LOCUS       sequence-413681-        5194 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway-compatible entry vector, with insert of the full length TAF5
            gene mutated to be resistant to TAF5 siRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5194)
  AUTHORS   Xiao MS, Damodaran AP, Kumari B, Dickson E, Xing K, On TA, Parab N, 
            King HE, Perez AR, Guiblet WM, Duncan G, Che A, Chari R, Andresson 
            T, Vidigal JA, Weatheritt RJ, Aregger M, Gonatopoulos-Pournatzis T
  TITLE     Genome-scale exon perturbation screens uncover exons critical for 
            cell fitness.
  JOURNAL   Mol Cell. 2024 Jul 11;84(13):2553-2572.e19. doi: 
            10.1016/j.molcel.2024.05.024. Epub 2024 Jun 24.
  PUBMED    38917794
REFERENCE   2  (bases 1 to 5194)
  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.molcel.2024.05.024"; journalName: "Mol Cell"; date: 
            "2024-07-11- 11"; volume: "84"; issue: "13"; pages: "2553-2572.e19"
COMMENT     Sequence Label: pDONR223-TAF5-FL_WobbleMut
FEATURES             Location/Qualifiers
     source          1..5194
                     /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(3073..3172)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3189..3208)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3190..3208)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3213..3229)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3213..3229)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3660..4451
                     /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      4544..5132
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5033..5052
                     /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 agcaggcttc gcggcgctgg cggaggagca gacggaggtg gcggtcaagc tagagcctga
      721 gggaccgcca acgctgctac ctccgcaggc gggggacggc gcaggcgagg gtagcggcgg
      781 cactaccaac aacggcccca acggcggcgg cgggaacgtt gcggcgtcgt cgtccactgg
      841 cggggatggc gggaccccca agcccacggt ggctgtctcc gccgctgccc cggcgggggc
      901 ggccccggtg cccgccgctg ctccggacgc cggcgctccg catgaccgac agactctact
      961 ggccgtgctg cagttcctac ggcagagcaa actccgcgag gccgaagagg cgctgcgccg
     1021 tgaggccggg ctgctggagg aggcagtggc gggctccgga gccccgggag aggtggacag
     1081 cgccggcgct gaggtgacca gcgcgcttct cagccgggtg accgcctcgg cccctggccc
     1141 tgcggccccc gaccctccgg gcactggcgc ttcgggggcc acggtcgtct caggttcagc
     1201 ctcaggtcct gcggctccgg gtaaagttgg aagtgttgct gtggaagacc agccagatgt
     1261 cagtgccgtg ttgtcagcct acaaccaaca aggagatccc acaatgtatg aagaatacta
     1321 tagtggactg aaacacttca ttgaatgttc cctggactgc catcgggcag agttgtccca
     1381 acttttttat cctctgtttg tgcacatgta cttggagcta gtctacaatc aacatgagaa
     1441 tgaagcaaag tcattctttg agaagttcca tggagatcag gaatgttatt accaggatga
     1501 cctacgagta ttatctagtc ttaccaaaaa ggaacacatg aaagggaatg agaccatgtt
     1561 ggattttcga acaagtaaat ttgttctgcg tatttcccgt gactcgtacc aactcttgaa
     1621 gaggcatctt caggagaaac agaacaatca gatatggaac atagttcagg agcacctcta
     1681 cattgacatc tttgatggga tgccgcgtag taagcaacag atagatgcga tggtgggaag
     1741 tttggcagga gaggctaaac gagaggcaaa caaatcaaag gtattttttg gtttattaaa
     1801 agaaccagaa attgaggtac ctttggatga cgaggatgaa gagggagaaa atgaagaagg
     1861 aaaacctaaa aagaagaagc ctaaaaaaga tagtattgga tccaaaagca aaaaacaaga
     1921 tcccaatgct ccacctcaga acagaatccc tcttcctgag ttgaaagatt cagataagtt
     1981 ggataagata atgaatatga aagaaaccac caaacgagtg cgccttgggc cggactgctt
     2041 accctccatt tgtttctata catttctcaa tgcttaccag ggtctcactg cagtggatgt
     2101 cactgatgat tctagtctga ttgctggagg ttttgcagat tcaactgtca gagtgtggtc
