LOCUS       sequence-192447-        5474 bp    DNA     circular SYN 15-DEC-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      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5474)
  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 5474)
  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_mP_GTSE1
FEATURES             Location/Qualifiers
     source          1..5474
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(18..48)
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     misc_feature    76..167
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     complement(108..127)
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     polyA_signal    181..229
                     /note="synthetic polyadenylation signal"
     CDS             join(334..402,403..1194)
                     /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      1394..1982
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1883..1902
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3626..3747
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3663..3682)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3717..3736
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(3791..5443)
                     /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      5440..5449
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 gctttaggtc gctagccctg gaagtcgagc ttccattata taccctctct cgaagatctc
       61 taagccagtt aggccagaga aatgttctgg cacctgcact tgcactgggg acagcctatt
      121 ttgctagttt gttttgtttc gttttgtttt gatggagagc gtatgttagt actatcgatt
      181 cacacaaaaa accaacacac agatgtaatg aaaataaaga tattttattg cggcctgtcc
      241 aatacctccc gtaccttaat attacttact tatccttgag agacgtacta gtaaccctga
      301 taaatgcttc aataatattg aaaaaggacg agtatgagca tccaacattt tcgtgtcgca
      361 ctcattccct tctttgcggc attttgcttg cctgtttttg cacaccccga aacgctggtg
      421 aaagtaaaag atgctgaaga tcaactgggt gcaagagtgg gctatatcga actggatctc
      481 aatagcggca agatccttga gtctttccgc cccgaagaac gattcccgat gatgagcact
      541 ttcaaagtac tgctatgtgg cgcggtgttg tcccgtatag acgccgggca agagcagctt
      601 ggtcgccgta tacactactc acaaaacgac ttggttgagt actcgccggt cacggaaaag
      661 catcttacgg atggcatgac ggtaagagaa ttgtgtagtg ctgccattac catgagcgac
      721 aacaccgcgg ccaacttact tctgacaacg atcggaggcc ctaaggagct gactgcattt
      781 cttcataata tgggtgatca tgtgacccgg cttgaccgct gggaaccaga gttgaacgaa
      841 gccataccga acgacgagcg tgataccacg atgccagtag caatggccac aactcttcgg
      901 aaactactca ctggcgaact tcttactcta gcatcacgac agcagctcat agactggatg
      961 gaggcggaca aagtagcagg accacttctt cgctcggccc tccctgctgg ctggttcatt
     1021 gctgacaaat cgggggccgg tgaacgcggc tctcgcggca tcattgctgc gctggggcct
     1081 gatggtaagc cctcacgaat cgtagtgatc tacacgacgg ggagtcaggc cactatggac
     1141 gaacgaaata ggcagatcgc tgagatcggt gcctcactga tcaagcactg gtaaccactg
     1201 cagtggttta gcatttgcgg ccgctgtcag accaagttta ctcatatata ctttagattg
     1261 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     1321 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     1381 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     1441 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     1501 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     1561 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     1621 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     1681 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     1741 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     1801 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     1861 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     1921 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     1981 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     2041 tggagccagt gtctgggggc gtggtggcgg gactggacct ctggggcctg tgttgggccc
     2101 tagctcggta actggtgacc cctttgccct ctgagcctgc ttttcatcct gttgtgaata
     2161 tcagatgaga tcatacacat ggaaatgctt gtcaactaaa acagcacaaa cacagtgtaa
     2221 tgaattatta ttatcccagt aaagtcatgg ttcgggaaaa tgagggaatg tgggctctgt
