LOCUS       sequence-193012-        4650 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Luciferase validation vector with CMV 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 4650)
  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 4650)
  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_CMV
FEATURES             Location/Qualifiers
     source          1..4650
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      630..639
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(682..712)
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     misc_feature    737..828
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     complement(769..788)
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     polyA_signal    842..890
                     /note="synthetic polyadenylation signal"
     CDS             join(995..1063,1064..1855)
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      2055..2643
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2544..2563
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2842..3221
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    3466..3587
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3503..3522)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3557..3576
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(3631..4650,1..633))
                     /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"
ORIGIN
        1 cggtagggct acgcccttgg gcaatccggt actgccacta ctgttcatga tcagggcgat
       61 ggttttgtcc cggtcgaagc tctcgggcac gaagtcgtac tcgttgaagc cgggtggcaa
      121 atgggaagtc acgaaggtgt acatgctttg gaagccctgg tagtcggtct tgctatccat
      181 gatgatgatc ttttgtatga tcggtagctt cttttgcacg ttgaggatct tttgcagccc
      241 tttcttgctc acgaatacga cggtgggctg gctgatgccc atgctgttca gcagctcgcg
      301 ctcgttgtag atgtcgttag ctggggccac agccacaccg atgaacaggg cacccaacac
      361 gggcatgaag aactgcaagc tattctcgct gcacaccacg atccgatggt ttgtattcag
      421 cccatagcgc ttcatagctt ctgccagccg aacgctcatc tcgaagtact cggcgtaggt
      481 aatgtccacc tcgatatgtg cgtcggtaaa ggcgatggtg ccgggcacca gggcgtagcg
      541 cttcatggct ttgtgcagct gctcgccggc ggtcccgtct tcgagtgggt agaatggcgc
      601 tgggcccttc ttaatgtttt tggcatcttc catggtggct ttaccaacag taccggattg
      661 ccaagcttta ggtcgctagc cctggaagtc gagcttccat tatataccct ctctcgaaga
      721 tccggccagt taggccagag aaatgttctg gcacctgcac ttgcactggg gacagcctat
      781 tttgctagtt tgttttgttt cgttttgttt tgatggagag cgtatgttag tactatcgat
      841 tcacacaaaa aaccaacaca cagatgtaat gaaaataaag atattttatt gcggcctgtc
      901 caatacctcc cgtaccttaa tattacttac ttatccttga gagacgtact agtaaccctg
      961 ataaatgctt caataatatt gaaaaaggac gagtatgagc atccaacatt ttcgtgtcgc
     1021 actcattccc ttctttgcgg cattttgctt gcctgttttt gcacaccccg aaacgctggt
     1081 gaaagtaaaa gatgctgaag atcaactggg tgcaagagtg ggctatatcg aactggatct
     1141 caatagcggc aagatccttg agtctttccg ccccgaagaa cgattcccga tgatgagcac
     1201 tttcaaagta ctgctatgtg gcgcggtgtt gtcccgtata gacgccgggc aagagcagct
     1261 tggtcgccgt atacactact cacaaaacga cttggttgag tactcgccgg tcacggaaaa
     1321 gcatcttacg gatggcatga cggtaagaga attgtgtagt gctgccatta ccatgagcga
     1381 caacaccgcg gccaacttac ttctgacaac gatcggaggc cctaaggagc tgactgcatt
     1441 tcttcataat atgggtgatc atgtgacccg gcttgaccgc tgggaaccag agttgaacga
     1501 agccataccg aacgacgagc gtgataccac gatgccagta gcaatggcca caactcttcg
     1561 gaaactactc actggcgaac ttcttactct agcatcacga cagcagctca tagactggat
     1621 ggaggcggac aaagtagcag gaccacttct tcgctcggcc ctccctgctg gctggttcat
