LOCUS       sequence-348004-        7049 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 1 module containing TP-CFP_P4 F.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7049)
  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 7049)
  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: pLBT0180
FEATURES             Location/Qualifiers
     source          1..7049
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    76..100
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      192..823
                     /label=oriV
                     /note="incP origin of replication"
     promoter        910..1014
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1015..1083,1084..1875)
                     /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     complement(1233..1252)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2046..2634
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2535..2554
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             2970..4118
                     /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    4274..4298
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     terminator      complement(4346..4598)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(join(4611..5324,5325..5327,5328..5330))
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4656..4677)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     5005..5024
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5264..5285
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     promoter        complement(5682..6027)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(5719..5739)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
ORIGIN
        1 gtggggccca ctgcatccac cccagtacat taaaaacgtc cgcaatgtgt tattaagttg
       61 tctaagcgtc aatttgttta caccacaata tatcctgcca ccagccagcc aacagctccc
      121 cgaccggcag ctcggcacaa aatcaccact cgatacaggc agcccatcag tcagatcagg
      181 atctcctttg cgacgctcac cgggctggtt gccctcgccg ctgggctggc ggccgtctat
      241 ggccctgcaa acgcgccaga aacgccgtcg aagccgtgtg cgagacaccg cggccgccgg
      301 cgttgtggat acctcgcgga aaacttggcc ctcactgaca gatgaggggc ggacgttgac
      361 acttgagggg ccgactcacc cggcgcggcg ttgacagatg aggggcaggc tcgatttcgg
      421 ccggcgacgt ggagctggcc agcctcgcaa atcggcgaaa acgcctgatt ttacgcgagt
      481 ttcccacaga tgatgtggac aagcctgggg ataagtgccc tgcggtattg acacttgagg
      541 ggcgcgacta ctgacagatg aggggcgcga tccttgacac ttgaggggca gagtgctgac
      601 agatgagggg cgcacctatt gacatttgag gggctgtcca caggcagaaa atccagcatt
      661 tgcaagggtt tccgcccgtt tttcggccac cgctaacctg tcttttaacc tgcttttaaa
      721 ccaatattta taaaccttgt ttttaaccag ggctgcgccc tgtgcgcgtg accgcgcacg
      781 ccgaaggggg gtgccccccc ttctcgaacc ctcccggccc gctaacgcgg gcctcccatc
      841 cccccagggg ctgcgcccct cggccgcgaa cggcctcacc ccaaaaatgg cagcgctggc
      901 caattcgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
      961 ctcatgagac aataaccttg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     1021 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     1081 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     1141 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     1201 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     1261 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     1321 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     1381 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     1441 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     1501 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     1561 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     1621 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     1681 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg ttctcgcggt
