LOCUS       sequence-190937-        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"
     primer_bind     129..149
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    328..449
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(365..384)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     419..438
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(493..2145)
                     /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      2142..2151
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     rep_origin      2250..2838
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2739..2758
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3028..3819,3820..3888))
                     /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     4046..4063
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 attgagatgc atgctttgca tacttctgcc tgctggggag cctggggact ttccacacct
       61 ggttgctgac taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga
      121 ctttccacac cctaactgac acacattcca cagctagtca tgccccgcgc ccaccggaag
      181 gagctgactg ggttgaaggc tctcaagggc atcggtcgac ggatccttat cgattttacc
      241 acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa
      301 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa
      361 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
      421 ggtttgtcca aactcatcaa tgtatcttat catgtctgct cgaagcggcc ggccgccccg
      481 actctagaat tattacacgg cgatcttgcc gcccttcttg gccttaatga gaatctcgcg
      541 gatcttgcgg gcgtccaact tgccggtcag tcctttaggc acctcgtcca cgaacacaac
      601 accaccgcgc agcttcttgg cggttgtaac ctggctggcc acatagtcca cgatctcctt
      661 ctcggtcatg gttttaccgt gttccagcac gacgactgcg gcgggcagct cgccggcatc
      721 gtcgtcgggc aggccggcga ccccggcgtc gaagatgttg gggtgttgca gcaggatgct
      781 ctccagttcg gctggggcta cctggtagcc cttgtatttg atcaggctct tcagccggtc
      841 cacgatgaag aagtgctcgt cctcgtccca gtaggcgatg tcgccgctgt gcagccagcc
      901 gtccttgtcg atgagagcgt ttgtagcctc ggggttgtta acgtagccgc tcatgatcat
      961 ggggccacgg acgcacagct cgccgcgctg gttcacaccc agtgtcttac cggtgtccaa
     1021 gtccaccacc ttagcctcga agaagggcac caccttgcct actgcgccag gcttgtcgtc
     1081 cccttcgggg gtgatcagaa tggcgctggt tgtttctgtc aggccgtagc cctggcggat
     1141 gcctggtagg tggaagcgtt tggccacggc ctcacctacc tccttgctga gcggcgcccc
     1201 gccgctggcg atctcgtgca agttgcttag gtcgtacttg tcgatgagag tgctcttagc
     1261 gaagaagcta aatagtgtgg gcaccagcag ggcagattga atcttatagt cttgcaagct
     1321 gcgcaagaat agctcctcct cgaagcggta catgagcacg acccgaaagc cgcagatcaa
     1381 gtagcccagc gtggtgaaca tgccgaagcc gtggtgaaat ggcaccacgc tgaggatagc
     1441 ggtgtcgggg atgatctggt tgccgaagat ggggtcgcgg gcatgactga atcggacaca
     1501 agcggtgcgg tgcggtaggg ctacgccctt gggcaatccg gtactgccac tactgttcat
     1561 gatcagggcg atggttttgt cccggtcgaa gctctcgggc acgaagtcgt actcgttgaa
     1621 gccgggtggc aaatgggaag tcacgaaggt gtacatgctt tggaagccct ggtagtcggt
     1681 cttgctatcc atgatgatga tcttttgtat gatcggtagc ttcttttgca cgttgaggat
     1741 cttttgcagc cctttcttgc tcacgaatac gacggtgggc tggctgatgc ccatgctgtt
     1801 cagcagctcg cgctcgttgt agatgtcgtt agctggggcc acagccacac cgatgaacag
     1861 ggcacccaac acgggcatga agaactgcaa gctattctcg ctgcacacca cgatccgatg
     1921 gtttgtattc agcccatagc gcttcatagc ttctgccagc cgaacgctca tctcgaagta
     1981 ctcggcgtag gtaatgtcca cctcgatatg tgcgtcggta aaggcgatgg tgccgggcac
     2041 cagggcgtag cgcttcatgg ctttgtgcag ctgctcgccg gcggtcccgt cttcgagtgg
     2101 gtagaatggc gctgggccct tcttaatgtt tttggcatct tccatggtgg ctttaccaac
     2161 agtaccggat tgccaagctt caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     2221 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     2281 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     2341 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     2401 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     2461 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     2521 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     2581 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     2641 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     2701 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     2761 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     2821 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     2881 tggccttttg ctcacatgtc atgagattat caaaaaggat cttcacctag atccttttaa
     2941 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagcg
     3001 gccgcaaatg ctaaaccact gcagtggtta ccagtgcttg atcagtgagg caccgatctc
     3061 agcgatctgc ctatttcgtt cgtccatagt ggcctgactc cccgtcgtgt agatcactac
     3121 gattcgtgag ggcttaccat caggccccag cgcagcaatg atgccgcgag agccgcgttc
     3181 accggccccc gatttgtcag caatgaacca gccagcaggg agggccgagc gaagaagtgg
     3241 tcctgctact ttgtccgcct ccatccagtc tatgagctgc tgtcgtgatg ctagagtaag
     3301 aagttcgcca gtgagtagtt tccgaagagt tgtggccatt gctactggca tcgtggtatc
     3361 acgctcgtcg ttcggtatgg cttcgttcaa ctctggttcc cagcggtcaa gccgggtcac
     3421 atgatcaccc atattatgaa gaaatgcagt cagctcctta gggcctccga tcgttgtcag
     3481 aagtaagttg gccgcggtgt tgtcgctcat ggtaatggca gcactacaca attctcttac
     3541 cgtcatgcca tccgtaagat gcttttccgt gaccggcgag tactcaacca agtcgttttg
     3601 tgagtagtgt atacggcgac caagctgctc ttgcccggcg tctatacggg acaacaccgc
     3661 gccacatagc agtactttga aagtgctcat catcgggaat cgttcttcgg ggcggaaaga
     3721 ctcaaggatc ttgccgctat tgagatccag ttcgatatag cccactcttg cacccagttg
     3781 atcttcagca tcttttactt tcaccagcgt ttcggggtgt gcaaaaacag gcaagcaaaa
     3841 tgccgcaaag aagggaatga gtgcgacacg aaaatgttgg atgctcatac tcgtcctttt
     3901 tcaatattat tgaagcattt atcagggtta ctagtacgtc tctcaagaca tgttctttcc
     3961 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     4021 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgctg
     4081 ccggcacctg tcctacgagt tgcatgataa agaagacagt cataagtgcg gcgacgacga
     4141 tggagcggag aatgggcgga actgggcgga gttaggggcg ggatgggcgg agttaggggc
     4201 gggactatgg ttgctgacta
//
