LOCUS       sequence-313673-        5060 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses truncated U1A in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5060)
  AUTHORS   Kawasaki S, Fujita Y, Nagaike T, Tomita K, Saito H
  TITLE     Synthetic mRNA devices that detect endogenous proteins and 
            distinguish mammalian cells.
  JOURNAL   Nucleic Acids Res. 2017 May 19. doi: 10.1093/nar/gkx298.
  PUBMED    28525643
REFERENCE   2  (bases 1 to 5060)
  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: "Nucleic 
            Acids Res. 2017 May 19. doi: 10.1093/nar/gkx298."
COMMENT     Sequence Label: pU1A-myc-T2A-tagRFP
FEATURES             Location/Qualifiers
     source          1..5060
                     /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             861..890
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             897..950
                     /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             957..1667
                     /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    1788..1909
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1825..1844)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1879..1898
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1916..2371)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2053..2074)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2265..2284
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2398..2502
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2504..2861
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2504..2695
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2527..2547)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2712..2847
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2896..3690
                     /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             2896..3690
                     /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(2950..2969)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3560..3579
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3878..3897)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3922..3969
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4298..4886
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4787..4806
                     /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 caccatggga tccgcagttc ccgagacccg ccctaaccac actatttata
      601 tcaacaacct caatgagaag atcaagaagg atgagctaaa aaagtccctg tacgccatct
      661 tctcccagtt tggccagatc ctggatatcc tggtatcacg gagcctgaag atgaggggcc
      721 aggcctttgt catcttcaag gaggtcagca gcgccaccaa cgccctgcgc tccatgcagg
      781 gtttcccttt ctatgacaaa cctatgcgta tccagtatgc caagaccgac tcagatatca
      841 ttgccaagat gaaaagatcc gaacaaaaac tcatctcaga agaggatctg agatctgaag
      901 gcagaggctc cctcctgacc tgcggagacg tggaagagaa ccccggacca agatccgtgt
      961 ctaagggcga agagctgatt aaggagaaca tgcacatgaa gctgtacatg gagggcaccg
     1021 tgaacaacca ccacttcaag tgcacatccg agggcgaagg caagccctac gagggcaccc
     1081 agaccatgag aatcaaggtg gtcgagggcg gccctctccc cttcgccttc gacatcctgg
     1141 ctaccagctt catgtacggc agcagaacct tcatcaacca cacccagggc atccccgact
     1201 tctttaagca gtccttccct gagggcttca catgggagag agtcaccaca tacgaagacg
     1261 ggggcgtgct gaccgctacc caggacacca gcctccagga cggctgcctc atctacaacg
     1321 tcaagatcag aggggtgaac ttcccatcca acggccctgt gatgcagaag aaaacactcg
     1381 gctgggaggc caacaccgag atgctgtacc ccgctgacgg cggcctggaa ggcagaagcg
     1441 acatggccct gaagctcgtg ggcgggggcc acctgatctg caacttcaag accacataca
     1501 gatccaagaa acccgctaag aacctcaaga tgcccggcgt ctactatgtg gaccacagac
     1561 tggaaagaat caaggaggcc gacaaagaga cctacgtcga gcagcacgag gtggctgtgg
     1621 ccagatactg cgacctccct agcaaactgg ggcacaaact taattagcgg ccgcgactct
     1681 agatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
     1741 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
     1801 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     1861 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt
     1921 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     1981 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     2041 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     2101 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     2161 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     2221 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     2281 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     2341 gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc
     2401 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     2461 taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc
     2521 agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     2581 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     2641 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     2701 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     2761 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     2821 agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga
     2881 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     2941 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     3001 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     3061 gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     3121 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     3181 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     3241 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     3301 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     3361 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     3421 gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     3481 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     3541 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     3601 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     3661 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     3721 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     3781 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     3841 tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac
     3901 cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt
     3961 cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga
     4021 gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt
     4081 cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag
     4141 gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     4201 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     4261 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     4321 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     4381 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     4441 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     4501 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     4561 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     4621 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     4681 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     4741 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     4801 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     4861 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     4921 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     4981 accgtattac cgccatgcat tagttattaa tagtaatcaa ttacggggtc attagttcat
     5041 agcccatata tggagttccg
//
