LOCUS       sequence-353743-        4019 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MPRAu backbone vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4019)
  AUTHORS   Griesemer D, Xue JR, Reilly SK, Ulirsch JC, Kukreja K, Davis JR, 
            Kanai M, Yang DK, Butts JC, Guney MH, Luban J, Montgomery SB, 
            Finucane HK, Novina CD, Tewhey R, Sabeti PC
  TITLE     Genome-wide functional screen of 3'UTR variants uncovers causal 
            variants for human disease and evolution.
  JOURNAL   Cell. 2021 Sep 30;184(20):5247-5260.e19. doi: 
            10.1016/j.cell.2021.08.025. Epub 2021 Sep 16.
  PUBMED    34534445
REFERENCE   2  (bases 1 to 4019)
  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"; doi: 
            "10.1016/j.cell.2021.08.025"; journalName: "Cell"; date: 
            "2021-09-30- 30"; volume: "184"; issue: "20"; pages: "5247-5260.e19"
COMMENT     Sequence Label: pmirGLO:Delta-luc::gfp Delta-AmpR::KanR
FEATURES             Location/Qualifiers
     source          1..4019
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(267..855)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(347..366)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    971..1254
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
     promoter        1311..1810
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(1346..1369)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     1743..1762
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      1851..1860
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1857..1859,1860..1862,1863..2576)
                     /codon_start=1
                     /product="Emerald GFP"
                     /label=EmGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1902..1923)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2163..2182)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2510..2531
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2759..2880
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2770..2789)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2824..2843
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3208..3312
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3313..4019,1..109)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSKPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(3386..3405)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc
       61 tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttctaat tcgaaatgac
      121 cgaccaagcg acgcccaacc ggtatcagct cactcaaagg cggtaatacg gttatccaca
      181 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
      241 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
      301 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
      361 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
      421 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
      481 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
      541 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
      601 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
      661 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
      721 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
      781 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
      841 aaaaaaggat ttcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
      901 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
      961 cttttatagt ccggaaatac aggaacgcac gctggatggc ccttcgctgg gatggtgaaa
     1021 ccatgaaaaa tggcagcttc agtggattaa gtgggggtaa tgtggcctgt accctctggt
     1081 tgcataggta ttcatacggt taaaatttat caggcgcgat tgcggcagtt tttcgggtgg
     1141 tttgttgcca tttttacctg tctgctgccg tgatcgcgct gaacgcgttt tagcggtgcg
     1201 tacaattaag ggattatggt aaatccactt actgtctgcc ctcgtagcca tcgagataaa
     1261 ccgcagtact ccggccacga tgcgtccggc gtagaggatc gagatctacc gggtagggga
     1321 ggcgcttttc ccaaggcagt ctggagcatg cgctttagca gccccgctgg gcacttggcg
     1381 ctacacaagt ggcctctggc ctcgcacaca ttccacatcc accggtaggc gccaaccggc
     1441 tccgttcttt ggtggcccct tcgcgccacc ttctactcct cccctagtca ggaagttccc
     1501 ccccgccccg cagctcgcgt cgtgcaggac gtgacaaatg gaagtagcac gtctcactag
     1561 tctcgtgcag atggacagca ccgctgagca atggaagcgg gtaggccttt ggggcagcgg
     1621 ccaatagcag ctttgctcct tcgctttctg ggctcagagg ctgggaaggg gtgggtccgg
     1681 gggcgggctc aggggcgggc tcaggggcgg ggcgggcgcc cgaaggtcct ccggaggccc
     1741 ggcattctgc acgcttcaaa agcgcacgtc tgccgcgctg ttctcctctt cctcatctcc
     1801 gggcctttcg acctgcagcc caagcttggc aatccggtac tgttggtaaa gccaccatgg
     1861 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
     1921 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
     1981 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
     2041 tgaccacctt gacctacggc gtgcagtgct tcgcccgcta ccccgaccac atgaagcagc
     2101 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
     2161 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
     2221 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
     2281 tggagtacaa ctacaacagc cacaaggtct atatcaccgc cgacaagcag aagaacggca
     2341 tcaaggtgaa cttcaagacc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc
     2401 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     2461 tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     2521 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaattct
     2581 agttgtttaa agcccaacgc tagcctcgag tctagagatc ggaagagcgt cgtcgacctg
     2641 caggcatgca agctgatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg
     2701 ccaccgctga gcaataacta gcataacccc ttggggcggc cgcttcgagc agacatgata
     2761 agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt
     2821 tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt
     2881 aacaacaaca attgcattca ttttatgttt caggttcagg gggagatgtg ggaggttttt
     2941 ttaagcaagt aaaacctcta caaatgtggt aaaatcgaat tttaacaaaa tattaacgct
     3001 tacaatttcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     3061 cgcggatcta tcctctttgc gcttgcgttt tcccttgtcc agatagccca gtagctgaca
     3121 ttcatccggg gtcagcaccg tttctgcgga ctggctttct acgtaatggt ttcttagacg
     3181 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     3241 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     3301 aaaaggaaga gtatgagcca tattcaacgg gaaacgtctt gctcgaagcc gcgattaaat
     3361 tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt cgggcaatca
     3421 ggtgcgacaa tctatcgatt gtatgggaag cccgatgcgc cagagttgtt tctgaaacat
     3481 ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa ctggctgacg
     3541 gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga tgcatggtta
     3601 ctcaccactg cgatccccgg gaaaacagca ttccaggtat tagaagaata tcctgattca
     3661 ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt
     3721 tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca atcacgaatg
     3781 aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg gcctgttgaa
     3841 caagtctgga aagaaatgca taagcttttg ccattctcac cggattcagt cgtcactcat
     3901 ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg ttgtattgat
     3961 gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg gaactgcct
//