     1681 tgctgacaaa tcgggggccg gtgaacgcgg ctctcgcggc atcattgctg cgctggggcc
     1741 tgatggtaag ccctcacgaa tcgtagtgat ctacacgacg gggagtcagg ccactatgga
     1801 cgaacgaaat aggcagatcg ctgagatcgg tgcctcactg atcaagcact ggtaaccact
     1861 gcagtggttt agcatttgcg gccgctgtca gaccaagttt actcatatat actttagatt
     1921 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     1981 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     2041 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     2101 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     2161 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     2221 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     2281 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     2341 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     2401 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     2461 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     2521 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     2581 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     2641 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     2701 atgtttacga cattttggaa agtcccgttg attttggtgc caaaacaaac tcccattgac
     2761 gtcaatgggg tggagacttg gaaatccccg tgagtcaaac cgctatccac gcccattggt
     2821 gtactgccaa aaccgcatca ccatggtaat agcgatgact aatacgtaga tgtactgcca
     2881 agtaggaaag tcccgtaagg tcatgtactg ggcataatgc caggcgggcc atttaccgtc
     2941 attgacgtca atagggggcg gacttggcat atgatacact tgatgtactg ccaagtgggc
     3001 agtttaccgt aaatactcca cccattgacg tcaatggaaa gtccctattg gcgttactat
     3061 gggaacatac gtcattattg acgtcaatgg gcgggggtcg ttgggcggtc agccaggcgg
     3121 gccatttacc gtaagttatg taacgcggaa ctccatatat gggctatgaa ctaatgaccc
     3181 cgtaattgat tactattaat aactagtcaa taatcaatgc caacatggcg gtcatattgg
     3241 acatgagcca atataaatgt acatattatg atatagatac aacgtatgca atggccaata
     3301 gccaatattg atttatgcta tataaccaat gaataatatg gctaatggcc aatattgagg
     3361 acccttatcg attttaccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
     3421 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
     3481 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     3541 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgctcg
     3601 aagcggccgg ccgccccgac tctagaatta ttacacggcg atcttgccgc ccttcttggc
     3661 cttaatgaga atctcgcgga tcttgcgggc gtccaacttg ccggtcagtc ctttaggcac
     3721 ctcgtccacg aacacaacac caccgcgcag cttcttggcg gttgtaacct ggctggccac
     3781 atagtccacg atctccttct cggtcatggt tttaccgtgt tccagcacga cgactgcggc
     3841 gggcagctcg ccggcatcgt cgtcgggcag gccggcgacc ccggcgtcga agatgttggg
     3901 gtgttgcagc aggatgctct ccagttcggc tggggctacc tggtagccct tgtatttgat
     3961 caggctcttc agccggtcca cgatgaagaa gtgctcgtcc tcgtcccagt aggcgatgtc
     4021 gccgctgtgc agccagccgt ccttgtcgat gagagcgttt gtagcctcgg ggttgttaac
     4081 gtagccgctc atgatcatgg ggccacggac gcacagctcg ccgcgctggt tcacacccag
     4141 tgtcttaccg gtgtccaagt ccaccacctt agcctcgaag aagggcacca ccttgcctac
     4201 tgcgccaggc ttgtcgtccc cttcgggggt gatcagaatg gcgctggttg tttctgtcag
     4261 gccgtagccc tggcggatgc ctggtaggtg gaagcgtttg gccacggcct cacctacctc
     4321 cttgctgagc ggcgccccgc cgctggcgat ctcgtgcaag ttgcttaggt cgtacttgtc
     4381 gatgagagtg ctcttagcga agaagctaaa tagtgtgggc accagcaggg cagattgaat
     4441 cttatagtct tgcaagctgc gcaagaatag ctcctcctcg aagcggtaca tgagcacgac
     4501 ccgaaagccg cagatcaagt agcccagcgt ggtgaacatg ccgaagccgt ggtgaaatgg
     4561 caccacgctg aggatagcgg tgtcggggat gatctggttg ccgaagatgg ggtcgcgggc
     4621 atgactgaat cggacacaag cggtgcggtg
//
