LOCUS       sequence-478593-        6064 bp    DNA     circular SYN 14-JUL-2025
DEFINITION  Contains EGFP-fLIN28A fluorogenic protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6064)
  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 6064)
  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-fLIN28A
FEATURES             Location/Qualifiers
     source          1..6064
                     /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(1617..1635)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1618..1635)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1655..1672)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1661..1885
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1931..2359
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2018..2037)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2228..2249
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2368..2388)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2373..2702
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2553..2688
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2615..2634
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2769..3563
                     /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(2823..2842)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3433..3452
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3737..3858
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3774..3793)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3828..3847
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3907..3923)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3907..3923)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3920..3942)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3931..3947
                     /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(3955..3961,3962..3979,3980..3985))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4000..4021
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4138..4155)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4309..4897)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4389..4408)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5068..5859,5860..5928))
                     /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     5691..5710
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5929..6033)
                     /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 ttccggtgga tccggttccg
     1141 gagacaggtg ctacaactgt ggaggtctag atcatcatgc caaggaatgc aagctgccac
     1201 cccagcccaa gaagtgccac ttctgccaga gcatcagcca tatggtagcc tcatgtccgc
     1261 tgaaggccca gggaggcgga ggctccggag gcggaggctc ccctcagctg ctgcacggtg
     1321 cgggcatctg taagtggttc aacgtgcgca tggggttcgg cttcctgtcc atgaccgccc
     1381 gcgccggggt cgcgctcgac cccccagtgg atgtctttgt gcaccagagt aagctgcaca
     1441 tggaagggtt ccggagcttg aaggagggtg aggcagtgga gttcaccttt aagaagtcag
     1501 ccaagggtct ggaatccatc cgtgtcaccg gacctggtgg agtattctgt attgggagtg
     1561 agggttcccg tcgccgccgt cgtcgccgcg gttaagaatt ctctagaggg ccctattcta
     1621 tagtgtcacc taaatgctag agctcgctga tcagcctcga ctgtgccttc tagttgccag
     1681 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     1741 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
     1801 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
     1861 gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg
     1921 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     1981 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     2041 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     2101 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     2161 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     2221 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     2281 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     2341 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     2401 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     2461 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     2521 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     2581 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     2641 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     2701 aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat
     2761 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     2821 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     2881 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     2941 atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     3001 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     3061 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     3121 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     3181 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     3241 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca
     3301 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     3361 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     3421 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     3481 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     3541 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac
     3601 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     3661 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     3721 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     3781 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     3841 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta
     3901 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     3961 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     4021 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     4081 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     4141 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     4201 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     4261 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     4321 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     4381 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     4441 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     4501 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     4561 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     4621 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     4681 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     4741 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     4801 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     4861 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     4921 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     4981 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     5041 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     5101 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     5161 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     5221 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     5281 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     5341 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     5401 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     5461 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     5521 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     5581 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     5641 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     5701 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     5761 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     5821 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     5881 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     5941 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     6001 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     6061 cgtc
//
