LOCUS       sequence-451375-        6294 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent reporter control for pMPRA3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6294)
  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 6294)
  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: pMPRA3-CMV-Clover
FEATURES             Location/Qualifiers
     source          1..6294
                     /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"
     promoter        1811..2168
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1811..2002
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1834..1854)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2019..2154
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2081..2100
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     regulatory      2193..2202
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3245..3293
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(3440..3457)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3611..4199)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3691..3710)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4399..5190,5191..5259))
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     mobile_element  5409..6176
                     /mobile_element_type="insertion sequence:IS1"
                     /label=IS1
                     /note="prokaryotic transposable element"
     misc_feature    6203..6294
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     6243..6262
                     /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 gataagcgca gcaccatggc ctgaaataac ctctgaaaga ggaacttggt
     1801 tagctacctt ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa
     1861 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     1921 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     1981 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     2041 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     2101 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     2161 tttgcaaaaa gctcgattct tctgacacta gcgccaccat ggcaccctgc atgctgctcc
     2221 tgctgttggc ggctgccctg gccccgactc agacccgcgc gggcccacat tcgctgaggt
     2281 atttccacac cgccgtgtcc cggcccggcc tcgggaagcc ccggttcatc tctgtcggct
     2341 acgtggacga cacgcagttc gtgcgcttcg acagcgacgc ggagaatccg aggtatgagc
     2401 cgcgggtgcg gtggatggag caggtggagc ccgagtattg ggagcggaac acgcagatcg
     2461 ccaagggcaa tgagcagatt ttccgagtga acctgaggac cgcgctgcgc tactacaacc
     2521 agagcgcggg cggctctcac acgttccaac ggatgtacgg ctgtgaggtg gggtcggact
     2581 ggcgcctcct ccgcgggtac gagcagtacg catacgacgg ctgcgattac atcgccctga
     2641 acgaagacct gaaaacgtgg acggcggccg acatggcggc gctgatcacc aaacacaagt
     2701 gggagcaggc tggtgatgca gagagagacc gggcctacct ggagggcacg tgcgtggagt
     2761 ggctccgcag atacctgcag ctcgggaacg cgacgctgcc gcgcacagat tccccaaagg
     2821 cccatgtgac ccgtcacagc agacctgaag ataaagtcac cctgaggtgc tgggccctgg
     2881 gcttctaccc tgctgacatc accctgacct ggcagttgaa tggggaggag ctgacccagg
     2941 acatggagct tgtggagacc aggcctgcag gggatggaac cttccagaag tgggcatctg
     3001 tggtggtgcc tcttgggaag gagcagtatt acacatgcca tgtgtaccat caggggctgc
     3061 ctgagcccct caccctgaga tgggagcctc ctccatccac tgtctccaac acggtaatca
     3121 ttgctgttct ggttgtcctt ggagctgcaa tagtcactgg agctgtggtg gcttttgtga
     3181 tgaagatgag aaggagaaac acaggtggaa aaggagggtg aactcttgaa ccggtcatgg
     3241 ccgcaataaa atatctttat tttcattaca tctgtgtgtt ggttttttgt gtgggatccg
     3301 tcgaccgatg cccttgagag ccttcaaccc agtcagctcc ttccggtggg cgcggggcat
     3361 gactatcgtc gccgcactta tgactgtctt ctttatcatg caactcgtag gacaggtgcc
     3421 ggcagcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     3481 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     3541 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     3601 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     3661 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     3721 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     3781 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     3841 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     3901 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     3961 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     4021 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     4081 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     4141 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     4201 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     4261 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     4321 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagc ggccgcaaat
     4381 gctaaaccac tgcagtggtt accagtgctt gatcagtgag gcaccgatct cagcgatctg
     4441 cctatttcgt tcgtccatag tggcctgact ccccgtcgtg tagatcacta cgattcgtga
     4501 gggcttacca tcaggcccca gcgcagcaat gatgccgcga gagccgcgtt caccggcccc
     4561 cgatttgtca gcaatgaacc agccagcagg gagggccgag cgaagaagtg gtcctgctac
     4621 tttgtccgcc tccatccagt ctatgagctg ctgtcgtgat gctagagtaa gaagttcgcc
     4681 agtgagtagt ttccgaagag ttgtggccat tgctactggc atcgtggtat cacgctcgtc
     4741 gttcggtatg gcttcgttca actctggttc ccagcggtca agccgggtca catgatcacc
     4801 catattatga agaaatgcag tcagctcctt agggcctccg atcgttgtca gaagtaagtt
     4861 ggccgcggtg ttgtcgctca tggtaatggc agcactacac aattctctta ccgtcatgcc
     4921 atccgtaaga tgcttttccg tgaccggcga gtactcaacc aagtcgtttt gtgagtagtg
     4981 tatacggcga ccaagctgct cttgcccggc gtctatacgg gacaacaccg cgccacatag
     5041 cagtactttg aaagtgctca tcatcgggaa tcgttcttcg gggcggaaag actcaaggat
     5101 cttgccgcta ttgagatcca gttcgatata gcccactctt gcacccagtt gatcttcagc
     5161 atcttttact ttcaccagcg tttcggggtg tgcaaaaaca ggcaagcaaa atgccgcaaa
     5221 gaagggaatg agtgcgacac gaaaatgttg gatgctcata ctcgtccttt ttcaatatta
     5281 ttgaagcatt tatcagggtt actagtacgt ctctcaagga taagtaagta atattaaggt
     5341 acgggaggta ttggacaggc cgcaataaaa tatctttatt ttcattacat ctgtgtgttg
     5401 gttttttggg taatgactcc aacttattga tagtgtttta tgttcagata atgcccgatg
     5461 actttgtcat gcagctccac cgattttgag aacgacagcg acttccgtcc cagccgtgcc
     5521 aggtgctgcc tcagattcag gttatgccgc tcaattcgct gcgtatatcg cttgctgatt
     5581 acgtgcagct ttcccttcag gcgggattca tacagcggcc agccatccgt catccatatc
     5641 accacgtcaa agggtgacag caggctcata agacgcccca gcgtcgccat agtgcgttca
     5701 ccgaatacgt gcgcaacaac cgtcttccgg agactgtcat acgcgtaaaa cagccagcgc
     5761 tggcgcgatt tagccccgac atagccccac tgttcgtcca tttccgcgca gacgatgacg
     5821 tcactgcccg gctgtatgcg cgaggttacc gactgcggcc tgagtttttt aagtgacgta
     5881 aaatcgtgtt gaggccaacg cccataatgc gggctgttgc ccggcatcca acgccattca
     5941 tggccatatc aatgattttc tggtgcgtac cgggttgaga agcggtgtaa gtgaactgca
     6001 gttgccatgt tttacggcag tgagagcaga gatagcgctg atgtccggcg gtgcttttgc
     6061 cgttacgcac caccccgtca gtagctgaac aggagggaca gctgatagaa acagaagcca
     6121 ctggagcacc tcaaaaacac catcatacac taaatcagta agttggcagc atcaccggtt
     6181 ttttgtgtga atcgatagta ctaacatacg ctctccatca aaacaaaacg aaacaaaaca
     6241 aactagcaaa ataggctgtc cccagtgcaa gtgcaggtgc cagaacattt ctct
//
