LOCUS       sequence-167543-        4180 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transient expression vector for frGFP in plants.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4180)
  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 4180)
  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-frGFP-nosT
FEATURES             Location/Qualifiers
     source          1..4180
                     /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..1179)
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     CDS             join(460..462,463..465,466..1179)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     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"
     primer_bind     1113..1134
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      1198..1450
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(1488..1504)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1523..1539)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1536..1558)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1547..1563
                     /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(1571..1577,1578..1595,1596..1601))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1616..1637
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1754..1771)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1925..2513)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2005..2024)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2684..3475,3476..3544))
                     /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     3307..3326
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3545..3649)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3717..3735
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3773..3795)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3895..3914
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4108..4130
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4122..4139
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4123..4139
                     /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 accatctctt tcaaggacga cggcaactac
      781 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
      841 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
      901 agccacaacg tctatatcac tgccgacaag cagaagaacg gcatcaaggc caacttcaag
      961 atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc
     1021 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc
     1081 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     1141 gccgggatca ctcacggcat ggacgagctg tacaagtaaa gcggccgccc ggctgcagat
     1201 cgttcaaaca tttggcaata aagtttctta agattgaatc ctgttgccgg tcttgcgatg
     1261 attatcatat aatttctgtt gaattacgtt aagcatgtaa taattaacat gtaatgcatg
     1321 acgttattta tgagatgggt ttttatgatt agagtcccgc aattatacat ttaatacgcg
     1381 atagaaaaca aaatatagcg cgcaaactag gataaattat cgcgcgcggt gtcatctatg
     1441 ttactagatc cgatgataag ctgtcaaaca tagaattcgt aatcatggtc atagctgttt
     1501 cctgtgtgaa ttcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     1561 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     1621 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     1681 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     1741 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     1801 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     1861 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     1921 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     1981 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     2041 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     2101 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     2161 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     2221 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     2281 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     2341 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     2401 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     2461 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     2521 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     2581 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     2641 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     2701 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     2761 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     2821 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     2881 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     2941 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     3001 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     3061 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     3121 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     3181 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     3241 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     3301 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     3361 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     3421 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     3481 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     3541 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     3601 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     3661 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
     3721 ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac
     3781 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag
     3841 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat
     3901 cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa
     3961 ggagaaaata ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc
     4021 gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc
     4081 gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg
     4141 ccaagcttgc atgcctgcag gtcgatctag aggatccccc
//
