LOCUS       sequence-332429-        2280 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses CHMP4B targeting sgRNA in non-lentiviral BPK1520 plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2280)
  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 2280)
  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: BPK1520 sgCHMP4B
FEATURES             Location/Qualifiers
     source          1..2280
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(48..636)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(128..147)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(807..1598,1599..1667))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1430..1449
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1668..1772)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1879..2127
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1879..1899
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2050..2069
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2149..2224
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
ORIGIN
        1 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
       61 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
      121 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
      181 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
      241 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
      301 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
      361 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
      421 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
      481 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
      541 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
      601 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
      661 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
      721 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
      781 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
      841 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
      901 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
      961 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     1021 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     1081 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     1141 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     1201 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     1261 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     1321 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     1381 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     1441 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     1501 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     1561 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     1621 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     1681 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     1741 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     1801 gtcgctagct gtacaaaaaa gcaggcttta aaggaaccaa ttcagtcgac tggatccggt
     1861 accaaggtcg ggcaggaaga gggcctattt cccatgattc cttcatattt gcatatacga
     1921 tacaaggctg ttagagagat aattagaatt aatttgactg taaacacaaa gatattagta
     1981 caaaatacgt gacgtagaaa gtaataattt cttgggtagt ttgcagtttt aaaattatgt
     2041 tttaaaatgg actatcatat gcttaccgta acttgaaagt atttcgattt cttggcttta
     2101 tatatcttgt ggaaaggacg aaacaccgca gcgctggacc ccattacagt tttagagcta
     2161 gaaatagcaa gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg
     2221 gtgctttttt taagcttggg ccgctcgagg tacctctcta catatgacat gtgagcaaaa
//
