LOCUS       sequence-312948-        5390 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses LIN28A in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5390)
  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 5390)
  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: pLin28A-myc-T2A-tagRFP
FEATURES             Location/Qualifiers
     source          1..5390
                     /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             1191..1220
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1227..1280
                     /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             1287..1997
                     /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    2118..2239
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2155..2174)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2209..2228
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2246..2701)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2383..2404)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2595..2614
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2728..2832
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2834..3191
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2834..3025
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2857..2877)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3042..3177
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3226..4020
                     /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             3226..4020
                     /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(3280..3299)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3890..3909
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4208..4227)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4252..4299
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4628..5216
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5117..5136
                     /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 ggctccgtgt ccaaccagca gtttgcaggt ggctgcgcca
      601 aggcggcaga agaggcgccc gaggaggcgc cggaggacgc ggcccgggcg gcggacgagc
      661 ctcagctgct gcacggtgcg ggcatctgta agtggttcaa cgtgcgcatg gggttcggct
      721 tcctgtccat gaccgcccgc gccggggtcg cgctcgaccc cccagtggat gtctttgtgc
      781 accagagtaa gctgcacatg gaagggttcc ggagcttgaa ggagggtgag gcagtggagt
      841 tcacctttaa gaagtcagcc aagggtctgg aatccatccg tgtcaccgga cctggtggag
      901 tattctgtat tgggagtgag aggcggccaa aaggaaagag catgcagaag cgcagatcaa
      961 aaggagacag gtgctacaac tgtggaggtc tagatcatca tgccaaggaa tgcaagctgc
     1021 caccccagcc caagaagtgc cacttctgcc agagcatcag ccatatggta gcctcatgtc
     1081 cgctgaaggc ccagcagggc cctagtgcac agggaaagcc aacctacttt cgagaggaag
     1141 aagaagaaat ccacagccct accctgctcc cggaggcaca gaatggatcc gaacaaaaac
     1201 tcatctcaga agaggatctg agatctgaag gcagaggctc cctcctgacc tgcggagacg
     1261 tggaagagaa ccccggacca agatccgtgt ctaagggcga agagctgatt aaggagaaca
     1321 tgcacatgaa gctgtacatg gagggcaccg tgaacaacca ccacttcaag tgcacatccg
     1381 agggcgaagg caagccctac gagggcaccc agaccatgag aatcaaggtg gtcgagggcg
     1441 gccctctccc cttcgccttc gacatcctgg ctaccagctt catgtacggc agcagaacct
     1501 tcatcaacca cacccagggc atccccgact tctttaagca gtccttccct gagggcttca
     1561 catgggagag agtcaccaca tacgaagacg ggggcgtgct gaccgctacc caggacacca
     1621 gcctccagga cggctgcctc atctacaacg tcaagatcag aggggtgaac ttcccatcca
     1681 acggccctgt gatgcagaag aaaacactcg gctgggaggc caacaccgag atgctgtacc
     1741 ccgctgacgg cggcctggaa ggcagaagcg acatggccct gaagctcgtg ggcgggggcc
     1801 acctgatctg caacttcaag accacataca gatccaagaa acccgctaag aacctcaaga
     1861 tgcccggcgt ctactatgtg gaccacagac tggaaagaat caaggaggcc gacaaagaga
     1921 cctacgtcga gcagcacgag gtggctgtgg ccagatactg cgacctccct agcaaactgg
     1981 ggcacaaact taattagcgg ccgcgactct agatcataat cagccatacc acatttgtag
     2041 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga
     2101 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     2161 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     2221 aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg
     2281 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     2341 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     2401 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     2461 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     2521 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     2581 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     2641 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     2701 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     2761 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     2821 aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg
     2881 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     2941 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3001 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     3061 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     3121 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     3181 gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     3241 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     3301 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     3361 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg
     3421 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     3481 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     3541 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     3601 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     3661 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     3721 gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc
     3781 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     3841 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     3901 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     3961 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     4021 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     4081 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     4141 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga
     4201 ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa
     4261 aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc
     4321 agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt
     4381 cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg
     4441 tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag attgatttaa
     4501 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     4561 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     4621 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     4681 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     4741 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     4801 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     4861 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     4921 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     4981 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     5041 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     5101 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     5161 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     5221 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     5281 ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat tagttattaa
     5341 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
//
