LOCUS       sequence-338275-        4769 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressing EGFP mRNA fused with 1 tandem repeat of a 50-base 
            sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4769)
  AUTHORS   Chen M, Ma Z, Wu X, Mao S, Yang Y, Tan J, Krueger CJ, Chen AK
  TITLE     A molecular beacon-based approach for live-cell imaging of RNA 
            transcripts with minimal target engineering at the single-molecule 
            level.
  JOURNAL   Sci Rep. 2017 May 8;7(1):1550. doi: 10.1038/s41598-017-01740-1.
  PUBMED    28484218
REFERENCE   2  (bases 1 to 4769)
  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: "Sci Rep."; 
            date: "2017-05-8"; volume: "7(1):1550. doi"; pages: " 
            10.1038/s41598-017-01740-1"
COMMENT     Sequence Label: pEGFP-N1-1x
FEATURES             Location/Qualifiers
     source          1..4769
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      649..658
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(655..657,658..660,661..1374)
                     /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(700..721)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(961..980)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1308..1329
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1497..1618
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1534..1553)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1588..1607
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1625..2080)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1762..1783)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1974..1993
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2107..2211
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2213..2570
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2213..2404
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2236..2256)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2421..2556
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2605..3399
                     /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             2605..3399
                     /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(2659..2678)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3269..3288
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3587..3606)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3631..3678
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4007..4595
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4496..4515
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggactcag atctcgagct caagcttcga attcgtcgac aggagttgtg tttgtggacg
      601 aagagcacca gccagctgat cgacctcgag ccgagggatc caccggtcgc caccatggtg
      661 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
      721 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
      781 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
      841 accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac
      901 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
      961 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
     1021 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     1081 gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc
     1141 aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac
     1201 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     1261 agcacccagt ccaagctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
     1321 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaaagcggc
     1381 cgcgactcta gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa
     1441 acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact
     1501 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     1561 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaag
     1621 gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     1681 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
     1741 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
     1801 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
     1861 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
     1921 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
     1981 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
     2041 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga
     2101 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     2161 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tcctgaggcg
     2221 gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     2281 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     2341 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     2401 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     2461 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
     2521 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa
     2581 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     2641 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     2701 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     2761 ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg
     2821 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     2881 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     2941 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     3001 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     3061 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     3121 aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     3181 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     3241 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     3301 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     3361 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     3421 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     3481 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     3541 gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa acacggaagg
     3601 agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg
     3661 gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata
     3721 ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc
     3781 ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca
     3841 tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     3901 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     3961 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     4021 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     4081 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     4141 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     4201 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     4261 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     4321 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     4381 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     4441 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     4501 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     4561 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     4621 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     4681 ctgtggataa ccgtattacc gccatgcatt agttattaat agtaatcaat tacggggtca
     4741 ttagttcata gcccatatat ggagttccg
//
