LOCUS       sequence-262517-        1374 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  All-in-one Sleeping Beauty vector for Expression for Tasmanian devil
            TNFRSF9-mAzurite fusion protein. Multiple cloning sites to swap 
            genes-of-interest (i.e. TNFRSF9)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1374)
  AUTHORS   Flies AS, Darby JM, Lennard PR, Murphy PR, Ong CEB, Pinfold TL, De 
            Luca A, Lyons AB, Woods GM, Patchett AL
  TITLE     A novel system to map protein interactions reveals evolutionarily 
            conserved immune evasion pathways on transmissible cancers.
  JOURNAL   Sci Adv. 2020 Jul 1;6(27). pii: 6/27/eaba5031. doi: 
            10.1126/sciadv.aba5031. Print 2020 Jul.
  PUBMED    32937435
REFERENCE   2  (bases 1 to 1374)
  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: "Sci Adv."; 
            date: "2020-07-1"; volume: "6(27). pii: 6/27/eaba5031. doi"; pages: 
            " 10.1126/sciadv.aba5031. Print 2020 Jul"
COMMENT     Sequence Label: pAF160
FEATURES             Location/Qualifiers
     source          1..1374
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             580..600
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             634..651
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             join(658..660,661..1374)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLSHGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNIYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FRTAAGITHGMDELYK"
     primer_bind     complement(700..721)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(961..980)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
ORIGIN
        1 atggtaaaca gctacaatag actggaggcg atatcaactc ttgttatact actctgggta
       61 actctggtcc agggttcctg tggcagctgt ccagctggta ctttctggaa tggtacagat
      121 tgtggaaaaa acaaaaatca agcttgcatc ccatgtcccc caaatagttt ctctagtaca
      181 gctggcgaac agaggacatg taacatatgc aggagatgtg aaggtatttt tagaacaaag
      241 agagaatgta ctccaacaag taatacagag tgtgattgta tgtctgggta tcattgttcc
      301 ggagtaggat gtaataggtg tcaaccagat tgcaagcaag gtcaagaact tctaggaaaa
      361 ggttgtagag actgtagctt tggtaccttt aatgatcaga cacgggggaa ctgcaaacgc
      421 tggacaaact gctctttgaa tggactagtt gtgcttgtaa atggaaccaa taaaagagat
      481 gtgatatgtg gatccaccca aacagttgcc actctggaca caatctccat gacaattcca
      541 gcttctctcg tgcacattga taagaaccat cagcccgggg agaaccttta cttccaggga
      601 tccgctggct ccgctgctgg ttctggcgaa ttccatcatc accatcacca tgtcgacgtg
      661 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
      721 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
      781 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
      841 accaccctga gccacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac
      901 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
      961 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
     1021 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     1081 gagtacaact tcaacagcca caacatctat atcatggccg acaagcagaa gaacggcatc
     1141 aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac
     1201 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     1261 agcacccagt ccaagctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
     1321 gagttccgca ccgccgccgg gatcactcac ggcatggacg agctgtacaa gtag
//
