LOCUS       sequence-332493-        4101 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry vector for human VPS4A ORF.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4101)
  AUTHORS   Neggers JE, Paolella BR, Asfaw A, Rothberg MV, Skipper TA, Yang A, 
            Kalekar RL, Krill-Burger JM, Dharia NV, Kugener G, Kalfon J, Yuan C,
            Dumont N, Gonzalez A, Abdusamad M, Li YY, Spurr LF, Wu WW, Durbin 
            AD, Wolpin BM, Piccioni F, Root DE, Boehm JS, Cherniack AD, 
            Tsherniak A, Hong AL, Hahn WC, Stegmaier K, Golub TR, Vazquez F, 
            Aguirre AJ
  TITLE     Synthetic Lethal Interaction between the ESCRT Paralog Enzymes VPS4A
            and VPS4B in Cancers Harboring Loss of Chromosome 18q or 16q.
  JOURNAL   Cell Rep. 2020 Dec 15;33(11):108493. doi: 
            10.1016/j.celrep.2020.108493.
  PUBMED    33326793
REFERENCE   2  (bases 1 to 4101)
  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"; journalName: "Cell Rep.";
            date: "2020-12-15"; volume: "33(11):108493. doi"; pages: " 
            10.1016/j.celrep.2020.108493"
COMMENT     Sequence Label: pDONR 223 VPS4A
FEATURES             Location/Qualifiers
     source          1..4101
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             262..1053
                     /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      1146..1734
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1635..1654
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1888..1905
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2064..2091)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2183..2269)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2318..2340
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2332..2349
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2333..2349
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2365..2464
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(3776..3875)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3892..3911)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3893..3911)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3916..3932)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3916..3932)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 acataagcct gttcggttcg taagctgtaa tgcaagtagc gtatgcgctc acgcaactgg
       61 tccagaacct tgaccgaacg cagcggtggt aacggcgcag tggcggtttt catggcttgt
      121 tatgactgtt tttttggggt acagtctatg cctcgggcat ccaagcagca agcgcgttac
      181 gccgtgggtc gatgtttgat gttatggagc agcaacgatg ttacgcagca gggcagtcgc
      241 cctaaaacaa agttaaacat tatgagggaa gcggtgatcg ccgaagtatc gactcaacta
      301 tcagaggtag ttggcgtcat cgagcgccat ctcgaaccga cgttgctggc cgtacatttg
      361 tacggctccg cagtggatgg cggcctgaag ccacacagtg atattgattt gctggttacg
      421 gtgaccgtaa ggcttgatga aacaacgcgg cgagctttga tcaacgacct tttggaaact
      481 tcggcttccc ctggagagag cgagattctc cgcgctgtag aagtcaccat tgttgtgcac
      541 gacgacatca ttccgtggcg ttatccagct aagcgcgaac tgcaatttgg agaatggcag
      601 cgcaatgaca ttcttgcagg tatcttcgag ccagccacga tcgacattga tctggctatc
      661 ttgctgacaa aagcaagaga acatagcgtt gccttggtag gtccagcggc ggaggaactc
      721 tttgatccgg ttcctgaaca ggatctattt gaggcgctaa atgaaacctt aacgctatgg
      781 aactcgccgc ccgactgggc tggcgatgag cgaaatgtag tgcttacgtt gtcccgcatt
      841 tggtacagcg cagtaaccgg caaaatcgcg ccgaaggatg tcgctgccga ctgggcaatg
      901 gagcgcctgc cggcccagta tcagcccgtc atacttgaag ctagacaggc ttatcttgga
      961 caagaagaag atcgcttggc ctcgcgcgca gatcagttgg aagaatttgt ccactacgtg
     1021 aaaggcgaga tcaccaaggt agtcggcaaa taaccctcga gccacccatg accaaaatcc
