LOCUS       sequence-199060-        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"
     regulatory      153..162
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             159..1811
                     /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    1855..1976
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1866..1885)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1920..1939
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2084..2104)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(2372..2960)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2452..2471)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3160..3951,3952..4020))
                     /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    4125..4173
                     /note="synthetic polyadenylation signal"
     misc_feature    4187..4278
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4227..4246
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gcttagagat ctccctaggg acttatataa gggggtgggg gcgcgttcgt
       61 cctcagtcgc gatcgaacac tcgagccgag cagacgtgcc tacggacccg ggctagcgac
      121 ctaaagcttg gcaatccggt actgttggta aagccaccat ggaagatgcc aaaaacatta
      181 agaagggccc agcgccattc tacccactcg aagacgggac cgccggcgag cagctgcaca
      241 aagccatgaa gcgctacgcc ctggtgcccg gcaccatcgc ctttaccgac gcacatatcg
      301 aggtggacat tacctacgcc gagtacttcg agatgagcgt tcggctggca gaagctatga
      361 agcgctatgg gctgaataca aaccatcgga tcgtggtgtg cagcgagaat agcttgcagt
      421 tcttcatgcc cgtgttgggt gccctgttca tcggtgtggc tgtggcccca gctaacgaca
      481 tctacaacga gcgcgagctg ctgaacagca tgggcatcag ccagcccacc gtcgtattcg
      541 tgagcaagaa agggctgcaa aagatcctca acgtgcaaaa gaagctaccg atcatacaaa
      601 agatcatcat catggatagc aagaccgact accagggctt ccaaagcatg tacaccttcg
      661 tgacttccca tttgccaccc ggcttcaacg agtacgactt cgtgcccgag agcttcgacc
      721 gggacaaaac catcgccctg atcatgaaca gtagtggcag taccggattg cccaagggcg
      781 tagccctacc gcaccgcacc gcttgtgtcc gattcagtca tgcccgcgac cccatcttcg
      841 gcaaccagat catccccgac accgctatcc tcagcgtggt gccatttcac cacggcttcg
      901 gcatgttcac cacgctgggc tacttgatct gcggctttcg ggtcgtgctc atgtaccgct
      961 tcgaggagga gctattcttg cgcagcttgc aagactataa gattcaatct gccctgctgg
     1021 tgcccacact atttagcttc ttcgctaaga gcactctcat cgacaagtac gacctaagca
     1081 acttgcacga gatcgccagc ggcggggcgc cgctcagcaa ggaggtaggt gaggccgtgg
     1141 ccaaacgctt ccacctacca ggcatccgcc agggctacgg cctgacagaa acaaccagcg
     1201 ccattctgat cacccccgaa ggggacgaca agcctggcgc agtaggcaag gtggtgccct
     1261 tcttcgaggc taaggtggtg gacttggaca ccggtaagac actgggtgtg aaccagcgcg
     1321 gcgagctgtg cgtccgtggc cccatgatca tgagcggcta cgttaacaac cccgaggcta
     1381 caaacgctct catcgacaag gacggctggc tgcacagcgg cgacatcgcc tactgggacg
     1441 aggacgagca cttcttcatc gtggaccggc tgaagagcct gatcaaatac aagggctacc
     1501 aggtagcccc agccgaactg gagagcatcc tgctgcaaca ccccaacatc ttcgacgccg
     1561 gggtcgccgg cctgcccgac gacgatgccg gcgagctgcc cgccgcagtc gtcgtgctgg
     1621 aacacggtaa aaccatgacc gagaaggaga tcgtggacta tgtggccagc caggttacaa
     1681 ccgccaagaa gctgcgcggt ggtgttgtgt tcgtggacga ggtgcctaaa ggactgaccg
     1741 gcaagttgga cgcccgcaag atccgcgaga ttctcattaa ggccaagaag ggcggcaaga
     1801 tcgccgtgta ataattctag agtcggggcg gccggccgct tcgagcagac atgataagat
     1861 acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg
     1921 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca
     1981 acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa
     2041 gcaagtaaaa cctctacaaa tgtggtaaaa tcgataaggc tgtggaatgt gtgtcagtta
     2101 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     2161 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     2221 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     2281 actccgccca gttccgccca ttctccgctc catcgtgagc aaaaggccag caaaaggcca
     2341 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     2401 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     2461 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     2521 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     2581 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     2641 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     2701 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     2761 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     2821 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     2881 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     2941 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     3001 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     3061 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     3121 ggtctgacag cggccgcaaa tgctaaacca ctgcagtggt taccagtgct tgatcagtga
     3181 ggcaccgatc tcagcgatct gcctatttcg ttcgtccata gtggcctgac tccccgtcgt
     3241 gtagatcact acgattcgtg agggcttacc atcaggcccc agcgcagcaa tgatgccgcg
     3301 agagccgcgt tcaccggccc ccgatttgtc agcaatgaac cagccagcag ggagggccga
     3361 gcgaagaagt ggtcctgcta ctttgtccgc ctccatccag tctatgagct gctgtcgtga
     3421 tgctagagta agaagttcgc cagtgagtag tttccgaaga gttgtggcca ttgctactgg
     3481 catcgtggta tcacgctcgt cgttcggtat ggcttcgttc aactctggtt cccagcggtc
     3541 aagccgggtc acatgatcac ccatattatg aagaaatgca gtcagctcct tagggcctcc
     3601 gatcgttgtc agaagtaagt tggccgcggt gttgtcgctc atggtaatgg cagcactaca
     3661 caattctctt accgtcatgc catccgtaag atgcttttcc gtgaccggcg agtactcaac
     3721 caagtcgttt tgtgagtagt gtatacggcg accaagctgc tcttgcccgg cgtctatacg
     3781 ggacaacacc gcgccacata gcagtacttt gaaagtgctc atcatcggga atcgttcttc
     3841 ggggcggaaa gactcaagga tcttgccgct attgagatcc agttcgatat agcccactct
     3901 tgcacccagt tgatcttcag catcttttac tttcaccagc gtttcggggt gtgcaaaaac
     3961 aggcaagcaa aatgccgcaa agaagggaat gagtgcgaca cgaaaatgtt ggatgctcat
     4021 actcgtcctt tttcaatatt attgaagcat ttatcagggt tactagtacg tctctcaagg
     4081 ataagtaagt aatattaagg tacgggaggt attggacagg ccgcaataaa atatctttat
     4141 tttcattaca tctgtgtgtt ggttttttgt gtgaatcgat agtactaaca tacgctctcc
     4201 atcaaaacaa aacgaaacaa aacaaactag caaaataggc tgtccccagt gcaagtgcag
     4261 gtgccagaac atttctct
//
