LOCUS       sequence-193160-        6376 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Luciferase validation vector with IFI27 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 6376)
  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 6376)
  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 INF enhancer vector_ORI_I...
FEATURES             Location/Qualifiers
     source          1..6376
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     56..75
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(345..1136,1137..1205))
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1363..1380
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    4021..4142
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4058..4077)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4112..4131
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(4186..5838)
                     /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      5835..5844
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     rep_origin      join(5943..6376,1..155)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
       61 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
      121 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
      181 ggttcctggc cttttgctgg ccttttgctc acatgtcatg agattatcaa aaaggatctt
      241 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
      301 aacttggtct gacagcggcc gcaaatgcta aaccactgca gtggttacca gtgcttgatc
      361 agtgaggcac cgatctcagc gatctgccta tttcgttcgt ccatagtggc ctgactcccc
      421 gtcgtgtaga tcactacgat tcgtgagggc ttaccatcag gccccagcgc agcaatgatg
      481 ccgcgagagc cgcgttcacc ggcccccgat ttgtcagcaa tgaaccagcc agcagggagg
      541 gccgagcgaa gaagtggtcc tgctactttg tccgcctcca tccagtctat gagctgctgt
      601 cgtgatgcta gagtaagaag ttcgccagtg agtagtttcc gaagagttgt ggccattgct
      661 actggcatcg tggtatcacg ctcgtcgttc ggtatggctt cgttcaactc tggttcccag
      721 cggtcaagcc gggtcacatg atcacccata ttatgaagaa atgcagtcag ctccttaggg
      781 cctccgatcg ttgtcagaag taagttggcc gcggtgttgt cgctcatggt aatggcagca
      841 ctacacaatt ctcttaccgt catgccatcc gtaagatgct tttccgtgac cggcgagtac
      901 tcaaccaagt cgttttgtga gtagtgtata cggcgaccaa gctgctcttg cccggcgtct
      961 atacgggaca acaccgcgcc acatagcagt actttgaaag tgctcatcat cgggaatcgt
     1021 tcttcggggc ggaaagactc aaggatcttg ccgctattga gatccagttc gatatagccc
     1081 actcttgcac ccagttgatc ttcagcatct tttactttca ccagcgtttc ggggtgtgca
     1141 aaaacaggca agcaaaatgc cgcaaagaag ggaatgagtg cgacacgaaa atgttggatg
     1201 ctcatactcg tcctttttca atattattga agcatttatc agggttacta gtacgtctct
     1261 caagacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     1321 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     1381 gaagcggaag agcgctgccg gcacctgtcc tacgagttgc atgataaaga agacagtcat
     1441 aagtgcggcg acgaccctcc cagatggaaa gttagggtgg acgcctgctg tgagaagaca
     1501 ggacagggca gcaaggctgc gggtgtcccc cattttctct cccacgggtc cctggacagg
     1561 acccccgccc ttcccagcag cctcctcgcc cccagcacca gcccctccca ggaacactca
     1621 cccaggggca aaactacggc agagccagag gccaccctga ccactttggc cacactggtc
     1681 actgctgatg aggtgagagc agaggcctcc atgcctgctc gggttaattc cgtggcctag
     1741 agagtaagag atcctcaact tcagccagac ctgccgaagg gatggagctc tgaggacggg
     1801 atgggcacac aacctgactt gcttcccact gtcctgaaaa atgtgacggg ctccaggggg
     1861 ctagggaggc tgctctgagc ttgtggagca gcaggccttg aaaggatcct ggggtgaggg
     1921 agggagggat gttggggagt tggggctacc aaaggacctg tgttaggtca gggccctgag
     1981 ggactttatt tccccagggg ggctgctttc tttctcctga aagtcccagc gccaatgtct
     2041 tagacacttc cgggactccc ttatctctcc acggagcaat tgaaaaagca ccaaaactaa
     2101 gaaatcagaa ccaatcgcat ttttaaggca tatacagcag gcgaggtgcc tgctcttatg
     2161 taatctcctc actaacgtga aaaagatggg accctcctcc tccgcatgtt acagatgagg
     2221 gagcgcgatc tggctcagtg atttgctcca ggtcagaagc tttccatagg gaagcaacac
     2281 cccgcccagg catggacttg caatggcttc gacttggttc ctcctaacac ttatctcttc
     2341 cccttgccca ctttacccca agccgccctc aaggcctgag gaaaacctgg gcaaggacaa
     2401 agctaattgt ccttacattc cagagtccag ggacaggtgg ggctcgagtc ccttcccatc
     2461 tgaggctccc tgatgagaac aaaaacaccg ggcagagagt ggatgcttaa ggcctcccct
     2521 ccgctccctg agcccacagc ccacccaagg tccggggcca cgggaagggt agctcctgcc
     2581 caccctccca gtcttaccca aagaaggaga gttcctgaga atgtgaagct gacctcaccg
     2641 tcttaagctt ggatgtgttc ccaaagggtg tgatcccagg ccttttaagc agctccaccc
     2701 cgactgaagc actgcccctt gacacaggca acaaatgaga taggtttcat tttccaggaa
