LOCUS       sequence-169135-        3966 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway Donor vector containing HRH4 , part of the Target 
            Accelerator Plasmid Collection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3966)
  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 3966)
  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_HRH4_WT
FEATURES             Location/Qualifiers
     source          1..3966
                     /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(1845..1944)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1961..1980)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1962..1980)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1985..2001)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1985..2001)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2432..3223
                     /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      3316..3904
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3805..3824
                     /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 atgccagata ctaatagcac aatcaattta tcactaagca ctcgtgttac
      721 tttagcattt tttatgtcct tagtagcttt tgctataatg ctaggaaatg ctttggtcat
      781 tttagctttt gtggtggaca aaaaccttag acatcgaagt agttattttt ttcttaactt
      841 ggccatctct gacttctttg tgggtgtgat ctccattcct ttgtacatcc ctcacacgct
      901 gttcgaatgg gattttggaa aggaaatctg tgtattttgg ctcactactg actatctgtt
      961 atgtacagca tctgtatata acattgtcct catcagctat gatcgatacc tgtcagtctc
     1021 aaatgctgtg tcttatagaa ctcaacatac tggggtcttg aagattgtta ctctgatggt
     1081 ggccgtttgg gtgctggcct tcttagtgaa tgggccaatg attctagttt cagagtcttg
     1141 gaaggatgaa ggtagtgaat gtgaacctgg atttttttcg gaatggtaca tccttgccat
     1201 cacatcattc ttggaattcg tgatcccagt catcttagtc gcttatttca acatgaatat
     1261 ttattggagc ctgtggaagc gtgatcatct cagtaggtgc caaagccatc ctggactgac
     1321 tgctgtctct tccaacatct gtggacactc attcagaggt agactatctt caaggagatc
     1381 tctttctgca tcgacagaag ttcctgcatc ctttcattca gagagacaga ggagaaagag
     1441 tagtctcatg ttttcctcaa gaaccaagat gaatagcaat acaattgctt ccaaaatggg
     1501 ttccttctcc caatcagatt ctgtagctct tcaccaaagg gaacatgttg aactgcttag
     1561 agccaggaga ttagccaagt cactggccat tctcttaggg gtttttgctg tttgctgggc
     1621 tccatattct ctgttcacaa ttgtcctttc attttattcc tcagcaacag gtcctaaatc
     1681 agtttggtat agaattgcat tttggcttca gtggttcaat tcctttgtca atcctctttt
     1741 gtatccattg tgtcacaagc gctttcaaaa ggctttcttg aaaatatttt gtataaaaaa
     1801 gcaacctcta ccatcacaac acagtcggtc agtatcttct tgattgccaa ctttcttgta
     1861 caaagttggc attataagaa agcattgctt atcaatttgt tgcaacgaac aggtcactat
     1921 cagtcaaaat aaaatcatta tttgccatcc agctgatatc ccctatagtg agtcgtatta
     1981 catggtcata gctgtttcct ggcagctctg gcccgtgtct caaaatctct gatgttacat
     2041 tgcacaagat aaaaatatat catcatgcct cctctagacc agccaggaca gaaatgcctc
     2101 gacttcgctg ctacccaagg ttgccgggtg acgcacaccg tggaaacgga tgaaggcacg
     2161 aacccagtgg acataagcct gttcggttcg taagctgtaa tgcaagtagc gtatgcgctc
     2221 acgcaactgg tccagaacct tgaccgaacg cagcggtggt aacggcgcag tggcggtttt
     2281 catggcttgt tatgactgtt tttttggggt acagtctatg cctcgggcat ccaagcagca
     2341 agcgcgttac gccgtgggtc gatgtttgat gttatggagc agcaacgatg ttacgcagca
     2401 gggcagtcgc cctaaaacaa agttaaacat tatgagggaa gcggtgatcg ccgaagtatc
     2461 gactcaacta tcagaggtag ttggcgtcat cgagcgccat ctcgaaccga cgttgctggc
     2521 cgtacatttg tacggctccg cagtggatgg cggcctgaag ccacacagtg atattgattt
     2581 gctggttacg gtgaccgtaa ggcttgatga aacaacgcgg cgagctttga tcaacgacct
     2641 tttggaaact tcggcttccc ctggagagag cgagattctc cgcgctgtag aagtcaccat
     2701 tgttgtgcac gacgacatca ttccgtggcg ttatccagct aagcgcgaac tgcaatttgg
     2761 agaatggcag cgcaatgaca ttcttgcagg tatcttcgag ccagccacga tcgacattga
     2821 tctggctatc ttgctgacaa aagcaagaga acatagcgtt gccttggtag gtccagcggc
     2881 ggaggaactc tttgatccgg ttcctgaaca ggatctattt gaggcgctaa atgaaacctt
     2941 aacgctatgg aactcgccgc ccgactgggc tggcgatgag cgaaatgtag tgcttacgtt
     3001 gtcccgcatt tggtacagcg cagtaaccgg caaaatcgcg ccgaaggatg tcgctgccga
     3061 ctgggcaatg gagcgcctgc cggcccagta tcagcccgtc atacttgaag ctagacaggc
     3121 ttatcttgga caagaagaag atcgcttggc ctcgcgcgca gatcagttgg aagaatttgt
     3181 ccactacgtg aaaggcgaga tcaccaaggt agtcggcaaa taaccctcga gccacccatg
     3241 accaaaatcc cttaacgtga gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa
     3301 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     3361 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     3421 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     3481 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     3541 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     3601 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     3661 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     3721 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     3781 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     3841 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     3901 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     3961 catgtt
//
