LOCUS       sequence-374391-        2199 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Triple reporter plasmid in pELPS with eDHFR (PET reporter gene) 
            fused to yellow fluorescent protein (YFP) and a T2A cleavage site 
            followed by Renilla luciferase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 2199)
  AUTHORS   Sellmyer MA, Richman SA, Lohith K, Hou C, Weng CC, Mach RH, O'Connor
            RS, Milone MC, Farwell MD
  TITLE     Imaging CAR T Cell Trafficking with eDHFR as a PET Reporter Gene.
  JOURNAL   Mol Ther. 2020 Jan 8;28(1):42-51. doi: 10.1016/j.ymthe.2019.10.007. 
            Epub 2019 Oct 15.
  PUBMED    31668558
REFERENCE   2  (bases 1 to 2199)
  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: 
            "10.1016/j.ymthe.2019.10.007"; journalName: "Mol Ther"; date: 
            "2020-01-8- 8"; volume: "28"; issue: "1"; pages: "42-51"
COMMENT     Sequence Label: pELPS-eDHFR-YFP-T2A-Renilla
FEATURES             Location/Qualifiers
     source          1..2199
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             join(484..486,487..1197)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(526..547)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(787..806)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1134..1155
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1207..1260
                     /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             1267..2199
                     /codon_start=1
                     /product="luciferase from the anthozoan coelenterate 
                     Renilla reniformis (sea pansy)"
                     /label=Rluc
                     /translation="TSKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAENA
                     VIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAWF
                     ELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDIA
                     LIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPREI
                     PLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKVK
                     GLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
ORIGIN
        1 atgataagtt tgattgctgc tctggctgtg gaccgggtaa tcggtatgga aaacgccatg
       61 ccctggaacc tgcctgccga tttggcttgg ttcaagcgca ataccctgaa caaaccagta
      121 atcatgggaa ggcatacatg ggaaagcatt ggaagaccac ttcccggtag aaagaatatt
      181 atcctgtcta gccagcccgg cacggatgat agggtgacat gggtaaagag cgtcgatgag
      241 gcgattgcgg cgtgtggtga cgtgccggaa attatggtta tcggaggcgg cagggtctac
      301 gaacagttcc tgccgaaggc acagaagctg tacctcaccc acatcgatgc agaggtggaa
      361 ggagacacgc actttccaga ttacgagcct gatgactggg agagtgtttt tagcgaattc
      421 catgacgcag acgcccaaaa ctctcactcc tactgctttg agattctcga acgaagggca
      481 tgcgtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
      541 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
      601 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
      661 ctcgtgacca ccttcggcta cggcctgcag tgcttcgccc gctaccccga ccacatgaag
      721 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
      781 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
      841 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
      901 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
      961 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
     1021 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     1081 tacctgagct accagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc
     1141 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagggc
     1201 agcggagagg gcagaggaag tcttctaaca tgcggtgacg tggaggagaa tcccggccct
     1261 gctagcactt cgaaagttta tgatccagaa caaaggaaac ggatgataac tggtccgcag
     1321 tggtgggcca gatgtaaaca aatgaatgtt cttgattcat ttattaatta ttatgattca
     1381 gaaaaacatg cagaaaatgc tgttattttt ttacatggta acgcggcctc ttcttattta
     1441 tggcgacatg ttgtgccaca tattgagcca gtagcgcggt gtattatacc agaccttatt
     1501 ggtatgggca aatcaggcaa atctggtaat ggttcttata ggttacttga tcattacaaa
     1561 tatcttactg catggtttga acttcttaat ttaccaaaga agatcatttt tgtcggccat
     1621 gattggggtg cttgtttggc atttcattat agctatgagc atcaagataa gatcaaagca
     1681 atagttcacg ctgaaagtgt agtagatgtg attgaatcat gggatgaatg gcctgatatt
     1741 gaagaagata ttgcgttgat caaatctgaa gaaggagaaa aaatggtttt ggagaataac
     1801 ttcttcgtgg aaaccatgtt gccatcaaaa atcatgagaa agttagaacc agaagaattt
     1861 gcagcatatc ttgaaccatt caaagagaaa ggtgaagttc gtcgtccaac attatcatgg
     1921 cctcgtgaaa tcccgttagt aaaaggtggt aaacctgacg ttgtacaaat tgttaggaat
     1981 tataatgctt atctacgtgc aagtgatgat ttaccaaaaa tgtttattga atcggaccca
     2041 ggattctttt ccaatgctat tgttgaaggt gccaagaagt ttcctaatac tgaatttgtc
     2101 aaagtaaaag gtcttcattt ttcgcaagaa gatgcacctg atgaaatggg aaaatatatc
     2161 aaatcgttcg ttgagcgagt tctcaaaaat gaacaataa
//
