LOCUS       sequence-168739-        4299 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway Donor vector containing SPAG8 , part of the Target 
            Accelerator Plasmid Collection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4299)
  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 4299)
  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_SPAG8_WT
FEATURES             Location/Qualifiers
     source          1..4299
                     /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(1699..1798)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1815..1834)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1816..1834)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1839..1855)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1839..1855)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2286..3077
                     /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      3170..3758
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3659..3678
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3912..3929
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4088..4115)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4207..4293)
                     /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 aagttggcac catggatacc aacgggtcta cggagggatc gcggtcgcgg tcgcgatctt
      241 tagacataca gcccagctcc gaaggactgg ggcccacttc ggaaccgttt ccttcttcag
      301 atgacagtcc caggtcggcc ctggcagctg caaccgcagc agctgcagcg gctgcatcag
      361 ctgctgcagc tactgcagcc ttcaccactg ccaaagcagc tgcattatct acaaagaccc
      421 cagcgccctg ttctgagttc atggagccgt cctctgaccc cagccttctt ggggagccct
      481 gtgcgggacc cggctttacc cacaatatag cccatgggag tcttggcttt gagcccgtct
      541 atgtttcctg tattgctcag gacacttgca ctacaactga ccatagttct aatcctggcc
      601 ctgttccagg ctctagctct gggcctgttc ttggttccag ctcaggtgct ggccatggct
      661 ctggctctgg ctctggtcct ggctgtggct ctgtccctgg ctctggctct ggtcctggtc
      721 ctggctctgg tcctggctct ggtcctggtc atggctctgg ctctcatcct ggtcctgcct
      781 ctgggcctgg tccagacact ggccctgact ctgagctcag cccctgtatt cctccagggt
      841 tcagaaacct ggtggcagat cgggtcctta actatacctc ctggagtcag cactgcccct
      901 gggagcccca gaaacaacca ccttgggaat ttttgcaagt cttagaaccg ggtgcccgag
      961 gattatggaa acccccagac attaaaggga agcttatggt ttgctatgaa actttgccgc
     1021 ggggccagtg cctcctctac aactgggagg aagagagagc caccaaccac ctggatcaag
     1081 tcccaagcat gcaggatggc tctgagagtt ttttcttccg acacggacac cggggactgc
     1141 tgactatgca actaaagtca cccatgccct ccagcaccac ccagaaagac tcgtaccagc
     1201 caccaggaaa cgtctattgg ccacttcgag ggaagcgtga agccatgctg gagatgctcc
     1261 tgcagcatca gatctgtaaa gaggtgcagg cagaacagga acccacaagg aagctcttcg
     1321 aggttgagtc tgtgacacac catgactacc gaatggagct ggcacaagca gggactcctg
     1381 ccccaacaaa gcctcacgac taccgccagg agcaacctga gaccttctgg atacagaggg
     1441 caccacagct gccgacatgg tggccattgc ccacccaggt accagcagca gaagactatc
     1501 tgacttggaa agaatggggg tttacaggag tccaggaggt cctttccgct ctcctaagag
     1561 ccacacctgg tgaatactca gtaaacattt gcggaatgaa tgaacaccct gtctgcagca
     1621 ggacatggac aaataggctc tgccatcagg aaatgggaag caagaaaacg gtaactcaag
     1681 aggacagagg ctggtgattg ccaactttct tgtacaaagt tgtcattata agaaagcatt
     1741 gcttatcaat ttgttgcaac gaacaggtca ctatcagtca aaataaaatc attatttgcc
     1801 atccagctga tatcccctat agtgagtcgt attacatggt catagctgtt tcctggcagc
     1861 tctggcccgt gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat
     1921 gcctcctcta gaccagccag gacagaaatg cctcgacttc gctgctaccc aaggttgccg
     1981 ggtgacgcac accgtggaaa cggatgaagg cacgaaccca gtggacataa gcctgttcgg
     2041 ttcgtaagct gtaatgcaag tagcgtatgc gctcacgcaa ctggtccaga accttgaccg
     2101 aacgcagcgg tggtaacggc gcagtggcgg ttttcatggc ttgttatgac tgtttttttg
     2161 gggtacagtc tatgcctcgg gcatccaagc agcaagcgcg ttacgccgtg ggtcgatgtt
     2221 tgatgttatg gagcagcaac gatgttacgc agcagggcag tcgccctaaa acaaagttaa
     2281 acattatgag ggaagcggtg atcgccgaag tatcgactca actatcagag gtagttggcg
     2341 tcatcgagcg ccatctcgaa ccgacgttgc tggccgtaca tttgtacggc tccgcagtgg
     2401 atggcggcct gaagccacac agtgatattg atttgctggt tacggtgacc gtaaggcttg
     2461 atgaaacaac gcggcgagct ttgatcaacg accttttgga aacttcggct tcccctggag
     2521 agagcgagat tctccgcgct gtagaagtca ccattgttgt gcacgacgac atcattccgt
     2581 ggcgttatcc agctaagcgc gaactgcaat ttggagaatg gcagcgcaat gacattcttg
     2641 caggtatctt cgagccagcc acgatcgaca ttgatctggc tatcttgctg acaaaagcaa
     2701 gagaacatag cgttgccttg gtaggtccag cggcggagga actctttgat ccggttcctg
     2761 aacaggatct atttgaggcg ctaaatgaaa ccttaacgct atggaactcg ccgcccgact
     2821 gggctggcga tgagcgaaat gtagtgctta cgttgtcccg catttggtac agcgcagtaa
     2881 ccggcaaaat cgcgccgaag gatgtcgctg ccgactgggc aatggagcgc ctgccggccc
     2941 agtatcagcc cgtcatactt gaagctagac aggcttatct tggacaagaa gaagatcgct
     3001 tggcctcgcg cgcagatcag ttggaagaat ttgtccacta cgtgaaaggc gagatcacca
     3061 aggtagtcgg caaataaccc tcgagccacc catgaccaaa atcccttaac gtgagttacg
     3121 cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     3181 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     3241 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     3301 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     3361 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     3421 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     3481 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     3541 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     3601 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     3661 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     3721 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     3781 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     3841 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     3901 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc
     3961 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
     4021 aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc tagccaggaa gagtttgtag
     4081 aaacgcaaaa aggccatccg tcaggatggc cttctgctta gtttgatgcc tggcagttta
     4141 tggcgggcgt cctgcccgcc accctccggg ccgttgcttc acaacgttca aatccgctcc
     4201 cggcggattt gtcctactca ggagagcgtt caccgacaaa caacagataa aacgaaaggc
     4261 ccagtcttcc gactgagcct ttcgttttat ttgatgcct
//
