LOCUS       sequence-164896-        6516 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway Donor vector containing IRS1 , part of the Target 
            Accelerator Plasmid Collection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6516)
  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 6516)
  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_IRS1_WT
FEATURES             Location/Qualifiers
     source          1..6516
                     /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"
     regulatory      3273..3282
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(3916..4015)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4032..4051)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4033..4051)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4056..4072)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4056..4072)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             4503..5294
                     /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      5387..5975
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5876..5895
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6129..6146
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(6305..6332)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(6424..6510)
                     /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 ggcgagccct ccggagagcg atggcttctc ggacgtgcgc aaggtgggct
      241 acctgcgcaa acccaagagc atgcacaaac gcttcttcgt actgcgcgcg gccagcgagg
      301 ctgggggccc ggcgcgcctc gagtactacg agaacgagaa gaagtggcgg cacaagtcga
      361 gcgcccccaa acgctcgatc ccccttgaga gctgcttcaa catcaacaag cgggctgact
      421 ccaagaacaa gcacctggtg gctctctaca cccgggacga gcactttgcc atcgcggcgg
      481 acagcgaggc cgagcaagac agctggtacc aggctctcct acagctgcac aaccgtgcta
      541 agggccacca cgacggagct gcggccctcg gggcgggagg tggtgggggc agctgcagcg
      601 gcagctccgg ccttggtgag gctggggagg acttgagcta cggtgacgtg cccccaggac
      661 ccgcattcaa agaggtctgg caagtgatcc tgaagcccaa gggcctgggt cagacaaaga
      721 acctgattgg tatctaccgc ctttgcctga ccagcaagac catcagcttc gtgaagctga
      781 actcggaggc agcggccgtg gtgctgcagc tgatgaacat caggcgctgt ggccactcgg
      841 aaaacttctt cttcatcgag gtgggccgtt ctgccgtgac ggggcccggg gagttctgga
      901 tgcaggtgga tgactctgtg gtggcccaga acatgcacga gaccatcctg gaggccatgc
      961 gggccatgag tgatgagttc cgccctcgca gcaagagcca gtcctcgtcc aactgctcta
     1021 accccatcag cgtccccctg cgccggcacc atctcaacaa tcccccgccc agccaggtgg
     1081 ggctgacccg ccgatcacgc actgagagca tcaccgccac ctccccggcc agcatggtgg
     1141 gcgggaagcc aggctccttc cgtgtccgcg cctccagtga cggcgaaggc accatgtccc
     1201 gcccagcctc ggtggacggc agccctgtga gtcccagcac caacagaacc cacgcccacc
     1261 ggcatcgggg cagcgcccgg ctgcaccccc cgctcaacca cagccgctcc atccccatgc
     1321 cggcttcccg ctgctcgcct tcggccacca gcccggtcag tctgtcgtcc agtagcacca
     1381 gtggccatgg ctccacctcg gattgtctct tcccacggcg atctagtgct tcggtgtctg
     1441 gttcccccag cgatggcggt ttcatctcct cggatgagta tggctccagt ccctgcgatt
     1501 tccggagttc cttccgcagt gtcactccgg attccctggg ccacacccca ccagcccgcg
     1561 gtgaggagga gctaagcaac tatatctgca tgggtggcaa ggggccctcc accctgaccg
     1621 cccccaacgg tcactacatt ttgtctcggg gtggcaatgg ccaccgctgc accccaggaa
     1681 caggcttggg cacgagtcca gccttggctg gggatgaagc agccagtgct gcagatctgg
     1741 ataatcggtt ccgaaagaga actcactcgg caggcacatc ccctaccatt acccaccaga
     1801 agaccccgtc ccagtcctca gtggcttcca ttgaggagta cacagagatg atgcctgcct
     1861 acccaccagg aggtggcagt ggaggccgac tgccgggaca caggcactcc gccttcgtgc
     1921 ccacccgctc ctacccagag gagggtctgg aaatgcaccc cttggagcgt cggggggggc
     1981 accaccgccc agacagctcc accctccaca cggatgatgg ctacatgccc atgtccccag
     2041 gggtggcccc agtgcccagt ggccgaaagg gcagtggaga ctatatgccc atgagcccca
     2101 agagcgtatc tgccccacag cagatcatca atcccatcag acgccatccc cagagagtgg
     2161 accccaatgg ctacatgatg atgtccccca gcggtggctg ctctcctgac attggaggtg
     2221 gccccagcag cagcagcagc agcagcaacg ccgtcccttc cgggaccagc tatggaaagc
     2281 tgtggacaaa cggggtaggg ggccaccact ctcatgtctt gcctcacccc aaacccccag
     2341 tggagagcag cggtggtaag ctcttacctt gcacaggtga ctacatgaac atgtcaccag
     2401 tgggggactc caacaccagc agcccctccg actgctacta cggccctgag gacccccagc
     2461 acaagccagt cctctcctac tactcattgc caagatcctt taagcacacc cagcgccccg
     2521 gggagccgga ggagggtgcc cggcatcagc acctccgcct ttccactagc tctggtcgcc
     2581 ttctctatgc tgcaacagca gatgattctt cctcttccac cagcagcgac agcctgggtg
     2641 ggggatactg cggggctagg ctggagccca gccttccaca tccccaccat caggttctgc
     2701 agccccatct gcctcgaaag gtggacacag ctgctcagac caatagccgc ctggcccggc
