LOCUS       sequence-168523-        3492 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway Donor vector containing TNF , part of the Target Accelerator
            Plasmid Collection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3492)
  AUTHORS   Kim E, Ilic N, Shrestha Y, Zou L, Kamburov A, Zhu C, Yang X, Lubonja
            R, Tran N, Nguyen C, Lawrence MS, Piccioni F, Bagul M, Doench JG, 
            Chouinard CR, Wu X, Hogstrom L, Natoli T, Tamayo P, Horn H, Corsello
            SM, Lage K, Root DE, Subramanian A, Golub TR, Getz G, Boehm JS, Hahn
            WC
  TITLE     Systematic Functional Interrogation of Rare Cancer Variants 
            Identifies Oncogenic Alleles.
  JOURNAL   Cancer Discov. 2016 Jul;6(7):714-26. doi: 
            10.1158/2159-8290.CD-16-0160. Epub 2016 May 4.
  PUBMED    27147599
REFERENCE   2  (bases 1 to 3492)
  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: 
            "10.1158/2159-8290.CD-16-0160"; journalName: "Cancer Discov"; date: 
            "2016-07"; volume: "6"; issue: "7"; pages: "714-26"
COMMENT     Sequence Label: pDONR223_TNF_p.S157R
FEATURES             Location/Qualifiers
     source          1..3492
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     43..65
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     57..74
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     58..74
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    90..189
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(892..991)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1008..1027)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1009..1027)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1032..1048)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1032..1048)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1479..2270
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      2363..2951
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2852..2871
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3105..3122
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3281..3308)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3400..3486)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ggcagttccc tactctcgcg ttaacgctag catggatgtt ttcccagtca cgacgttgta
       61 aaacgacggc cagtcttaag ctcgggcccc aaataatgat tttattttga ctgatagtga
      121 cctgttcgtt gcaacaaatt gatgagcaat gcttttttat aatgccaact ttgtacaaaa
      181 aagttggcat gagcactgaa agcatgatcc gggacgtgga gctggccgag gaggcgctcc
      241 ccaagaagac aggggggccc cagggctcca ggcggtgctt gttcctcagc ctcttctcct
      301 tcctgatcgt ggcaggcgcc accacgctct tctgcctgct gcactttgga gtgatcggcc
      361 cccagaggga agagttcccc agggacctct ctctaatcag ccctctggcc caggcagtca
      421 gatcatcttc tcgaaccccg agtgacaagc ctgtagccca tgttgtagca aaccctcaag
      481 ctgaggggca gctccagtgg ctgaaccgcc gggccaatgc cctcctggcc aatggcgtgg
      541 agctgagaga taaccagctg gtggtgccat cagagggcct gtacctcatc tactcccagg
      601 tcctcttcaa gggccaaggc tgcccctcca cccatgtgct cctcacccac accatcaggc
      661 gcatcgccgt ctcctaccag accaaggtca acctcctctc tgccatcaag agcccctgcc
      721 agagggagac cccagagggg gctgaggcca agccctggta tgagcccatc tatctgggag
      781 gggtcttcca gctggagaag ggtgaccgac tcagcgctga gatcaatcgg cccgactatc
      841 tcgactttgc cgagtctggg caggtctact ttgggatcat tgccctgtga ttgccaactt
      901 tcttgtacaa agttggcatt ataagaaagc attgcttatc aatttgttgc aacgaacagg
      961 tcactatcag tcaaaataaa atcattattt gccatccagc tgatatcccc tatagtgagt
     1021 cgtattacat ggtcatagct gtttcctggc agctctggcc cgtgtctcaa aatctctgat
     1081 gttacattgc acaagataaa aatatatcat catgcctcct ctagaccagc caggacagaa
     1141 atgcctcgac ttcgctgcta cccaaggttg ccgggtgacg cacaccgtgg aaacggatga
     1201 aggcacgaac ccagtggaca taagcctgtt cggttcgtaa gctgtaatgc aagtagcgta
     1261 tgcgctcacg caactggtcc agaaccttga ccgaacgcag cggtggtaac ggcgcagtgg
     1321 cggttttcat ggcttgttat gactgttttt ttggggtaca gtctatgcct cgggcatcca
     1381 agcagcaagc gcgttacgcc gtgggtcgat gtttgatgtt atggagcagc aacgatgtta
     1441 cgcagcaggg cagtcgccct aaaacaaagt taaacattat gagggaagcg gtgatcgccg
     1501 aagtatcgac tcaactatca gaggtagttg gcgtcatcga gcgccatctc gaaccgacgt
     1561 tgctggccgt acatttgtac ggctccgcag tggatggcgg cctgaagcca cacagtgata
     1621 ttgatttgct ggttacggtg accgtaaggc ttgatgaaac aacgcggcga gctttgatca
     1681 acgacctttt ggaaacttcg gcttcccctg gagagagcga gattctccgc gctgtagaag
     1741 tcaccattgt tgtgcacgac gacatcattc cgtggcgtta tccagctaag cgcgaactgc
     1801 aatttggaga atggcagcgc aatgacattc ttgcaggtat cttcgagcca gccacgatcg
     1861 acattgatct ggctatcttg ctgacaaaag caagagaaca tagcgttgcc ttggtaggtc
     1921 cagcggcgga ggaactcttt gatccggttc ctgaacagga tctatttgag gcgctaaatg
     1981 aaaccttaac gctatggaac tcgccgcccg actgggctgg cgatgagcga aatgtagtgc
     2041 ttacgttgtc ccgcatttgg tacagcgcag taaccggcaa aatcgcgccg aaggatgtcg
     2101 ctgccgactg ggcaatggag cgcctgccgg cccagtatca gcccgtcata cttgaagcta
     2161 gacaggctta tcttggacaa gaagaagatc gcttggcctc gcgcgcagat cagttggaag
     2221 aatttgtcca ctacgtgaaa ggcgagatca ccaaggtagt cggcaaataa ccctcgagcc
     2281 acccatgacc aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg
     2341 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     2401 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     2461 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     2521 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     2581 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     2641 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     2701 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     2761 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     2821 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     2881 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     2941 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     3001 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     3061 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     3121 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     3181 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     3241 atacgcgtac cgctagccag gaagagtttg tagaaacgca aaaaggccat ccgtcaggat
     3301 ggccttctgc ttagtttgat gcctggcagt ttatggcggg cgtcctgccc gccaccctcc
     3361 gggccgttgc ttcacaacgt tcaaatccgc tcccggcgga tttgtcctac tcaggagagc
     3421 gttcaccgac aaacaacaga taaaacgaaa ggcccagtct tccgactgag cctttcgttt
     3481 tatttgatgc ct
//
