LOCUS       sequence-197080-        5483 bp    DNA     linear   UNA 15-APR-2025
DEFINITION  Luciferase validation vector with GTSE1 enhancer inserted downstream
            of the reporter gene. The candidate's enhancer activity is reflected
            by luciferase activity.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 5483)
  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 5483)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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_GTSE1
FEATURES             Location/Qualifiers
     source          1..5483
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        28..58
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      101..110
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             107..1759
                     /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    1803..1924
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1814..1833)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1868..1887
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(3577..4165)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3657..3676)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4365..5156,5157..5225))
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     polyA_signal    5330..5378
                     /note="synthetic polyadenylation signal"
     misc_feature    5392..5483
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5432..5451
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gcttagagat cttcgagaga gggtatataa tggaagctcg acttccaggg
       61 ctagcgacct aaagcttggc aatccggtac tgttggtaaa gccaccatgg aagatgccaa
      121 aaacattaag aagggcccag cgccattcta cccactcgaa gacgggaccg ccggcgagca
      181 gctgcacaaa gccatgaagc gctacgccct ggtgcccggc accatcgcct ttaccgacgc
      241 acatatcgag gtggacatta cctacgccga gtacttcgag atgagcgttc ggctggcaga
      301 agctatgaag cgctatgggc tgaatacaaa ccatcggatc gtggtgtgca gcgagaatag
      361 cttgcagttc ttcatgcccg tgttgggtgc cctgttcatc ggtgtggctg tggccccagc
      421 taacgacatc tacaacgagc gcgagctgct gaacagcatg ggcatcagcc agcccaccgt
      481 cgtattcgtg agcaagaaag ggctgcaaaa gatcctcaac gtgcaaaaga agctaccgat
      541 catacaaaag atcatcatca tggatagcaa gaccgactac cagggcttcc aaagcatgta
      601 caccttcgtg acttcccatt tgccacccgg cttcaacgag tacgacttcg tgcccgagag
      661 cttcgaccgg gacaaaacca tcgccctgat catgaacagt agtggcagta ccggattgcc
      721 caagggcgta gccctaccgc accgcaccgc ttgtgtccga ttcagtcatg cccgcgaccc
      781 catcttcggc aaccagatca tccccgacac cgctatcctc agcgtggtgc catttcacca
      841 cggcttcggc atgttcacca cgctgggcta cttgatctgc ggctttcggg tcgtgctcat
      901 gtaccgcttc gaggaggagc tattcttgcg cagcttgcaa gactataaga ttcaatctgc
      961 cctgctggtg cccacactat ttagcttctt cgctaagagc actctcatcg acaagtacga
     1021 cctaagcaac ttgcacgaga tcgccagcgg cggggcgccg ctcagcaagg aggtaggtga
     1081 ggccgtggcc aaacgcttcc acctaccagg catccgccag ggctacggcc tgacagaaac
     1141 aaccagcgcc attctgatca cccccgaagg ggacgacaag cctggcgcag taggcaaggt
     1201 ggtgcccttc ttcgaggcta aggtggtgga cttggacacc ggtaagacac tgggtgtgaa
     1261 ccagcgcggc gagctgtgcg tccgtggccc catgatcatg agcggctacg ttaacaaccc
     1321 cgaggctaca aacgctctca tcgacaagga cggctggctg cacagcggcg acatcgccta
     1381 ctgggacgag gacgagcact tcttcatcgt ggaccggctg aagagcctga tcaaatacaa
     1441 gggctaccag gtagccccag ccgaactgga gagcatcctg ctgcaacacc ccaacatctt
     1501 cgacgccggg gtcgccggcc tgcccgacga cgatgccggc gagctgcccg ccgcagtcgt
     1561 cgtgctggaa cacggtaaaa ccatgaccga gaaggagatc gtggactatg tggccagcca
     1621 ggttacaacc gccaagaagc tgcgcggtgg tgttgtgttc gtggacgagg tgcctaaagg
     1681 actgaccggc aagttggacg cccgcaagat ccgcgagatt ctcattaagg ccaagaaggg
     1741 cggcaagatc gccgtgtaat aattctagag tcggggcggc cggccgcttc gagcagacat
     1801 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     1861 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     1921 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt
     1981 tttttaaagc aagtaaaacc tctacaaatg tggtaaaatc gataaggatc cggagctggg
     2041 attcaaagcc aagtcaggcc ggacacggtg gctcacgcct ataatcccag cactttggga
