LOCUS       sequence-190933-        4638 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 4638)
  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 4638)
  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_ORI_CMV
FEATURES             Location/Qualifiers
     source          1..4638
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     2..19
                     /label=L4440
                     /note="L4440 vector, forward primer"
     enhancer        297..676
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    922..1043
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(959..978)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1013..1032
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(1087..2739)
                     /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"
     regulatory      2736..2745
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     rep_origin      2844..3432
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3333..3352
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3622..4413,4414..4482))
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
ORIGIN
        1 cagcgagtca gtgagcgagg aagcggaaga gcgctgccgg cacctgtcct acgagttgca
       61 tgataaagaa gacagtcata agtgcggcga cgatagtcat gccccgcgcc caccggaagg
      121 agctgactgg gttgaaggct ctcaagggca tcggtcgact tacgacattt tggaaagtcc
      181 cgttgatttt ggtgccaaaa caaactccca ttgacgtcaa tggggtggag acttggaaat
      241 ccccgtgagt caaaccgcta tccacgccca ttggtgtact gccaaaaccg catcaccatg
      301 gtaatagcga tgactaatac gtagatgtac tgccaagtag gaaagtcccg taaggtcatg
      361 tactgggcat aatgccaggc gggccattta ccgtcattga cgtcaatagg gggcggactt
      421 ggcatatgat acacttgatg tactgccaag tgggcagttt accgtaaata ctccacccat
      481 tgacgtcaat ggaaagtccc tattggcgtt actatgggaa catacgtcat tattgacgtc
      541 aatgggcggg ggtcgttggg cggtcagcca ggcgggccat ttaccgtaag ttatgtaacg
      601 cggaactcca tatatgggct atgaactaat gaccccgtaa ttgattacta ttaataacta
      661 gtcaataatc aatgccaaca tggcggtcat attggacatg agccaatata aatgtacata
      721 ttatgatata gatacaacgt atgcaatggc caatagccaa tattgattta tgctatataa
      781 ccaatgaata atatggctaa tggccaatat ttgaggatcc ttatcgattt taccacattt
      841 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa
      901 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc
      961 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     1021 tccaaactca tcaatgtatc ttatcatgtc tgctcgaagc ggccggccgc cccgactcta
     1081 gaattattac acggcgatct tgccgccctt cttggcctta atgagaatct cgcggatctt
     1141 gcgggcgtcc aacttgccgg tcagtccttt aggcacctcg tccacgaaca caacaccacc
     1201 gcgcagcttc ttggcggttg taacctggct ggccacatag tccacgatct ccttctcggt
     1261 catggtttta ccgtgttcca gcacgacgac tgcggcgggc agctcgccgg catcgtcgtc
     1321 gggcaggccg gcgaccccgg cgtcgaagat gttggggtgt tgcagcagga tgctctccag
     1381 ttcggctggg gctacctggt agcccttgta tttgatcagg ctcttcagcc ggtccacgat
     1441 gaagaagtgc tcgtcctcgt cccagtaggc gatgtcgccg ctgtgcagcc agccgtcctt
     1501 gtcgatgaga gcgtttgtag cctcggggtt gttaacgtag ccgctcatga tcatggggcc
     1561 acggacgcac agctcgccgc gctggttcac acccagtgtc ttaccggtgt ccaagtccac
     1621 caccttagcc tcgaagaagg gcaccacctt gcctactgcg ccaggcttgt cgtccccttc
     1681 gggggtgatc agaatggcgc tggttgtttc tgtcaggccg tagccctggc ggatgcctgg
     1741 taggtggaag cgtttggcca cggcctcacc tacctccttg ctgagcggcg ccccgccgct
