LOCUS       sequence-312939-        5609 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses U1A in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5609)
  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 5609)
  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: pU1A_full-myc-T2A-tagRFP
FEATURES             Location/Qualifiers
     source          1..5609
                     /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      553..562
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1410..1439
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1446..1499
                     /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             1506..2216
                     /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    2337..2458
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2374..2393)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2428..2447
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2465..2920)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2602..2623)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2814..2833
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2947..3051
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3053..3410
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3053..3244
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3076..3096)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3261..3396
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3445..4239
                     /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             3445..4239
                     /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(3499..3518)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4109..4128
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4427..4446)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4471..4518
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4847..5435
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5336..5355
                     /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 caccatggcg gcagttcccg agacccgccc taaccacact atttatatca
      601 acaacctcaa tgagaagatc aagaaggatg agctaaaaaa gtccctgtac gccatcttct
      661 cccagtttgg ccagatcctg gatatcctgg tatcacggag cctgaagatg aggggccagg
      721 cctttgtcat cttcaaggag gtcagcagcg ccaccaacgc cctgcgctcc atgcagggtt
      781 tccctttcta tgacaaacct atgcgtatcc agtatgccaa gaccgactca gatatcattg
      841 ccaagatgaa aggcaccttc gtggagcggg accgcaagcg ggagaagagg aagcccaaga
      901 gccaggagac cccggccacc aagaaggctg tgcaaggcgg gggagccacc cccgtggtgg
      961 gggctgtcca ggggcctgtc ccgggcatgc cgccgatgac tcaggcgccc cgcattatgc
     1021 accacatgcc gggccagccg ccctacatgc cgccccctgg tatgatcccc ccgccaggcc
     1081 ttgcacctgg ccagatccca ccaggggcca tgcccccgca gcagcttatg ccaggacaga
     1141 tgccccctgc ccagcctctt tctgagaatc caccgaatca catcttgttc ctcaccaacc
     1201 tgccagagga gaccaacgag ctcatgctgt ccatgctttt caatcagttc cctggcttca
     1261 aggaggtccg tctggtaccc gggcggcatg acatcgcctt cgtggagttt gacaatgagg
     1321 tacaggcagg ggcagctcgc gatgccctgc agggctttaa gatcacgcag aacaacgcca
     1381 tgaagatctc ctttgccaag aagggatccg aacaaaaact catctcagaa gaggatctga
     1441 gatctgaagg cagaggctcc ctcctgacct gcggagacgt ggaagagaac cccggaccaa
     1501 gatccgtgtc taagggcgaa gagctgatta aggagaacat gcacatgaag ctgtacatgg
     1561 agggcaccgt gaacaaccac cacttcaagt gcacatccga gggcgaaggc aagccctacg
     1621 agggcaccca gaccatgaga atcaaggtgg tcgagggcgg ccctctcccc ttcgccttcg
     1681 acatcctggc taccagcttc atgtacggca gcagaacctt catcaaccac acccagggca
     1741 tccccgactt ctttaagcag tccttccctg agggcttcac atgggagaga gtcaccacat
     1801 acgaagacgg gggcgtgctg accgctaccc aggacaccag cctccaggac ggctgcctca
     1861 tctacaacgt caagatcaga ggggtgaact tcccatccaa cggccctgtg atgcagaaga
     1921 aaacactcgg ctgggaggcc aacaccgaga tgctgtaccc cgctgacggc ggcctggaag
     1981 gcagaagcga catggccctg aagctcgtgg gcgggggcca cctgatctgc aacttcaaga
     2041 ccacatacag atccaagaaa cccgctaaga acctcaagat gcccggcgtc tactatgtgg
     2101 accacagact ggaaagaatc aaggaggccg acaaagagac ctacgtcgag cagcacgagg
     2161 tggctgtggc cagatactgc gacctcccta gcaaactggg gcacaaactt aattagcggc
     2221 cgcgactcta gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa
     2281 acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact
     2341 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     2401 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaag
     2461 gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     2521 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
     2581 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
     2641 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
     2701 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
     2761 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
     2821 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
     2881 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga
     2941 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     3001 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tcctgaggcg
     3061 gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     3121 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     3181 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     3241 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     3301 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
     3361 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa
     3421 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     3481 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     3541 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     3601 ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg
     3661 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     3721 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     3781 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     3841 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     3901 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     3961 aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     4021 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     4081 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     4141 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     4201 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     4261 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     4321 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     4381 gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa acacggaagg
     4441 agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg
     4501 gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata
     4561 ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc
     4621 ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca
     4681 tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     4741 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     4801 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     4861 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     4921 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     4981 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     5041 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     5101 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     5161 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     5221 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     5281 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     5341 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     5401 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     5461 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     5521 ctgtggataa ccgtattacc gccatgcatt agttattaat agtaatcaat tacggggtca
     5581 ttagttcata gcccatatat ggagttccg
//