     1741 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     1801 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     1861 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     1921 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     1981 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     2041 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     2101 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     2161 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     2221 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     2281 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     2341 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     2401 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     2461 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     2521 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     2581 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     2641 agcaacgcgg cctttttacg gttcctggca gatcctagat gtggcgcaac gatgccggcg
     2701 acaagcagga gcgcaccgac ttcttccgca tcaagtgttt tggctctcag gccgaggccc
     2761 acggcaagta tttgggcaag gggtcgctgg tattcgtgca gggcaagatt cggaatacca
     2821 agtacgagaa ggacggccag acggtctacg ggaccgactt cattgccgat aaggtggatt
     2881 atctggacac caaggcacca ggcgggtcaa atcaggaata agggcacatt gccccggcgt
     2941 gagtcggggc aatcccgcaa ggagggtgaa tgaatcggac gtttgaccgg aaggcataca
     3001 ggcaagaact gatcgacgcg gggttttccg ccgaggatgc cgaaaccatc gcaagccgca
     3061 ccgtcatgcg tgcgccccgc gaaaccttcc agtccgtcgg ctcgatggtc cagcaagcta
     3121 cggccaagat cgagcgcgac agcgtgcaac tggctccccc tgccctgccc gcgccatcgg
     3181 ccgccgtgga gcgttcgcgt cgtcttgaac aggaggcggc aggtttggcg aagtcgatga
     3241 ccatcgacac gcgaggaact atgacgacca agaagcgaaa aaccgccggc gaggacctgg
     3301 caaaacaggt cagcgaggcc aagcaggccg cgttgctgaa acacacgaag cagcagatca
     3361 aggaaatgca gctttccttg ttcgatattg cgccgtggcc ggacacgatg cgagcgatgc
     3421 caaacgacac ggcccgctct gccctgttca ccacgcgcaa caagaaaatc ccgcgcgagg
     3481 cgctgcaaaa caaggtcatt ttccacgtca acaaggacgt gaagatcacc tacaccggcg
     3541 tcgagctgcg ggccgacgat gacgaactgg tgtggcagca ggtgttggag tacgcgaagc
     3601 gcacccctat cggcgagccg atcaccttca cgttctacga gctttgccag gacctgggct
     3661 ggtcgatcaa tggccggtat tacacgaagg ccgaggaatg cctgtcgcgc ctacaggcga
     3721 cggcgatggg cttcacgtcc gaccgcgttg ggcacctgga atcggtgtcg ctgctgcacc
     3781 gcttccgcgt cctggaccgt ggcaagaaaa cgtcccgttg ccaggtcctg atcgacgagg
     3841 aaatcgtcgt gctgtttgct ggcgaccact acacgaaatt catatgggag aagtaccgca
     3901 agctgtcgcc gacggcccga cggatgttcg actatttcag ctcgcaccgg gagccgtacc
     3961 cgctcaagct ggaaaccttc cgcctcatgt gcggatcgga ttccacccgc gtgaagaagt
     4021 ggcgcgagca ggtcggcgaa gcctgcgaag agttgcgagg cagcggcctg gtggaacacg
     4081 cctgggtcaa tgatgacctg gtgcattgca aacgctaggg ccttgtgggg tcagttccgg
     4141 ctgggggttc agcagccagc gcctgatctg gggaaccctg tggttggcat gcacatacaa
     4201 atggacgaac ggataaacct tttcacgccc ttttaaatat ccgattattc taataaacgc
     4261 tcttttctct taggtttacc cgccaatata tcctgtcaaa cactgatagt ttaaaccact
