LOCUS       sequence-183852-        4226 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Luciferase validation vector with SV40 enhancer inserted downstream 
            of the reporter gene. The candidate's enhancer activity is reflected
            by luciferase activity.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4226)
  AUTHORS   Muerdter F, Boryn LM, Woodfin AR, Neumayr C, Rath M, Zabidi MA, 
            Pagani M, Haberle V, Kazmar T, Catarino RR, Schernhuber K, Arnold 
            CD, Stark A
  TITLE     Resolving systematic errors in widely used enhancer activity assays 
            in human cells.
  JOURNAL   Nat Methods. 2017 Dec 11. pii: nmeth.4534. doi: 10.1038/nmeth.4534.
  PUBMED    29256496
REFERENCE   2  (bases 1 to 4226)
  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: "Nat 
            Methods. 2017 Dec 11. pii: nmeth.4534. doi: 10.1038/nmeth.4534."
COMMENT     Sequence Label: STARR-seq luciferase validation vector_mP_SV40
FEATURES             Location/Qualifiers
     source          1..4226
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        28..58
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      101..110
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             107..1759
                     /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"
     polyA_signal    1803..1924
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1814..1833)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1868..1887
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2032..2052)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(2320..2908)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2400..2419)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3108..3899,3900..3968))
                     /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    4073..4121
                     /note="synthetic polyadenylation signal"
     misc_feature    4135..4226
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4175..4194
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gcttagagat cttcgagaga gggtatataa tggaagctcg acttccaggg
       61 ctagcgacct aaagcttggc aatccggtac tgttggtaaa gccaccatgg aagatgccaa
      121 aaacattaag aagggcccag cgccattcta cccactcgaa gacgggaccg ccggcgagca
      181 gctgcacaaa gccatgaagc gctacgccct ggtgcccggc accatcgcct ttaccgacgc
      241 acatatcgag gtggacatta cctacgccga gtacttcgag atgagcgttc ggctggcaga
      301 agctatgaag cgctatgggc tgaatacaaa ccatcggatc gtggtgtgca gcgagaatag
      361 cttgcagttc ttcatgcccg tgttgggtgc cctgttcatc ggtgtggctg tggccccagc
      421 taacgacatc tacaacgagc gcgagctgct gaacagcatg ggcatcagcc agcccaccgt
      481 cgtattcgtg agcaagaaag ggctgcaaaa gatcctcaac gtgcaaaaga agctaccgat
      541 catacaaaag atcatcatca tggatagcaa gaccgactac cagggcttcc aaagcatgta
      601 caccttcgtg acttcccatt tgccacccgg cttcaacgag tacgacttcg tgcccgagag
      661 cttcgaccgg gacaaaacca tcgccctgat catgaacagt agtggcagta ccggattgcc
      721 caagggcgta gccctaccgc accgcaccgc ttgtgtccga ttcagtcatg cccgcgaccc
      781 catcttcggc aaccagatca tccccgacac cgctatcctc agcgtggtgc catttcacca
      841 cggcttcggc atgttcacca cgctgggcta cttgatctgc ggctttcggg tcgtgctcat
      901 gtaccgcttc gaggaggagc tattcttgcg cagcttgcaa gactataaga ttcaatctgc
      961 cctgctggtg cccacactat ttagcttctt cgctaagagc actctcatcg acaagtacga
     1021 cctaagcaac ttgcacgaga tcgccagcgg cggggcgccg ctcagcaagg aggtaggtga
     1081 ggccgtggcc aaacgcttcc acctaccagg catccgccag ggctacggcc tgacagaaac
     1141 aaccagcgcc attctgatca cccccgaagg ggacgacaag cctggcgcag taggcaaggt
     1201 ggtgcccttc ttcgaggcta aggtggtgga cttggacacc ggtaagacac tgggtgtgaa
     1261 ccagcgcggc gagctgtgcg tccgtggccc catgatcatg agcggctacg ttaacaaccc
     1321 cgaggctaca aacgctctca tcgacaagga cggctggctg cacagcggcg acatcgccta
     1381 ctgggacgag gacgagcact tcttcatcgt ggaccggctg aagagcctga tcaaatacaa
     1441 gggctaccag gtagccccag ccgaactgga gagcatcctg ctgcaacacc ccaacatctt
     1501 cgacgccggg gtcgccggcc tgcccgacga cgatgccggc gagctgcccg ccgcagtcgt
     1561 cgtgctggaa cacggtaaaa ccatgaccga gaaggagatc gtggactatg tggccagcca
     1621 ggttacaacc gccaagaagc tgcgcggtgg tgttgtgttc gtggacgagg tgcctaaagg
     1681 actgaccggc aagttggacg cccgcaagat ccgcgagatt ctcattaagg ccaagaaggg
     1741 cggcaagatc gccgtgtaat aattctagag tcggggcggc cggccgcttc gagcagacat
     1801 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     1861 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     1921 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt
     1981 tttttaaagc aagtaaaacc tctacaaatg tggtaaaatc gataaggctg tggaatgtgt
     2041 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     2101 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     2161 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     2221 cgcccctaac tccgcccagt tccgcccatt ctccgctcca tcgtgagcaa aaggccagca
     2281 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     2341 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     2401 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     2461 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     2521 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     2581 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     2641 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     2701 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     2761 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     2821 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     2881 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     2941 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     3001 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     3061 gtaaacttgg tctgacagcg gccgcaaatg ctaaaccact gcagtggtta ccagtgcttg
     3121 atcagtgagg caccgatctc agcgatctgc ctatttcgtt cgtccatagt ggcctgactc
     3181 cccgtcgtgt agatcactac gattcgtgag ggcttaccat caggccccag cgcagcaatg
     3241 atgccgcgag agccgcgttc accggccccc gatttgtcag caatgaacca gccagcaggg
     3301 agggccgagc gaagaagtgg tcctgctact ttgtccgcct ccatccagtc tatgagctgc
     3361 tgtcgtgatg ctagagtaag aagttcgcca gtgagtagtt tccgaagagt tgtggccatt
     3421 gctactggca tcgtggtatc acgctcgtcg ttcggtatgg cttcgttcaa ctctggttcc
     3481 cagcggtcaa gccgggtcac atgatcaccc atattatgaa gaaatgcagt cagctcctta
     3541 gggcctccga tcgttgtcag aagtaagttg gccgcggtgt tgtcgctcat ggtaatggca
     3601 gcactacaca attctcttac cgtcatgcca tccgtaagat gcttttccgt gaccggcgag
     3661 tactcaacca agtcgttttg tgagtagtgt atacggcgac caagctgctc ttgcccggcg
     3721 tctatacggg acaacaccgc gccacatagc agtactttga aagtgctcat catcgggaat
     3781 cgttcttcgg ggcggaaaga ctcaaggatc ttgccgctat tgagatccag ttcgatatag
     3841 cccactcttg cacccagttg atcttcagca tcttttactt tcaccagcgt ttcggggtgt
     3901 gcaaaaacag gcaagcaaaa tgccgcaaag aagggaatga gtgcgacacg aaaatgttgg
     3961 atgctcatac tcgtcctttt tcaatattat tgaagcattt atcagggtta ctagtacgtc
     4021 tctcaaggat aagtaagtaa tattaaggta cgggaggtat tggacaggcc gcaataaaat
     4081 atctttattt tcattacatc tgtgtgttgg ttttttgtgt gaatcgatag tactaacata
     4141 cgctctccat caaaacaaaa cgaaacaaaa caaactagca aaataggctg tccccagtgc
     4201 aagtgcaggt gccagaacat ttctct
//
