LOCUS       sequence-183840-        4246 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 4246)
  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 4246)
  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_empty
FEATURES             Location/Qualifiers
     source          1..4246
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        25..55
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      98..107
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             104..1756
                     /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    1800..1921
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1811..1830)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1865..1884
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2169..2186)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2340..2928)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2420..2439)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3128..3919,3920..3988))
                     /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    4093..4141
                     /note="synthetic polyadenylation signal"
     misc_feature    4155..4246
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4195..4214
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gccggatctt cgagagaggg tatataatgg aagctcgact tccagggcta
       61 gcgacctaaa gcttggcaat ccggtactgt tggtaaagcc accatggaag atgccaaaaa
      121 cattaagaag ggcccagcgc cattctaccc actcgaagac gggaccgccg gcgagcagct
      181 gcacaaagcc atgaagcgct acgccctggt gcccggcacc atcgccttta ccgacgcaca
      241 tatcgaggtg gacattacct acgccgagta cttcgagatg agcgttcggc tggcagaagc
      301 tatgaagcgc tatgggctga atacaaacca tcggatcgtg gtgtgcagcg agaatagctt
      361 gcagttcttc atgcccgtgt tgggtgccct gttcatcggt gtggctgtgg ccccagctaa
      421 cgacatctac aacgagcgcg agctgctgaa cagcatgggc atcagccagc ccaccgtcgt
      481 attcgtgagc aagaaagggc tgcaaaagat cctcaacgtg caaaagaagc taccgatcat
      541 acaaaagatc atcatcatgg atagcaagac cgactaccag ggcttccaaa gcatgtacac
      601 cttcgtgact tcccatttgc cacccggctt caacgagtac gacttcgtgc ccgagagctt
      661 cgaccgggac aaaaccatcg ccctgatcat gaacagtagt ggcagtaccg gattgcccaa
      721 gggcgtagcc ctaccgcacc gcaccgcttg tgtccgattc agtcatgccc gcgaccccat
      781 cttcggcaac cagatcatcc ccgacaccgc tatcctcagc gtggtgccat ttcaccacgg
      841 cttcggcatg ttcaccacgc tgggctactt gatctgcggc tttcgggtcg tgctcatgta
      901 ccgcttcgag gaggagctat tcttgcgcag cttgcaagac tataagattc aatctgccct
      961 gctggtgccc acactattta gcttcttcgc taagagcact ctcatcgaca agtacgacct
     1021 aagcaacttg cacgagatcg ccagcggcgg ggcgccgctc agcaaggagg taggtgaggc
     1081 cgtggccaaa cgcttccacc taccaggcat ccgccagggc tacggcctga cagaaacaac
     1141 cagcgccatt ctgatcaccc ccgaagggga cgacaagcct ggcgcagtag gcaaggtggt
     1201 gcccttcttc gaggctaagg tggtggactt ggacaccggt aagacactgg gtgtgaacca
     1261 gcgcggcgag ctgtgcgtcc gtggccccat gatcatgagc ggctacgtta acaaccccga
     1321 ggctacaaac gctctcatcg acaaggacgg ctggctgcac agcggcgaca tcgcctactg
     1381 ggacgaggac gagcacttct tcatcgtgga ccggctgaag agcctgatca aatacaaggg
     1441 ctaccaggta gccccagccg aactggagag catcctgctg caacacccca acatcttcga
     1501 cgccggggtc gccggcctgc ccgacgacga tgccggcgag ctgcccgccg cagtcgtcgt
     1561 gctggaacac ggtaaaacca tgaccgagaa ggagatcgtg gactatgtgg ccagccaggt
     1621 tacaaccgcc aagaagctgc gcggtggtgt tgtgttcgtg gacgaggtgc ctaaaggact
     1681 gaccggcaag ttggacgccc gcaagatccg cgagattctc attaaggcca agaagggcgg
     1741 caagatcgcc gtgtaataat tctagagtcg gggcggccgg ccgcttcgag cagacatgat
     1801 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     1861 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     1921 taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt
     1981 ttaaagcaag taaaacctct acaaatgtgg taaaatcgat aaggatccgt cgaccgatgc
     2041 ccttgagagc cttcaaccca gtcagctcct tccggtgggc gcggggcatg actatcgtcg
     2101 ccgcacttat gactgtcttc tttatcatgc aactcgtagg acaggtgccg gcagcgctct
     2161 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     2221 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     2281 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     2341 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     2401 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     2461 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     2521 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     2581 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     2641 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     2701 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     2761 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     2821 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     2881 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     2941 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     3001 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     3061 tcaatctaaa gtatatatga gtaaacttgg tctgacagcg gccgcaaatg ctaaaccact
     3121 gcagtggtta ccagtgcttg atcagtgagg caccgatctc agcgatctgc ctatttcgtt
     3181 cgtccatagt ggcctgactc cccgtcgtgt agatcactac gattcgtgag ggcttaccat
     3241 caggccccag cgcagcaatg atgccgcgag agccgcgttc accggccccc gatttgtcag
     3301 caatgaacca gccagcaggg agggccgagc gaagaagtgg tcctgctact ttgtccgcct
     3361 ccatccagtc tatgagctgc tgtcgtgatg ctagagtaag aagttcgcca gtgagtagtt
     3421 tccgaagagt tgtggccatt gctactggca tcgtggtatc acgctcgtcg ttcggtatgg
     3481 cttcgttcaa ctctggttcc cagcggtcaa gccgggtcac atgatcaccc atattatgaa
     3541 gaaatgcagt cagctcctta gggcctccga tcgttgtcag aagtaagttg gccgcggtgt
     3601 tgtcgctcat ggtaatggca gcactacaca attctcttac cgtcatgcca tccgtaagat
     3661 gcttttccgt gaccggcgag tactcaacca agtcgttttg tgagtagtgt atacggcgac
     3721 caagctgctc ttgcccggcg tctatacggg acaacaccgc gccacatagc agtactttga
     3781 aagtgctcat catcgggaat cgttcttcgg ggcggaaaga ctcaaggatc ttgccgctat
     3841 tgagatccag ttcgatatag cccactcttg cacccagttg atcttcagca tcttttactt
     3901 tcaccagcgt ttcggggtgt gcaaaaacag gcaagcaaaa tgccgcaaag aagggaatga
     3961 gtgcgacacg aaaatgttgg atgctcatac tcgtcctttt tcaatattat tgaagcattt
     4021 atcagggtta ctagtacgtc tctcaaggat aagtaagtaa tattaaggta cgggaggtat
     4081 tggacaggcc gcaataaaat atctttattt tcattacatc tgtgtgttgg ttttttgtgt
     4141 gaatcgatag tactaacata cgctctccat caaaacaaaa cgaaacaaaa caaactagca
     4201 aaataggctg tccccagtgc aagtgcaggt gccagaacat ttctct
//
