtggag
      961 cacgacactc tggtctactc caaaaatgtc aaagatacag tctcagaaga tcaaagggct
     1021 attgagactt ttcaacaaag gataatttcg ggaaacctcc tcggattcca ttgcccagct
     1081 atctgtcact tcatcgaaag gacagtagaa aaggaaggtg gctcctacaa atgccatcat
     1141 tgcgataaag gaaaggctat cattcaagat ctctctgccg acagtggtcc caaagatgga
     1201 cccccaccca cgaggagcat cgtggaaaaa gaagaggttc caaccacgtc tacaaagcaa
     1261 gtggattgat gtgacatctc cactgacgta agggatgacg cacaatccca ctatccttcg
     1321 caagaccctt cctctatata aggaagttca tttcatttgg agaggacacg ctcgagtata
     1381 agagctcatt tttacaacaa ttaccaacaa caacaaacaa caaacaacat tacaattaca
     1441 tttacaatta tcgatacaat ggcttcctct atgctctctt ccgctactat ggttgcctct
     1501 ccggctcagg ccactatggt cgctcctttc aacggactta agtcctccgc tgccttccca
     1561 gccacccgca aggctaacaa cgacattact tccatcacaa gcaacggcgg aagagttaac
     1621 tgcatgcagg tcatttatat ttcttctttc acttttttat tattccatat gatttttttc
     1681 ggttctttct tcgaatctac ataaactaat atcattggaa aaatcgaaaa aataggtgtg
     1741 gcctccgatt ggaaagaaga agtttgagac tctctcttac cttcctgacc ttaccgattc
     1801 cgaaggttgc ttctccactc aaaatcccga ccgcgaagag gtccggtggc tccgggaaga
     1861 gcagagatgg attcgcgagg agcaacgatg gattcgtgaa gaacagagat ggatacgcga
     1921 acgtgaatcg cttctacaag agatttcgga tctacagctc agaattcaat ccctagagtc
     1981 acgaaattcg caattgggga attctattcc cgatacgatt tcgaatatcg ctgctttgct
     2041 tcaggttttg aaggagaaga atcggatttc tgagagtgga ttgagcgcaa cgccgatggt
     2101 attggagagt acgagagaac aaattgttga ggaggtggaa gaagaagaga agcgagtgat
     2161 tattgctgaa gagaaagtta gggtttcgga gccggtgaag aagatcaaga ggaggatatt
     2221 gaaagttgga agcgaaggcg acgatgttca agctttgcag gaagctctgt tgaaattagg
     2281 attctattcg ggcgaagagg atatggagtt ctcgagcttt tcaagtggga ctgcaagtgc
     2341 tgttaagact tggcaagcat cgcttggggt ccgtgaggat ggggtaatga cagcagagct
     2401 tcttcagagg ttgttcatgg atgaggacgt agagacagat aaggatgaag caagtacaat
     2461 gaagaaagag gaagctggta atggggcggt atttacttca gtgacacaag tccctgagaa
     2521 gaagcaatca atcgtgaaag atcaaagtga cagagaagtt gacgttactc aaaatcgggt
     2581 ttttcttctt ggagaaaaca gatgggaaga tccctccagg ctcattggca ggaacaaacc
     2641 ggtagacaga agtgaatcaa caaacaccaa aacgaggtgc atcacttgtc gaggggaggg
     2701 tcgattgatg tgcctagagt gcgatggaac cggtgagcca aacattgagc cgcagttcat
     2761 ggagtgggtt ggtgaagata cgaagtgtcc gtactgtgaa ggtcttggct atacagtttg
     2821 cgatgtctgc gacggcaaaa aaaacttatc ttcgggcagc gtgcagctcg ccgaccacta
     2881 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     2941 ctaccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     3001 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt acccatacga
     3061 tgttcctgac tatgcgggct atccctatga cgtcccggac tatgcaggat cctatccata
     3121 tgacgttcca gattacgctg cttaagcttc tctagctaga gtcgatcgac aagctcgagt
     3181 ttctccataa taatgtgtga gtagttccca gataagggaa ttagggttcc tatagggttt
     3241 cgctcatgtg ttgagcatat aagaaaccct tagtatgtat ttgtatttgt aaaatacttc
     3301 tatcaataaa atttctaatt cctaaaacca aaatccagta ctaaaatcca gatcgcttta
     3361 cttgtcttct gcacgaagtg gtttaaacta tcagtgtttg acaggatata ttggcgggta
     3421 aacctaagag aaaagagcgt ttattagaat aatcggatat ttaaaagggc gtgaaaaggt
     3481 ttatccgttc gtccatttgt atgtgcatgc caaccacagg gttccccaga tcaggcgctg
     3541 gctgctgaac ccccagccgg aactgacccc acaaggccct agcgtttgca atgcaccagg
     3601 tcatcattga cccaggcgtg ttccaccagg ccgctgcctc gcaactcttc gcaggcttcg
     3661 ccgacctgct cgcgccactt cttcacgcgg gtggaatccg atccgcacat gaggcggaag
     3721 gtttccagct tgagcgggta cggctcccgg tgcgagctga aatagtcgaa catccgtcgg
     3781 gccgtcggcg acagcttgcg gtacttctcc catatgaatt tcgtgtagtg gtcgccagca
     3841 aacagcacga cgatttcctc gtcgatcagg acctggcaac gggacgtttt cttgccacgg
     3901 tccaggacgc ggaagcggtg cagcagcgac accgattcca ggtgcccaac gcggtcggac
     3961 gtgaagccca tcgccgtcgc ctgtaggcgc gacaggcatt cctcggcctt cgtgtaatac
     4021 cggccattga tcgaccagcc caggtcctgg caaagctcgt agaacgtgaa ggtgatcggc
     4081 tcgccgatag gggtgcgctt cgcgtactcc aacacctgct gccacaccag ttcgtcatcg
     4141 tcggcccgca gctcgacgcc ggtgtaggtg atcttcacgt ccttgttgac gtggaaaatg
