LOCUS       sequence-193007-        5436 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Luciferase validation vector with IGF1R 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 5436)
  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 5436)
  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_IGF1R
FEATURES             Location/Qualifiers
     source          1..5436
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1546..1555
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1598..1628)
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     misc_feature    1653..1744
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     complement(1685..1704)
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     polyA_signal    1758..1806
                     /note="synthetic polyadenylation signal"
     CDS             join(1911..1979,1980..2771)
                     /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      2971..3559
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3460..3479
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5168..5289
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5205..5224)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5259..5278
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(5333..5436,1..1549))
                     /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"
ORIGIN
        1 caacaccacc gcgcagcttc ttggcggttg taacctggct ggccacatag tccacgatct
       61 ccttctcggt catggtttta ccgtgttcca gcacgacgac tgcggcgggc agctcgccgg
      121 catcgtcgtc gggcaggccg gcgaccccgg cgtcgaagat gttggggtgt tgcagcagga
      181 tgctctccag ttcggctggg gctacctggt agcccttgta tttgatcagg ctcttcagcc
      241 ggtccacgat gaagaagtgc tcgtcctcgt cccagtaggc gatgtcgccg ctgtgcagcc
      301 agccgtcctt gtcgatgaga gcgtttgtag cctcggggtt gttaacgtag ccgctcatga
      361 tcatggggcc acggacgcac agctcgccgc gctggttcac acccagtgtc ttaccggtgt
      421 ccaagtccac caccttagcc tcgaagaagg gcaccacctt gcctactgcg ccaggcttgt
      481 cgtccccttc gggggtgatc agaatggcgc tggttgtttc tgtcaggccg tagccctggc
      541 ggatgcctgg taggtggaag cgtttggcca cggcctcacc tacctccttg ctgagcggcg
      601 ccccgccgct ggcgatctcg tgcaagttgc ttaggtcgta cttgtcgatg agagtgctct
      661 tagcgaagaa gctaaatagt gtgggcacca gcagggcaga ttgaatctta tagtcttgca
      721 agctgcgcaa gaatagctcc tcctcgaagc ggtacatgag cacgacccga aagccgcaga
      781 tcaagtagcc cagcgtggtg aacatgccga agccgtggtg aaatggcacc acgctgagga
      841 tagcggtgtc ggggatgatc tggttgccga agatggggtc gcgggcatga ctgaatcgga
      901 cacaagcggt gcggtgcggt agggctacgc ccttgggcaa tccggtactg ccactactgt
      961 tcatgatcag ggcgatggtt ttgtcccggt cgaagctctc gggcacgaag tcgtactcgt
     1021 tgaagccggg tggcaaatgg gaagtcacga aggtgtacat gctttggaag ccctggtagt
     1081 cggtcttgct atccatgatg atgatctttt gtatgatcgg tagcttcttt tgcacgttga
     1141 ggatcttttg cagccctttc ttgctcacga atacgacggt gggctggctg atgcccatgc
     1201 tgttcagcag ctcgcgctcg ttgtagatgt cgttagctgg ggccacagcc acaccgatga
     1261 acagggcacc caacacgggc atgaagaact gcaagctatt ctcgctgcac accacgatcc
     1321 gatggtttgt attcagccca tagcgcttca tagcttctgc cagccgaacg ctcatctcga
     1381 agtactcggc gtaggtaatg tccacctcga tatgtgcgtc ggtaaaggcg atggtgccgg
     1441 gcaccagggc gtagcgcttc atggctttgt gcagctgctc gccggcggtc ccgtcttcga
     1501 gtgggtagaa tggcgctggg cccttcttaa tgtttttggc atcttccatg gtggctttac
     1561 caacagtacc ggattgccaa gctttaggtc gctagccctg gaagtcgagc ttccattata
     1621 taccctctct cgaagatccg gccagttagg ccagagaaat gttctggcac ctgcacttgc
     1681 actggggaca gcctattttg ctagtttgtt ttgtttcgtt ttgttttgat ggagagcgta
     1741 tgttagtact atcgattcac acaaaaaacc aacacacaga tgtaatgaaa ataaagatat
     1801 tttattgcgg cctgtccaat acctcccgta ccttaatatt acttacttat ccttgagaga
     1861 cgtactagta accctgataa atgcttcaat aatattgaaa aaggacgagt atgagcatcc
     1921 aacattttcg tgtcgcactc attcccttct ttgcggcatt ttgcttgcct gtttttgcac
     1981 accccgaaac gctggtgaaa gtaaaagatg ctgaagatca actgggtgca agagtgggct
     2041 atatcgaact ggatctcaat agcggcaaga tccttgagtc tttccgcccc gaagaacgat
