LOCUS       sequence-183851-        4220 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Luciferase validation vector with SV40 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 4220)
  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 4220)
  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_SV40
FEATURES             Location/Qualifiers
     source          1..4220
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(65..133,134..925)
                     /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      complement(1115..1703)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1195..1214)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     regulatory      1802..1811
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1808..3460
                     /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    3504..3625
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3515..3534)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3569..3588
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3804..3824)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     primer_bind     complement(4110..4127)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 catgtcttga gagacgtact agtaaccctg ataaatgctt caataatatt gaaaaaggac
       61 gagtatgagc atccaacatt ttcgtgtcgc actcattccc ttctttgcgg cattttgctt
      121 gcctgttttt gcacaccccg aaacgctggt gaaagtaaaa gatgctgaag atcaactggg
      181 tgcaagagtg ggctatatcg aactggatct caatagcggc aagatccttg agtctttccg
      241 ccccgaagaa cgattcccga tgatgagcac tttcaaagta ctgctatgtg gcgcggtgtt
      301 gtcccgtata gacgccgggc aagagcagct tggtcgccgt atacactact cacaaaacga
      361 cttggttgag tactcgccgg tcacggaaaa gcatcttacg gatggcatga cggtaagaga
      421 attgtgtagt gctgccatta ccatgagcga caacaccgcg gccaacttac ttctgacaac
      481 gatcggaggc cctaaggagc tgactgcatt tcttcataat atgggtgatc atgtgacccg
      541 gcttgaccgc tgggaaccag agttgaacga agccataccg aacgacgagc gtgataccac
      601 gatgccagta gcaatggcca caactcttcg gaaactactc actggcgaac ttcttactct
      661 agcatcacga cagcagctca tagactggat ggaggcggac aaagtagcag gaccacttct
      721 tcgctcggcc ctccctgctg gctggttcat tgctgacaaa tcgggggccg gtgaacgcgg
      781 ctctcgcggc atcattgctg cgctggggcc tgatggtaag ccctcacgaa tcgtagtgat
      841 ctacacgacg gggagtcagg ccactatgga cgaacgaaat aggcagatcg ctgagatcgg
      901 tgcctcactg atcaagcact ggtaaccact gcagtggttt agcatttgcg gccgctgtca
      961 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     1021 atctaggtga agatcctttt tgataatctc atgacatgtg agcaaaaggc cagcaaaagg
     1081 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     1141 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     1201 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     1261 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     1321 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     1381 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     1441 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     1501 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     1561 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     1621 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     1681 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     1741 agtggaacga aaactcacgt taagggattt tgaagcttgg caatccggta ctgttggtaa
     1801 agccaccatg gaagatgcca aaaacattaa gaagggccca gcgccattct acccactcga
     1861 agacgggacc gccggcgagc agctgcacaa agccatgaag cgctacgccc tggtgcccgg
     1921 caccatcgcc tttaccgacg cacatatcga ggtggacatt acctacgccg agtacttcga
     1981 gatgagcgtt cggctggcag aagctatgaa gcgctatggg ctgaatacaa accatcggat
     2041 cgtggtgtgc agcgagaata gcttgcagtt cttcatgccc gtgttgggtg ccctgttcat
     2101 cggtgtggct gtggccccag ctaacgacat ctacaacgag cgcgagctgc tgaacagcat
     2161 gggcatcagc cagcccaccg tcgtattcgt gagcaagaaa gggctgcaaa agatcctcaa
     2221 cgtgcaaaag aagctaccga tcatacaaaa gatcatcatc atggatagca agaccgacta
     2281 ccagggcttc caaagcatgt acaccttcgt gacttcccat ttgccacccg gcttcaacga
     2341 gtacgacttc gtgcccgaga gcttcgaccg ggacaaaacc atcgccctga tcatgaacag
     2401 tagtggcagt accggattgc ccaagggcgt agccctaccg caccgcaccg cttgtgtccg
     2461 attcagtcat gcccgcgacc ccatcttcgg caaccagatc atccccgaca ccgctatcct
     2521 cagcgtggtg ccatttcacc acggcttcgg catgttcacc acgctgggct acttgatctg
     2581 cggctttcgg gtcgtgctca tgtaccgctt cgaggaggag ctattcttgc gcagcttgca
     2641 agactataag attcaatctg ccctgctggt gcccacacta tttagcttct tcgctaagag
     2701 cactctcatc gacaagtacg acctaagcaa cttgcacgag atcgccagcg gcggggcgcc
     2761 gctcagcaag gaggtaggtg aggccgtggc caaacgcttc cacctaccag gcatccgcca
     2821 gggctacggc ctgacagaaa caaccagcgc cattctgatc acccccgaag gggacgacaa
     2881 gcctggcgca gtaggcaagg tggtgccctt cttcgaggct aaggtggtgg acttggacac
     2941 cggtaagaca ctgggtgtga accagcgcgg cgagctgtgc gtccgtggcc ccatgatcat
     3001 gagcggctac gttaacaacc ccgaggctac aaacgctctc atcgacaagg acggctggct
     3061 gcacagcggc gacatcgcct actgggacga ggacgagcac ttcttcatcg tggaccggct
     3121 gaagagcctg atcaaataca agggctacca ggtagcccca gccgaactgg agagcatcct
     3181 gctgcaacac cccaacatct tcgacgccgg ggtcgccggc ctgcccgacg acgatgccgg
     3241 cgagctgccc gccgcagtcg tcgtgctgga acacggtaaa accatgaccg agaaggagat
     3301 cgtggactat gtggccagcc aggttacaac cgccaagaag ctgcgcggtg gtgttgtgtt
     3361 cgtggacgag gtgcctaaag gactgaccgg caagttggac gcccgcaaga tccgcgagat
     3421 tctcattaag gccaagaagg gcggcaagat cgccgtgtaa taattctaga gtcggggcgg
     3481 ccggccgctt cgagcagaca tgataagata cattgatgag tttggacaaa ccacaactag
     3541 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     3601 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     3661 tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtaaaat
     3721 cgataaggat ccgtcgaccg atgcccttga gagccttcaa cccagtcagc tccttccggt
     3781 gggcgcgggg catgactagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     3841 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     3901 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     3961 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     4021 ttctccgctc catcgtcgtc gccgcactta tgactgtctt ctttatcatg caactcgtag
     4081 gacaggtgcc ggcagcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     4141 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     4201 ggggataacg caggaaagaa
//
