LOCUS       sequence-193159-        5468 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 5468)
  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 5468)
  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_ORI_GTSE1
FEATURES             Location/Qualifiers
     source          1..5468
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     173..192
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(462..1253,1254..1322))
                     /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     1480..1497
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    3230..3351
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3267..3286)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3321..3340
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(3395..5047)
                     /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      5044..5053
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     rep_origin      join(5152..5468,1..272)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
       61 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
      121 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
      181 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
      241 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
      301 tcctggcctt ttgctggcct tttgctcaca tgtcatgaga ttatcaaaaa ggatcttcac
      361 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
      421 ttggtctgac agcggccgca aatgctaaac cactgcagtg gttaccagtg cttgatcagt
      481 gaggcaccga tctcagcgat ctgcctattt cgttcgtcca tagtggcctg actccccgtc
      541 gtgtagatca ctacgattcg tgagggctta ccatcaggcc ccagcgcagc aatgatgccg
      601 cgagagccgc gttcaccggc ccccgatttg tcagcaatga accagccagc agggagggcc
      661 gagcgaagaa gtggtcctgc tactttgtcc gcctccatcc agtctatgag ctgctgtcgt
      721 gatgctagag taagaagttc gccagtgagt agtttccgaa gagttgtggc cattgctact
      781 ggcatcgtgg tatcacgctc gtcgttcggt atggcttcgt tcaactctgg ttcccagcgg
      841 tcaagccggg tcacatgatc acccatatta tgaagaaatg cagtcagctc cttagggcct
      901 ccgatcgttg tcagaagtaa gttggccgcg gtgttgtcgc tcatggtaat ggcagcacta
      961 cacaattctc ttaccgtcat gccatccgta agatgctttt ccgtgaccgg cgagtactca
     1021 accaagtcgt tttgtgagta gtgtatacgg cgaccaagct gctcttgccc ggcgtctata
     1081 cgggacaaca ccgcgccaca tagcagtact ttgaaagtgc tcatcatcgg gaatcgttct
     1141 tcggggcgga aagactcaag gatcttgccg ctattgagat ccagttcgat atagcccact
     1201 cttgcaccca gttgatcttc agcatctttt actttcacca gcgtttcggg gtgtgcaaaa
     1261 acaggcaagc aaaatgccgc aaagaaggga atgagtgcga cacgaaaatg ttggatgctc
     1321 atactcgtcc tttttcaata ttattgaagc atttatcagg gttactagta cgtctctcaa
     1381 gacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     1441 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     1501 gcggaagagc gctgccggca cctgtcctac gagttgcatg ataaagaaga cagtcataag
     1561 tgcggcgacg atagtcatgc cccgcgccca ccggaaggag ctgactgggt tgaaggctct
     1621 caagggcatc ggtcgaccat ggagccagtg tctgggggcg tggtggcggg actggacctc
     1681 tggggcctgt gttgggccct agctcggtaa ctggtgaccc ctttgccctc tgagcctgct
     1741 tttcatcctg ttgtgaatat cagatgagat catacacatg gaaatgcttg tcaactaaaa
     1801 cagcacaaac acagtgtaat gaattattat tatcccagta aagtcatggt tcgggaaaat
     1861 gagggaatgt gggctctgtt gcgcgtgcct gggaagctaa atccctgcgg tgttgttcct
     1921 gtgggcaagt gtctagcgca tgcaggtgcc gtctgacaag tcgacttcgc tctgtgaagg
     1981 tcgtgctcgg ccctgatggg atatacagcc agggtggggc ggcgcagtga agatggctct
     2041 tctctttctc ccccgtgtcc atccatcaga ccacagcctt tgcctgggca ggggtccccg
     2101 tgtaggggtg aatgcctctc atgtggctgc ctgcagcggg gtggcccttt gtgtacttgt
     2161 ctgagccagt gccacctaca ctcaagttgg gacagtggag atactcggag aattgcccat
     2221 tcggtgacga tcccccaaca gtaggtctcc ccctgatcac tttacctgac cgcacacttc
     2281 ctttcttgga cttaggtgac tccccggaca gctcaacacc aaagctttcg cgggcacagc
