LOCUS       sequence-352051-        3356 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MPRA v3 minimal promoter with GFP donor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3356)
  AUTHORS   Tewhey R, Kotliar D, Park DS, Liu B, Winnicki S, Reilly SK, Andersen
            KG, Mikkelsen TS, Lander ES, Schaffner SF, Sabeti PC
  TITLE     Direct Identification of Hundreds of Expression-Modulating Variants 
            using a Multiplexed Reporter Assay.
  JOURNAL   Cell. 2016 Jun 2;165(6):1519-29. doi: 10.1016/j.cell.2016.04.027.
  PUBMED    27259153
REFERENCE   2  (bases 1 to 3356)
  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.2016.04"; journalName: "Cell"; date: "2016-06-2- 2";
            volume: "165"; issue: "6"; pages: "1519-29"
COMMENT     Sequence Label: pMPRAv3:minP-GFP
FEATURES             Location/Qualifiers
     source          1..3356
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(16..604)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(96..115)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(804..1595,1596..1664))
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     polyA_signal    1769..1817
                     /note="synthetic polyadenylation signal"
     misc_feature    1831..1922
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     1871..1890
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        1999..2030
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      2057..2066
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2063..2065,2066..2068,2069..2782)
                     /codon_start=1
                     /product="Emerald GFP"
                     /label=EmGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2108..2129)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2369..2388)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2716..2737
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2832..2953
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2843..2862)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2897..2916
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3201..3218)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
       61 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
      121 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
      181 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
      241 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
      301 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
      361 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
      421 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
      481 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
      541 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
      601 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
      661 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
      721 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagcggccg
      781 caaatgctaa accactgcag tggttaccag tgcttgatca gtgaggcacc gatctcagcg
      841 atctgcctat ttcgttcgtc catagtggcc tgactccccg tcgtgtagat cactacgatt
      901 cgtgagggct taccatcagg ccccagcgca gcaatgatgc cgcgagagcc gcgttcaccg
      961 gcccccgatt tgtcagcaat gaaccagcca gcagggaggg ccgagcgaag aagtggtcct
     1021 gctactttgt ccgcctccat ccagtctatg agctgctgtc gtgatgctag agtaagaagt
     1081 tcgccagtga gtagtttccg aagagttgtg gccattgcta ctggcatcgt ggtatcacgc
     1141 tcgtcgttcg gtatggcttc gttcaactct ggttcccagc ggtcaagccg ggtcacatga
     1201 tcacccatat tatgaagaaa tgcagtcagc tccttagggc ctccgatcgt tgtcagaagt
     1261 aagttggccg cggtgttgtc gctcatggta atggcagcac tacacaattc tcttaccgtc
     1321 atgccatccg taagatgctt ttccgtgacc ggcgagtact caaccaagtc gttttgtgag
     1381 tagtgtatac ggcgaccaag ctgctcttgc ccggcgtcta tacgggacaa caccgcgcca
     1441 catagcagta ctttgaaagt gctcatcatc gggaatcgtt cttcggggcg gaaagactca
     1501 aggatcttgc cgctattgag atccagttcg atatagccca ctcttgcacc cagttgatct
     1561 tcagcatctt ttactttcac cagcgtttcg gggtgtgcaa aaacaggcaa gcaaaatgcc
     1621 gcaaagaagg gaatgagtgc gacacgaaaa tgttggatgc tcatactcgt cctttttcaa
     1681 tattattgaa gcatttatca gggttactag tacgtctctc aaggataagt aagtaatatt
     1741 aaggtacggg aggtattgga caggccgcaa taaaatatct ttattttcat tacatctgtg
     1801 tgttggtttt ttgtgtgaat cgatagtact aacatacgct ctccatcaaa acaaaacgaa
     1861 acaaaacaaa ctagcaaaat aggctgtccc cagtgcaagt gcaggtgcca gaacatttct
     1921 ctggcctaac tggccggtac ctgagctcgc tagcctcgag gatatcaaga tctggcctcg
     1981 gcggccaagc ttagacacta gagggtatat aatggaagct cgacttccag cttggcaatc
     2041 cggtactgtt ggtaaagcca ccatggtgag caagggcgag gagctgttca ccggggtggt
     2101 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
     2161 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
     2221 gctgcccgtg ccctggccca ccctcgtgac caccttgacc tacggcgtgc agtgcttcgc
     2281 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
     2341 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
     2401 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
     2461 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca aggtctatat
     2521 caccgccgac aagcagaaga acggcatcaa ggtgaacttc aagacccgcc acaacatcga
     2581 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     2641 cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa
     2701 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
     2761 catggacgag ctgtacaagt aatgataata attctagagt cggggcggcc ggccgcttcg
     2821 agcagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     2881 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     2941 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt
     3001 gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtaaaatcg ataaggatcc
     3061 gtcgaccgat gcccttgaga gccttcaacc cagtcagctc cttccggtgg gcgcggggca
     3121 tgactatcgt cgccgcactt atgactgtct tctttatcat gcaactcgta ggacaggtgc
     3181 cggcagcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     3241 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     3301 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggc
//
