LOCUS       sequence-190943-        4702 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 4702)
  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 4702)
  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_CMV
FEATURES             Location/Qualifiers
     source          1..4702
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      556..1144
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1045..1064
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        1343..1722
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    1967..2088
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2004..2023)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2058..2077
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(2132..3784)
                     /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      3781..3790
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    3940..4031
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     complement(3972..3991)
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     polyA_signal    4045..4093
                     /note="synthetic polyadenylation signal"
     CDS             join(4198..4266,4267..4702,1..356)
                     /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 aagccatacc gaacgacgag cgtgatacca cgatgccagt agcaatggcc acaactcttc
       61 ggaaactact cactggcgaa cttcttactc tagcatcacg acagcagctc atagactgga
      121 tggaggcgga caaagtagca ggaccacttc ttcgctcggc cctccctgct ggctggttca
      181 ttgctgacaa atcgggggcc ggtgaacgcg gctctcgcgg catcattgct gcgctggggc
      241 ctgatggtaa gccctcacga atcgtagtga tctacacgac ggggagtcag gccactatgg
      301 acgaacgaaa taggcagatc gctgagatcg gtgcctcact gatcaagcac tggtaaccac
      361 tgcagtggtt tagcatttgc ggccgctgtc agaccaagtt tactcatata tactttagat
      421 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
      481 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
      541 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
      601 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
      661 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
      721 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
      781 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
      841 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
      901 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
      961 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     1021 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     1081 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     1141 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     1201 catgtttacg acattttgga aagtcccgtt gattttggtg ccaaaacaaa ctcccattga
     1261 cgtcaatggg gtggagactt ggaaatcccc gtgagtcaaa ccgctatcca cgcccattgg
     1321 tgtactgcca aaaccgcatc accatggtaa tagcgatgac taatacgtag atgtactgcc
     1381 aagtaggaaa gtcccgtaag gtcatgtact gggcataatg ccaggcgggc catttaccgt
     1441 cattgacgtc aatagggggc ggacttggca tatgatacac ttgatgtact gccaagtggg
     1501 cagtttaccg taaatactcc acccattgac gtcaatggaa agtccctatt ggcgttacta
     1561 tgggaacata cgtcattatt gacgtcaatg ggcgggggtc gttgggcggt cagccaggcg
     1621 ggccatttac cgtaagttat gtaacgcgga actccatata tgggctatga actaatgacc
     1681 ccgtaattga ttactattaa taactagtca ataatcaatg ccaacatggc ggtcatattg
     1741 gacatgagcc aatataaatg tacatattat gatatagata caacgtatgc aatggccaat
     1801 agccaatatt gatttatgct atataaccaa tgaataatat ggctaatggc caatattgag
     1861 gatccttatc gattttacca catttgtaga ggttttactt gctttaaaaa acctcccaca
