LOCUS       sequence-190939-        5483 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Luciferase validation vector with AGAP1 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 5483)
  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 5483)
  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_AGAP1
FEATURES             Location/Qualifiers
     source          1..5483
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    1086..1207
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1123..1142)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1177..1196
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(1251..2903)
                     /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      2900..2909
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     rep_origin      3008..3596
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3497..3516
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3786..4577,4578..4646))
                     /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     4804..4821
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 atgcagcccc tactgcaccg tcaggaccca cagtggctgg gcaagcctca gtcctggagt
       61 cccgcaggct ggcctgctcc agaaaccagc gcttcccaga agccagtgca gcctccctcg
      121 cagcccagcc ctccgggcct ctccccacag gcatgctagc aacatcccag caggcgcaaa
      181 gcattccaga gtgaaggcgg ccacccctga aaacacccct ctatcttcaa ggcctacatt
      241 tctccagtat tctgacagct tttagtacgg gcaagctatg ttaaaggaaa gcctccctga
      301 ctaaccttgg aagtgtaatt caatgccggt gacgattcag ccgtggatgt tactcttagg
      361 cgtgccacct gtgactcatg ctcagagggt agaatggggg ccacagccac accctctccc
      421 cttccttcca agggctgcag caaagtatcc tggcacacta aggaactttt aaaagtcatt
      481 gataaggctc attaatgtat caaaatgtca cctactcaat gtgaaccatt aaaaattatt
      541 agtacaagta ttagtagaaa tagtcattaa agtgcagaaa taccaggaga aatatcaagt
      601 agaaatattc atcaccttgg agctacgccg gcagccccct tacctcctgc tccaagccct
      661 tcagccacgt gggtcaggag ctgtgccctg tggtctcagc cctcaggtca aagccctgaa
      721 acctcagggt gagcaaggag agtctgcgtg gagaatcaat ttcactcaaa cacttggaga
      781 aaaacagcaa tgttaactca caaataatag aaagtatatt caaacacgtg atgcatgtat
      841 gaagcagaaa gtaaaagtca cacatatttt taaaataaga cgttgcagct gtgtcctctc
      901 atcttgaagg ggatgtattt cctctcctct gcaatctggg gagacgcaca agccgcacta
      961 caaacccgcc caggggtggg atccttatcg attttaccac atttgtagag gttttacttg
     1021 ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg
     1081 ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     1141 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     1201 tatcttatca tgtctgctcg aagcggccgg ccgccccgac tctagaatta ttacacggcg
     1261 atcttgccgc ccttcttggc cttaatgaga atctcgcgga tcttgcgggc gtccaacttg
     1321 ccggtcagtc ctttaggcac ctcgtccacg aacacaacac caccgcgcag cttcttggcg
     1381 gttgtaacct ggctggccac atagtccacg atctccttct cggtcatggt tttaccgtgt
     1441 tccagcacga cgactgcggc gggcagctcg ccggcatcgt cgtcgggcag gccggcgacc
     1501 ccggcgtcga agatgttggg gtgttgcagc aggatgctct ccagttcggc tggggctacc
     1561 tggtagccct tgtatttgat caggctcttc agccggtcca cgatgaagaa gtgctcgtcc
     1621 tcgtcccagt aggcgatgtc gccgctgtgc agccagccgt ccttgtcgat gagagcgttt
     1681 gtagcctcgg ggttgttaac gtagccgctc atgatcatgg ggccacggac gcacagctcg
     1741 ccgcgctggt tcacacccag tgtcttaccg gtgtccaagt ccaccacctt agcctcgaag
     1801 aagggcacca ccttgcctac tgcgccaggc ttgtcgtccc cttcgggggt gatcagaatg
     1861 gcgctggttg tttctgtcag gccgtagccc tggcggatgc ctggtaggtg gaagcgtttg
     1921 gccacggcct cacctacctc cttgctgagc ggcgccccgc cgctggcgat ctcgtgcaag
     1981 ttgcttaggt cgtacttgtc gatgagagtg ctcttagcga agaagctaaa tagtgtgggc
     2041 accagcaggg cagattgaat cttatagtct tgcaagctgc gcaagaatag ctcctcctcg
     2101 aagcggtaca tgagcacgac ccgaaagccg cagatcaagt agcccagcgt ggtgaacatg
     2161 ccgaagccgt ggtgaaatgg caccacgctg aggatagcgg tgtcggggat gatctggttg
     2221 ccgaagatgg ggtcgcgggc atgactgaat cggacacaag cggtgcggtg cggtagggct
