LOCUS       sequence-75539-e         789 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Backbone for transient transfection of Massively Parallel Reporter 
            Assays (MPRA) libraries, pPGK-H2Kk for physical selection of 
            transfected cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 789)
  AUTHORS   Melnikov A, Zhang X, Rogov P, Wang L, Mikkelsen TS
  TITLE     Massively parallel reporter assays in cultured Mammalian cells.
  JOURNAL   J Vis Exp. 2014 Aug 17;(90). doi: 10.3791/51719.
  PUBMED    25177895
REFERENCE   2  (bases 1 to 789)
  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: "J Vis 
            Exp."; date: "2014-08-17"; volume: "(90). doi"; pages: " 
            10.3791/51719"
COMMENT     Sequence Label: pMPRA2
FEATURES             Location/Qualifiers
     source          1..789
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    38..162
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(203..508)
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     complement(305..324)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
ORIGIN
        1 cggccggccg ccccgactag cttggccgcc gaggccaacc actttgtaca agaaagctga
       61 acgagaaacg taaaatgata taaatatcaa tatattaaat tagattttgc ataaaaaaca
      121 gactacataa tactgtaaaa cacaacatat ccagtcacta tggtcgacct gcagactggc
      181 tgtgtataag ggagcctgac atttatattc cccagaacat caggttaatg gcgtttttga
      241 tgtcattttc gcggtggctg agatcagcca cttcttcccc gataacggag accggcacac
      301 tggccatatc ggtggtcatc atgcgccagc tttcatcccc gatatgcacc accgggtaaa
      361 gttcacggga gactttatct gacagcagac gtgcactggc cagggggatc accatccgtc
      421 gcccgggcgt gtcaataata tcactctgta catccacaaa cagacgataa cggctctctc
      481 ttttataggt gtaaacctta aactgcattt caccagcccc tgttctcgtc agcaaaagag
      541 ccgttcattt caataaaccg ggcgacctca gccatccctt cctgattttc cgctttccag
      601 cgttcggcac gcagacgacg ggcttcattc tgcatggttg tgcttaccag accggagata
      661 ttgacatcat atatgccttg agcaactgat agctgtcgct gtcaactgtc actgtaatac
      721 gctgcttcat agcatacctc tttttgacat acttcgggta tacatatcag tatatattct
      781 tataccgca
//
