LOCUS       sequence-75517-e         966 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 966)
  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 966)
  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..966
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    46..170
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(195..200,201..218,219..225)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             279..938
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(345..364)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 ttctctggcc taactggcca gtgtggtgga attctgcaga tatcaacaag tttgtacaaa
       61 aaagctgaac gagaaacgta aaatgatata aatatcaata tattaaatta gattttgcat
      121 aaaaaacaga ctacataata ctgtaaaaca caacatatcc agtcactatg gcggccgcat
      181 taggcacccc aggctttaca ctttatgctt ccggctcgta taatgtgtgg attttgagtt
      241 aggatccgtc gagattttca ggagctaagg aagctaaaat ggagaaaaaa atcactggat
      301 ataccaccgt tgatatatcc caatggcatc gtaaagaaca ttttgaggca tttcagtcag
      361 ttgctcaatg tacctataac cagaccgttc agctggatat tacggccttt ttaaagaccg
      421 taaagaaaaa taagcacaag ttttatccgg cctttattca cattcttgcc cgcctgatga
      481 atgctcatcc ggaattccgt atggcaatga aagacggtga gctggtgata tgggatagtg
      541 ttcacccttg ttacaccgtt ttccatgagc aaactgaaac gttttcatcg ctctggagtg
      601 aataccacga cgatttccgg cagtttctac acatatattc gcaagatgtg gcgtgttacg
      661 gtgaaaacct ggcctatttc cctaaagggt ttattgagaa tatgtttttc gtctcagcca
      721 atccctgggt gagtttcacc agttttgatt taaacgtggc caatatggac aacttcttcg
      781 cccccgtttt caccatgggc aaatattata cgcaaggcga caaggtgctg atgccgctgg
      841 cgattcaggt tcatcatgcc gtttgtgatg gcttccatgt cggcagaatg cttaatgaat
      901 tacaacagta ctgcgatgag tggcagggcg gggcgtaaag atctggatcc ggcttactaa
      961 aagcca
//