     1801 ggcgatctcg tgcaagttgc ttaggtcgta cttgtcgatg agagtgctct tagcgaagaa
     1861 gctaaatagt gtgggcacca gcagggcaga ttgaatctta tagtcttgca agctgcgcaa
     1921 gaatagctcc tcctcgaagc ggtacatgag cacgacccga aagccgcaga tcaagtagcc
     1981 cagcgtggtg aacatgccga agccgtggtg aaatggcacc acgctgagga tagcggtgtc
     2041 ggggatgatc tggttgccga agatggggtc gcgggcatga ctgaatcgga cacaagcggt
     2101 gcggtgcggt agggctacgc ccttgggcaa tccggtactg ccactactgt tcatgatcag
     2161 ggcgatggtt ttgtcccggt cgaagctctc gggcacgaag tcgtactcgt tgaagccggg
     2221 tggcaaatgg gaagtcacga aggtgtacat gctttggaag ccctggtagt cggtcttgct
     2281 atccatgatg atgatctttt gtatgatcgg tagcttcttt tgcacgttga ggatcttttg
     2341 cagccctttc ttgctcacga atacgacggt gggctggctg atgcccatgc tgttcagcag
     2401 ctcgcgctcg ttgtagatgt cgttagctgg ggccacagcc acaccgatga acagggcacc
     2461 caacacgggc atgaagaact gcaagctatt ctcgctgcac accacgatcc gatggtttgt
     2521 attcagccca tagcgcttca tagcttctgc cagccgaacg ctcatctcga agtactcggc
     2581 gtaggtaatg tccacctcga tatgtgcgtc ggtaaaggcg atggtgccgg gcaccagggc
     2641 gtagcgcttc atggctttgt gcagctgctc gccggcggtc ccgtcttcga gtgggtagaa
     2701 tggcgctggg cccttcttaa tgtttttggc atcttccatg gtggctttac caacagtacc
     2761 ggattgccaa gcttcaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     2821 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     2881 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     2941 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     3001 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     3061 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     3121 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     3181 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     3241 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     3301 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     3361 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     3421 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     3481 tttgctcaca tgtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     3541 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agcggccgca
     3601 aatgctaaac cactgcagtg gttaccagtg cttgatcagt gaggcaccga tctcagcgat
     3661 ctgcctattt cgttcgtcca tagtggcctg actccccgtc gtgtagatca ctacgattcg
     3721 tgagggctta ccatcaggcc ccagcgcagc aatgatgccg cgagagccgc gttcaccggc
     3781 ccccgatttg tcagcaatga accagccagc agggagggcc gagcgaagaa gtggtcctgc
     3841 tactttgtcc gcctccatcc agtctatgag ctgctgtcgt gatgctagag taagaagttc
     3901 gccagtgagt agtttccgaa gagttgtggc cattgctact ggcatcgtgg tatcacgctc
     3961 gtcgttcggt atggcttcgt tcaactctgg ttcccagcgg tcaagccggg tcacatgatc
     4021 acccatatta tgaagaaatg cagtcagctc cttagggcct ccgatcgttg tcagaagtaa
     4081 gttggccgcg gtgttgtcgc tcatggtaat ggcagcacta cacaattctc ttaccgtcat
     4141 gccatccgta agatgctttt ccgtgaccgg cgagtactca accaagtcgt tttgtgagta
     4201 gtgtatacgg cgaccaagct gctcttgccc ggcgtctata cgggacaaca ccgcgccaca
     4261 tagcagtact ttgaaagtgc tcatcatcgg gaatcgttct tcggggcgga aagactcaag
     4321 gatcttgccg ctattgagat ccagttcgat atagcccact cttgcaccca gttgatcttc
     4381 agcatctttt actttcacca gcgtttcggg gtgtgcaaaa acaggcaagc aaaatgccgc
     4441 aaagaaggga atgagtgcga cacgaaaatg ttggatgctc atactcgtcc tttttcaata
     4501 ttattgaagc atttatcagg gttactagta cgtctctcaa gacatgttct ttcctgcgtt
     4561 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     4621 cagccgaacg accgagcg
//
