LOCUS       sequence-169313-        3315 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway Donor vector containing ASCL4 , part of the Target 
            Accelerator Plasmid Collection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3315)
  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 3315)
  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_ASCL4_WT
FEATURES             Location/Qualifiers
     source          1..3315
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1194..1293)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1310..1329)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1311..1329)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1334..1350)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1334..1350)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1781..2572
                     /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      2665..3253
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3154..3173
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agttggcacc atgatggaga cgcgtaaacc ggcggaacgg ctggccttgc catactcgct
      721 gcgcaccgcg cccctgggcg ttccggggac cctgcccgga ctcccgcgga gggaccccct
      781 cagggtcgcc ctgcgtctgg acgccgcgtg ctgggagtgg gcgcgcagcg gctgcgcacg
      841 gggatggcag tacttgcccg tgccgctgga cagcgccttc gagcccgcct tcctccgcaa
      901 gcgcaacgag cgcgagcggc agcgggtgcg ctgcgtgaac gagggctatg cgcgcctccg
      961 agaccacctg ccccgggagc tggcagacaa gcgcctcagc aaagtggaga cgctccgcgc
     1021 tgccatcgac tacatcaagc acctgcagga gctgctggag cgccaggcct gggggctcga
     1081 gggcgcggcc ggcgccgtcc cccagcgcag ggcggaatgc aacagcgacg gggagtccaa
     1141 ggcctcttcg gcgccttcgc ccagcagcga gcccgaggag gggggcagct agttgccaac
     1201 tttcttgtac aaagttggca ttataagaaa gcattgctta tcaatttgtt gcaacgaaca
     1261 ggtcactatc agtcaaaata aaatcattat ttgccatcca gctgatatcc cctatagtga
     1321 gtcgtattac atggtcatag ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg
     1381 atgttacatt gcacaagata aaaatatatc atcatgcctc ctctagacca gccaggacag
     1441 aaatgcctcg acttcgctgc tacccaaggt tgccgggtga cgcacaccgt ggaaacggat
     1501 gaaggcacga acccagtgga cataagcctg ttcggttcgt aagctgtaat gcaagtagcg
     1561 tatgcgctca cgcaactggt ccagaacctt gaccgaacgc agcggtggta acggcgcagt
     1621 ggcggttttc atggcttgtt atgactgttt ttttggggta cagtctatgc ctcgggcatc
     1681 caagcagcaa gcgcgttacg ccgtgggtcg atgtttgatg ttatggagca gcaacgatgt
     1741 tacgcagcag ggcagtcgcc ctaaaacaaa gttaaacatt atgagggaag cggtgatcgc
     1801 cgaagtatcg actcaactat cagaggtagt tggcgtcatc gagcgccatc tcgaaccgac
     1861 gttgctggcc gtacatttgt acggctccgc agtggatggc ggcctgaagc cacacagtga
     1921 tattgatttg ctggttacgg tgaccgtaag gcttgatgaa acaacgcggc gagctttgat
     1981 caacgacctt ttggaaactt cggcttcccc tggagagagc gagattctcc gcgctgtaga
     2041 agtcaccatt gttgtgcacg acgacatcat tccgtggcgt tatccagcta agcgcgaact
     2101 gcaatttgga gaatggcagc gcaatgacat tcttgcaggt atcttcgagc cagccacgat
     2161 cgacattgat ctggctatct tgctgacaaa agcaagagaa catagcgttg ccttggtagg
     2221 tccagcggcg gaggaactct ttgatccggt tcctgaacag gatctatttg aggcgctaaa
     2281 tgaaacctta acgctatgga actcgccgcc cgactgggct ggcgatgagc gaaatgtagt
     2341 gcttacgttg tcccgcattt ggtacagcgc agtaaccggc aaaatcgcgc cgaaggatgt
     2401 cgctgccgac tgggcaatgg agcgcctgcc ggcccagtat cagcccgtca tacttgaagc
     2461 tagacaggct tatcttggac aagaagaaga tcgcttggcc tcgcgcgcag atcagttgga
     2521 agaatttgtc cactacgtga aaggcgagat caccaaggta gtcggcaaat aaccctcgag
     2581 ccacccatga ccaaaatccc ttaacgtgag ttacgcgtcg ttccactgag cgtcagaccc
     2641 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     2701 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     2761 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     2821 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     2881 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     2941 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     3001 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     3061 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     3121 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     3181 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     3241 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     3301 ttttgctcac atgtt
//