     4201 accttgtttt gcagcgcctc gcgcgggatt ttcttgttgc gcgtggtgaa cagggcagag
     4261 cgggccgtgt cgtttggcat cgctcgcatc gtgtccggcc acggcgcaat atcgaacaag
     4321 gaaagctgca tttccttgat ctgctgcttc gtgtgtttca gcaacgcggc ctgcttggcc
     4381 tcgctgacct gttttgccag gtcctcgccg gcggtttttc gcttcttggt cgtcatagtt
     4441 cctcgcgtgt cgatggtcat cgacttcgcc aaacctgccg cctcctgttc aagacgacgc
     4501 gaacgctcca cggcggccga tggcgcgggc agggcagggg gagccagttg cacgctgtcg
     4561 cgctcgatct tggccgtagc ttgctggacc atcgagccga cggactggaa ggtttcgcgg
     4621 ggcgcacgca tgacggtgcg gcttgcgatg gtttcggcat cctcggcgga aaaccccgcg
     4681 tcgatcagtt cttgcctgta tgccttccgg tcaaacgtcc gattcattca ccctccttgc
     4741 gggattgccc cgactcacgc cggggcaatg tgcccttatt cctgatttga cccgcctggt
     4801 gccttggtgt ccagataatc caccttatcg gcaatgaagt cggtcccgta gaccgtctgg
     4861 ccgtccttct cgtacttggt attccgaatc ttgccctgca cgaataccag cgaccccttg
     4921 cccaaatact tgccgtgggc ctcggcctga gagccaaaac acttgatgcg gaagaagtcg
     4981 gtgcgctcct gcttgtcgcc ggcatcgttg cgccacatct aggatctgcc aggaaccgta
     5041 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     5101 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     5161 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     5221 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     5281 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     5341 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     5401 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     5461 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     5521 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     5581 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     5641 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     5701 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     5761 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     5821 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     5881 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     5941 ccagtgctgc aatgataccg cgagaaccac gctcaccggc tccagattta tcagcaataa
     6001 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     6061 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     6121 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     6181 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     6241 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     6301 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     6361 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     6421 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     6481 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     6541 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     6601 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     6661 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcaag
     6721 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     6781 ttccgcgcac gaattggcca gcgctgccat ttttggggtg aggccgttcg cggccgaggg
     6841 gcgcagcccc tggggggatg ggaggcccgc gttagcgggc cgggagggtt cgagaagggg
     6901 gggcaccccc cttcggcgtg cgcggtcacg cgcacagggc gcagccctgg ttaaaaacaa
     6961 ggtttataaa tattggttta aaagcaggtt aaaagacagg ttagcggtgg ccgaaaaacg
     7021 ggcggaaacc cttgcaaatg ctggattttc tgcctgtgga cagcccctca aatgtcaata
     7081 ggtgcgcccc tcatctgtca gcactctgcc cctcaagtgt caaggatcgc gcccctcatc
     7141 tgtcagtagt cgcgcccctc aagtgtcaat accgcagggc acttatcccc aggcttgtcc
     7201 acatcatctg tgggaaactc gcgtaaaatc aggcgttttc gccgatttgc gaggctggcc
     7261 agctccacgt cgccggccga aatcgagcct gcccctcatc tgtcaacgcc gcgccgggtg
     7321 agtcggcccc tcaagtgtca acgtccgccc ctcatctgtc agtgagggcc aagttttccg
     7381 cgaggtatcc acaacgccgg cggccgcggt gtctcgcaca cggcttcgac ggcgtttctg
     7441 gcgcgtttgc agggccatag acggccgcca gcccagcggc gagggcaacc agcccggtga
     7501 gcgtcgcaaa ggagatcctg atctgactga tgggctgcct gtatcgagtg gtgattttgt
     7561 gccgagctgc cggtcgggga gctgttggct ggctggtggc aggatatatt gtggtgtaaa
     7621 caaattgacg cttagacaac ttaataacac attgcggacg tttttaatgt actggggtgg
     7681 atgcagtggg ccccac
//
