LOCUS       sequence-190938-        5488 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 5488)
  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 5488)
  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_SCP1_IGF1R
FEATURES             Location/Qualifiers
     source          1..5488
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    901..1022
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(938..957)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     992..1011
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(1066..2718)
                     /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      2715..2724
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    2874..2965
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     complement(2906..2925)
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     polyA_signal    2979..3027
                     /note="synthetic polyadenylation signal"
     CDS             join(3132..3200,3201..3992)
                     /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      4192..4780
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4681..4700
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gaatacttac tcccccgtcg gatgttaatg agcaaattgc ccttgaagat ggtgcatcct
       61 tggagcatct gagcagaagt aacagaatca atggtctttg ttttcttttc ttcctcacag
      121 accttcgggc aaggaccttc acaagggatg cagtacatgc tgttgaagaa gaaaaaaaaa
      181 atcaaaaaca cacataattg agtctgtctt cttgcattaa aaaaaccata aggatatgct
      241 tttcattgtt attacagcat gcattagaaa aagcaactgt tttgaagtgt tcatgtggtc
      301 acatggtatc tcacccccaa gcccacccaa gataagccac tcacattgca agcaaaagga
      361 gaaacagcct acaaaggaag agcgtgttgc ttgcttccag acaaaataca tgctgcatcc
      421 tcaaggcatt tgtgtgtcat aggctgttga ctggtgagcc agctggttcc atgtgggaga
      481 gacagtgctg cagaaaggaa gcttgggggg ttctggagga aaacagagct gctttctgat
      541 actgtgaccc tggcctgtgt acctggagga gtcgctagga gagaaggcag ggacagcaaa
      601 gggagtgcag tgtctctgcc taacagtagt ctagctcctg cagtcagtgt tttggcaaag
      661 cccagtgagg gataccaaaa ggcatgtggt atagcacaga tccattattt tcttgctaaa
      721 gcacaaccga aattatttct tgacggtgtt ttgcagaaac ttgtaaattg atccaaatgt
      781 tgcttttgct gcaggatcct tatcgatttt accacatttg tagaggtttt acttgcttta
      841 aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt
      901 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
      961 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     1021 tatcatgtct gctcgaagcg gccggccgcc ccgactctag aattattaca cggcgatctt
     1081 gccgcccttc ttggccttaa tgagaatctc gcggatcttg cgggcgtcca acttgccggt
     1141 cagtccttta ggcacctcgt ccacgaacac aacaccaccg cgcagcttct tggcggttgt
     1201 aacctggctg gccacatagt ccacgatctc cttctcggtc atggttttac cgtgttccag
     1261 cacgacgact gcggcgggca gctcgccggc atcgtcgtcg ggcaggccgg cgaccccggc
     1321 gtcgaagatg ttggggtgtt gcagcaggat gctctccagt tcggctgggg ctacctggta
     1381 gcccttgtat ttgatcaggc tcttcagccg gtccacgatg aagaagtgct cgtcctcgtc
     1441 ccagtaggcg atgtcgccgc tgtgcagcca gccgtccttg tcgatgagag cgtttgtagc
     1501 ctcggggttg ttaacgtagc cgctcatgat catggggcca cggacgcaca gctcgccgcg
     1561 ctggttcaca cccagtgtct taccggtgtc caagtccacc accttagcct cgaagaaggg
     1621 caccaccttg cctactgcgc caggcttgtc gtccccttcg ggggtgatca gaatggcgct
     1681 ggttgtttct gtcaggccgt agccctggcg gatgcctggt aggtggaagc gtttggccac
     1741 ggcctcacct acctccttgc tgagcggcgc cccgccgctg gcgatctcgt gcaagttgct
     1801 taggtcgtac ttgtcgatga gagtgctctt agcgaagaag ctaaatagtg tgggcaccag
     1861 cagggcagat tgaatcttat agtcttgcaa gctgcgcaag aatagctcct cctcgaagcg
     1921 gtacatgagc acgacccgaa agccgcagat caagtagccc agcgtggtga acatgccgaa
     1981 gccgtggtga aatggcacca cgctgaggat agcggtgtcg gggatgatct ggttgccgaa
     2041 gatggggtcg cgggcatgac tgaatcggac acaagcggtg cggtgcggta gggctacgcc
     2101 cttgggcaat ccggtactgc cactactgtt catgatcagg gcgatggttt tgtcccggtc
     2161 gaagctctcg ggcacgaagt cgtactcgtt gaagccgggt ggcaaatggg aagtcacgaa