     2281 acgcccttgg gcaatccggt actgccacta ctgttcatga tcagggcgat ggttttgtcc
     2341 cggtcgaagc tctcgggcac gaagtcgtac tcgttgaagc cgggtggcaa atgggaagtc
     2401 acgaaggtgt acatgctttg gaagccctgg tagtcggtct tgctatccat gatgatgatc
     2461 ttttgtatga tcggtagctt cttttgcacg ttgaggatct tttgcagccc tttcttgctc
     2521 acgaatacga cggtgggctg gctgatgccc atgctgttca gcagctcgcg ctcgttgtag
     2581 atgtcgttag ctggggccac agccacaccg atgaacaggg cacccaacac gggcatgaag
     2641 aactgcaagc tattctcgct gcacaccacg atccgatggt ttgtattcag cccatagcgc
     2701 ttcatagctt ctgccagccg aacgctcatc tcgaagtact cggcgtaggt aatgtccacc
     2761 tcgatatgtg cgtcggtaaa ggcgatggtg ccgggcacca gggcgtagcg cttcatggct
     2821 ttgtgcagct gctcgccggc ggtcccgtct tcgagtgggt agaatggcgc tgggcccttc
     2881 ttaatgtttt tggcatcttc catggtggct ttaccaacag taccggattg ccaagcttca
     2941 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     3001 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     3061 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     3121 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     3181 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     3241 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     3301 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     3361 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     3421 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     3481 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     3541 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     3601 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgtcat
     3661 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     3721 aatctaaagt atatatgagt aaacttggtc tgacagcggc cgcaaatgct aaaccactgc
     3781 agtggttacc agtgcttgat cagtgaggca ccgatctcag cgatctgcct atttcgttcg
     3841 tccatagtgg cctgactccc cgtcgtgtag atcactacga ttcgtgaggg cttaccatca
     3901 ggccccagcg cagcaatgat gccgcgagag ccgcgttcac cggcccccga tttgtcagca
     3961 atgaaccagc cagcagggag ggccgagcga agaagtggtc ctgctacttt gtccgcctcc
     4021 atccagtcta tgagctgctg tcgtgatgct agagtaagaa gttcgccagt gagtagtttc
     4081 cgaagagttg tggccattgc tactggcatc gtggtatcac gctcgtcgtt cggtatggct
     4141 tcgttcaact ctggttccca gcggtcaagc cgggtcacat gatcacccat attatgaaga
     4201 aatgcagtca gctccttagg gcctccgatc gttgtcagaa gtaagttggc cgcggtgttg
     4261 tcgctcatgg taatggcagc actacacaat tctcttaccg tcatgccatc cgtaagatgc
     4321 ttttccgtga ccggcgagta ctcaaccaag tcgttttgtg agtagtgtat acggcgacca
     4381 agctgctctt gcccggcgtc tatacgggac aacaccgcgc cacatagcag tactttgaaa
     4441 gtgctcatca tcgggaatcg ttcttcgggg cggaaagact caaggatctt gccgctattg
     4501 agatccagtt cgatatagcc cactcttgca cccagttgat cttcagcatc ttttactttc
     4561 accagcgttt cggggtgtgc aaaaacaggc aagcaaaatg ccgcaaagaa gggaatgagt
     4621 gcgacacgaa aatgttggat gctcatactc gtcctttttc aatattattg aagcatttat
     4681 cagggttact agtacgtctc tcaagacatg ttctttcctg cgttatcccc tgattctgtg
     4741 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     4801 cgcagcgagt cagtgagcga ggaagcggaa gagcgctgcc ggcacctgtc ctacgagttg
     4861 catgataaag aagacagtca taagtgcggc gacgatagtc atgccccgcg cccaccggaa
     4921 ggagctgact gggttgaagg ctctcaaggg catcggtcga cccaggtatt ttctgccttc
     4981 tcatagaaca aacgtgactc gtgggcagtt ccatacaatg cgtaagtgtt agtttaccac
     5041 tcccaggctg agtgacattg agtaagtggc tcaacttctc tgagcattca ttctgtctgt
     5101 gaaatgggga ttatgctatt atccctccta aggtggttgt gagaattgag atttttcatt
     5161 acataaatat ttactgagag ctactctagg ggtctgggat acactggtga ccaaaaaaag
     5221 aaattcccaa catggggtgc ttatgttcca gtgggctgcc acagaaaggg gtcggcctag
     5281 tgcagaggaa tgctcatttc actgagttca aaatacttcc tcttccatca attcctgtag
     5341 ccaagaaaga tactcgaatt taatcccatt atttcccaga aacatgagaa caagccagaa
     5401 aacatttgtc cccaggaaac caaaattcag gatccccatc tgctctcatc tttctctggg
     5461 atacttgttg taagtctgtc aaa
//
