LOCUS       sequence-312936-        5135 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses PP7CP in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5135)
  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 5135)
  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: pPP7CP-myc-T2A-tagRFP
FEATURES             Location/Qualifiers
     source          1..5135
                     /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             936..965
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             972..1025
                     /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             1032..1742
                     /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    1863..1984
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1900..1919)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1954..1973
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1991..2446)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2128..2149)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2340..2359
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2473..2577
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2579..2936
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2579..2770
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2602..2622)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2787..2922
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2971..3765
                     /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             2971..3765
                     /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(3025..3044)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3635..3654
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3953..3972)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3997..4044
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4373..4961
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4862..4881
                     /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 cgccaccatg ctagcctcca aaaccatcgt tctttcggtc ggcgaggcta
      601 ctcgcactct gactgagatc cagtccaccg cagaccgtca gatcttcgaa gagaaggtcg
      661 ggcctctggt gggtcggctg cgcctcacgg cttcgctccg tcaaaacgga gccaagaccg
      721 cgtatcgagt caacctaaaa ctggatcagg cggacgtcgt tgattccgga cttccgaaag
      781 tgcgctacac tcaggtatgg tcgcacgacg tgacaatcgt tgcgaatagc accgaggcct
      841 cgcgcaaatc gttgtacgat ttgaccaagt ccctcgtcgc gacctcgcag gtcgaagatc
      901 ttgtcgtcaa ccttgtgccg ctgggccgtg gatccgaaca aaaactcatc tcagaagagg
      961 atctgagatc tgaaggcaga ggctccctcc tgacctgcgg agacgtggaa gagaaccccg
     1021 gaccaagatc cgtgtctaag ggcgaagagc tgattaagga gaacatgcac atgaagctgt
     1081 acatggaggg caccgtgaac aaccaccact tcaagtgcac atccgagggc gaaggcaagc
     1141 cctacgaggg cacccagacc atgagaatca aggtggtcga gggcggccct ctccccttcg
     1201 ccttcgacat cctggctacc agcttcatgt acggcagcag aaccttcatc aaccacaccc
     1261 agggcatccc cgacttcttt aagcagtcct tccctgaggg cttcacatgg gagagagtca
     1321 ccacatacga agacgggggc gtgctgaccg ctacccagga caccagcctc caggacggct
     1381 gcctcatcta caacgtcaag atcagagggg tgaacttccc atccaacggc cctgtgatgc
     1441 agaagaaaac actcggctgg gaggccaaca ccgagatgct gtaccccgct gacggcggcc
     1501 tggaaggcag aagcgacatg gccctgaagc tcgtgggcgg gggccacctg atctgcaact
     1561 tcaagaccac atacagatcc aagaaacccg ctaagaacct caagatgccc ggcgtctact
     1621 atgtggacca cagactggaa agaatcaagg aggccgacaa agagacctac gtcgagcagc
     1681 acgaggtggc tgtggccaga tactgcgacc tccctagcaa actggggcac aaacttaatt
     1741 agcggccgcg actctagatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt
     1801 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg
     1861 ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     1921 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     1981 cttaaggcgt aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     2041 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     2101 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
     2161 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
     2221 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
     2281 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
     2341 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
     2401 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt
     2461 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     2521 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtcct
     2581 gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     2641 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     2701 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     2761 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     2821 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     2881 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc
     2941 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     3001 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     3061 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     3121 accgacctgt ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg
     3181 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     3241 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     3301 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     3361 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     3421 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     3481 ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     3541 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     3601 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     3661 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     3721 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     3781 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     3841 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     3901 agcgcgggga tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac
     3961 ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa
     4021 cgcacggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg
     4081 tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac
     4141 cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc
     4201 ctgccatagc ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat
     4261 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     4321 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     4381 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     4441 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     4501 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     4561 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     4621 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     4681 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     4741 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     4801 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     4861 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     4921 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     4981 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     5041 ctgattctgt ggataaccgt attaccgcca tgcattagtt attaatagta atcaattacg
     5101 gggtcattag ttcatagccc atatatggag ttccg
//
