LOCUS       sequence-150626-        5070 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BiFC vector for transient expression of eGN155-MXMT in plants.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5070)
  AUTHORS   Fujii Y, Kodama Y
  TITLE     In planta comparative analysis of improved green fluorescent 
            proteins with reference to fluorescence intensity and bimolecular 
            fluorescence complementation ability.
  JOURNAL   Plant Biotech. 2015; 32(1): 81-87.
REFERENCE   2  (bases 1 to 5070)
  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 
            Biotech."; date: "2015"; volume: " 32(1)"; pages: " 81-87"
COMMENT     Sequence Label: 35S-eGN155-MXMT-NosT
FEATURES             Location/Qualifiers
     source          1..5070
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        21..365
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     308..328
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     misc_feature    385..440
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5'-leader sequence (Gallie et al., 1988)"
     CDS             join(460..462,463..465,466..924)
                     /codon_start=1
                     /product="improved N-terminal fragment of mVenus for use in
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN155(I152L)
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMA"
     primer_bind     complement(505..526)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(766..785)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     terminator      2088..2340
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(2378..2394)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2413..2429)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2426..2448)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2437..2453
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(2461..2467,2468..2485,2486..2491))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2506..2527
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2644..2661)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2815..3403)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2895..2914)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3574..4365,4366..4434))
                     /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     4197..4216
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4435..4539)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4607..4625
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4663..4685)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4785..4804
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4998..5020
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5012..5029
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5013..5029
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 ctcagaagac cgagggctat tgagactttt caacaaaggg taatatcggg aaacctcctc
       61 ggattccatt gcccagctat ctgtcactta tcgaaaggac agtagaaaag gaaggtggct
      121 cctacaaatg ccatcattgc gataaaggaa aggctatcgt tcaagatgcc tctaccgaca
      181 gtggtcccaa agatggaccc ccacccacga ggaacatcgt ggaaaaagaa gacgttccaa
      241 ccacgtcttc aaagcaagtg gattgatgtg atatctccac tgacgtaagg gatgacgcac
      301 aatcccacta tccttcgcaa gacccttcct ctatataagg aagttcattt catttggaga
      361 ggacaggctt cttgagatcc ttcaacaatt accaacaaca acaaacaaca aacaacatta
      421 caattactat ttacaattac agtcgactct agaggatcca tggtgagcaa gggcgaggag
      481 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
      541 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc
      601 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac
      661 ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
      721 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
      781 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
      841 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
      901 agccacaacg tctatatcat ggccctgtac aagatggagc tccaagaagt cctgcatatg
      961 aatgaaggtg aaggcgatac aagctacgcc aagaatgcat cctacaatct ggctcttgcc
     1021 aaggtgaaac ctttccttga acaatgcata cgagaattgt tgcgggccaa cttgcccaac
     1081 atcaacaagt gcattaaagt tgcggatttg ggatgcgctt ctggaccaaa cacactttta
     1141 acagtgcggg acattgtgca aagtattgac aaagttggcc aggaagagaa gaatgaatta
     1201 gaacgtccca ccattcagat ttttctgaat gatcttttcc aaaatgattt caattcggtt
     1261 ttcaagttgc tgccaagctt ctaccgcaaa ctcgagaaag aaaatggacg caagatagga
     1321 tcgtgcctaa taagcgcaat gcctggctct ttctacggca gactcttccc cgaggagtcc
     1381 atgcattttt tgcactcttg ttacagtgtt cattggttat ctcaggttcc cagcggtttg
     1441 gtgattgaat tggggattgg tgcaaacaaa gggagtattt actcttccaa aggatgtcgt
     1501 ccgcccgtcc agaaggcata tttggatcaa tttacgaaag attttaccac atttctaagg
     1561 attcattcga aagagttgtt ttcacgtggc cgaatgctcc ttacctgcat ttgtaaagta
     1621 gatgaattcg acgaaccgaa tcccctagac ttacttgaca tggcaataaa cgacttgatt
     1681 gttgagggac ttctggagga agaaaaattg gatagtttca atattccatt ctttacacct
     1741 tcagcagaag aagtaaagtg catagttgag gaggaaggtt cttgcgaaat tttatatctg
     1801 gagactttta aggcccatta tgatgctgcc ttctctattg atgatgatta cccagtaaga
     1861 tcccatgaac aaattaaagc agagtatgtg gcatcattaa ttagatcagt ttacgaaccc
     1921 atcctcgcaa gtcattttgg agaagctatt atgcctgact tattccacag gcttgcgaag
     1981 catgcagcaa aggttctcca catgggcaaa ggctgctata ataatcttat catttctctc
     2041 gccaaaaagc cagagaagtc agacgtgtaa gcggccgccc ggctgcagat cgttcaaaca
     2101 tttggcaata aagtttctta agattgaatc ctgttgccgg tcttgcgatg attatcatat
     2161 aatttctgtt gaattacgtt aagcatgtaa taattaacat gtaatgcatg acgttattta
     2221 tgagatgggt ttttatgatt agagtcccgc aattatacat ttaatacgcg atagaaaaca
     2281 aaatatagcg cgcaaactag gataaattat cgcgcgcggt gtcatctatg ttactagatc
     2341 cgatgataag ctgtcaaaca tagaattcgt aatcatggtc atagctgttt cctgtgtgaa
     2401 ttcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     2461 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     2521 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     2581 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     2641 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     2701 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     2761 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     2821 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     2881 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     2941 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     3001 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     3061 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     3121 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     3181 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     3241 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     3301 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     3361 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     3421 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     3481 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     3541 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     3601 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     3661 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     3721 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     3781 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     3841 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     3901 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     3961 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     4021 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     4081 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     4141 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     4201 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     4261 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     4321 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     4381 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     4441 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     4501 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     4561 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     4621 gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     4681 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     4741 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat
     4801 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
     4861 ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg
     4921 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg
     4981 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg ccaagcttgc
     5041 atgcctgcag gtcgatctag aggatccccc
//
