LOCUS       sequence-162795-        3732 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway Donor vector containing SPP1, used as an over expression 
            control for gene expression analysis. Part of the Target Accelerator
            Plasmid Collection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3732)
  AUTHORS   Berger AH, Brooks AN, Wu X, Shrestha Y, Chouinard C, Piccioni F, 
            Bagul M, Kamburov A, Imielinski M, Hogstrom L, Zhu C, Yang X, Pantel
            S, Sakai R, Watson J, Kaplan N, Campbell JD, Singh S, Root DE, 
            Narayan R, Natoli T, Lahr DL, Tirosh I, Tamayo P, Getz G, Wong B, 
            Doench J, Subramanian A, Golub TR, Meyerson M, Boehm JS
  TITLE     High-throughput Phenotyping of Lung Cancer Somatic Mutations.
  JOURNAL   Cancer Cell. 2016 Aug 8;30(2):214-28. doi: 
            10.1016/j.ccell.2016.06.022. Epub 2016 Jul 28.
  PUBMED    27478040
REFERENCE   2  (bases 1 to 3732)
  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: 
            "10.1016/j.ccell.2016.06.022"; journalName: "Cancer Cell"; date: 
            "2016-08-8- 8"; volume: "30"; issue: "2"; pages: "214-28"
COMMENT     Sequence Label: pDONR223_SPP1_WT_V5
FEATURES             Location/Qualifiers
     source          1..3732
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     121..137
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     121..137
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     142..161
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        142..160
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    178..277
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1220..1319)
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1335..1352)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1335..1351)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1344..1366)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     terminator      1415..1501
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1593..1620
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(1779..1796)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1950..2538)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2030..2049)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2631..3422)
                     /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"
ORIGIN
        1 accttgggta gcagcgaagt cgaggcattt ctgtcctggc tggtctagag gaggcatgat
       61 gatatatttt tatcttgtgc aatgtaacat cagagatttt gagacacggg ccagagctgc
      121 caggaaacag ctatgaccat gtaatacgac tcactatagg ggatatcagc tggatggcaa
      181 ataatgattt tattttgact gatagtgacc tgttcgttgc aacaaattga taagcaatgc
      241 tttcttataa tgccaacttt gtacaagaaa gttgggtaat tgacctcaga agatgcacta
      301 tctaattcat gagaaatacg aaatttcagg tgtttatctt cttccttact tttggggtct
      361 acaaccagca tatcttcatg gctgtgaaat tcatggctgt ggaattcacg gctgactttg
      421 gaaagttcct gactatcaat cacatcggaa tgctcattgc tctcatcatt ggctttccgc
      481 ttatataatc tggactgctt gtggctgtgg gtttcagcac tctggtcatc cagctgactc
      541 gtttcataac tgtccttccc acggctgtcc caatcagaag gcgcgttcag gtcctgggca
      601 acggggatgg ccttgtatgc accattcaac tcctcgcttt ccatgtgtga ggtgatgtcc
      661 tcgtctgtag catcagggta ctggatgtca ggtctgcgaa acttcttaga ttttgacctc
      721 agtccataaa ccacactatc acctcggcca tcatatgtgt ctactgtggg gacaactgga
      781 gtgaaaactt cggttgctgg caggtccgtg ggaaaatcag tgaccagttc atcagattca
      841 tcagaatggt gagactcatc agactggtga gaatcatcag tgtcatctac atcatcagag
      901 tcgttcgagt caatggagtc ctggctgtcc acatggtcat catcatcttc atcatccata
      961 tcatccatgt ggtcatggct ttcgttggac ttacttggaa gggtctcttg tttaaagtca
     1021 ttggtttctt cagaggacac agcattctgt ggggctagga gattctgctt ctgagatggg
     1081 tcagggttta gccatgtggc cacagcatct gggtatttgt tgtaaagctg cttttcctca
     1141 gaacttccag aatcagcctg tttaactggt atggcacagg tgatgcctag gaggcaaaag
     1201 caaatcactg caattctcat gccaactttt ttgtacaaag ttggcattat aaaaaagcat
     1261 tgctcatcaa tttgttgcaa cgaacaggtc actatcagtc aaaataaaat cattatttgg
     1321 ggcccgagct taagactggc cgtcgtttta caacgtcgtg actgggaaaa catccatgct
     1381 agcgttaacg cgagagtagg gaactgccag gcatcaaata aaacgaaagg ctcagtcgga
     1441 agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctcctga gtaggacaaa
     1501 tccgccggga gcggatttga acgttgtgaa gcaacggccc ggagggtggc gggcaggacg
     1561 cccgccataa actgccaggc atcaaactaa gcagaaggcc atcctgacgg atggcctttt
     1621 tgcgtttcta caaactcttc ctggctagcg gtacgcgtat taattgcgtt gcgctcactg
     1681 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     1741 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     1801 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     1861 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     1921 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     1981 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     2041 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     2101 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     2161 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     2221 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     2281 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     2341 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     2401 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     2461 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     2521 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     2581 aacgacgcgt aactcacgtt aagggatttt ggtcatgggt ggctcgaggg ttatttgccg
     2641 actaccttgg tgatctcgcc tttcacgtag tggacaaatt cttccaactg atctgcgcgc
     2701 gaggccaagc gatcttcttc ttgtccaaga taagcctgtc tagcttcaag tatgacgggc
     2761 tgatactggg ccggcaggcg ctccattgcc cagtcggcag cgacatcctt cggcgcgatt
     2821 ttgccggtta ctgcgctgta ccaaatgcgg gacaacgtaa gcactacatt tcgctcatcg
     2881 ccagcccagt cgggcggcga gttccatagc gttaaggttt catttagcgc ctcaaataga
     2941 tcctgttcag gaaccggatc aaagagttcc tccgccgctg gacctaccaa ggcaacgcta
     3001 tgttctcttg cttttgtcag caagatagcc agatcaatgt cgatcgtggc tggctcgaag
     3061 atacctgcaa gaatgtcatt gcgctgccat tctccaaatt gcagttcgcg cttagctgga
     3121 taacgccacg gaatgatgtc gtcgtgcaca acaatggtga cttctacagc gcggagaatc
     3181 tcgctctctc caggggaagc cgaagtttcc aaaaggtcgt tgatcaaagc tcgccgcgtt
     3241 gtttcatcaa gccttacggt caccgtaacc agcaaatcaa tatcactgtg tggcttcagg
     3301 ccgccatcca ctgcggagcc gtacaaatgt acggccagca acgtcggttc gagatggcgc
     3361 tcgatgacgc caactacctc tgatagttga gtcgatactt cggcgatcac cgcttccctc
     3421 ataatgttta actttgtttt agggcgactg ccctgctgcg taacatcgtt gctgctccat
     3481 aacatcaaac atcgacccac ggcgtaacgc gcttgctgct tggatgcccg aggcatagac
     3541 tgtaccccaa aaaaacagtc ataacaagcc atgaaaaccg ccactgcgcc gttaccaccg
     3601 ctgcgttcgg tcaaggttct ggaccagttg cgtgagcgca tacgctactt gcattacagc
     3661 ttacgaaccg aacaggctta tgtccactgg gttcgtgcct tcatccgttt ccacggtgtg
     3721 cgtcacccgg ca
//
