LOCUS       sequence-193155-        5496 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 5496)
  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 5496)
  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_AGAP1
FEATURES             Location/Qualifiers
     source          1..5496
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1290..1299
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1342..1372)
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     misc_feature    1397..1488
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     complement(1429..1448)
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     polyA_signal    1502..1550
                     /note="synthetic polyadenylation signal"
     CDS             join(1655..1723,1724..2515)
                     /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      2715..3303
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3204..3223
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4972..5093
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5009..5028)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5063..5082
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(5137..5496,1..1293))
                     /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 gtgctcgtcc tcgtcccagt aggcgatgtc gccgctgtgc agccagccgt ccttgtcgat
       61 gagagcgttt gtagcctcgg ggttgttaac gtagccgctc atgatcatgg ggccacggac
      121 gcacagctcg ccgcgctggt tcacacccag tgtcttaccg gtgtccaagt ccaccacctt
      181 agcctcgaag aagggcacca ccttgcctac tgcgccaggc ttgtcgtccc cttcgggggt
      241 gatcagaatg gcgctggttg tttctgtcag gccgtagccc tggcggatgc ctggtaggtg
      301 gaagcgtttg gccacggcct cacctacctc cttgctgagc ggcgccccgc cgctggcgat
      361 ctcgtgcaag ttgcttaggt cgtacttgtc gatgagagtg ctcttagcga agaagctaaa
      421 tagtgtgggc accagcaggg cagattgaat cttatagtct tgcaagctgc gcaagaatag
      481 ctcctcctcg aagcggtaca tgagcacgac ccgaaagccg cagatcaagt agcccagcgt
      541 ggtgaacatg ccgaagccgt ggtgaaatgg caccacgctg aggatagcgg tgtcggggat
      601 gatctggttg ccgaagatgg ggtcgcgggc atgactgaat cggacacaag cggtgcggtg
      661 cggtagggct acgcccttgg gcaatccggt actgccacta ctgttcatga tcagggcgat
      721 ggttttgtcc cggtcgaagc tctcgggcac gaagtcgtac tcgttgaagc cgggtggcaa
      781 atgggaagtc acgaaggtgt acatgctttg gaagccctgg tagtcggtct tgctatccat
      841 gatgatgatc ttttgtatga tcggtagctt cttttgcacg ttgaggatct tttgcagccc
      901 tttcttgctc acgaatacga cggtgggctg gctgatgccc atgctgttca gcagctcgcg
      961 ctcgttgtag atgtcgttag ctggggccac agccacaccg atgaacaggg cacccaacac
     1021 gggcatgaag aactgcaagc tattctcgct gcacaccacg atccgatggt ttgtattcag
     1081 cccatagcgc ttcatagctt ctgccagccg aacgctcatc tcgaagtact cggcgtaggt
     1141 aatgtccacc tcgatatgtg cgtcggtaaa ggcgatggtg ccgggcacca gggcgtagcg
     1201 cttcatggct ttgtgcagct gctcgccggc ggtcccgtct tcgagtgggt agaatggcgc
     1261 tgggcccttc ttaatgtttt tggcatcttc catggtggct ttaccaacag taccggattg
     1321 ccaagcttta ggtcgctagc cctggaagtc gagcttccat tatataccct ctctcgaaga
     1381 tccggccagt taggccagag aaatgttctg gcacctgcac ttgcactggg gacagcctat
     1441 tttgctagtt tgttttgttt cgttttgttt tgatggagag cgtatgttag tactatcgat
     1501 tcacacaaaa aaccaacaca cagatgtaat gaaaataaag atattttatt gcggcctgtc
     1561 caatacctcc cgtaccttaa tattacttac ttatccttga gagacgtact agtaaccctg
     1621 ataaatgctt caataatatt gaaaaaggac gagtatgagc atccaacatt ttcgtgtcgc
     1681 actcattccc ttctttgcgg cattttgctt gcctgttttt gcacaccccg aaacgctggt
     1741 gaaagtaaaa gatgctgaag atcaactggg tgcaagagtg ggctatatcg aactggatct
     1801 caatagcggc aagatccttg agtctttccg ccccgaagaa cgattcccga tgatgagcac
     1861 tttcaaagta ctgctatgtg gcgcggtgtt gtcccgtata gacgccgggc aagagcagct
     1921 tggtcgccgt atacactact cacaaaacga cttggttgag tactcgccgg tcacggaaaa
     1981 gcatcttacg gatggcatga cggtaagaga attgtgtagt gctgccatta ccatgagcga
     2041 caacaccgcg gccaacttac ttctgacaac gatcggaggc cctaaggagc tgactgcatt
     2101 tcttcataat atgggtgatc atgtgacccg gcttgaccgc tgggaaccag agttgaacga
