LOCUS       sequence-332492-        4122 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry vector for human VPS4B ORF.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4122)
  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 4122)
  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 VPS4B
FEATURES             Location/Qualifiers
     source          1..4122
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(1..28)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(120..206)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     255..277
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     269..286
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     270..286
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    302..401
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             1224..1235
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     protein_bind    complement(1734..1833)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1850..1869)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1851..1869)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1874..1890)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1874..1890)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2321..3112
                     /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      3205..3793
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3694..3713
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3947..3964
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 aaaaggccat ccgtcaggat ggccttctgc ttagtttgat gcctggcagt ttatggcggg
       61 cgtcctgccc gccaccctcc gggccgttgc ttcacaacgt tcaaatccgc tcccggcgga
      121 tttgtcctac tcaggagagc gttcaccgac aaacaacaga taaaacgaaa ggcccagtct
      181 tccgactgag cctttcgttt tatttgatgc ctggcagttc cctactctcg cgttaacgct
      241 agcatggatg ttttcccagt cacgacgttg taaaacgacg gccagtctta agctcgggcc
      301 ccaaataatg attttatttt gactgatagt gacctgttcg ttgcaacaaa ttgatgagca
      361 atgctttttt ataatgccaa ctttgtacaa aaaagttggc atgtcatcca cttcgcccaa
      421 cctccagaaa gcgatagatc tggctagcaa agcagcgcaa gaagacaagg ctgggaacta
      481 cgaagaagcc cttcagctct atcagcatgc tgtgcagtat tttcttcatg tcgttaaata
      541 tgaagcacag ggtgataaag ccaagcaaag tatcagggca aagtgtacag aatatcttga
      601 tagagcagaa aaactaaagg agtacctgaa aaataaagag aaaaaagcac agaagccagt
      661 gaaagaagga cagccgagtc cagcagatga gaaggggaat gacagtgatg gggaaggaga
      721 atctgatgat cctgaaaaaa agaaactaca gaatcaactt caaggtgcca ttgttataga
      781 acgaccaaat gtgaaatgga gtgacgttgc tggacttgaa ggagccaaag aagcactgaa
      841 agaggctgtg atactgccta ttaaatttcc tcatcttttt acaggcaaga gaacaccttg
      901 gaggggaatc ctattatttg ggccgcctgg aacaggaaag tcctacttag ccaaagctgt
      961 agcaacagaa gccaacaact caacattttt ttcaatatct tcctctgatc ttgtttctaa
     1021 gtggctaggt gaaagtgaaa aactggttaa gaatttattc caacttgcca gagagaacaa
     1081 gccctccatt atcttcattg atgaaattga ttctctctgt ggttcaagaa gtgaaaatga
     1141 aagtgaagcc gcacgtagaa ttaagacgga gttcctagtg caaatgcaag gggttggtgt
     1201 agacaatgat ggaattttgg ttctgggagc tacaaatata ccctgggttc tggattctgc
     1261 cattaggcga agatttgaga aacgaattta tattcccttg ccggaacccc atgcccgagc
     1321 agcaatgttt aaactgcacc tagggaccac tcagaacagt ctcacggaag cagactttcg
     1381 ggaacttggg aggaaaacag atggttattc aggggcagat ataagtatca ttgtacgtga
     1441 tgcccttatg cagcctgtta ggaaagtaca gtcagctact cattttaaaa aggttcgcgg
     1501 accttcccga gctgatccta accatcttgt agatgatctg ctaacacctt gctctccagg
     1561 tgaccctggt gccattgaaa tgacatggat ggatgtccct ggagataaac ttttggagcc
     1621 agttgtttcc atgtcggata tgttgcggtc actatctaac acaaaaccta cagtcaatga
     1681 acatgacttg ttgaaattaa agaagtttac agaagatttt ggtcaagaag gctacccaac
     1741 tttcttgtac aaagttggca ttataagaaa gcattgctta tcaatttgtt gcaacgaaca
     1801 ggtcactatc agtcaaaata aaatcattat ttgccatcca gctgatatcc cctatagtga
     1861 gtcgtattac atggtcatag ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg
     1921 atgttacatt gcacaagata aaaatatatc atcatgcctc ctctagacca gccaggacag
     1981 aaatgcctcg acttcgctgc tacccaaggt tgccgggtga cgcacaccgt ggaaacggat
     2041 gaaggcacga acccagtgga cataagcctg ttcggttcgt aagctgtaat gcaagtagcg
     2101 tatgcgctca cgcaactggt ccagaacctt gaccgaacgc agcggtggta acggcgcagt
     2161 ggcggttttc atggcttgtt atgactgttt ttttggggta cagtctatgc ctcgggcatc
     2221 caagcagcaa gcgcgttacg ccgtgggtcg atgtttgatg ttatggagca gcaacgatgt
     2281 tacgcagcag ggcagtcgcc ctaaaacaaa gttaaacatt atgagggaag cggtgatcgc
     2341 cgaagtatcg actcaactat cagaggtagt tggcgtcatc gagcgccatc tcgaaccgac
     2401 gttgctggcc gtacatttgt acggctccgc agtggatggc ggcctgaagc cacacagtga
     2461 tattgatttg ctggttacgg tgaccgtaag gcttgatgaa acaacgcggc gagctttgat
     2521 caacgacctt ttggaaactt cggcttcccc tggagagagc gagattctcc gcgctgtaga
     2581 agtcaccatt gttgtgcacg acgacatcat tccgtggcgt tatccagcta agcgcgaact
     2641 gcaatttgga gaatggcagc gcaatgacat tcttgcaggt atcttcgagc cagccacgat
     2701 cgacattgat ctggctatct tgctgacaaa agcaagagaa catagcgttg ccttggtagg
     2761 tccagcggcg gaggaactct ttgatccggt tcctgaacag gatctatttg aggcgctaaa
     2821 tgaaacctta acgctatgga actcgccgcc cgactgggct ggcgatgagc gaaatgtagt
     2881 gcttacgttg tcccgcattt ggtacagcgc agtaaccggc aaaatcgcgc cgaaggatgt
     2941 cgctgccgac tgggcaatgg agcgcctgcc ggcccagtat cagcccgtca tacttgaagc
     3001 tagacaggct tatcttggac aagaagaaga tcgcttggcc tcgcgcgcag atcagttgga
     3061 agaatttgtc cactacgtga aaggcgagat caccaaggta gtcggcaaat aaccctcgag
     3121 ccacccatga ccaaaatccc ttaacgtgag ttacgcgtcg ttccactgag cgtcagaccc
     3181 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     3241 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     3301 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     3361 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     3421 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     3481 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     3541 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     3601 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     3661 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     3721 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     3781 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     3841 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     3901 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     3961 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     4021 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
     4081 taatacgcgt accgctagcc aggaagagtt tgtagaaacg ca
//