     2101 ggcagaggcg ggcggatcac gaggccagga gatcgagacc atcctggcta acacggtgaa
     2161 accccatctc tactaaaaaa aatacaaaaa taattagccg ggtgtggtgg cgggtgcctg
     2221 tagtcccagc tactagggag gctgaggcag gagaatggcg tgaacccggg aggtggagtt
     2281 tgcagtgagc cgagatcgtg ccactgcact ccagcctggg cgacagagca agactccatc
     2341 tcaaaaagaa aaaaaaagcc aagtcagaat caaagtgcca cgccagtgct cttcaactgc
     2401 ggtacaagtc acagtcaccc aggagcttta ggaaatccag ctgctgggcc tcagcctaga
     2461 tgaaccaaat cagaacccct ggggtgggct gggcacgggc agtcctaaga gcgctggagg
     2521 gacttctgct gtgcagcctg gcatggcaac cacaccagtc cagggctggg agctgccgta
     2581 ctgcatcgcg gaagtctcag atgtcttaag tctgcagtag atctaaaagg ggctggtaca
     2641 catcaggagc tcggcctgca cccaagagac tggcgtggaa gccagatgcg caggccaggc
     2701 tgcagggtta ccaagtggtt gaggtaagca gatgcgtgag tcattctaca gcaataaaga
     2761 gaggaagcgg accaacgtca ctcacctgcc aactgacgtg cacgactgcg gccgctgtgc
     2821 ccgcgaaagc tttggtgttg agctgtccgg ggagtcacct aagtccaaga aaggaagtgt
     2881 gcggtcaggt aaagtgatca gggggagacc tactgttggg ggatcgtcac cgaatgggca
     2941 attctccgag tatctccact gtcccaactt gagtgtaggt ggcactggct cagacaagta
     3001 cacaaagggc caccccgctg caggcagcca catgagaggc attcacccct acacggggac
     3061 ccctgcccag gcaaaggctg tggtctgatg gatggacacg ggggagaaag agaagagcca
     3121 tcttcactgc gccgccccac cctggctgta tatcccatca gggccgagca cgaccttcac
     3181 agagcgaagt cgacttgtca gacggcacct gcatgcgcta gacacttgcc cacaggaaca
     3241 acaccgcagg gatttagctt cccaggcacg cgcaacagag cccacattcc ctcattttcc
     3301 cgaaccatga ctttactggg ataataataa ttcattacac tgtgtttgtg ctgttttagt
     3361 tgacaagcat ttccatgtgt atgatctcat ctgatattca caacaggatg aaaagcaggc
     3421 tcagagggca aaggggtcac cagttaccga gctagggccc aacacaggcc ccagaggtcc
     3481 agtcccgcca ccacgccccc agacactggc tccatgcatg tgagcaaaag gccagcaaaa
     3541 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     3601 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     3661 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     3721 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     3781 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     3841 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     3901 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     3961 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     4021 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     4081 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     4141 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     4201 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     4261 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     4321 aacttggtct gacagcggcc gcaaatgcta aaccactgca gtggttacca gtgcttgatc
     4381 agtgaggcac cgatctcagc gatctgccta tttcgttcgt ccatagtggc ctgactcccc
     4441 gtcgtgtaga tcactacgat tcgtgagggc ttaccatcag gccccagcgc agcaatgatg
     4501 ccgcgagagc cgcgttcacc ggcccccgat ttgtcagcaa tgaaccagcc agcagggagg
     4561 gccgagcgaa gaagtggtcc tgctactttg tccgcctcca tccagtctat gagctgctgt
     4621 cgtgatgcta gagtaagaag ttcgccagtg agtagtttcc gaagagttgt ggccattgct
     4681 actggcatcg tggtatcacg ctcgtcgttc ggtatggctt cgttcaactc tggttcccag
     4741 cggtcaagcc gggtcacatg atcacccata ttatgaagaa atgcagtcag ctccttaggg
     4801 cctccgatcg ttgtcagaag taagttggcc gcggtgttgt cgctcatggt aatggcagca
     4861 ctacacaatt ctcttaccgt catgccatcc gtaagatgct tttccgtgac cggcgagtac
     4921 tcaaccaagt cgttttgtga gtagtgtata cggcgaccaa gctgctcttg cccggcgtct
     4981 atacgggaca acaccgcgcc acatagcagt actttgaaag tgctcatcat cgggaatcgt
     5041 tcttcggggc ggaaagactc aaggatcttg ccgctattga gatccagttc gatatagccc
     5101 actcttgcac ccagttgatc ttcagcatct tttactttca ccagcgtttc ggggtgtgca
     5161 aaaacaggca agcaaaatgc cgcaaagaag ggaatgagtg cgacacgaaa atgttggatg
     5221 ctcatactcg tcctttttca atattattga agcatttatc agggttacta gtacgtctct
     5281 caaggataag taagtaatat taaggtacgg gaggtattgg acaggccgca ataaaatatc
     5341 tttattttca ttacatctgt gtgttggttt tttgtgtgaa tcgatagtac taacatacgc
     5401 tctccatcaa aacaaaacga aacaaaacaa actagcaaaa taggctgtcc ccagtgcaag
     5461 tgcaggtgcc agaacatttc tct
//
