LOCUS       sequence-83417-e        3973 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent reporter control for pMPRA1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3973)
  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 3973)
  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: pMPRA1-CMV-Clover
FEATURES             Location/Qualifiers
     source          1..3973
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        14..392
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        393..596
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     547..567
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     641..660
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        641..659
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(725..727,728..730,731..1441)
                     /codon_start=1
                     /product="bright green-yellow fluorescent protein derived 
                     from GFP (Lam et al., 2012)"
                     /label=Clover
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGVACFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSHQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(770..791)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     1378..1399
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        complement(1457..1475)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1458..1475)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     polyA_signal    1533..1654
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1544..1563)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1598..1617
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1902..1919)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2073..2660)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2153..2172)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2860..3651,3652..3720))
                     /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    3820..3868
                     /note="synthetic polyadenylation signal"
     misc_feature    3882..3973
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     3922..3941
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gccacattga ttattgacta gttattaata gtaatcaatt acggggtcat
       61 tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg
      121 gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa
      181 cgccaatagg gactttccat tgacgtcaat gggtggacta tttacggtaa actgcccact
      241 tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta
      301 aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt
      361 acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg
      421 ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg
      481 ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc
      541 cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctctct
      601 ggctaactag agaacccact gcttactggc ttatcgaaat taatacgact cactataggg
      661 agacccaagc ttggtaccga gctcggatcc actagtaacg gccgccagtg tgctggaatt
      721 cggcatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
      781 ggacggcgac gtaaacggcc acaagttcag cgtccgcggc gagggcgagg gcgatgccac
      841 caacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
      901 caccctcgtg accaccttcg gctacggcgt ggcctgcttc agccgctacc ccgaccacat
      961 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
     1021 ctctttcaag gacgacggta cctacaagac ccgcgccgag gtgaagttcg agggcgacac
     1081 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     1141 gcacaagctg gagtacaact tcaacagcca caacgtctat atcacggccg acaagcagaa
     1201 gaacggcatc aaggctaact tcaagatccg ccacaacgtt gaggacggca gcgtgcagct
     1261 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     1321 ccactacctg agccatcagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
     1381 ggtcctgctg gagttcgtga ccgccgccgg gattacacat ggcatggacg agctgtacaa
     1441 gtctagaggg ccctattcta tagtgtcacc taaatgctag ggcctcggcg gccaagctag
     1501 tcggggcggc cggccgcttc gagcagacat gataagatac attgatgagt ttggacaaac
     1561 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     1621 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     1681 gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg
     1741 tggtaaaatc gataaggatc cgtcgaccga tgcccttgag agccttcaac ccagtcagct
     1801 ccttccggtg ggcgcggggc atgactatcg tcgccgcact tatgactgtc ttctttatca
     1861 tgcaactcgt aggacaggtg ccggcagcgc tcttccgctt cctcgctcac tgactcgctg
     1921 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     1981 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     2041 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     2101 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     2161 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     2221 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     2281 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     2341 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     2401 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     2461 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     2521 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     2581 tccggcaaac aaaccaccgc tggtagcggt ggttttttgt ttgcaagcag cagattacgc
     2641 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     2701 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     2761 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     2821 ggtctgacag cggccgcaaa tgctaaacca ctgcagtggt taccagtgct tgatcagtga
     2881 ggcaccgatc tcagcgatct gcctatttcg ttcgtccata gtggcctgac tccccgtcgt
     2941 gtagatcact acgattcgtg agggcttacc atcaggcccc agcgcagcaa tgatgccgcg
     3001 agagccgcgt tcaccggccc ccgatttgtc agcaatgaac cagccagcag ggagggccga
     3061 gcgaagaagt ggtcctgcta ctttgtccgc ctccatccag tctatgagct gctgtcgtga
     3121 tgctagagta agaagttcgc cagtgagtag tttccgaaga gttgtggcca ttgctactgg
     3181 catcgtggta tcacgctcgt cgttcggtat ggcttcgttc aactctggtt cccagcggtc
     3241 aagccgggtc acatgatcac ccatattatg aagaaatgca gtcagctcct tagggcctcc
     3301 gatcgttgtc agaagtaagt tggccgcggt gttgtcgctc atggtaatgg cagcactaca
     3361 caattctctt accgtcatgc catccgtaag atgcttttcc gtgaccggcg agtactcaac
     3421 caagtcgttt tgtgagtagt gtatacggcg accaagctgc tcttgcccgg cgtctatacg
     3481 ggacaacacc gcgccacata gcagtacttt gaaagtgctc atcatcggga atcgttcttc
     3541 ggggcggaaa gactcaagga tcttgccgct attgagatcc agttcgatat agcccactct
     3601 tgcacccagt tgatcttcag catcttttac tttcaccagc gtttcggggt gtgcaaaaac
     3661 aggcaagcaa aatgccgcaa agaagggaat gagtgcgaca cgaaaatgtt ggatgctcat
     3721 tcctttttca atattattga agcatttatc agggttacta gtacgtctct caaggataag
     3781 taagtaatat taaggtacgg gaggtattgg acaggccgca ataaaatatc tttattttca
     3841 ttacatctgt gtgttggttt tttgtgtgaa tcgatagtac taacatacgc tctccatcaa
     3901 aacaaaacga aacaaaacaa actagcaaaa taggctgtcc ccagtgcaag tgcaggtgcc
     3961 agaacatttc tct
//