     2101 tcccgatgat gagcactttc aaagtactgc tatgtggcgc ggtgttgtcc cgtatagacg
     2161 ccgggcaaga gcagcttggt cgccgtatac actactcaca aaacgacttg gttgagtact
     2221 cgccggtcac ggaaaagcat cttacggatg gcatgacggt aagagaattg tgtagtgctg
     2281 ccattaccat gagcgacaac accgcggcca acttacttct gacaacgatc ggaggcccta
     2341 aggagctgac tgcatttctt cataatatgg gtgatcatgt gacccggctt gaccgctggg
     2401 aaccagagtt gaacgaagcc ataccgaacg acgagcgtga taccacgatg ccagtagcaa
     2461 tggccacaac tcttcggaaa ctactcactg gcgaacttct tactctagca tcacgacagc
     2521 agctcataga ctggatggag gcggacaaag tagcaggacc acttcttcgc tcggccctcc
     2581 ctgctggctg gttcattgct gacaaatcgg gggccggtga acgcggctct cgcggcatca
     2641 ttgctgcgct ggggcctgat ggtaagccct cacgaatcgt agtgatctac acgacgggga
     2701 gtcaggccac tatggacgaa cgaaataggc agatcgctga gatcggtgcc tcactgatca
     2761 agcactggta accactgcag tggtttagca tttgcggccg ctgtcagacc aagtttactc
     2821 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     2881 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     2941 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     3001 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     3061 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     3121 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     3181 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     3241 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     3301 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     3361 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     3421 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     3481 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     3541 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     3601 ctggcctttt gctcacatgt ccaaagaagt gaagagatca taggctaagt cagtgggaaa
     3661 tcctttgcac aaatccaact cagaagcaac ttaccaggaa tgccatctat tgcaaataaa
     3721 ggtcattgta caggggtgag accgaaactt tatgtacaca atgacacaca cttgtatcct
     3781 tttgtctctc acaagtaaat ttggaaatct agcattcttt tggacttcat aaaattttaa
     3841 cgtgatatca ctaaacttcc ctttgctcag tcttctcagt aaattcaagt aaccaaaact
     3901 ctttagactt gaaagatgtg tggatttggc tacagctgga gtctgctgag tcacacgtct
     3961 gatccaatgg tctcttctgg ggtttgtatg tgtcagggct tgcctctcct atgcaagatg
     4021 tgactaacaa aaagatgagc gatgaccctc tttacaggaa taaatgacgt gccaatggct
     4081 aaaagacaga ttgcctttca atctgttctt tatctttcag ggaaactcaa agaagagacg
     4141 tcatgcgtga ggcttacaca ccatgttgtt tatctaagcc catattttag tggtgactca
     4201 gggacgggag catgaaaagc atcaacaaaa gaaaaccatg agatctagcc gtttttccag
     4261 ggggatggaa tacttactcc cccgtcggat gttaatgagc aaattgccct tgaagatggt
     4321 gcatccttgg agcatctgag cagaagtaac agaatcaatg gtctttgttt tcttttcttc
     4381 ctcacagacc ttcgggcaag gaccttcaca agggatgcag tacatgctgt tgaagaagaa
     4441 aaaaaaaatc aaaaacacac ataattgagt ctgtcttctt gcattaaaaa aaccataagg
     4501 atatgctttt cattgttatt acagcatgca ttagaaaaag caactgtttt gaagtgttca
     4561 tgtggtcaca tggtatctca cccccaagcc cacccaagat aagccactca cattgcaagc
     4621 aaaaggagaa acagcctaca aaggaagagc gtgttgcttg cttccagaca aaatacatgc
     4681 tgcatcctca aggcatttgt gtgtcatagg ctgttgactg gtgagccagc tggttccatg
     4741 tgggagagac agtgctgcag aaaggaagct tggggggttc tggaggaaaa cagagctgct
     4801 ttctgatact gtgaccctgg cctgtgtacc tggaggagtc gctaggagag aaggcaggga
     4861 cagcaaaggg agtgcagtgt ctctgcctaa cagtagtcta gctcctgcag tcagtgtttt
     4921 ggcaaagccc agtgagggat accaaaaggc atgtggtata gcacagatcc attattttct
     4981 tgctaaagca caaccgaaat tatttcttga cggtgttttg cagaaacttg taaattgatc
     5041 caaatgttgc ttttgctgca ggatccttat cgattttacc acatttgtag aggttttact
     5101 tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt
     5161 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     5221 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     5281 tgtatcttat catgtctgct cgaagcggcc ggccgccccg actctagaat tattacacgg
     5341 cgatcttgcc gcccttcttg gccttaatga gaatctcgcg gatcttgcgg gcgtccaact
     5401 tgccggtcag tcctttaggc acctcgtcca cgaaca
//