     2761 accgaaactg cagacatctt tctcccctca tgcgcagtcc agaggaaaat tcagttattt
     2821 acagaaatag gatttgccca tggcatcagt cattgtcagt taatataaaa tgctttgtca
     2881 tttaattcaa ggatgggagt gtgctctacg cagaatgaag tcacaagaag taagatcaaa
     2941 gttctataat aaaatgataa atattacttt ctaatgaaga aggtgatttt cagaataata
     3001 gtagtccttg cgataggcag atgttccgtt tttcaaaaaa agtctcaact gaaggcccac
     3061 tgtgctagat actgactagg cactgggaca ggactgtaaa caaaacagac caaaatatcc
     3121 atcctcacag agtttaccag caatgggggc aaactaaaca taataaataa ctaaatgtat
     3181 agaatgttat ttgatgaagc gctaaagcaa aaggaggtga agcagtgacc taggcagaat
     3241 tatggggggg cagtcattcc aggtggaggc cacagcagtg gcagagcccg tgtggcagga
     3301 gcagcaaggg ggccagtgtg gcctcagcag gtgggcaggg gagagaggca ggaggtgaac
     3361 tcagagttta acagggatcc aggccctgtg aggccacaaa agcactgcaa ggacttgatc
     3421 ttgtgtcctg aagccattgc aggggattat catctatacc tttttgggga aagggagatg
     3481 aagctgagaa atgacacagt caggctcact ctgcagctgc tttgagaaca ggcggaagag
     3541 aaagcaccag gtcctggaat gtacatttaa atcttccaca tgagtagtgt gcgagttgta
     3601 agattcaaaa tggagtcact tgggtaaaac ctcagcagac agagccagca aggctttaat
     3661 gggagggctc tcacgcaaga tgaggctctc ccgtttgcct gctaacagga actatcacaa
     3721 gaaatgtttc caaactgcag cttatcacat gggccacaca aggacagcta gaagcacagg
     3781 gagagcgagc cagttatgca aaaacacttg cctggcacag tgtctcacaa acccaatcca
     3841 aaactgctag tcatgccccg cgcccaccgg aaggagctga ctgggttgaa ggctctcaag
     3901 ggcatcggtc gacggatcct tatcgatttt accacatttg tagaggtttt acttgcttta
     3961 aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt
     4021 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     4081 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     4141 tatcatgtct gctcgaagcg gccggccgcc ccgactctag aattattaca cggcgatctt
     4201 gccgcccttc ttggccttaa tgagaatctc gcggatcttg cgggcgtcca acttgccggt
     4261 cagtccttta ggcacctcgt ccacgaacac aacaccaccg cgcagcttct tggcggttgt
     4321 aacctggctg gccacatagt ccacgatctc cttctcggtc atggttttac cgtgttccag
     4381 cacgacgact gcggcgggca gctcgccggc atcgtcgtcg ggcaggccgg cgaccccggc
     4441 gtcgaagatg ttggggtgtt gcagcaggat gctctccagt tcggctgggg ctacctggta
     4501 gcccttgtat ttgatcaggc tcttcagccg gtccacgatg aagaagtgct cgtcctcgtc
     4561 ccagtaggcg atgtcgccgc tgtgcagcca gccgtccttg tcgatgagag cgtttgtagc
     4621 ctcggggttg ttaacgtagc cgctcatgat catggggcca cggacgcaca gctcgccgcg
     4681 ctggttcaca cccagtgtct taccggtgtc caagtccacc accttagcct cgaagaaggg
     4741 caccaccttg cctactgcgc caggcttgtc gtccccttcg ggggtgatca gaatggcgct
     4801 ggttgtttct gtcaggccgt agccctggcg gatgcctggt aggtggaagc gtttggccac
     4861 ggcctcacct acctccttgc tgagcggcgc cccgccgctg gcgatctcgt gcaagttgct
     4921 taggtcgtac ttgtcgatga gagtgctctt agcgaagaag ctaaatagtg tgggcaccag
     4981 cagggcagat tgaatcttat agtcttgcaa gctgcgcaag aatagctcct cctcgaagcg
     5041 gtacatgagc acgacccgaa agccgcagat caagtagccc agcgtggtga acatgccgaa
     5101 gccgtggtga aatggcacca cgctgaggat agcggtgtcg gggatgatct ggttgccgaa
     5161 gatggggtcg cgggcatgac tgaatcggac acaagcggtg cggtgcggta gggctacgcc
     5221 cttgggcaat ccggtactgc cactactgtt catgatcagg gcgatggttt tgtcccggtc
     5281 gaagctctcg ggcacgaagt cgtactcgtt gaagccgggt ggcaaatggg aagtcacgaa
     5341 ggtgtacatg ctttggaagc cctggtagtc ggtcttgcta tccatgatga tgatcttttg
     5401 tatgatcggt agcttctttt gcacgttgag gatcttttgc agccctttct tgctcacgaa
     5461 tacgacggtg ggctggctga tgcccatgct gttcagcagc tcgcgctcgt tgtagatgtc
     5521 gttagctggg gccacagcca caccgatgaa cagggcaccc aacacgggca tgaagaactg
     5581 caagctattc tcgctgcaca ccacgatccg atggtttgta ttcagcccat agcgcttcat
     5641 agcttctgcc agccgaacgc tcatctcgaa gtactcggcg taggtaatgt ccacctcgat
     5701 atgtgcgtcg gtaaaggcga tggtgccggg caccagggcg tagcgcttca tggctttgtg
     5761 cagctgctcg ccggcggtcc cgtcttcgag tgggtagaat ggcgctgggc ccttcttaat
     5821 gtttttggca tcttccatgg tggctttacc aacagtaccg gattgccaag cttcaaaatc
     5881 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     5941 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     6001 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     6061 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     6121 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     6181 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     6241 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     6301 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     6361 gggagaaagg cggaca
//
