LOCUS       sequence-332430-        2279 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses CHMP1a targeting sgRNA in non-lentiviral BPK1520 plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2279)
  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 2279)
  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 sgCHMP1A
FEATURES             Location/Qualifiers
     source          1..2279
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_RNA        29..104
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     rep_origin      complement(208..796)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(288..307)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(967..1758,1759..1827))
                     /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     1590..1609
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1828..1932)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        join(2039..2279,1..8)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2039..2059
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2210..2229
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
ORIGIN
        1 gaaacaccga acctggacgt ccatacatgt tttagagcta gaaatagcaa gttaaaataa
       61 ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg gtgctttttt taagcttggg
      121 ccgctcgagg tacctctcta catatgacat gtgagcaaaa ggccagcaaa aggccaggaa
      181 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
      241 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
      301 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
      361 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
      421 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
      481 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
      541 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
      601 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
      661 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
      721 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
      781 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
      841 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
      901 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
      961 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     1021 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     1081 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     1141 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     1201 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     1261 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     1321 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     1381 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     1441 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     1501 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     1561 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     1621 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     1681 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     1741 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     1801 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     1861 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     1921 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgctagct gtacaaaaaa
     1981 gcaggcttta aaggaaccaa ttcagtcgac tggatccggt accaaggtcg ggcaggaaga
     2041 gggcctattt cccatgattc cttcatattt gcatatacga tacaaggctg ttagagagat
     2101 aattagaatt aatttgactg taaacacaaa gatattagta caaaatacgt gacgtagaaa
     2161 gtaataattt cttgggtagt ttgcagtttt aaaattatgt tttaaaatgg actatcatat
     2221 gcttaccgta acttgaaagt atttcgattt cttggcttta tatatcttgt ggaaaggac
//
