LOCUS       sequence-190944-        4298 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Luciferase validation vector; empty vector without insert to 
            determine baseline.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4298)
  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 4298)
  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_SCP1_empty
FEATURES             Location/Qualifiers
     source          1..4298
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      283..871
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     772..791
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1025..1042
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    1290..1411
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1327..1346)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1381..1400
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(1455..3107)
                     /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      3104..3113
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    3263..3354
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     complement(3295..3314)
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     polyA_signal    3368..3416
                     /note="synthetic polyadenylation signal"
     CDS             join(3521..3589,3590..4298,1..83)
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
ORIGIN
        1 acacgacggg gagtcaggcc actatggacg aacgaaatag gcagatcgct gagatcggtg
       61 cctcactgat caagcactgg taaccactgc agtggtttag catttgcggc cgctgtcaga
      121 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
      181 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
      241 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
      301 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
      361 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
      421 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
      481 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
      541 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
      601 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
      661 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
      721 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
      781 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
      841 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
      901 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
      961 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     1021 gcgcagcgag tcagtgagcg aggaagcgga agagcgctgc cggcacctgt cctacgagtt
     1081 gcatgataaa gaagacagtc ataagtgcgg cgacgatagt catgccccgc gcccaccgga
     1141 aggagctgac tgggttgaag gctctcaagg gcatcggtcg acggatcctt atcgatttta
     1201 ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga
     1261 aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca
     1321 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     1381 gtggtttgtc caaactcatc aatgtatctt atcatgtctg ctcgaagcgg ccggccgccc
     1441 cgactctaga attattacac ggcgatcttg ccgcccttct tggccttaat gagaatctcg
     1501 cggatcttgc gggcgtccaa cttgccggtc agtcctttag gcacctcgtc cacgaacaca
     1561 acaccaccgc gcagcttctt ggcggttgta acctggctgg ccacatagtc cacgatctcc
     1621 ttctcggtca tggttttacc gtgttccagc acgacgactg cggcgggcag ctcgccggca
     1681 tcgtcgtcgg gcaggccggc gaccccggcg tcgaagatgt tggggtgttg cagcaggatg
     1741 ctctccagtt cggctggggc tacctggtag cccttgtatt tgatcaggct cttcagccgg
     1801 tccacgatga agaagtgctc gtcctcgtcc cagtaggcga tgtcgccgct gtgcagccag
     1861 ccgtccttgt cgatgagagc gtttgtagcc tcggggttgt taacgtagcc gctcatgatc
     1921 atggggccac ggacgcacag ctcgccgcgc tggttcacac ccagtgtctt accggtgtcc
     1981 aagtccacca ccttagcctc gaagaagggc accaccttgc ctactgcgcc aggcttgtcg
     2041 tccccttcgg gggtgatcag aatggcgctg gttgtttctg tcaggccgta gccctggcgg
     2101 atgcctggta ggtggaagcg tttggccacg gcctcaccta cctccttgct gagcggcgcc
     2161 ccgccgctgg cgatctcgtg caagttgctt aggtcgtact tgtcgatgag agtgctctta
     2221 gcgaagaagc taaatagtgt gggcaccagc agggcagatt gaatcttata gtcttgcaag
     2281 ctgcgcaaga atagctcctc ctcgaagcgg tacatgagca cgacccgaaa gccgcagatc
     2341 aagtagccca gcgtggtgaa catgccgaag ccgtggtgaa atggcaccac gctgaggata
     2401 gcggtgtcgg ggatgatctg gttgccgaag atggggtcgc gggcatgact gaatcggaca
     2461 caagcggtgc ggtgcggtag ggctacgccc ttgggcaatc cggtactgcc actactgttc
     2521 atgatcaggg cgatggtttt gtcccggtcg aagctctcgg gcacgaagtc gtactcgttg
     2581 aagccgggtg gcaaatggga agtcacgaag gtgtacatgc tttggaagcc ctggtagtcg
     2641 gtcttgctat ccatgatgat gatcttttgt atgatcggta gcttcttttg cacgttgagg
     2701 atcttttgca gccctttctt gctcacgaat acgacggtgg gctggctgat gcccatgctg
     2761 ttcagcagct cgcgctcgtt gtagatgtcg ttagctgggg ccacagccac accgatgaac
     2821 agggcaccca acacgggcat gaagaactgc aagctattct cgctgcacac cacgatccga
     2881 tggtttgtat tcagcccata gcgcttcata gcttctgcca gccgaacgct catctcgaag
     2941 tactcggcgt aggtaatgtc cacctcgata tgtgcgtcgg taaaggcgat ggtgccgggc
     3001 accagggcgt agcgcttcat ggctttgtgc agctgctcgc cggcggtccc gtcttcgagt
     3061 gggtagaatg gcgctgggcc cttcttaatg tttttggcat cttccatggt ggctttacca
     3121 acagtaccgg attgccaagc tttaggtcgc tagcccggtc cgtaggcacg tctgctcggc
     3181 tcgagtgttc gatcgcgact gaggacgaac gcgcccccac ccccttatat aagtacccta
     3241 gggagatccg gccagttagg ccagagaaat gttctggcac ctgcacttgc actggggaca
     3301 gcctattttg ctagtttgtt ttgtttcgtt ttgttttgat ggagagcgta tgttagtact
     3361 atcgattcac acaaaaaacc aacacacaga tgtaatgaaa ataaagatat tttattgcgg
     3421 cctgtccaat acctcccgta ccttaatatt acttacttat ccttgagaga cgtactagta
     3481 accctgataa atgcttcaat aatattgaaa aaggacgagt atgagcatcc aacattttcg
     3541 tgtcgcactc attcccttct ttgcggcatt ttgcttgcct gtttttgcac accccgaaac
     3601 gctggtgaaa gtaaaagatg ctgaagatca actgggtgca agagtgggct atatcgaact
     3661 ggatctcaat agcggcaaga tccttgagtc tttccgcccc gaagaacgat tcccgatgat
     3721 gagcactttc aaagtactgc tatgtggcgc ggtgttgtcc cgtatagacg ccgggcaaga
     3781 gcagcttggt cgccgtatac actactcaca aaacgacttg gttgagtact cgccggtcac
     3841 ggaaaagcat cttacggatg gcatgacggt aagagaattg tgtagtgctg ccattaccat
     3901 gagcgacaac accgcggcca acttacttct gacaacgatc ggaggcccta aggagctgac
     3961 tgcatttctt cataatatgg gtgatcatgt gacccggctt gaccgctggg aaccagagtt
     4021 gaacgaagcc ataccgaacg acgagcgtga taccacgatg ccagtagcaa tggccacaac
     4081 tcttcggaaa ctactcactg gcgaacttct tactctagca tcacgacagc agctcataga
     4141 ctggatggag gcggacaaag tagcaggacc acttcttcgc tcggccctcc ctgctggctg
     4201 gttcattgct gacaaatcgg gggccggtga acgcggctct cgcggcatca ttgctgcgct
     4261 ggggcctgat ggtaagccct cacgaatcgt agtgatct
//