     2161 ggtaacaccc aaaaagcttc gtagtgtcaa acaagcatca gatcttagtc ttatagacaa
     2221 agaatcagat gatgtcttag aaagaatcat ggatgagaaa acagcaagtg agttgaagat
     2281 tttgtatggt cacagtgggc ctgtctacgg agccagcttc agtccggata ggaactatct
     2341 gctttcctct tcagaggacg gaactgttag attgtggagc cttcaaacat ttacttgttt
     2401 ggtgggatat aaaggacaca actatccagt atgggacaca caattttctc catatggata
     2461 ttattttgtg tcagggggcc atgaccgagt agctcggctc tgggctacag accactatca
     2521 gcctttaaga atatttgccg gccatcttgc tgatgtgaat tgtaccagat tccatccaaa
     2581 ttctaattat gttgctacgg gctctgcaga cagaactgtg cggctctggg acgtcctgaa
     2641 tggtaactgt gtaaggatct tcactggaca caagggacca attcattcct tgacattttc
     2701 tcccaatggg agattcctgg ctacaggagc aacagatggc agagtgcttc tttgggatat
     2761 tggacatggt ttgatggttg gagaattaaa aggccacact gatacagtct gttcacttag
     2821 gtttagtaga gatggtgaaa ttttggccag cggcagcatg gacaacacag ttcgattatg
     2881 ggatgctatc aaagcctttg aagatttaga gaccgatgac tttactacag ccactgggca
     2941 tataaattta cctgagaatt cacaggagtt attgttggga acatatatga ccaaatcaac
     3001 accagttgta caccttcatt ttactcgaag aaacctggtt ctagctgcag gagcttatag
     3061 tccacaatga taacccagct ttcttgtaca aagttggcat tataagaaag cattgcttat
     3121 caatttgttg caacgaacag gtcactatca gtcaaaataa aatcattatt tgccatccag
     3181 ctgatatccc ctatagtgag tcgtattaca tggtcatagc tgtttcctgg cagctctggc
     3241 ccgtgtctca aaatctctga tgttacattg cacaagataa aaatatatca tcatgcctcc
     3301 tctagaccag ccaggacaga aatgcctcga cttcgctgct acccaaggtt gccgggtgac
     3361 gcacaccgtg gaaacggatg aaggcacgaa cccagtggac ataagcctgt tcggttcgta
     3421 agctgtaatg caagtagcgt atgcgctcac gcaactggtc cagaaccttg accgaacgca
     3481 gcggtggtaa cggcgcagtg gcggttttca tggcttgtta tgactgtttt tttggggtac
     3541 agtctatgcc tcgggcatcc aagcagcaag cgcgttacgc cgtgggtcga tgtttgatgt
     3601 tatggagcag caacgatgtt acgcagcagg gcagtcgccc taaaacaaag ttaaacatta
     3661 tgagggaagc ggtgatcgcc gaagtatcga ctcaactatc agaggtagtt ggcgtcatcg
     3721 agcgccatct cgaaccgacg ttgctggccg tacatttgta cggctccgca gtggatggcg
     3781 gcctgaagcc acacagtgat attgatttgc tggttacggt gaccgtaagg cttgatgaaa
     3841 caacgcggcg agctttgatc aacgaccttt tggaaacttc ggcttcccct ggagagagcg
     3901 agattctccg cgctgtagaa gtcaccattg ttgtgcacga cgacatcatt ccgtggcgtt
     3961 atccagctaa gcgcgaactg caatttggag aatggcagcg caatgacatt cttgcaggta
     4021 tcttcgagcc agccacgatc gacattgatc tggctatctt gctgacaaaa gcaagagaac
     4081 atagcgttgc cttggtaggt ccagcggcgg aggaactctt tgatccggtt cctgaacagg
     4141 atctatttga ggcgctaaat gaaaccttaa cgctatggaa ctcgccgccc gactgggctg
     4201 gcgatgagcg aaatgtagtg cttacgttgt cccgcatttg gtacagcgca gtaaccggca
     4261 aaatcgcgcc gaaggatgtc gctgccgact gggcaatgga gcgcctgccg gcccagtatc
     4321 agcccgtcat acttgaagct agacaggctt atcttggaca agaagaagat cgcttggcct
     4381 cgcgcgcaga tcagttggaa gaatttgtcc actacgtgaa aggcgagatc accaaggtag
     4441 tcggcaaata accctcgagc cacccatgac caaaatccct taacgtgagt tacgcgtcgt
     4501 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     4561 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     4621 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     4681 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     4741 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     4801 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     4861 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     4921 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     4981 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     5041 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     5101 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     5161 tcctggcctt ttgctggcct tttgctcaca tgtt
//