     2281 tgcgcgtgcc tgggaagcta aatccctgcg gtgttgttcc tgtgggcaag tgtctagcgc
     2341 atgcaggtgc cgtctgacaa gtcgacttcg ctctgtgaag gtcgtgctcg gccctgatgg
     2401 gatatacagc cagggtgggg cggcgcagtg aagatggctc ttctctttct cccccgtgtc
     2461 catccatcag accacagcct ttgcctgggc aggggtcccc gtgtaggggt gaatgcctct
     2521 catgtggctg cctgcagcgg ggtggccctt tgtgtacttg tctgagccag tgccacctac
     2581 actcaagttg ggacagtgga gatactcgga gaattgccca ttcggtgacg atcccccaac
     2641 agtaggtctc cccctgatca ctttacctga ccgcacactt cctttcttgg acttaggtga
     2701 ctccccggac agctcaacac caaagctttc gcgggcacag cggccgcagt cgtgcacgtc
     2761 agttggcagg tgagtgacgt tggtccgctt cctctcttta ttgctgtaga atgactcacg
     2821 catctgctta cctcaaccac ttggtaaccc tgcagcctgg cctgcgcatc tggcttccac
     2881 gccagtctct tgggtgcagg ccgagctcct gatgtgtacc agcccctttt agatctactg
     2941 cagacttaag acatctgaga cttccgcgat gcagtacggc agctcccagc cctggactgg
     3001 tgtggttgcc atgccaggct gcacagcaga agtccctcca gcgctcttag gactgcccgt
     3061 gcccagccca ccccaggggt tctgatttgg ttcatctagg ctgaggccca gcagctggat
     3121 ttcctaaagc tcctgggtga ctgtgacttg taccgcagtt gaagagcact ggcgtggcac
     3181 tttgattctg acttggcttt ttttttcttt ttgagatgga gtcttgctct gtcgcccagg
     3241 ctggagtgca gtggcacgat ctcggctcac tgcaaactcc acctcccggg ttcacgccat
     3301 tctcctgcct cagcctccct agtagctggg actacaggca cccgccacca cacccggcta
     3361 attatttttg tatttttttt agtagagatg gggtttcacc gtgttagcca ggatggtctc
     3421 gatctcctgg cctcgtgatc cgcccgcctc tgcctcccaa agtgctggga ttataggcgt
     3481 gagccaccgt gtccggcctg acttggcttt gaatcccagc tcccttatcg attttaccac
     3541 atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca
     3601 taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata
     3661 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     3721 tttgtccaaa ctcatcaatg tatcttatca tgtctgctcg aagcggccgg ccgccccgac
     3781 tctagaatta ttacacggcg atcttgccgc ccttcttggc cttaatgaga atctcgcgga
     3841 tcttgcgggc gtccaacttg ccggtcagtc ctttaggcac ctcgtccacg aacacaacac
     3901 caccgcgcag cttcttggcg gttgtaacct ggctggccac atagtccacg atctccttct
     3961 cggtcatggt tttaccgtgt tccagcacga cgactgcggc gggcagctcg ccggcatcgt
     4021 cgtcgggcag gccggcgacc ccggcgtcga agatgttggg gtgttgcagc aggatgctct
     4081 ccagttcggc tggggctacc tggtagccct tgtatttgat caggctcttc agccggtcca
     4141 cgatgaagaa gtgctcgtcc tcgtcccagt aggcgatgtc gccgctgtgc agccagccgt
     4201 ccttgtcgat gagagcgttt gtagcctcgg ggttgttaac gtagccgctc atgatcatgg
     4261 ggccacggac gcacagctcg ccgcgctggt tcacacccag tgtcttaccg gtgtccaagt
     4321 ccaccacctt agcctcgaag aagggcacca ccttgcctac tgcgccaggc ttgtcgtccc
     4381 cttcgggggt gatcagaatg gcgctggttg tttctgtcag gccgtagccc tggcggatgc
     4441 ctggtaggtg gaagcgtttg gccacggcct cacctacctc cttgctgagc ggcgccccgc
     4501 cgctggcgat ctcgtgcaag ttgcttaggt cgtacttgtc gatgagagtg ctcttagcga
     4561 agaagctaaa tagtgtgggc accagcaggg cagattgaat cttatagtct tgcaagctgc
     4621 gcaagaatag ctcctcctcg aagcggtaca tgagcacgac ccgaaagccg cagatcaagt
     4681 agcccagcgt ggtgaacatg ccgaagccgt ggtgaaatgg caccacgctg aggatagcgg
     4741 tgtcggggat gatctggttg ccgaagatgg ggtcgcgggc atgactgaat cggacacaag
     4801 cggtgcggtg cggtagggct acgcccttgg gcaatccggt actgccacta ctgttcatga
     4861 tcagggcgat ggttttgtcc cggtcgaagc tctcgggcac gaagtcgtac tcgttgaagc
     4921 cgggtggcaa atgggaagtc acgaaggtgt acatgctttg gaagccctgg tagtcggtct
     4981 tgctatccat gatgatgatc ttttgtatga tcggtagctt cttttgcacg ttgaggatct
     5041 tttgcagccc tttcttgctc acgaatacga cggtgggctg gctgatgccc atgctgttca
     5101 gcagctcgcg ctcgttgtag atgtcgttag ctggggccac agccacaccg atgaacaggg
     5161 cacccaacac gggcatgaag aactgcaagc tattctcgct gcacaccacg atccgatggt
     5221 ttgtattcag cccatagcgc ttcatagctt ctgccagccg aacgctcatc tcgaagtact
     5281 cggcgtaggt aatgtccacc tcgatatgtg cgtcggtaaa ggcgatggtg ccgggcacca
     5341 gggcgtagcg cttcatggct ttgtgcagct gctcgccggc ggtcccgtct tcgagtgggt
     5401 agaatggcgc tgggcccttc ttaatgtttt tggcatcttc catggtggct ttaccaacag
     5461 taccggattg ccaa
//