     2161 agccataccg aacgacgagc gtgataccac gatgccagta gcaatggcca caactcttcg
     2221 gaaactactc actggcgaac ttcttactct agcatcacga cagcagctca tagactggat
     2281 ggaggcggac aaagtagcag gaccacttct tcgctcggcc ctccctgctg gctggttcat
     2341 tgctgacaaa tcgggggccg gtgaacgcgg ctctcgcggc atcattgctg cgctggggcc
     2401 tgatggtaag ccctcacgaa tcgtagtgat ctacacgacg gggagtcagg ccactatgga
     2461 cgaacgaaat aggcagatcg ctgagatcgg tgcctcactg atcaagcact ggtaaccact
     2521 gcagtggttt agcatttgcg gccgctgtca gaccaagttt actcatatat actttagatt
     2581 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     2641 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     2701 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     2761 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     2821 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     2881 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     2941 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     3001 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     3061 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     3121 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     3181 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     3241 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     3301 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     3361 atgtccaggt attttctgcc ttctcataga acaaacgtga ctcgtgggca gttccatacg
     3421 atgcataagt gttagtttac cactcccagg ctgagtgaca ttgagtaagt ggctcaactt
     3481 ctctgagcat tcattctgtc tgtgaaatgg ggattatgct attatccctc ctaaggtggt
     3541 tgtgagaatt gagatttttc attacataaa tatttactga gagctactct aggggtctgg
     3601 gatacactgg tgaccaaaaa aagaaattcc caacatgggg tgcttatgtt ccagtgggct
     3661 gccacagaaa ggggtcggcc tagtgcagag gaatgctcat ttcactgagt tcaaaatact
     3721 tcctcttcca tcaattcctg tagccaagaa agatactcga atttaatccc attatttccc
     3781 agaaacatga gaacaagcca gaaaacattt gtccccagga aaccaaaatt caggatcccc
     3841 atctgctctc atctttctct gggatacttg ttgtaagtct gtcaaaatgc agcccctact
     3901 gcaccgtcag gacccacagt ggctgggcaa gcctcagtcc tggagtcccg caggctggcc
     3961 tgctccagaa accagcgctt cccagaagcc agtgcagcct ccctcgcagc ccagccctcc
     4021 gggcctctcc ccacaggcat gctagcaaca tcccagcagg cgcaaagcat tccagagtga
     4081 aggcggccac ccctgaaaac acccctctat cttcaaggcc tacatttctc cagtattctg
     4141 acagctttta gtacgggcaa gctatgttaa aggaaagcct ccctgactaa ccttggaagt
     4201 gtaattcaat gccggtgacg attcagccgt ggatgttact cttaggcgtg ccacctgtga
     4261 ctcatgctca gagggtagaa tgggggccac agccacaccc tctccccttc cttccaaggg
     4321 ctgcagcaaa gtatcctggc acactaagga acttttaaaa gtcattgata aggctcatta
     4381 atgtatcaaa atgtcaccta ctcaatgtga accattaaaa attattagta caagtattag
     4441 tagaaatagt cattaaagtg cagaaatacc aggagaaata tcaagtagaa atattcatca
     4501 ccttggagct acgccggcag cccccttacc tcctgctcca agcccttcag ccacgtgggt
     4561 caggagctgt gccctgtggt ctcagccctc aggtcaaagc cctgaaacct cagggtgagc
     4621 aaggagagtc tgcgtggaga atcaatttca ctcaaacact tggagaaaaa cagcaatgtt
     4681 aactcacaaa taatagaaag tatattcaaa cacgtgatgc atgtatgaag cagaaagtaa
     4741 aagtcacaca tatttttaaa ataagacgtt gcagctgtgt cctctcatct tgaaggggat
     4801 gtatttcctc tcctctgcaa tctggggaga cgcacaagcc gcactacaaa cccgcccagg
     4861 ggtgggatcc ttatcgattt taccacattt gtagaggttt tacttgcttt aaaaaacctc
     4921 ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt
     4981 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     5041 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     5101 tgctcgaagc ggccggccgc cccgactcta gaattattac acggcgatct tgccgccctt
     5161 cttggcctta atgagaatct cgcggatctt gcgggcgtcc aacttgccgg tcagtccttt
     5221 aggcacctcg tccacgaaca caacaccacc gcgcagcttc ttggcggttg taacctggct
     5281 ggccacatag tccacgatct ccttctcggt catggtttta ccgtgttcca gcacgacgac
     5341 tgcggcgggc agctcgccgg catcgtcgtc gggcaggccg gcgaccccgg cgtcgaagat
     5401 gttggggtgt tgcagcagga tgctctccag ttcggctggg gctacctggt agcccttgta
     5461 tttgatcagg ctcttcagcc ggtccacgat gaagaa
//
