LOCUS       sequence-190940-        5423 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 5423)
  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 5423)
  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_IGF1R
FEATURES             Location/Qualifiers
     source          1..5423
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     375..394
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(664..1455,1456..1524))
                     /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     1682..1699
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    3387..3508
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3424..3443)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3478..3497
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(3552..5204)
                     /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      5201..5210
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     rep_origin      join(5309..5423,1..474)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
       61 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
      121 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
      181 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
      241 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
      301 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
      361 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
      421 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
      481 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgtcatga
      541 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
      601 tctaaagtat atatgagtaa acttggtctg acagcggccg caaatgctaa accactgcag
      661 tggttaccag tgcttgatca gtgaggcacc gatctcagcg atctgcctat ttcgttcgtc
      721 catagtggcc tgactccccg tcgtgtagat cactacgatt cgtgagggct taccatcagg
      781 ccccagcgca gcaatgatgc cgcgagagcc gcgttcaccg gcccccgatt tgtcagcaat
      841 gaaccagcca gcagggaggg ccgagcgaag aagtggtcct gctactttgt ccgcctccat
      901 ccagtctatg agctgctgtc gtgatgctag agtaagaagt tcgccagtga gtagtttccg
      961 aagagttgtg gccattgcta ctggcatcgt ggtatcacgc tcgtcgttcg gtatggcttc
     1021 gttcaactct ggttcccagc ggtcaagccg ggtcacatga tcacccatat tatgaagaaa
     1081 tgcagtcagc tccttagggc ctccgatcgt tgtcagaagt aagttggccg cggtgttgtc
     1141 gctcatggta atggcagcac tacacaattc tcttaccgtc atgccatccg taagatgctt
     1201 ttccgtgacc ggcgagtact caaccaagtc gttttgtgag tagtgtatac ggcgaccaag
     1261 ctgctcttgc ccggcgtcta tacgggacaa caccgcgcca catagcagta ctttgaaagt
     1321 gctcatcatc gggaatcgtt cttcggggcg gaaagactca aggatcttgc cgctattgag
     1381 atccagttcg atatagccca ctcttgcacc cagttgatct tcagcatctt ttactttcac
     1441 cagcgtttcg gggtgtgcaa aaacaggcaa gcaaaatgcc gcaaagaagg gaatgagtgc
     1501 gacacgaaaa tgttggatgc tcatactcgt cctttttcaa tattattgaa gcatttatca
     1561 gggttactag tacgtctctc aagacatgtt ctttcctgcg ttatcccctg attctgtgga
     1621 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     1681 cagcgagtca gtgagcgagg aagcggaaga gcgctgccgg cacctgtcct acgagttgca
     1741 tgataaagaa gacagtcata agtgcggcga cgatagtcat gccccgcgcc caccggaagg
     1801 agctgactgg gttgaaggct ctcaagggca tcggtcgacc caaagaagtg aagagatcat
     1861 aggctaagtc agtgggaaat cctttgcaca aatccaactc agaagcaact taccaggaat
     1921 gccatctatt gcaaataaag gtcattgtac aggggtgaga ccgaaacttt atgtacacaa
     1981 tgacacacac ttgtaccctt ttgtctctca caagtaaatt tggaaatcta gcattctttt
     2041 ggacttcata aaattttaac gtgatatcac taaacttccc tttgctcagt cttctcagta
     2101 aattcaagta accaaaactc tttagacttg aaagatgtgt ggatttggct acagctggag
     2161 tctgctgagt cacacgtctg atccaatggt ctcttctggg gtttgtatgt gtcagggctt
     2221 gcctctccta tgcaagatgt gactaacaaa aagatgagcg atgaccctct ttacaggaat
     2281 aaatgacgtg ccaatggcta aaagacagat tgcctttcaa tctgttcttt atctttcagg