     1921 cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc
     1981 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     2041 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgctc
     2101 gaagcggccg gccgccccga ctctagaatt attacacggc gatcttgccg cccttcttgg
     2161 ccttaatgag aatctcgcgg atcttgcggg cgtccaactt gccggtcagt cctttaggca
     2221 cctcgtccac gaacacaaca ccaccgcgca gcttcttggc ggttgtaacc tggctggcca
     2281 catagtccac gatctccttc tcggtcatgg ttttaccgtg ttccagcacg acgactgcgg
     2341 cgggcagctc gccggcatcg tcgtcgggca ggccggcgac cccggcgtcg aagatgttgg
     2401 ggtgttgcag caggatgctc tccagttcgg ctggggctac ctggtagccc ttgtatttga
     2461 tcaggctctt cagccggtcc acgatgaaga agtgctcgtc ctcgtcccag taggcgatgt
     2521 cgccgctgtg cagccagccg tccttgtcga tgagagcgtt tgtagcctcg gggttgttaa
     2581 cgtagccgct catgatcatg gggccacgga cgcacagctc gccgcgctgg ttcacaccca
     2641 gtgtcttacc ggtgtccaag tccaccacct tagcctcgaa gaagggcacc accttgccta
     2701 ctgcgccagg cttgtcgtcc ccttcggggg tgatcagaat ggcgctggtt gtttctgtca
     2761 ggccgtagcc ctggcggatg cctggtaggt ggaagcgttt ggccacggcc tcacctacct
     2821 ccttgctgag cggcgccccg ccgctggcga tctcgtgcaa gttgcttagg tcgtacttgt
     2881 cgatgagagt gctcttagcg aagaagctaa atagtgtggg caccagcagg gcagattgaa
     2941 tcttatagtc ttgcaagctg cgcaagaata gctcctcctc gaagcggtac atgagcacga
     3001 cccgaaagcc gcagatcaag tagcccagcg tggtgaacat gccgaagccg tggtgaaatg
     3061 gcaccacgct gaggatagcg gtgtcgggga tgatctggtt gccgaagatg gggtcgcggg
     3121 catgactgaa tcggacacaa gcggtgcggt gcggtagggc tacgcccttg ggcaatccgg
     3181 tactgccact actgttcatg atcagggcga tggttttgtc ccggtcgaag ctctcgggca
     3241 cgaagtcgta ctcgttgaag ccgggtggca aatgggaagt cacgaaggtg tacatgcttt
     3301 ggaagccctg gtagtcggtc ttgctatcca tgatgatgat cttttgtatg atcggtagct
     3361 tcttttgcac gttgaggatc ttttgcagcc ctttcttgct cacgaatacg acggtgggct
     3421 ggctgatgcc catgctgttc agcagctcgc gctcgttgta gatgtcgtta gctggggcca
     3481 cagccacacc gatgaacagg gcacccaaca cgggcatgaa gaactgcaag ctattctcgc
     3541 tgcacaccac gatccgatgg tttgtattca gcccatagcg cttcatagct tctgccagcc
     3601 gaacgctcat ctcgaagtac tcggcgtagg taatgtccac ctcgatatgt gcgtcggtaa
     3661 aggcgatggt gccgggcacc agggcgtagc gcttcatggc tttgtgcagc tgctcgccgg
     3721 cggtcccgtc ttcgagtggg tagaatggcg ctgggccctt cttaatgttt ttggcatctt
     3781 ccatggtggc tttaccaaca gtaccggatt gccaagcttt aggtcgctag cccggtccgt
     3841 aggcacgtct gctcggctcg agtgttcgat cgcgactgag gacgaacgcg cccccacccc
     3901 cttatataag taccctaggg agatccggcc agttaggcca gagaaatgtt ctggcacctg
     3961 cacttgcact ggggacagcc tattttgcta gtttgttttg tttcgttttg ttttgatgga
     4021 gagcgtatgt tagtactatc gattcacaca aaaaaccaac acacagatgt aatgaaaata
     4081 aagatatttt attgcggcct gtccaatacc tcccgtacct taatattact tacttatcct
     4141 tgagagacgt actagtaacc ctgataaatg cttcaataat attgaaaaag gacgagtatg
     4201 agcatccaac attttcgtgt cgcactcatt cccttctttg cggcattttg cttgcctgtt
     4261 tttgcacacc ccgaaacgct ggtgaaagta aaagatgctg aagatcaact gggtgcaaga
     4321 gtgggctata tcgaactgga tctcaatagc ggcaagatcc ttgagtcttt ccgccccgaa
     4381 gaacgattcc cgatgatgag cactttcaaa gtactgctat gtggcgcggt gttgtcccgt
     4441 atagacgccg ggcaagagca gcttggtcgc cgtatacact actcacaaaa cgacttggtt
     4501 gagtactcgc cggtcacgga aaagcatctt acggatggca tgacggtaag agaattgtgt
     4561 agtgctgcca ttaccatgag cgacaacacc gcggccaact tacttctgac aacgatcgga
     4621 ggccctaagg agctgactgc atttcttcat aatatgggtg atcatgtgac ccggcttgac
     4681 cgctgggaac cagagttgaa cg
//
