LOCUS       sequence-190935-        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"
     primer_bind     173..192
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     426..443
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    691..812
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(728..747)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     782..801
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(856..2508)
                     /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      2505..2514
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2557..2587)
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     misc_feature    2612..2703
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     complement(2644..2663)
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     polyA_signal    2717..2765
                     /note="synthetic polyadenylation signal"
     CDS             join(2870..2938,2939..3730)
                     /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      join(3930..4246,1..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 tgttctttcc tgcgttatcc cctgattctg
      361 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
      421 agcgcagcga gtcagtgagc gaggaagcgg aagagcgctg ccggcacctg tcctacgagt
      481 tgcatgataa agaagacagt cataagtgcg gcgacgatag tcatgccccg cgcccaccgg
      541 aaggagctga ctgggttgaa ggctctcaag ggcatcggtc gacggatcct tatcgatttt
      601 accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg
      661 aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac
      721 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
      781 tgtggtttgt ccaaactcat caatgtatct tatcatgtct gctcgaagcg gccggccgcc
      841 ccgactctag aattattaca cggcgatctt gccgcccttc ttggccttaa tgagaatctc
      901 gcggatcttg cgggcgtcca acttgccggt cagtccttta ggcacctcgt ccacgaacac
      961 aacaccaccg cgcagcttct tggcggttgt aacctggctg gccacatagt ccacgatctc
     1021 cttctcggtc atggttttac cgtgttccag cacgacgact gcggcgggca gctcgccggc
     1081 atcgtcgtcg ggcaggccgg cgaccccggc gtcgaagatg ttggggtgtt gcagcaggat
     1141 gctctccagt tcggctgggg ctacctggta gcccttgtat ttgatcaggc tcttcagccg
     1201 gtccacgatg aagaagtgct cgtcctcgtc ccagtaggcg atgtcgccgc tgtgcagcca
     1261 gccgtccttg tcgatgagag cgtttgtagc ctcggggttg ttaacgtagc cgctcatgat
     1321 catggggcca cggacgcaca gctcgccgcg ctggttcaca cccagtgtct taccggtgtc
     1381 caagtccacc accttagcct cgaagaaggg caccaccttg cctactgcgc caggcttgtc
     1441 gtccccttcg ggggtgatca gaatggcgct ggttgtttct gtcaggccgt agccctggcg
     1501 gatgcctggt aggtggaagc gtttggccac ggcctcacct acctccttgc tgagcggcgc
     1561 cccgccgctg gcgatctcgt gcaagttgct taggtcgtac ttgtcgatga gagtgctctt
     1621 agcgaagaag ctaaatagtg tgggcaccag cagggcagat tgaatcttat agtcttgcaa
     1681 gctgcgcaag aatagctcct cctcgaagcg gtacatgagc acgacccgaa agccgcagat
     1741 caagtagccc agcgtggtga acatgccgaa gccgtggtga aatggcacca cgctgaggat
     1801 agcggtgtcg gggatgatct ggttgccgaa gatggggtcg cgggcatgac tgaatcggac
     1861 acaagcggtg cggtgcggta gggctacgcc cttgggcaat ccggtactgc cactactgtt
     1921 catgatcagg gcgatggttt tgtcccggtc gaagctctcg ggcacgaagt cgtactcgtt
     1981 gaagccgggt ggcaaatggg aagtcacgaa ggtgtacatg ctttggaagc cctggtagtc
     2041 ggtcttgcta tccatgatga tgatcttttg tatgatcggt agcttctttt gcacgttgag
     2101 gatcttttgc agccctttct tgctcacgaa tacgacggtg ggctggctga tgcccatgct
     2161 gttcagcagc tcgcgctcgt tgtagatgtc gttagctggg gccacagcca caccgatgaa
     2221 cagggcaccc aacacgggca tgaagaactg caagctattc tcgctgcaca ccacgatccg
     2281 atggtttgta ttcagcccat agcgcttcat agcttctgcc agccgaacgc tcatctcgaa
     2341 gtactcggcg taggtaatgt ccacctcgat atgtgcgtcg gtaaaggcga tggtgccggg
     2401 caccagggcg tagcgcttca tggctttgtg cagctgctcg ccggcggtcc cgtcttcgag
     2461 tgggtagaat ggcgctgggc ccttcttaat gtttttggca tcttccatgg tggctttacc
     2521 aacagtaccg gattgccaag ctttaggtcg ctagccctgg aagtcgagct tccattatat
     2581 accctctctc gaagatccgg ccagttaggc cagagaaatg ttctggcacc tgcacttgca
     2641 ctggggacag cctattttgc tagtttgttt tgtttcgttt tgttttgatg gagagcgtat
     2701 gttagtacta tcgattcaca caaaaaacca acacacagat gtaatgaaaa taaagatatt
     2761 ttattgcggc ctgtccaata cctcccgtac cttaatatta cttacttatc cttgagagac
     2821 gtactagtaa ccctgataaa tgcttcaata atattgaaaa aggacgagta tgagcatcca
     2881 acattttcgt gtcgcactca ttcccttctt tgcggcattt tgcttgcctg tttttgcaca
     2941 ccccgaaacg ctggtgaaag taaaagatgc tgaagatcaa ctgggtgcaa gagtgggcta
     3001 tatcgaactg gatctcaata gcggcaagat ccttgagtct ttccgccccg aagaacgatt
     3061 cccgatgatg agcactttca aagtactgct atgtggcgcg gtgttgtccc gtatagacgc
     3121 cgggcaagag cagcttggtc gccgtataca ctactcacaa aacgacttgg ttgagtactc
     3181 gccggtcacg gaaaagcatc ttacggatgg catgacggta agagaattgt gtagtgctgc
     3241 cattaccatg agcgacaaca ccgcggccaa cttacttctg acaacgatcg gaggccctaa
     3301 ggagctgact gcatttcttc ataatatggg tgatcatgtg acccggcttg accgctggga
     3361 accagagttg aacgaagcca taccgaacga cgagcgtgat accacgatgc cagtagcaat
     3421 ggccacaact cttcggaaac tactcactgg cgaacttctt actctagcat cacgacagca
     3481 gctcatagac tggatggagg cggacaaagt agcaggacca cttcttcgct cggccctccc
     3541 tgctggctgg ttcattgctg acaaatcggg ggccggtgaa cgcggctctc gcggcatcat
     3601 tgctgcgctg gggcctgatg gtaagccctc acgaatcgta gtgatctaca cgacggggag
     3661 tcaggccact atggacgaac gaaataggca gatcgctgag atcggtgcct cactgatcaa
     3721 gcactggtaa ccactgcagt ggtttagcat ttgcggccgc tgtcagacca agtttactca
     3781 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     3841 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     3901 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     3961 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     4021 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     4081 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     4141 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     4201 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggct
//