     2341 ggccgcagtc gtgcacgtca gttggcaggt gagtgacgtt ggtccgcttc ctctctttat
     2401 tgctgtagaa tgactcacgc atctgcttac ctcaaccact tggtaaccct gcagcctggc
     2461 ctgcgcatct ggcttccacg ccagtctctt gggtgcaggc cgagctcctg atgtgtacca
     2521 gcccctttta gatctactgc agacttaaga catctgagac ttccgcgatg cagtacggca
     2581 gctcccagcc ctggactggt gtggttgcca tgccaggctg cacagcagaa gtccctccag
     2641 cgctcttagg actgcccgtg cccagcccac cccaggggtt ctgatttggt tcatctaggc
     2701 tgaggcccag cagctggatt tcctaaagct cctgggtgac tgtgacttgt accgcagttg
     2761 aagagcactg gcgtggcact ttgattctga cttggctttt tttttctttt tgagatggag
     2821 tcttgctctg tcgcccaggc tggagtgcag tggcacgatc tcggctcact gcaaactcca
     2881 cctcccgggt tcacgccatt ctcctgcctc agcctcccta gtagctggga ctacaggcac
     2941 ccgccaccac acccggctaa ttatttttgt atttttttta gtagagatgg ggtttcaccg
     3001 tgttagccag gatggtctcg atctcctggc ctcgtgatcc gcccgcctct gcctcccaaa
     3061 gtgctgggat tataggcgtg agccaccgtg tccggcctga cttggctttg aatcccagct
     3121 ccggatcctt atcgatttta ccacatttgt agaggtttta cttgctttaa aaaacctccc
     3181 acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat
     3241 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     3301 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     3361 ctcgaagcgg ccggccgccc cgactctaga attattacac ggcgatcttg ccgcccttct
     3421 tggccttaat gagaatctcg cggatcttgc gggcgtccaa cttgccggtc agtcctttag
     3481 gcacctcgtc cacgaacaca acaccaccgc gcagcttctt ggcggttgta acctggctgg
     3541 ccacatagtc cacgatctcc ttctcggtca tggttttacc gtgttccagc acgacgactg
     3601 cggcgggcag ctcgccggca tcgtcgtcgg gcaggccggc gaccccggcg tcgaagatgt
     3661 tggggtgttg cagcaggatg ctctccagtt cggctggggc tacctggtag cccttgtatt
     3721 tgatcaggct cttcagccgg tccacgatga agaagtgctc gtcctcgtcc cagtaggcga
     3781 tgtcgccgct gtgcagccag ccgtccttgt cgatgagagc gtttgtagcc tcggggttgt
     3841 taacgtagcc gctcatgatc atggggccac ggacgcacag ctcgccgcgc tggttcacac
     3901 ccagtgtctt accggtgtcc aagtccacca ccttagcctc gaagaagggc accaccttgc
     3961 ctactgcgcc aggcttgtcg tccccttcgg gggtgatcag aatggcgctg gttgtttctg
     4021 tcaggccgta gccctggcgg atgcctggta ggtggaagcg tttggccacg gcctcaccta
     4081 cctccttgct gagcggcgcc ccgccgctgg cgatctcgtg caagttgctt aggtcgtact
     4141 tgtcgatgag agtgctctta gcgaagaagc taaatagtgt gggcaccagc agggcagatt
     4201 gaatcttata gtcttgcaag ctgcgcaaga atagctcctc ctcgaagcgg tacatgagca
     4261 cgacccgaaa gccgcagatc aagtagccca gcgtggtgaa catgccgaag ccgtggtgaa
     4321 atggcaccac gctgaggata gcggtgtcgg ggatgatctg gttgccgaag atggggtcgc
     4381 gggcatgact gaatcggaca caagcggtgc ggtgcggtag ggctacgccc ttgggcaatc
     4441 cggtactgcc actactgttc atgatcaggg cgatggtttt gtcccggtcg aagctctcgg
     4501 gcacgaagtc gtactcgttg aagccgggtg gcaaatggga agtcacgaag gtgtacatgc
     4561 tttggaagcc ctggtagtcg gtcttgctat ccatgatgat gatcttttgt atgatcggta
     4621 gcttcttttg cacgttgagg atcttttgca gccctttctt gctcacgaat acgacggtgg
     4681 gctggctgat gcccatgctg ttcagcagct cgcgctcgtt gtagatgtcg ttagctgggg
     4741 ccacagccac accgatgaac agggcaccca acacgggcat gaagaactgc aagctattct
     4801 cgctgcacac cacgatccga tggtttgtat tcagcccata gcgcttcata gcttctgcca
     4861 gccgaacgct catctcgaag tactcggcgt aggtaatgtc cacctcgata tgtgcgtcgg
     4921 taaaggcgat ggtgccgggc accagggcgt agcgcttcat ggctttgtgc agctgctcgc
     4981 cggcggtccc gtcttcgagt gggtagaatg gcgctgggcc cttcttaatg tttttggcat
     5041 cttccatggt ggctttacca acagtaccgg attgccaagc ttcaaaatcc cttaacgtga
     5101 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     5161 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     5221 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     5281 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     5341 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     5401 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     5461 gtcgggct
//
