LOCUS       sequence-312937-        5120 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses L7Ae in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5120)
  AUTHORS   Ono H, Kawasaki S, Saito H
  TITLE     Orthogonal Protein-Responsive mRNA Switches for Mammalian Synthetic 
            Biology.
  JOURNAL   ACS Synth Biol. 2020 Jan 17;9(1):169-174. doi: 
            10.1021/acssynbio.9b00343. Epub 2019 Dec 13.
  PUBMED    31765565
REFERENCE   2  (bases 1 to 5120)
  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/acssynbio.9b00343"; journalName: "ACS Synth Biol"; date: 
            "2020-01-17- 17"; volume: "9"; issue: "1"; pages: "169-174"
COMMENT     Sequence Label: pL7Ae-myc-T2A-tagRFP
FEATURES             Location/Qualifiers
     source          1..5120
                     /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"
     CDS             921..950
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             957..1010
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             1017..1727
                     /codon_start=1
                     /product="monomeric derivative of red fluorescent protein 
                     from Entacmaea quadricolor (Merzlyak et al., 2007)"
                     /label=TagRFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMR
                     IKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGV
                     LTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRSDM
                     ALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAVA
                     RYCDLPSKLGHKLN"
     polyA_signal    1848..1969
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1885..1904)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1939..1958
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1976..2431)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2113..2134)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2325..2344
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2458..2562
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2564..2921
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2564..2755
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2587..2607)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2772..2907
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2956..3750
                     /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             2956..3750
                     /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(3010..3029)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3620..3639
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3938..3957)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3982..4029
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4358..4946
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4847..4866
                     /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 gctagggctc
      541 tgcaggtcga ccccaccatg tacgtgagat ttgaggttcc tgaggacatg cagaacgaag
      601 ctctgagtct gctggagaag gttagggaga gcggtaaggt aaagaaaggt accaacgaga
      661 cgacaaaggc tgtggagagg ggactggcaa agctcgttta catcgcagag gatgttgacc
      721 cgcctgagat cgttgctcat ctgcccctcc tctgcgagga gaagaatgtg ccgtacattt
      781 acgttaaaag caagaacgac cttggaaggg ctgtgggcat tgaggtgcca tgcgcttcgg
      841 cagcgataat caacgaggga gagctgagaa aggagcttgg aagccttgtg gagaagatta
      901 aaggccttca gaagggatcc gaacaaaaac tcatctcaga agaggatctg agatctgaag
      961 gcagaggctc cctcctgacc tgcggagacg tggaagagaa ccccggacca agatccgtgt
     1021 ctaagggcga agagctgatt aaggagaaca tgcacatgaa gctgtacatg gagggcaccg
     1081 tgaacaacca ccacttcaag tgcacatccg agggcgaagg caagccctac gagggcaccc
     1141 agaccatgag aatcaaggtg gtcgagggcg gccctctccc cttcgccttc gacatcctgg
     1201 ctaccagctt catgtacggc agcagaacct tcatcaacca cacccagggc atccccgact
     1261 tctttaagca gtccttccct gagggcttca catgggagag agtcaccaca tacgaagacg
     1321 ggggcgtgct gaccgctacc caggacacca gcctccagga cggctgcctc atctacaacg
     1381 tcaagatcag aggggtgaac ttcccatcca acggccctgt gatgcagaag aaaacactcg
     1441 gctgggaggc caacaccgag atgctgtacc ccgctgacgg cggcctggaa ggcagaagcg
     1501 acatggccct gaagctcgtg ggcgggggcc acctgatctg caacttcaag accacataca
     1561 gatccaagaa acccgctaag aacctcaaga tgcccggcgt ctactatgtg gaccacagac
     1621 tggaaagaat caaggaggcc gacaaagaga cctacgtcga gcagcacgag gtggctgtgg
     1681 ccagatactg cgacctccct agcaaactgg ggcacaaact taattagcgg ccgcgactct
     1741 agatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
     1801 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
     1861 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     1921 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt
     1981 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     2041 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     2101 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     2161 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     2221 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     2281 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     2341 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     2401 gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc
     2461 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     2521 taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc
     2581 agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     2641 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     2701 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     2761 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     2821 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     2881 agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga
     2941 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     3001 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     3061 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     3121 gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     3181 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     3241 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     3301 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     3361 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     3421 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     3481 gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     3541 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     3601 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     3661 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     3721 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     3781 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     3841 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     3901 tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac
     3961 cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt
     4021 cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga
     4081 gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt
     4141 cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag
     4201 gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     4261 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     4321 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     4381 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     4441 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     4501 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     4561 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     4621 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     4681 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     4741 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     4801 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     4861 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     4921 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     4981 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     5041 accgtattac cgccatgcat tagttattaa tagtaatcaa ttacggggtc attagttcat
     5101 agcccatata tggagttccg
//
