LOCUS       sequence-73910-e        4633 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pSV40mod-luc2 donor plasmid for MPRA library construction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4633)
  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 4633)
  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: pMPRAdonor3
FEATURES             Location/Qualifiers
     source          1..4633
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        58..254
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      105..240
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      278..287
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             284..1936
                     /codon_start=1
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="synthetic luc2 version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(2005..2021)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2005..2021)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2018..2040)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2029..2045
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(2053..2059,2060..2077,2078..2083))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2098..2119
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2236..2253)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2407..2995)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2487..2506)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3166..3957,3958..4026))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3789..3808
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4027..4131)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4199..4217
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4255..4277)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4377..4396
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4590..4612
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4604..4621
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4605..4621
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 ggtacctgag ctcgctagcc tcgaggatat caagatctgg cctcggcggc caagctttgc
       61 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
      121 cgcccagttc cgcccattct ccgccccatc gctgactaat tttttttatt tatgcagagg
      181 ccgaagtcgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
      241 taggcttttg caaaggcaat ccggtactgt tggtaaagcc accatggaag atgccaaaaa
      301 cattaagaag ggcccagcgc cattctaccc actcgaagac gggaccgccg gcgagcagct
      361 gcacaaagcc atgaagcgct acgccctggt gcccggcacc atcgccttta ccgacgcaca
      421 tatcgaggtg gacattacct acgccgagta cttcgagatg agcgttcggc tggcagaagc
      481 tatgaagcgc tatgggctga atacaaacca tcggatcgtg gtgtgcagcg agaatagctt
      541 gcagttcttc atgcccgtgt tgggtgccct gttcatcggt gtggctgtgg ccccagctaa
      601 cgacatctac aacgagcgcg agctgctgaa cagcatgggc atcagccagc ccaccgtcgt
      661 attcgtgagc aagaaagggc tgcaaaagat cctcaacgtg caaaagaagc taccgatcat
      721 acaaaagatc atcatcatgg atagcaagac cgactaccag ggcttccaaa gcatgtacac
      781 cttcgtgact tcccatttgc cacccggctt caacgagtac gacttcgtgc ccgagagctt
      841 cgaccgggac aaaaccatcg ccctgatcat gaacagtagt ggcagtaccg gattgcccaa
      901 gggcgtagcc ctaccgcacc gcaccgcttg tgtccgattc agtcatgccc gcgaccccat
      961 cttcggcaac cagatcatcc ccgacaccgc tatcctcagc gtggtgccat ttcaccacgg
     1021 cttcggcatg ttcaccacgc tgggctactt gatctgcggc tttcgggtcg tgctcatgta
     1081 ccgcttcgag gaggagctat tcttgcgcag cttgcaagac tataagattc aatctgccct
     1141 gctggtgccc acactattta gcttcttcgc taagagcact ctcatcgaca agtacgacct
     1201 aagcaacttg cacgagatcg ccagcggcgg ggcgccgctc agcaaggagg taggtgaggc
     1261 cgtggccaaa cgcttccacc taccaggcat ccgccagggc tacggcctga cagaaacaac
     1321 cagcgccatt ctgatcaccc ccgaagggga cgacaagcct ggcgcagtag gcaaggtggt
     1381 gcccttcttc gaggctaagg tggtggactt ggacaccggt aagacactgg gtgtgaacca
     1441 gcgcggcgag ctgtgcgtcc gtggccccat gatcatgagc ggctacgtta acaaccccga
     1501 ggctacaaac gctctcatcg acaaggacgg ctggctgcac agcggcgaca tcgcctactg
     1561 ggacgaggac gagcacttct tcatcgtgga ccggctgaag agcctgatca aatacaaggg
     1621 ctaccaggta gccccagccg aactggagag catcctgctg caacacccca acatcttcga
     1681 cgccggggtc gccggcctgc ccgacgacga tgccggcgag ctgcccgccg cagtcgtcgt
     1741 gctggaacac ggtaaaacca tgaccgagaa ggagatcgtg gactatgtgg ccagccaggt
     1801 tacaaccgcc aagaagctgc gcggtggtgt tgtgttcgtg gacgaggtgc ctaaaggact
     1861 gaccggcaag ttggacgccc gcaagatccg cgagattctc attaaggcca agaagggcgg
     1921 caagatcgcc gtgtaataat tctagatatc ggatcccggg cccgtcgact gcagaggcct
     1981 gcatgcaagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     2041 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     2101 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     2161 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     2221 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     2281 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     2341 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     2401 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     2461 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     2521 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     2581 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     2641 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     2701 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     2761 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     2821 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     2881 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     2941 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     3001 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     3061 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     3121 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     3181 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     3241 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     3301 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     3361 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     3421 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     3481 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     3541 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     3601 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     3661 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     3721 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     3781 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     3841 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     3901 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     3961 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     4021 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     4081 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     4141 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     4201 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg
     4261 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     4321 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc
     4381 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt
     4441 aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg
     4501 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag
     4561 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag
     4621 tgaattcgag ctc
//
