LOCUS       sequence-478590-        6022 bp    DNA     circular SYN 14-JUL-2025
DEFINITION  Contains EGFP-fL7Ae fluorogenic protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6022)
  AUTHORS   Zhou WJ, Wu MY, Shao XJ, Tang LJ, Wang F, Jiang JH
  TITLE     Fluorogenic Interacting Protein Stabilization for Orthogonal RNA 
            Imaging.
  JOURNAL   Angew Chem Int Ed Engl. 2025 Apr 10:e202502350. doi: 
            10.1002/anie.202502350.
  PUBMED    40208703
REFERENCE   2  (bases 1 to 6022)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Angew Chem 
            Int Ed Engl. 2025 Apr 10:e202502350. doi: 10.1002/anie.202502350."
COMMENT     Sequence Label: EGFP-fL7Ae
FEATURES             Location/Qualifiers
     source          1..6022
                     /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"
     CDS             1056..1082
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             1092..1118
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     promoter        complement(1575..1593)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1576..1593)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1613..1630)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1619..1843
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1889..2317
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1976..1995)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2186..2207
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2326..2346)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2331..2660
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2511..2646
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2573..2592
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2727..3521
                     /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(2781..2800)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3391..3410
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3695..3816
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3732..3751)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3786..3805
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3865..3881)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3865..3881)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3878..3900)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3889..3905
                     /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(3913..3919,3920..3937,3938..3943))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3958..3979
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4096..4113)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4267..4855)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4347..4366)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5026..5817,5818..5886))
                     /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     5649..5668
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5887..5991)
                     /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 gctccctgcc ccccctggag cgcctgaccc
     1081 tgggaggctc cctgcccccc ctggagcgcc tgaccctggg aggcggaggc tccggaggcg
     1141 gaggctccgc cggcaccggc ggcatgtacg tgcgcttcga ggtgcccgag gacatgcaga
     1201 acgaggccct gagcctgctg gaaaaagtgc gcgagagcgg caaagtgaag aagggcacca
     1261 acgaaaccac caaggccgtg gaacggggcc tggccaagct ggtgtatatc gccgaggacg
     1321 tggacccccc cgagattgtg gcccatctgc ccctgctgtg cgaagagaag aacgtgccct
     1381 acatctacgt gaagtccaag aacgacctgg gcagagccgt gggcatcgag gtgccatgtg
     1441 cctctgccgc catcatcaac gagggcgagc tgcggaaaga actgggcagc ctggtggaaa
     1501 agatcaaggg cctgcagaag ggttcccgtc gccgccgtcg tcgccgcggt taagaattct
     1561 ctagagggcc ctattctata gtgtcaccta aatgctagag ctcgctgatc agcctcgact
     1621 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     1681 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     1741 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     1801 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
     1861 accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
     1921 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     1981 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     2041 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     2101 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     2161 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     2221 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     2281 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
     2341 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     2401 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     2461 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     2521 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     2581 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     2641 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc
     2701 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     2761 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     2821 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     2881 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca
     2941 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     3001 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     3061 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     3121 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     3181 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     3241 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     3301 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     3361 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     3421 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     3481 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
     3541 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     3601 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     3661 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
     3721 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     3781 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
     3841 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     3901 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     3961 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     4021 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     4081 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     4141 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     4201 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     4261 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     4321 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     4381 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     4441 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     4501 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     4561 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     4621 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     4681 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     4741 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     4801 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     4861 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     4921 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     4981 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     5041 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     5101 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     5161 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     5221 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     5281 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     5341 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     5401 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     5461 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     5521 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     5581 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     5641 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     5701 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     5761 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     5821 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     5881 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     5941 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     6001 ccgaaaagtg ccacctgacg tc
//