     2761 ccacgaggct gtccctgggg gatcccaagg ccagcacctt acctcgggcc cgagagcagc
     2821 agcagcagca gcagcccttg ctgcaccctc cagagcccaa gagcccgggg gaatatgtca
     2881 atattgaatt tgggagtgat cagtctggct acttgtctgg cccggtggct ttccacagct
     2941 caccttctgt caggtgtcca tcccagctcc agccagctcc cagagaggaa gagactggca
     3001 ctgaggagta catgaagatg gacctggggc cgggccggag ggcagcctgg caggagagca
     3061 ctggggtcga gatgggcaga ctgggccctg cacctcccgg ggctgctagc atttgcaggc
     3121 ctacccgggc agtgcccagc agccggggtg actacatgac catgcagatg agttgtcccc
     3181 gtcagagcta cgtggacacc tcgccagctg cccctgtaag ctatgctgac atgcgaacag
     3241 gcattgctgc agaggaggtg agcctgccca gggccaccat ggctgctgcc tcctcatcct
     3301 cagcagcctc tgcttccccg actgggcctc aaggggcagc agagctggct gcccactcgt
     3361 ccctgctggg gggcccacaa ggacctgggg gcatgagcgc cttcacccgg gtgaacctca
     3421 gtcctaaccg caaccagagt gccaaagtga tccgtgcaga cccacaaggg tgccggcgga
     3481 ggcatagctc cgagactttc tcctcaacac ccagtgccac ccgggtgggc aacacagtgc
     3541 cctttggagc gggggcagca gtagggggcg gtggcggtag cagcagcagc agcgaggatg
     3601 tgaaacgcca cagctctgct tcctttgaga atgtgtggct gaggcctggg gagcttgggg
     3661 gagcccccaa ggagccagcc aaactgtgtg gggctgctgg gggtttggag aatggtctta
     3721 actacataga cctggatttg gtcaaggact tcaaacagtg ccctcaggag tgcacccctg
     3781 aaccgcagcc tcccccaccc ccaccccctc atcaacccct gggcagcggt gagagcagct
     3841 ccacccgccg ctcaagtgag gatttaagcg cctatgccag catcagtttc cagaagcagc
     3901 cagaggaccg tcagtaccca actttcttgt acaaagttgg cattataaga aagcattgct
     3961 tatcaatttg ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt atttgccatc
     4021 cagctgatat cccctatagt gagtcgtatt acatggtcat agctgtttcc tggcagctct
     4081 ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgcc
     4141 tcctctagac cagccaggac agaaatgcct cgacttcgct gctacccaag gttgccgggt
     4201 gacgcacacc gtggaaacgg atgaaggcac gaacccagtg gacataagcc tgttcggttc
     4261 gtaagctgta atgcaagtag cgtatgcgct cacgcaactg gtccagaacc ttgaccgaac
     4321 gcagcggtgg taacggcgca gtggcggttt tcatggcttg ttatgactgt ttttttgggg
     4381 tacagtctat gcctcgggca tccaagcagc aagcgcgtta cgccgtgggt cgatgtttga
     4441 tgttatggag cagcaacgat gttacgcagc agggcagtcg ccctaaaaca aagttaaaca
     4501 ttatgaggga agcggtgatc gccgaagtat cgactcaact atcagaggta gttggcgtca
     4561 tcgagcgcca tctcgaaccg acgttgctgg ccgtacattt gtacggctcc gcagtggatg
     4621 gcggcctgaa gccacacagt gatattgatt tgctggttac ggtgaccgta aggcttgatg
     4681 aaacaacgcg gcgagctttg atcaacgacc ttttggaaac ttcggcttcc cctggagaga
     4741 gcgagattct ccgcgctgta gaagtcacca ttgttgtgca cgacgacatc attccgtggc
     4801 gttatccagc taagcgcgaa ctgcaatttg gagaatggca gcgcaatgac attcttgcag
     4861 gtatcttcga gccagccacg atcgacattg atctggctat cttgctgaca aaagcaagag
     4921 aacatagcgt tgccttggta ggtccagcgg cggaggaact ctttgatccg gttcctgaac
     4981 aggatctatt tgaggcgcta aatgaaacct taacgctatg gaactcgccg cccgactggg
     5041 ctggcgatga gcgaaatgta gtgcttacgt tgtcccgcat ttggtacagc gcagtaaccg
     5101 gcaaaatcgc gccgaaggat gtcgctgccg actgggcaat ggagcgcctg ccggcccagt
     5161 atcagcccgt catacttgaa gctagacagg cttatcttgg acaagaagaa gatcgcttgg
     5221 cctcgcgcgc agatcagttg gaagaatttg tccactacgt gaaaggcgag atcaccaagg
     5281 tagtcggcaa ataaccctcg agccacccat gaccaaaatc ccttaacgtg agttacgcgt
     5341 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     5401 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     5461 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     5521 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     5581 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     5641 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     5701 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     5761 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     5821 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     5881 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     5941 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     6001 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     6061 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     6121 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
     6181 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
     6241 cgggcagtga gcgcaacgca attaatacgc gtaccgctag ccaggaagag tttgtagaaa
     6301 cgcaaaaagg ccatccgtca ggatggcctt ctgcttagtt tgatgcctgg cagtttatgg
     6361 cgggcgtcct gcccgccacc ctccgggccg ttgcttcaca acgttcaaat ccgctcccgg
     6421 cggatttgtc ctactcagga gagcgttcac cgacaaacaa cagataaaac gaaaggccca
     6481 gtcttccgac tgagcctttc gttttatttg atgcct
//
