LOCUS       sequence-269306-        1212 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  mMoClo TUPV4, with EF1alpha promoter and EBFP2-P2A-BlastR in the 
            MCS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1212)
  AUTHORS   Donahue PS, Draut JW, Muldoon JJ, Edelstein HI, Bagheri N, Leonard 
            JN
  TITLE     The COMET toolkit for composing customizable genetic programs in 
            mammalian cells.
  JOURNAL   Nat Commun. 2020 Feb 7;11(1):779. doi: 10.1038/s41467-019-14147-5.
  PUBMED    32034124
REFERENCE   2  (bases 1 to 1212)
  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: "Nat 
            Commun."; date: "2020-02-7"; volume: "11(1):779. doi"; pages: " 
            10.1038/s41467-019-14147-5"
COMMENT     Sequence Label: pJM453 EF1alpha EBFP2-P2A-BlastR (TUPV4)
FEATURES             Location/Qualifiers
     source          1..1212
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             join(1..3,4..6,7..717)
                     /codon_start=1
                     /product="enhanced blue variant of GFP (Ai et al., 2007)"
                     /label=EBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLSHGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGVDFKEDGNILGHKLEYNFNSHNIYIMAVKQKNGIK
                     VNFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDSHYLSTQSVLSKDPNEKRDHMVLL
                     EFRTAAGITLGMDELYK"
     primer_bind     complement(46..67)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             745..801
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             814..1212
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
ORIGIN
        1 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
       61 ggcgacgtaa acggccacaa gttcagcgtg aggggcgagg gcgagggcga tgccaccaac
      121 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
      181 ctcgtgacca ccctgagcca cggcgtgcag tgcttcgccc gctaccccga ccacatgaag
      241 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
      301 ttcaaggacg acggcaccta caagacccgc gccgaggtga agttcgaggg cgacacccta
      361 gtgaaccgca tcgagctgaa gggcgtcgac ttcaaggagg acggcaacat cctggggcac
      421 aagctggagt acaacttcaa cagccacaac atctatatca tggccgtcaa gcagaagaac
      481 ggcatcaagg tgaacttcaa gatccgccac aacgtggagg acggcagcgt gcagctcgcc
      541 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacagccac
      601 tacctgagca cccagtccgt gctgagcaaa gaccccaacg agaagcgcga tcacatggtc
      661 ctgctggagt tccgcaccgc cgccgggatc actctcggca tggacgagct gtacaaggaa
      721 ttcggtagca agcttggtag tggagcaact aacttttccc tgctcaaaca agccggggat
      781 gtagaggaga accccggccc cgcccccctt aagatggcca agcctttgtc tcaagaagaa
      841 tccaccctca ttgaaagagc aacggctaca atcaacagca tccccatctc tgaggactac
      901 agcgtcgcca gcgcagctct ctctagcgac ggccgcatct tcactggtgt caatgtatat
      961 cattttactg ggggaccttg tgcagaactc gtggtgctgg gcactgctgc tgctgcggca
     1021 gctggcaacc tgacttgtat cgtcgcgatc ggaaatgaga acaggggcat cttgagcccc
     1081 tgcggacggt gccgacaggt gcttctcgat ctgcatcctg ggatcaaagc catagtgaag
     1141 gacagtgatg gacagccgac ggcagttggg attcgtgaat tgctgccctc tggttatgtg
     1201 tgggagggct aa
//