     1081 cttaacgtga gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     1141 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     1201 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     1261 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     1321 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     1381 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     1441 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     1501 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     1561 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     1621 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     1681 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     1741 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     1801 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     1861 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     1921 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     1981 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatacgcg taccgctagc
     2041 caggaagagt ttgtagaaac gcaaaaaggc catccgtcag gatggccttc tgcttagttt
     2101 gatgcctggc agtttatggc gggcgtcctg cccgccaccc tccgggccgt tgcttcacaa
     2161 cgttcaaatc cgctcccggc ggatttgtcc tactcaggag agcgttcacc gacaaacaac
     2221 agataaaacg aaaggcccag tcttccgact gagcctttcg ttttatttga tgcctggcag
     2281 ttccctactc tcgcgttaac gctagcatgg atgttttccc agtcacgacg ttgtaaaacg
     2341 acggccagtc ttaagctcgg gccccaaata atgattttat tttgactgat agtgacctgt
     2401 tcgttgcaac aaattgatga gcaatgcttt tttataatgc caactttgta caaaaaagtt
     2461 ggcatgacaa cgtcaaccct ccagaaagcc attgatctgg tgacgaaagc cacagaggag
     2521 gacaaagcca agaactacga ggaggcgctg cggctgtacc agcatgcggt ggagtacttc
     2581 ctccacgcta tcaagtatga ggcccacagc gacaaggcca aggagagcat tcgagccaag
     2641 tgcgtgcagt acctagaccg ggccgagaag ctgaaggatt atttacgaag caaagagaaa
     2701 cacggcaaga agccagtcaa agagaaccag agtgagggca agggcagtga cagtgacagt
     2761 gaaggggata atccggagaa aaagaaactg caagaacagc tgatgggtgc cgtcgtgatg
     2821 gagaagccca acatacggtg gaacgacgtg gccgggctgg agggggccaa ggaggccctc
     2881 aaagaagctg tcattttgcc aatcaaattc ccacacttgt tcacaggcaa gcgcaccccc
     2941 tggcggggga ttctgctgtt cggaccccct ggcacaggga aatcctacct ggccaaagcc
     3001 gtggcaacag aggccaacaa ctccaccttc ttctctgtgt cctcctcaga tctgatgtcc
     3061 aagtggctgg gggagagtga aaagctggtc aagaacctgt ttgagctggc caggcagcac
     3121 aagccctcca tcatcttcat cgatgaggtg gattccctct gcgggtcccg aaatgaaaat
     3181 gagagtgagg ccgcccggag gatcaaaacg gagttcttgg tccagatgca gggggtgggg
     3241 aataacaatg atgggactct ggttcttgga gccacaaaca tcccatgggt gttggattcg
     3301 gccatcagga ggaggtttga aaaacgaatt tatatcccct tgccggagga agctgcccgc
     3361 gcccagatgt tccggttgca tctcgggagc actccccaca acctcacgga tgcaaacatc
     3421 cacgagctgg cccggaagac ggaaggctac tcgggcgcgg acatcagcat catcgtgcgg
     3481 gactctctca tgcagcccgt gaggaaggtg cagtcggcca cacacttcaa aaaggtctgt
     3541 ggcccctctc gcaccaaccc cagcatgatg attgatgacc tcctgactcc atgctcacca
     3601 ggggacccag gagccatgga gatgacttgg atggatgtcc ctggggacaa actcttagag
     3661 cctgtggttt gcatgtcgga catgctgcgg tctctggcca ccacccggcc cacggtgaat
     3721 gcagacgacc tcctgaaagt gaagaaattc tcagaggact ttgggcaaga gagttaccca
     3781 actttcttgt acaaagttgg cattataaga aagcattgct tatcaatttg ttgcaacgaa
     3841 caggtcacta tcagtcaaaa taaaatcatt atttgccatc cagctgatat cccctatagt
     3901 gagtcgtatt acatggtcat agctgtttcc tggcagctct ggcccgtgtc tcaaaatctc
     3961 tgatgttaca ttgcacaaga taaaaatata tcatcatgcc tcctctagac cagccaggac
     4021 agaaatgcct cgacttcgct gctacccaag gttgccgggt gacgcacacc gtggaaacgg
     4081 atgaaggcac gaacccagtg g
//