     4321 tcgtgcagaa gacaatctga gcgtcgatct agtaacatag atgacaccgc gcgcgataat
     4381 ttatcctagt ttgcgcgcta tattttgttt tctatcgcgt attaaatgta taattgcggg
     4441 actctaatca taaaaaccca tctcataaat aacgtcatgc attacatgtt aattattaca
     4501 tgcttaacgt aattcaacag aaattatatg ataatcatcg caagaccggc aacaggattc
     4561 aatcttaaga aactttattg ccaaatgttt gaacgatctg cttgacaagc ttacttgtac
     4621 agctcgtcca tgccgagagt gatcccggcg gcggtcacga actccagcag gaccatgtga
     4681 tcgcgcttct cgttggggtc tttgctcagg gcggactggg tgctcaggta gtggttgtcg
     4741 ggcagcagca cggggccgtc gccgatgggg gtgttctgct ggtagtggtc ggcgagctgc
     4801 acgctgccgt cctcgatgtt gtggcggatc ttgaagttgg ccttgatgcc gttcttctgc
     4861 ttgtcggcgg tgatatagac gttgtggctg atgtagttgt actccagctt gtgccccagg
     4921 atgttgccgt cctccttgaa gtcgatgccc ttcagctcga tgcggttcac cagggtgtcg
     4981 ccctcgaact tcacctcggc gcgggtcttg tagttgccgt cgtccttgaa gaagatggtg
     5041 cgctcctgga cgtagccttc gggcatggcg gacttgaaga agtcgtgctg cttcatgtgg
     5101 tcggggtagc ggctgaagca ctgcacgccc caggtcaggg tggtcacgag ggtgggccag
     5161 ggcacgggca gcttgccggt ggtgcagatg aacttcaggg tcagcttgcc gtaggtggca
     5221 tcgccctcgc cctcgccgga cacgctgaac ttgtggccgt ttacgtcgcc gtccagctcg
     5281 accaggatgg gcaccacccc ggtgaacagc tcctcgccct tgctcaccat ttcggaatcg
     5341 gtaaggtcag gaaggtaaga gagagtctca aacttcttct ttccaatcgg aggccacacc
     5401 tattttttcg atttttccaa tgatattagt ttatgtagat tcgaagaaag aaccgaaaaa
     5461 aatcatatgg aataataaaa aagtgaaaga agaaatataa atgacctgca tgcagttaac
     5521 tcttccgccg ttgcttgtga tggaagtaat gtcgttgtta gccttgcggg tggctgggaa
     5581 ggcagcggag gacttaagtc cgttgaaagg agcgaccata gtggcctgag ccggagaggc
     5641 aaccatagta gcggaagaga gcatagagga agccatgggc gtgtcctctc caaatgaaat
     5701 gaacttcctt atatagagga agggtcttgc gaaggatagt gggattgtgc gtcatccctt
     5761 acgtcagtgg agatgtcaca tcaatccact tgctttgtag acgtggttgg aacctcttct
     5821 ttttccacga tgctcctcgt gggtgggggt ccatctttgg gaccactgtc ggcagagaga
     5881 tcttgaatga tagcctttcc tttatcgcaa tgatggcatt tgtaggagcc accttccttt
     5941 tctactgtcc tttcgatgaa gtgacagata gctgggcaat ggaatccgag gaggtttccc
     6001 gaaattatcc tttgttgaaa agtctcaata gccctttgat cttctgagac tgtatctttg
     6061 acatttttgg agtagaccag agtgtcgtgc tccaccatgt tgacgaagat tttcttcttg
     6121 tcattgagtc gtaaaagact ctgtatgaac tgttcgccag tcttgacggc gagttctgtt
     6181 agatcctcga tttgaatctt agactccatg catggcctta gattcagtag gaactacctt
     6241 tttagagact ccaatctcta ttacttgcct tggtttatga agcaagcctt gaatcgtcca
     6301 tactggaata gtacttctga tcttgagaaa tatgtctttc tctgtgttct tgatgcaatt
     6361 agtcctgaat cttttgactg catctttaac cttcttggga aggtatttga tctcctggag
     6421 attgttactc gggtagatcg tcttgatgag acttgctgcg taggcctctc taaccatctg
     6481 tgggtcagca ttctttctga aattgaagag gctaaccttc tcattatcag tggtgaacat
     6541 agtgtcgtca ccttcacctt cgaacttcct tcctagatcg taaagataga ggaaatcgtc
     6601 cattgtaatc tccggggcaa aggagatctc ttttggggct ggatcactgc tgggcctttt
     6661 ggttcctagc gtgagccagt gggctttttg ctttggtggg cttgttaggg ccttagcaaa
     6721 gctcttgggc ttgagttgag cttctccttt ggggatgaag ttcaacctgt ctgtttgctg
     6781 acttgttgtg tacgcgtcag ctgctgctct tgcctctgta atagtggcaa atttcttgtg
     6841 tgcaactccg ggaacgccgt ttgttgccgc ctttgtacaa ccccagtcat cgtatatacc
     6901 ggcatgtgga ccgttataca caacgtagta gttgatatga gggtgttgaa tacccgattc
     6961 tgctctgaga ggagcaactg tgctgttaag ctcagatttt tgtgggattg gaattcctcc
     7021 ccatgggaat tcgtaattgt cttcacaga
//
