LOCUS       sequence-199061-        4278 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 4278)
  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 4278)
  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_SCP1_SV40
FEATURES             Location/Qualifiers
     source          1..4278
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(140..160)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(428..1016)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(508..527)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1216..2007,2008..2076))
                     /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    2181..2229
                     /note="synthetic polyadenylation signal"
     misc_feature    2243..2334
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     2283..2302
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     regulatory      2487..2496
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2493..4145
                     /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    join(4189..4278,1..32)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4200..4219)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4254..4273
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt
       61 ttcaggttca gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg
      121 gtaaaatcga taaggctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
      181 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
      241 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
      301 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
      361 ccgctccatc gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
      421 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
      481 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
      541 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
      601 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
      661 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
      721 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
      781 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
      841 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
      901 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
      961 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     1021 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     1081 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     1141 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagcggc cgcaaatgct
     1201 aaaccactgc agtggttacc agtgcttgat cagtgaggca ccgatctcag cgatctgcct
     1261 atttcgttcg tccatagtgg cctgactccc cgtcgtgtag atcactacga ttcgtgaggg
     1321 cttaccatca ggccccagcg cagcaatgat gccgcgagag ccgcgttcac cggcccccga
     1381 tttgtcagca atgaaccagc cagcagggag ggccgagcga agaagtggtc ctgctacttt
     1441 gtccgcctcc atccagtcta tgagctgctg tcgtgatgct agagtaagaa gttcgccagt
     1501 gagtagtttc cgaagagttg tggccattgc tactggcatc gtggtatcac gctcgtcgtt
     1561 cggtatggct tcgttcaact ctggttccca gcggtcaagc cgggtcacat gatcacccat
     1621 attatgaaga aatgcagtca gctccttagg gcctccgatc gttgtcagaa gtaagttggc
     1681 cgcggtgttg tcgctcatgg taatggcagc actacacaat tctcttaccg tcatgccatc
     1741 cgtaagatgc ttttccgtga ccggcgagta ctcaaccaag tcgttttgtg agtagtgtat
     1801 acggcgacca agctgctctt gcccggcgtc tatacgggac aacaccgcgc cacatagcag
     1861 tactttgaaa gtgctcatca tcgggaatcg ttcttcgggg cggaaagact caaggatctt
     1921 gccgctattg agatccagtt cgatatagcc cactcttgca cccagttgat cttcagcatc
     1981 ttttactttc accagcgttt cggggtgtgc aaaaacaggc aagcaaaatg ccgcaaagaa
     2041 gggaatgagt gcgacacgaa aatgttggat gctcatactc gtcctttttc aatattattg
     2101 aagcatttat cagggttact agtacgtctc tcaaggataa gtaagtaata ttaaggtacg
     2161 ggaggtattg gacaggccgc aataaaatat ctttattttc attacatctg tgtgttggtt
     2221 ttttgtgtga atcgatagta ctaacatacg ctctccatca aaacaaaacg aaacaaaaca
     2281 aactagcaaa ataggctgtc cccagtgcaa gtgcaggtgc cagaacattt ctctggccta
     2341 actggcttag agatctccct agggacttat ataagggggt gggggcgcgt tcgtcctcag
     2401 tcgcgatcga acactcgagc cgagcagacg tgcctacgga cccgggctag cgacctaaag
     2461 cttggcaatc cggtactgtt ggtaaagcca ccatggaaga tgccaaaaac attaagaagg
     2521 gcccagcgcc attctaccca ctcgaagacg ggaccgccgg cgagcagctg cacaaagcca
     2581 tgaagcgcta cgccctggtg cccggcacca tcgcctttac cgacgcacat atcgaggtgg
     2641 acattaccta cgccgagtac ttcgagatga gcgttcggct ggcagaagct atgaagcgct
     2701 atgggctgaa tacaaaccat cggatcgtgg tgtgcagcga gaatagcttg cagttcttca
     2761 tgcccgtgtt gggtgccctg ttcatcggtg tggctgtggc cccagctaac gacatctaca
     2821 acgagcgcga gctgctgaac agcatgggca tcagccagcc caccgtcgta ttcgtgagca
     2881 agaaagggct gcaaaagatc ctcaacgtgc aaaagaagct accgatcata caaaagatca
     2941 tcatcatgga tagcaagacc gactaccagg gcttccaaag catgtacacc ttcgtgactt
     3001 cccatttgcc acccggcttc aacgagtacg acttcgtgcc cgagagcttc gaccgggaca
     3061 aaaccatcgc cctgatcatg aacagtagtg gcagtaccgg attgcccaag ggcgtagccc
     3121 taccgcaccg caccgcttgt gtccgattca gtcatgcccg cgaccccatc ttcggcaacc
     3181 agatcatccc cgacaccgct atcctcagcg tggtgccatt tcaccacggc ttcggcatgt
     3241 tcaccacgct gggctacttg atctgcggct ttcgggtcgt gctcatgtac cgcttcgagg
     3301 aggagctatt cttgcgcagc ttgcaagact ataagattca atctgccctg ctggtgccca
     3361 cactatttag cttcttcgct aagagcactc tcatcgacaa gtacgaccta agcaacttgc
     3421 acgagatcgc cagcggcggg gcgccgctca gcaaggaggt aggtgaggcc gtggccaaac
     3481 gcttccacct accaggcatc cgccagggct acggcctgac agaaacaacc agcgccattc
     3541 tgatcacccc cgaaggggac gacaagcctg gcgcagtagg caaggtggtg cccttcttcg
     3601 aggctaaggt ggtggacttg gacaccggta agacactggg tgtgaaccag cgcggcgagc
     3661 tgtgcgtccg tggccccatg atcatgagcg gctacgttaa caaccccgag gctacaaacg
     3721 ctctcatcga caaggacggc tggctgcaca gcggcgacat cgcctactgg gacgaggacg
     3781 agcacttctt catcgtggac cggctgaaga gcctgatcaa atacaagggc taccaggtag
     3841 ccccagccga actggagagc atcctgctgc aacaccccaa catcttcgac gccggggtcg
     3901 ccggcctgcc cgacgacgat gccggcgagc tgcccgccgc agtcgtcgtg ctggaacacg
     3961 gtaaaaccat gaccgagaag gagatcgtgg actatgtggc cagccaggtt acaaccgcca
     4021 agaagctgcg cggtggtgtt gtgttcgtgg acgaggtgcc taaaggactg accggcaagt
     4081 tggacgcccg caagatccgc gagattctca ttaaggccaa gaagggcggc aagatcgccg
     4141 tgtaataatt ctagagtcgg ggcggccggc cgcttcgagc agacatgata agatacattg
     4201 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
     4261 gtgatgctat tgctttat
//
