LOCUS       sequence-363944-        5971 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EGFP-tDeg fluorogenic protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5971)
  AUTHORS   Zhou WJ, Li H, Zhang KK, Wang F, Chu X, Jiang JH
  TITLE     Genetically Encoded Sensor Enables Endogenous RNA Imaging with 
            Conformation-Switching Induced Fluorogenic Proteins.
  JOURNAL   J Am Chem Soc. 2021 Sep 8;143(35):14394-14401. doi: 
            10.1021/jacs.1c07719. Epub 2021 Aug 25.
  PUBMED    34431301
REFERENCE   2  (bases 1 to 5971)
  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"; doi: 
            "10.1021/jacs.1c07719"; journalName: "J Am Chem Soc"; date: 
            "2021-09-8- 8"; volume: "143"; issue: "35"; pages: "14394-14401"
COMMENT     Sequence Label: miniCMV-EGFP-tDeg
FEATURES             Location/Qualifiers
     source          1..5971
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        235..273
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     270..294
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             join(309..311,312..314,315..1025)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(309..311,312..314,315..1025)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(354..375)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(615..634)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     962..983
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        complement(1524..1542)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1525..1542)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1562..1579)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1568..1792
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1838..2266
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1925..1944)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2135..2156
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2275..2295)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2280..2609
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2460..2595
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2522..2541
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2676..3470
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2730..2749)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3340..3359
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3644..3765
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3681..3700)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3735..3754
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3814..3830)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3814..3830)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3827..3849)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3838..3854
                     /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(3862..3868,3869..3886,3887..3892))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3907..3928
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4045..4062)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4216..4804)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4296..4315)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4975..5766,5767..5835))
                     /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     5598..5617
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5836..5940)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttggtagg
      241 cgtgtacggt gggaggccta tataagcaga gctcgtttag tgaaccgtca gatcgcgccg
      301 acatcaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
      361 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
      421 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
      481 ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc
      541 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
      601 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
      661 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
      721 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc
      781 agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc
      841 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
      901 acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc
      961 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
     1021 acaagggagg cggaggctcc ggaggcggag gctccatggc ttctaacttt actcagttcg
     1081 ttctcgtcga caatggcgga actggcgacg tgactgtcgc cccaagcaac ttcgctaacg
     1141 ggatcgctga atggatcagc tctaactcgc gttcacaggc ttacaaagta acctgtagcg
     1201 ttcgtcagag ctctgcgcag aatcgcaaat acaccatcaa agtcgaggtg cctaaaggcg
     1261 cctggcgttc gtacttaaat atggaactaa ccattccaat tttcgccacg aattccgact
     1321 gcgagcttat tgttaaggca atgcaaggtc tcctaaaaga tggaaacccg attccctcag
     1381 caatcgcagc aaactccggc atctacggag gcggaggctc cggcggaaga tccggtggtg
     1441 gttctggtcc tcgtccccgt ggtactcgtg gtaaaggtcg ccgtattcgt cgccgcggtt
     1501 aagaattctc tagagggccc tattctatag tgtcacctaa atgctagagc tcgctgatca
     1561 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     1621 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     1681 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     1741 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag
     1801 gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta
     1861 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     1921 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     1981 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     2041 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     2101 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     2161 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     2221 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg
     2281 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     2341 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     2401 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     2461 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     2521 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
     2581 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg
     2641 gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     2701 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     2761 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     2821 tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt
     2881 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     2941 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     3001 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     3061 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     3121 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     3181 aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg
     3241 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     3301 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     3361 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     3421 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     3481 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     3541 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     3601 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc
     3661 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     3721 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc
     3781 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     3841 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     3901 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     3961 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     4021 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     4081 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     4141 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     4201 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     4261 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     4321 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     4381 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     4441 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     4501 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     4561 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     4621 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     4681 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     4741 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     4801 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     4861 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     4921 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     4981 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     5041 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     5101 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     5161 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     5221 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     5281 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     5341 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     5401 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     5461 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     5521 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     5581 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     5641 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     5701 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     5761 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     5821 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     5881 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     5941 cacatttccc cgaaaagtgc cacctgacgt c
//
