LOCUS       sequence-190934-        6007 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Luciferase validation vector with ISG15 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 6007)
  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 6007)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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..6007
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1758..1775)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             join(1933..2001,2002..2793)
                     /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      complement(2983..3571)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3063..3082)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     regulatory      3670..3679
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3676..5328
                     /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    5372..5493
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5383..5402)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5437..5456
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 gaggatcgct ttagcctggc aggcggaggt tgcaatgaga tgagatcgtg cctctgcact
       61 ccagcctggg cgacagagtg agagaccctg tctcaaaaac acaaaaacaa caacaacaaa
      121 aaaacaccaa tctgagcaaa tactgcccta aaccgagtgt tgttatctct gggtagtttg
      181 gagttcttgt ttctcaatta accatgggga tgttttccaa gtttactaat tttgcaagtt
      241 ggtaaatgga aaatgaaacc attagtccat gtgatgacag ctttagtgca tcctgtgaag
      301 gatctggaat gcgcgatatt taggtgtttc cagggtgttg ggtgggggtg gggatgccgt
      361 ccgctgtccg gagtccccac cacttttgct tttccctgtc tttcggtcat tcggttttgt
      421 ttcttccgct cactctgggg catgcctcgg gaaagggaaa ccgaaactga agccaaattt
      481 ggccaccagc gcaggctcgg cggcacgccc cctgacgtgt gtgcctcagg cttaataata
      541 gggccggtgc tgcctgcgga agccggcggc tgagaggcag cgaactcatc tttgccagta
      601 caggagctcg tgccgtggcc cacagcccac agcccacagc catggtaagg cagatgtcac
      661 aggtgggggg aggtgggctc tgtgccagcc aattttcgtc tccctccccc agccaaggtc
      721 tcccaggggt gcagggagag cggagctgct cagagcttgg ccaggttcta agtgtgctcc
      781 tgaaagcagg tcacccctga gatcctcagg gtggggcaca gaggggcacc ctagcaggta
      841 aagggaggcc acgggatggc ggtgggcagc tggccttcta gtaacgagcc ctcagtgcct
      901 tctgtgcctg gggtccctgc cgacgggatg tagaggacag acaggaggga gcactgtccc
      961 tgggtacagg agctcgccct gcagccagtg ccttgtgtgt ggtgggcctg gggctggcgc
     1021 cgcagtctct gaacctgtgt gacgcctgca gggctgggac ctgacggtga agatgctggc
     1081 gggcaacgaa ttccaggtgt ccctgagcag ctccatgtcg gtgtcagagc tgaaggcgca
     1141 gatcacccag aagatcggcg tgcacgcctt ccagcagcgt ctggctgtcc acccgagcgg
     1201 tgtggcgctg caggacaggg tcccccttgc cagccagggc ctgggccccg gcagcacggt
     1261 cctgctggtg gtggacaaat gcgacgaacc tctgaacatc ctggtgagga ataacaaggg
     1321 ccgcagcagc acctacgagg tgcggctgac gcagaccgtg gcccacctga agcagcaagt
     1381 gagcgggctg gagggtgtgc aggacgacct gttctggctg accttcgagg ggaagcccct
     1441 ggaggaccag ctcccgctgg gggagtacgg cctcaagccc ctgagcaccg tgttcatgaa
     1501 tctgcgcctg cggggaggcg gcacagagcc tggcgggcgg agctaagggc ctccaccagc
     1561 atccgagcag gatcaagggc cggaaataaa ggctgttgta aagagaaatg gctgcctctg
     1621 tgtctgtgct tgcccctcct ccagcgtccg cctcagaccc ctggtgatgg ggcgggagaa
     1681 gtctcgtcgc cgcacttatg actgtcttct ttatcatgca actcgtagga caggtgccgg
     1741 cagcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     1801 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     1861 ggaaagaaca tgtcttgaga gacgtactag taaccctgat aaatgcttca ataatattga
     1921 aaaaggacga gtatgagcat ccaacatttt cgtgtcgcac tcattccctt ctttgcggca
     1981 ttttgcttgc ctgtttttgc acaccccgaa acgctggtga aagtaaaaga tgctgaagat
     2041 caactgggtg caagagtggg ctatatcgaa ctggatctca atagcggcaa gatccttgag
     2101 tctttccgcc ccgaagaacg attcccgatg atgagcactt tcaaagtact gctatgtggc
     2161 gcggtgttgt cccgtataga cgccgggcaa gagcagcttg gtcgccgtat acactactca
     2221 caaaacgact tggttgagta ctcgccggtc acggaaaagc atcttacgga tggcatgacg
     2281 gtaagagaat tgtgtagtgc tgccattacc atgagcgaca acaccgcggc caacttactt
     2341 ctgacaacga tcggaggccc taaggagctg actgcatttc ttcataatat gggtgatcat
     2401 gtgacccggc ttgaccgctg ggaaccagag ttgaacgaag ccataccgaa cgacgagcgt
     2461 gataccacga tgccagtagc aatggccaca actcttcgga aactactcac tggcgaactt