     2341 gaaactcaaa gaagagacgt catgcgtgag gcttacacac catgttgttt atctaagccc
     2401 atattttagt ggtgactcag ggacgggagc atgaaaagca tcaacaaaag aaaaccatga
     2461 gatctagccg tttttccagg gggatggaat acttactccc ccgtcggatg ttaatgagca
     2521 aattgccctt gaagatggtg catccttgga gcatctgagc agaagtaaca gaatcaatgg
     2581 tctttgtttt cttttcttcc tcacagacct tcgggcaagg accttcacaa gggatgcagt
     2641 acatgctgtt gaagaagaaa aaaaaaatca aaaacacaca taattgagtc tgtcttcttg
     2701 cattaaaaaa accataagga tatgcttttc attgttatta cagcatgcat tagaaaaagc
     2761 aactgttttg aagtgttcat gtggtcacat ggtatctcac ccccaagccc acccaagata
     2821 agccactcac attgcaagca aaaggagaaa cagcctacaa aggaagagcg tgttgcttgc
     2881 ttccagacaa aatacatgct gcatcctcaa ggcatttgtg tgtcataggc tgttgactgg
     2941 tgagccagct ggttccatgt gggagagaca gtgctgcaga aaggaagctt ggggggttct
     3001 ggaggaaaac agagctgctt tctgatactg tgaccctggc ctgtgtacct ggaggagtcg
     3061 ctaggagaga aggcagggac agcaaaggga gtgcagtgtc tctgcctaac agtagtctag
     3121 ctcctgcagt cagtgttttg gcaaagccca gtgagggata ccaaaaggca tgtggtatag
     3181 cacagatcca ttattttctt gctaaagcac aaccgaaatt atttcttgac ggtgttttgc
     3241 agaaacttgt aaattgatcc aaatgttgct tttgctgcag gatccttatc gattttacca
     3301 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     3361 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     3421 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     3481 gtttgtccaa actcatcaat gtatcttatc atgtctgctc gaagcggccg gccgccccga
     3541 ctctagaatt attacacggc gatcttgccg cccttcttgg ccttaatgag aatctcgcgg
     3601 atcttgcggg cgtccaactt gccggtcagt cctttaggca cctcgtccac gaacacaaca
     3661 ccaccgcgca gcttcttggc ggttgtaacc tggctggcca catagtccac gatctccttc
     3721 tcggtcatgg ttttaccgtg ttccagcacg acgactgcgg cgggcagctc gccggcatcg
     3781 tcgtcgggca ggccggcgac cccggcgtcg aagatgttgg ggtgttgcag caggatgctc
     3841 tccagttcgg ctggggctac ctggtagccc ttgtatttga tcaggctctt cagccggtcc
     3901 acgatgaaga agtgctcgtc ctcgtcccag taggcgatgt cgccgctgtg cagccagccg
     3961 tccttgtcga tgagagcgtt tgtagcctcg gggttgttaa cgtagccgct catgatcatg
     4021 gggccacgga cgcacagctc gccgcgctgg ttcacaccca gtgtcttacc ggtgtccaag
     4081 tccaccacct tagcctcgaa gaagggcacc accttgccta ctgcgccagg cttgtcgtcc
     4141 ccttcggggg tgatcagaat ggcgctggtt gtttctgtca ggccgtagcc ctggcggatg
     4201 cctggtaggt ggaagcgttt ggccacggcc tcacctacct ccttgctgag cggcgccccg
     4261 ccgctggcga tctcgtgcaa gttgcttagg tcgtacttgt cgatgagagt gctcttagcg
     4321 aagaagctaa atagtgtggg caccagcagg gcagattgaa tcttatagtc ttgcaagctg
     4381 cgcaagaata gctcctcctc gaagcggtac atgagcacga cccgaaagcc gcagatcaag
     4441 tagcccagcg tggtgaacat gccgaagccg tggtgaaatg gcaccacgct gaggatagcg
     4501 gtgtcgggga tgatctggtt gccgaagatg gggtcgcggg catgactgaa tcggacacaa
     4561 gcggtgcggt gcggtagggc tacgcccttg ggcaatccgg tactgccact actgttcatg
     4621 atcagggcga tggttttgtc ccggtcgaag ctctcgggca cgaagtcgta ctcgttgaag
     4681 ccgggtggca aatgggaagt cacgaaggtg tacatgcttt ggaagccctg gtagtcggtc
     4741 ttgctatcca tgatgatgat cttttgtatg atcggtagct tcttttgcac gttgaggatc
     4801 ttttgcagcc ctttcttgct cacgaatacg acggtgggct ggctgatgcc catgctgttc
     4861 agcagctcgc gctcgttgta gatgtcgtta gctggggcca cagccacacc gatgaacagg
     4921 gcacccaaca cgggcatgaa gaactgcaag ctattctcgc tgcacaccac gatccgatgg
     4981 tttgtattca gcccatagcg cttcatagct tctgccagcc gaacgctcat ctcgaagtac
     5041 tcggcgtagg taatgtccac ctcgatatgt gcgtcggtaa aggcgatggt gccgggcacc
     5101 agggcgtagc gcttcatggc tttgtgcagc tgctcgccgg cggtcccgtc ttcgagtggg
     5161 tagaatggcg ctgggccctt cttaatgttt ttggcatctt ccatggtggc tttaccaaca
     5221 gtaccggatt gccaagcttc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     5281 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     5341 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     5401 caactctttt tccgaaggta act
//
