LOCUS       sequence-466897-        5034 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BiFC vector for transient expression of frGN155-MXMT in plants..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5034)
  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 5034)
  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-frGN155-MXMT-NosT
FEATURES             Location/Qualifiers
     source          1..5034
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        22..367
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     310..330
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     misc_feature    387..442
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5'-leader sequence (Gallie et al., 1988)"
     CDS             join(462..464,465..467,468..926)
                     /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
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITA"
     primer_bind     complement(507..528)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(768..787)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     terminator      2090..2342
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(2380..2396)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2380..2396)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2393..2415)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2404..2420
                     /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(2428..2434,2435..2452,2453..2458))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2473..2494
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2611..2628)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2782..3370)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2862..2881)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3541..4332,4333..4401))
                     /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     4164..4183
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4402..4506)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4574..4592
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4630..4652)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4752..4771
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4965..4987
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4979..4996
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4980..4996
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 ctcagaagac cagagggcta ttgagacttt tcaacaaagg gtaatatcgg gaaacctcct
       61 cggattccat tgcccagcta tctgtcactt catcgaaagg acagtagaaa aggaaggtgg
      121 ctcctacaaa tgccatcatt gcgataaagg aaaggctatc gttcaagatg cctctaccga
      181 cagtggtccc aaagatggac ccccacccac gaggaacatc gtggaaaaag aagacgttcc
      241 aaccacgtct tcaaagcaag tggattgatg tgatatctcc actgacgtaa gggatgacgc
      301 acaatcccac tatccttcgc aagacccttc ctctatataa ggaagttcat ttcatttgga
      361 gaggacaggc ttcttgagat ccttcaacaa ttaccaacaa caacaaacaa caaacaacat
      421 tacaattact atttacaatt acagtcgact ctagaggatc catggtgagc aagggcgagg
      481 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
      541 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt
      601 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct
      661 acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
      721 ccgccatgcc cgaaggctac gtccaggagc gcaccatctc tttcaaggac gacggcaact
      781 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
      841 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca
      901 acagccacaa cgtctatatc actgccctgt acaagatgga gctccaagaa gtcctgcata
      961 tgaatgaagg tgaaggcgat acaagctacg ccaagaatgc atcctacaat ctggctcttg
     1021 ccaaggtgaa acctttcctt gaacaatgca tacgagaatt gttgcgggcc aacttgccca
     1081 acatcaacaa gtgcattaaa gttgcggatt tgggatgcgc ttctggacca aacacacttt
     1141 taacagtgcg ggacattgtg caaagtattg acaaagttgg ccaggaagag aagaatgaat
     1201 tagaacgtcc caccattcag atttttctga atgatctttt ccaaaatgat ttcaattcgg
     1261 ttttcaagtt gctgccaagc ttctaccgca aactcgagaa agaaaatgga cgcaagatag
     1321 gatcgtgcct aataagcgca atgcctggct ctttctacgg cagactcttc cccgaggagt
     1381 ccatgcattt tttgcactct tgttacagtg ttcattggtt atctcaggtt cccagcggtt
     1441 tggtgattga attggggatt ggtgcaaaca aagggagtat ttactcttcc aaaggatgtc
     1501 gtccgcccgt ccagaaggca tatttggatc aatttacgaa agattttacc acatttctaa
     1561 ggattcattc gaaagagttg ttttcacgtg gccgaatgct ccttacctgc atttgtaaag
     1621 tagatgaatt cgacgaaccg aatcccctag acttacttga catggcaata aacgacttga
     1681 ttgttgaggg acttctggag gaagaaaaat tggatagttt caatattcca ttctttacac
     1741 cttcagcaga agaagtaaag tgcatagttg aggaggaagg ttcttgcgaa attttatatc
     1801 tggagacttt taaggcccat tatgatgctg ccttctctat tgatgatgat tacccagtaa
     1861 gatcccatga acaaattaaa gcagagtatg tggcatcatt aattagatca gtttacgaac
     1921 ccatcctcgc aagtcatttt ggagaagcta ttatgcctga cttattccac aggcttgcga
     1981 agcatgcagc aaaggttctc cacatgggca aaggctgcta taataatctt atcatttctc
     2041 tcgccaaaaa gccagagaag tcagacgtgt aagcggccgc ccggctgcag atcgttcaaa
     2101 catttggcaa taaagtttct taagattgaa tcctgttgcc ggtcttgcga tgattatcat
     2161 ataatttctg ttgaattacg ttaagcatgt aataattaac atgtaatgca tgacgttatt
     2221 tatgagatgg gtttttatga ttagagtccc gcaattatac atttaatacg cgatagaaaa
     2281 caaaatatag cgcgcaaact aggataaatt atcgcgcgcg gtgtcatcta tgttactaga
     2341 tccgatgata agctgtcaaa catgagaatt cgtaatcatg tcatagctgt ttcctgtgtg
     2401 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     2461 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     2521 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     2581 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     2641 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     2701 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     2761 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     2821 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     2881 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     2941 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     3001 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     3061 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     3121 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     3181 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     3241 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     3301 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     3361 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     3421 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     3481 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     3541 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     3601 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     3661 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     3721 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     3781 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     3841 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     3901 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     3961 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     4021 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     4081 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     4141 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     4201 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     4261 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     4321 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     4381 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     4441 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     4501 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac
     4561 attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga
     4621 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga
     4681 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg
     4741 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat
     4801 accgcacaga tgcgtaagga gaaaataccg catcaggcgc cattcgccat tcaggctgcg
     4861 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
     4921 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
     4981 taaaacgacg gccagtgcca agcttgcatg cctgcaggct ctagaggatc cccc
//