     2521 cttactctag catcacgaca gcagctcata gactggatgg aggcggacaa agtagcagga
     2581 ccacttcttc gctcggccct ccctgctggc tggttcattg ctgacaaatc gggggccggt
     2641 gaacgcggct ctcgcggcat cattgctgcg ctggggcctg atggtaagcc ctcacgaatc
     2701 gtagtgatct acacgacggg gagtcaggcc actatggacg aacgaaatag gcagatcgct
     2761 gagatcggtg cctcactgat caagcactgg taaccactgc agtggtttag catttgcggc
     2821 cgctgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     2881 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gacatgtgag caaaaggcca
     2941 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     3001 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     3061 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     3121 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     3181 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     3241 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     3301 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     3361 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     3421 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     3481 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     3541 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     3601 tgacgctcag tggaacgaaa actcacgtta agggattttg aagcttggca atccggtact
     3661 gttggtaaag ccaccatgga agatgccaaa aacattaaga agggcccagc gccattctac
     3721 ccactcgaag acgggaccgc cggcgagcag ctgcacaaag ccatgaagcg ctacgccctg
     3781 gtgcccggca ccatcgcctt taccgacgca catatcgagg tggacattac ctacgccgag
     3841 tacttcgaga tgagcgttcg gctggcagaa gctatgaagc gctatgggct gaatacaaac
     3901 catcggatcg tggtgtgcag cgagaatagc ttgcagttct tcatgcccgt gttgggtgcc
     3961 ctgttcatcg gtgtggctgt ggccccagct aacgacatct acaacgagcg cgagctgctg
     4021 aacagcatgg gcatcagcca gcccaccgtc gtattcgtga gcaagaaagg gctgcaaaag
     4081 atcctcaacg tgcaaaagaa gctaccgatc atacaaaaga tcatcatcat ggatagcaag
     4141 accgactacc agggcttcca aagcatgtac accttcgtga cttcccattt gccacccggc
     4201 ttcaacgagt acgacttcgt gcccgagagc ttcgaccggg acaaaaccat cgccctgatc
     4261 atgaacagta gtggcagtac cggattgccc aagggcgtag ccctaccgca ccgcaccgct
     4321 tgtgtccgat tcagtcatgc ccgcgacccc atcttcggca accagatcat ccccgacacc
     4381 gctatcctca gcgtggtgcc atttcaccac ggcttcggca tgttcaccac gctgggctac
     4441 ttgatctgcg gctttcgggt cgtgctcatg taccgcttcg aggaggagct attcttgcgc
     4501 agcttgcaag actataagat tcaatctgcc ctgctggtgc ccacactatt tagcttcttc
     4561 gctaagagca ctctcatcga caagtacgac ctaagcaact tgcacgagat cgccagcggc
     4621 ggggcgccgc tcagcaagga ggtaggtgag gccgtggcca aacgcttcca cctaccaggc
     4681 atccgccagg gctacggcct gacagaaaca accagcgcca ttctgatcac ccccgaaggg
     4741 gacgacaagc ctggcgcagt aggcaaggtg gtgcccttct tcgaggctaa ggtggtggac
     4801 ttggacaccg gtaagacact gggtgtgaac cagcgcggcg agctgtgcgt ccgtggcccc
     4861 atgatcatga gcggctacgt taacaacccc gaggctacaa acgctctcat cgacaaggac
     4921 ggctggctgc acagcggcga catcgcctac tgggacgagg acgagcactt cttcatcgtg
     4981 gaccggctga agagcctgat caaatacaag ggctaccagg tagccccagc cgaactggag
     5041 agcatcctgc tgcaacaccc caacatcttc gacgccgggg tcgccggcct gcccgacgac
     5101 gatgccggcg agctgcccgc cgcagtcgtc gtgctggaac acggtaaaac catgaccgag
     5161 aaggagatcg tggactatgt ggccagccag gttacaaccg ccaagaagct gcgcggtggt
     5221 gttgtgttcg tggacgaggt gcctaaagga ctgaccggca agttggacgc ccgcaagatc
     5281 cgcgagattc tcattaaggc caagaagggc ggcaagatcg ccgtgtaata attctagagt
     5341 cggggcggcc ggccgcttcg agcagacatg ataagataca ttgatgagtt tggacaaacc
     5401 acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta
     5461 tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg
     5521 tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt
     5581 ggtaaaatcg ataaggatcc gtcgaccgat gcccttgaga gccttcaacc cagtcagctc
     5641 cttccggtgg gcgcggggca tgactatcct ggatgctatg ccctcatcag atagataatt
     5701 tgcaaatatt tcttcccatt ctatggattg tcttttcact ttctcaatag tgtcccagag
     5761 ttcatttttg tagaaaataa aagataggtc tcttttatta aaaaacaatc tgaggctccg
     5821 ggtgcagtgg ctcacgcctg taatcccagc agtttcagag gccgaggcag gtggatcact
     5881 tgagcccagg agttcgagat cagcctgggc gacatggcga gacccccatc tctactaaaa
     5941 atacaaaaaa ttagccgggc ctggtggtgc acccctgtgg tcccagctac gtgggaggct
     6001 gaggtga
//
