LOCUS       sequence-384640-        6077 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of mEGFP mRNA with 8xbiRhoBAST in 3'UTR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6077)
  AUTHORS   Buhler B, Schokolowski J, Benderoth A, Englert D, Grun F, Jaschke A,
            Sunbul M
  TITLE     Avidity-based bright and photostable light-up aptamers for 
            single-molecule mRNA imaging.
  JOURNAL   Nat Chem Biol. 2023 Apr;19(4):478-487. doi: 
            10.1038/s41589-022-01228-8. Epub 2023 Jan 19.
  PUBMED    36658339
REFERENCE   2  (bases 1 to 6077)
  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: 
            "10.1038/s41589-022-01228-8"; journalName: "Nat Chem Biol"; date: 
            "2023-04"; volume: "19"; issue: "4"; pages: "478-487"
COMMENT     Sequence Label: mEGFP-8xbiRhoBAST
FEATURES             Location/Qualifiers
     source          1..6077
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        133..436
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        437..640
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     591..611
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      676..685
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(682..684,685..687,688..1398)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(727..748)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(988..1007)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1335..1356
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2761..2813
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    2937..3058
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2974..2993)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3028..3047
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3065..3520)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3202..3223)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3414..3433
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3547..3651
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3653..4010
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3653..3844
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3676..3696)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3861..3996
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4045..4839
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4045..4839
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4099..4118)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4709..4728
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5027..5046)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5071..5118
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5447..6035
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5936..5955
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
       61 accgccatgc attagttatt aatagtaatc aattacgggg tcattagttc atagcccata
      121 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      181 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      241 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      301 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      361 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      421 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
      481 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
      541 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
      601 cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat
      661 ccgctagcga attccgccac catggtgagc aagggcgagg agctgttcac cggggtggtg
      721 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
      781 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
      841 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc
      901 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
      961 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     1021 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     1081 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
     1141 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag
     1201 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     1261 gtgctgctgc ccgacaacca ctacctgagc acccagtcca agctgagcaa agaccccaac
     1321 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     1381 atggacgagc tgtacaagtc cggactcaga tctcgacggg tctagaggta tgtgggagga
     1441 acctccgcga aagcggtgaa ggagaggcgc aaggttaacc gcctcaggtt ccaggaggat
     1501 tcgtcctcca aaggcctccg cgaaagcggt gaaggagcgc cacaaggtta actggcgcag
     1561 gccttaccca cataccgcta gaggagcccg gaaggaacct ccgcgaaagc ggtgaaggag
     1621 aggcgcaagg ttaaccgcct caggttccag cggagttcgc tccgcaaagg cctccgcgaa
     1681 agcggtgaag gagcgccaca aggttaactg gcgcaggcct tatccgggct ccgctagagg
     1741 tatgtgggag gaacctccgc gaaagcggtg aaggagaggc gcaaggttaa ccgcctcagg
     1801 ttccaggagg attcgtcctc caaaggcctc cgcgaaagcg gtgaaggagc gccacaaggt
     1861 taactggcgc aggccttacc cacataccgc tagaggagcc cggaaggaac ctccgcgaaa
     1921 gcggtgaagg agaggcgcaa ggttaaccgc ctcaggttcc agcggagttc gctccgcaaa
     1981 ggcctccgcg aaagcggtga aggagcgcca caaggttaac tggcgcaggc cttatccggg
     2041 ctccgctaga ggtatgtggg aggaacctcc gcgaaagcgg tgaaggagag gcgcaaggtt
     2101 aaccgcctca ggttccagga ggattcgtcc tccaaaggcc tccgcgaaag cggtgaagga
     2161 gcgccacaag gttaactggc gcaggcctta cccacatacc gctagaggag cccggaagga
     2221 acctccgcga aagcggtgaa ggagaggcgc aaggttaacc gcctcaggtt ccagcggagt
     2281 tcgctccgca aaggcctccg cgaaagcggt gaaggagcgc cacaaggtta actggcgcag
     2341 gccttatccg ggctccgcta gaggtatgtg ggaggaacct ccgcgaaagc ggtgaaggag
     2401 aggcgcaagg ttaaccgcct caggttccag gaggattcgt cctccaaagg cctccgcgaa
     2461 agcggtgaag gagcgccaca aggttaactg gcgcaggcct tacccacata ccgctagagg
     2521 agcccggaag gaacctccgc gaaagcggtg aaggagaggc gcaaggttaa ccgcctcagg
     2581 ttccagcgga gttcgctccg caaaggcctc cgcgaaagcg gtgaaggagc gccacaaggt
     2641 taactggcgc aggccttatc cgggctccgc tagagcggac ttcggtccgc tttttactag
     2701 gacctgcagg catgcaagct tgacgtcggt taccgatatc catatggcgg ccgcatcgat
     2761 ctcgagctca agcttcgaat tctgcagtcg acggtaccgc gggcccggga tccaccggat
     2821 ctagataact gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa
     2881 acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact
     2941 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     3001 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaac
     3061 gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     3121 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
     3181 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
     3241 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
     3301 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
     3361 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
     3421 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
     3481 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga
     3541 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     3601 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tcctgaggcg
     3661 gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     3721 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     3781 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     3841 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     3901 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
     3961 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa
     4021 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     4081 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     4141 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     4201 ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg
     4261 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     4321 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     4381 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     4441 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     4501 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     4561 aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     4621 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     4681 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     4741 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     4801 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     4861 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     4921 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     4981 gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa acacggaagg
     5041 agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg
     5101 gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata
     5161 ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc
     5221 ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca
     5281 tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     5341 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     5401 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     5461 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     5521 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     5581 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     5641 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     5701 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     5761 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     5821 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     5881 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     5941 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     6001 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     6061 ggttcctggc cttttgc
//
