LOCUS       sequence-193009-        4226 bp    DNA     circular SYN 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      synthetic DNA construct
  ORGANISM  synthetic DNA construct
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="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(271..859)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(351..370)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1059..1850,1851..1919))
                     /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    2024..2072
                     /note="synthetic polyadenylation signal"
     misc_feature    2086..2177
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     2126..2145
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        2205..2235
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      2278..2287
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2284..3936
                     /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    3980..4101
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3991..4010)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4045..4064
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(join(4209..4226,1..3))
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
       61 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
      121 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
      181 ctccgcccag ttccgcccat tctccgctcc atcgtgagca aaaggccagc aaaaggccag
      241 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
      301 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
      361 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
      421 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
      481 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
      541 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
      601 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
      661 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
      721 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
      781 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
      841 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
      901 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
      961 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     1021 gtctgacagc ggccgcaaat gctaaaccac tgcagtggtt accagtgctt gatcagtgag
     1081 gcaccgatct cagcgatctg cctatttcgt tcgtccatag tggcctgact ccccgtcgtg
     1141 tagatcacta cgattcgtga gggcttacca tcaggcccca gcgcagcaat gatgccgcga
     1201 gagccgcgtt caccggcccc cgatttgtca gcaatgaacc agccagcagg gagggccgag
     1261 cgaagaagtg gtcctgctac tttgtccgcc tccatccagt ctatgagctg ctgtcgtgat
     1321 gctagagtaa gaagttcgcc agtgagtagt ttccgaagag ttgtggccat tgctactggc
     1381 atcgtggtat cacgctcgtc gttcggtatg gcttcgttca actctggttc ccagcggtca
     1441 agccgggtca catgatcacc catattatga agaaatgcag tcagctcctt agggcctccg
     1501 atcgttgtca gaagtaagtt ggccgcggtg ttgtcgctca tggtaatggc agcactacac
     1561 aattctctta ccgtcatgcc atccgtaaga tgcttttccg tgaccggcga gtactcaacc
     1621 aagtcgtttt gtgagtagtg tatacggcga ccaagctgct cttgcccggc gtctatacgg
     1681 gacaacaccg cgccacatag cagtactttg aaagtgctca tcatcgggaa tcgttcttcg
     1741 gggcggaaag actcaaggat cttgccgcta ttgagatcca gttcgatata gcccactctt
     1801 gcacccagtt gatcttcagc atcttttact ttcaccagcg tttcggggtg tgcaaaaaca
     1861 ggcaagcaaa atgccgcaaa gaagggaatg agtgcgacac gaaaatgttg gatgctcata
     1921 ctcgtccttt ttcaatatta ttgaagcatt tatcagggtt actagtacgt ctctcaagga
     1981 taagtaagta atattaaggt acgggaggta ttggacaggc cgcaataaaa tatctttatt
     2041 ttcattacat ctgtgtgttg gttttttgtg tgaatcgata gtactaacat acgctctcca
     2101 tcaaaacaaa acgaaacaaa acaaactagc aaaataggct gtccccagtg caagtgcagg
     2161 tgccagaaca tttctctggc ctaactggct tagagatctt cgagagaggg tatataatgg
     2221 aagctcgact tccagggcta gcgacctaaa gcttggcaat ccggtactgt tggtaaagcc
     2281 accatggaag atgccaaaaa cattaagaag ggcccagcgc cattctaccc actcgaagac
     2341 gggaccgccg gcgagcagct gcacaaagcc atgaagcgct acgccctggt gcccggcacc
     2401 atcgccttta ccgacgcaca tatcgaggtg gacattacct acgccgagta cttcgagatg
     2461 agcgttcggc tggcagaagc tatgaagcgc tatgggctga atacaaacca tcggatcgtg
     2521 gtgtgcagcg agaatagctt gcagttcttc atgcccgtgt tgggtgccct gttcatcggt
     2581 gtggctgtgg ccccagctaa cgacatctac aacgagcgcg agctgctgaa cagcatgggc
     2641 atcagccagc ccaccgtcgt attcgtgagc aagaaagggc tgcaaaagat cctcaacgtg
     2701 caaaagaagc taccgatcat acaaaagatc atcatcatgg atagcaagac cgactaccag
     2761 ggcttccaaa gcatgtacac cttcgtgact tcccatttgc cacccggctt caacgagtac
     2821 gacttcgtgc ccgagagctt cgaccgggac aaaaccatcg ccctgatcat gaacagtagt
     2881 ggcagtaccg gattgcccaa gggcgtagcc ctaccgcacc gcaccgcttg tgtccgattc
     2941 agtcatgccc gcgaccccat cttcggcaac cagatcatcc ccgacaccgc tatcctcagc
     3001 gtggtgccat ttcaccacgg cttcggcatg ttcaccacgc tgggctactt gatctgcggc
     3061 tttcgggtcg tgctcatgta ccgcttcgag gaggagctat tcttgcgcag cttgcaagac
     3121 tataagattc aatctgccct gctggtgccc acactattta gcttcttcgc taagagcact
     3181 ctcatcgaca agtacgacct aagcaacttg cacgagatcg ccagcggcgg ggcgccgctc
     3241 agcaaggagg taggtgaggc cgtggccaaa cgcttccacc taccaggcat ccgccagggc
     3301 tacggcctga cagaaacaac cagcgccatt ctgatcaccc ccgaagggga cgacaagcct
     3361 ggcgcagtag gcaaggtggt gcccttcttc gaggctaagg tggtggactt ggacaccggt
     3421 aagacactgg gtgtgaacca gcgcggcgag ctgtgcgtcc gtggccccat gatcatgagc
     3481 ggctacgtta acaaccccga ggctacaaac gctctcatcg acaaggacgg ctggctgcac
     3541 agcggcgaca tcgcctactg ggacgaggac gagcacttct tcatcgtgga ccggctgaag
     3601 agcctgatca aatacaaggg ctaccaggta gccccagccg aactggagag catcctgctg
     3661 caacacccca acatcttcga cgccggggtc gccggcctgc ccgacgacga tgccggcgag
     3721 ctgcccgccg cagtcgtcgt gctggaacac ggtaaaacca tgaccgagaa ggagatcgtg
     3781 gactatgtgg ccagccaggt tacaaccgcc aagaagctgc gcggtggtgt tgtgttcgtg
     3841 gacgaggtgc ctaaaggact gaccggcaag ttggacgccc gcaagatccg cgagattctc
     3901 attaaggcca agaagggcgg caagatcgcc gtgtaataat tctagagtcg gggcggccgg
     3961 ccgcttcgag cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
     4021 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
     4081 ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag
     4141 ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg taaaatcgat
     4201 aaggctgtgg aatgtgtgtc agttag
//