     2221 ggtgtacatg ctttggaagc cctggtagtc ggtcttgcta tccatgatga tgatcttttg
     2281 tatgatcggt agcttctttt gcacgttgag gatcttttgc agccctttct tgctcacgaa
     2341 tacgacggtg ggctggctga tgcccatgct gttcagcagc tcgcgctcgt tgtagatgtc
     2401 gttagctggg gccacagcca caccgatgaa cagggcaccc aacacgggca tgaagaactg
     2461 caagctattc tcgctgcaca ccacgatccg atggtttgta ttcagcccat agcgcttcat
     2521 agcttctgcc agccgaacgc tcatctcgaa gtactcggcg taggtaatgt ccacctcgat
     2581 atgtgcgtcg gtaaaggcga tggtgccggg caccagggcg tagcgcttca tggctttgtg
     2641 cagctgctcg ccggcggtcc cgtcttcgag tgggtagaat ggcgctgggc ccttcttaat
     2701 gtttttggca tcttccatgg tggctttacc aacagtaccg gattgccaag ctttaggtcg
     2761 ctagcccggt ccgtaggcac gtctgctcgg ctcgagtgtt cgatcgcgac tgaggacgaa
     2821 cgcgccccca cccccttata taagtaccct agggagatcc ggccagttag gccagagaaa
     2881 tgttctggca cctgcacttg cactggggac agcctatttt gctagtttgt tttgtttcgt
     2941 tttgttttga tggagagcgt atgttagtac tatcgattca cacaaaaaac caacacacag
     3001 atgtaatgaa aataaagata ttttattgcg gcctgtccaa tacctcccgt accttaatat
     3061 tacttactta tccttgagag acgtactagt aaccctgata aatgcttcaa taatattgaa
     3121 aaaggacgag tatgagcatc caacattttc gtgtcgcact cattcccttc tttgcggcat
     3181 tttgcttgcc tgtttttgca caccccgaaa cgctggtgaa agtaaaagat gctgaagatc
     3241 aactgggtgc aagagtgggc tatatcgaac tggatctcaa tagcggcaag atccttgagt
     3301 ctttccgccc cgaagaacga ttcccgatga tgagcacttt caaagtactg ctatgtggcg
     3361 cggtgttgtc ccgtatagac gccgggcaag agcagcttgg tcgccgtata cactactcac
     3421 aaaacgactt ggttgagtac tcgccggtca cggaaaagca tcttacggat ggcatgacgg
     3481 taagagaatt gtgtagtgct gccattacca tgagcgacaa caccgcggcc aacttacttc
     3541 tgacaacgat cggaggccct aaggagctga ctgcatttct tcataatatg ggtgatcatg
     3601 tgacccggct tgaccgctgg gaaccagagt tgaacgaagc cataccgaac gacgagcgtg
     3661 ataccacgat gccagtagca atggccacaa ctcttcggaa actactcact ggcgaacttc
     3721 ttactctagc atcacgacag cagctcatag actggatgga ggcggacaaa gtagcaggac
     3781 cacttcttcg ctcggccctc cctgctggct ggttcattgc tgacaaatcg ggggccggtg
     3841 aacgcggctc tcgcggcatc attgctgcgc tggggcctga tggtaagccc tcacgaatcg
     3901 tagtgatcta cacgacgggg agtcaggcca ctatggacga acgaaatagg cagatcgctg
     3961 agatcggtgc ctcactgatc aagcactggt aaccactgca gtggtttagc atttgcggcc
     4021 gctgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     4081 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     4141 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     4201 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     4261 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     4321 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     4381 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     4441 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     4501 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     4561 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     4621 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     4681 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     4741 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     4801 tttacggttc ctggcctttt gctggccttt tgctcacatg tccaaagaag tgaagagatc
     4861 ataggctaag tcagtgggaa atcctttgca caaatccaac tcagaagcaa cttaccagga
     4921 atgccatcta ttgcaaataa aggtcattgt acaggggtga gaccgaaact ttatgtacac
     4981 aatgacacac acttgtatcc ttttgtctct cacaagtaaa tttggaaatc tagcattctt
     5041 ttggacttca taaaatttta acgtgatatc actaaacttc cctttgctca gtcttctcag
     5101 taaattcaag taaccaaaac tctttagact tgaaagatgt gtggatttgg ctacagctgg
     5161 agtctgctga gtcacacgtc tgatccaatg gtctcttctg gggtttgtat gtgtcagggc
     5221 ttgcctctcc tatgcaagat gtgactaaca aaaagatgag cgatgaccct ctttacagga
     5281 ataaatgacg tgccaatggc taaaagacag attgcctttc aatctgttct ttatctttca
     5341 gggaaactca aagaagagac gtcatgcgtg aggcttacac accatgttgt ttatctaagc
     5401 ccatatttta gtggtgactc agggacggga gcatgaaaag catcaacaaa agaaaaccat
     5461 gagatctagc cgtttttcca gggggatg
//
