LOCUS       sequence-193011-        5548 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 5548)
  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 5548)
  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_AGAP1
FEATURES             Location/Qualifiers
     source          1..5548
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    1408..1529
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1419..1438)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1473..1492
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(3198..3786)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3278..3297)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3986..4777,4778..4846))
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     polyA_signal    4951..4999
                     /note="synthetic polyadenylation signal"
     misc_feature    5013..5104
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5053..5072
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     regulatory      5254..5263
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(5260..5548,1..1364)
                     /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 tcatcggtgt ggctgtggcc ccagctaacg acatctacaa cgagcgcgag ctgctgaaca
       61 gcatgggcat cagccagccc accgtcgtat tcgtgagcaa gaaagggctg caaaagatcc
      121 tcaacgtgca aaagaagcta ccgatcatac aaaagatcat catcatggat agcaagaccg
      181 actaccaggg cttccaaagc atgtacacct tcgtgacttc ccatttgcca cccggcttca
      241 acgagtacga cttcgtgccc gagagcttcg accgggacaa aaccatcgcc ctgatcatga
      301 acagtagtgg cagtaccgga ttgcccaagg gcgtagccct accgcaccgc accgcttgtg
      361 tccgattcag tcatgcccgc gaccccatct tcggcaacca gatcatcccc gacaccgcta
      421 tcctcagcgt ggtgccattt caccacggct tcggcatgtt caccacgctg ggctacttga
      481 tctgcggctt tcgggtcgtg ctcatgtacc gcttcgagga ggagctattc ttgcgcagct
      541 tgcaagacta taagattcaa tctgccctgc tggtgcccac actatttagc ttcttcgcta
      601 agagcactct catcgacaag tacgacctaa gcaacttgca cgagatcgcc agcggcgggg
      661 cgccgctcag caaggaggta ggtgaggccg tggccaaacg cttccaccta ccaggcatcc
      721 gccagggcta cggcctgaca gaaacaacca gcgccattct gatcaccccc gaaggggacg
      781 acaagcctgg cgcagtaggc aaggtggtgc ccttcttcga ggctaaggtg gtggacttgg
      841 acaccggtaa gacactgggt gtgaaccagc gcggcgagct gtgcgtccgt ggccccatga
      901 tcatgagcgg ctacgttaac aaccccgagg ctacaaacgc tctcatcgac aaggacggct
      961 ggctgcacag cggcgacatc gcctactggg acgaggacga gcacttcttc atcgtggacc
     1021 ggctgaagag cctgatcaaa tacaagggct accaggtagc cccagccgaa ctggagagca
     1081 tcctgctgca acaccccaac atcttcgacg ccggggtcgc cggcctgccc gacgacgatg
     1141 ccggcgagct gcccgccgca gtcgtcgtgc tggaacacgg taaaaccatg accgagaagg
     1201 agatcgtgga ctatgtggcc agccaggtta caaccgccaa gaagctgcgc ggtggtgttg
     1261 tgttcgtgga cgaggtgcct aaaggactga ccggcaagtt ggacgcccgc aagatccgcg
     1321 agattctcat taaggccaag aagggcggca agatcgccgt gtaataattc tagagtcggg
     1381 gcggccggcc gcttcgagca gacatgataa gatacattga tgagtttgga caaaccacaa
     1441 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
     1501 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
     1561 aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta
     1621 aaatcgataa ggatcccacc cctgggcggg tttgtagtgc ggcttgtgcg tctccccaga
     1681 ttgcagagga gaggaaatac atccccttca agatgagagg acacagctgc aacgtcttat
     1741 tttaaaaata tgtgtgactt ttactttctg cttcatacat gcatcacgtg tttgaatata
     1801 ctttctatta tttgtgagtt aacattgctg tttttctcca agtgtttgag tgaaattgat
     1861 tctccacgca gactctcctt gctcaccctg aggtttcagg gctttgacct gagggctgag
     1921 accacagggc acagctcctg acccacgtgg ctgaagggct tggagcagga ggtaaggggg
     1981 ctgccggcgt agctccaagg tgatgaatat ttctacttga tatttctcct ggtatttctg
     2041 cactttaatg actatttcta ctaatacttg tactaataat ttttaatggt tcacattgag
     2101 taggtgacat tttgatacat taatgagcct tatcaatgac ttttaaaagt tccttagtgt
     2161 gccaggatac tttgctgcag cccttggaag gaaggggaga gggtgtggct gtggccccca
     2221 ttctaccctc tgagcatgag tcacaggtgg cacgcctaag agtaacatcc acggctgaat
     2281 cgtcaccggc attgaactac acttccaagg ttagtcaggg aggctttcct ttaacatagc
     2341 ttgcccgtac taaaagctgt cagaatactg gagaaatgta ggccttgaag atagaggggt
     2401 gttttcaggg gtggctgcct tcactctgga atgctttgcg cctgctggga tgttgctagc
     2461 atgcctgtgg ggagaggccc ggagggctgg gctgcgaggg aggctgcact ggcttctggg
     2521 aagcgctggt ttctggagca ggccagcctg cgggactcca ggactgaggc ttgcccagcc
     2581 actgtgggtc ctgacggtgc agtaggggct gcattttgac agacttacaa caagtatccc
     2641 agagaaagat gagagcagat ggggatcctg aattttggtt tcctggggac aaatgttttc
     2701 tggcttgttc tcatgtttct gggaaataat gggattaaat tcgagtatct ttcttggcta
     2761 caggaattga tggaagagga agtattttga actcagtgaa atgagcattc ctctgcacta
     2821 ggccgacccc tttctgtggc agcccactgg aacataagca ccccatgttg ggaatttctt
     2881 tttttggtca ccagtgtatc ccagacccct agagtagctc tcagtaaata tttatgtaat
     2941 gaaaaatctc aattctcaca accaccttag gagggataat agcataatcc ccatttcaca
     3001 gacagaatga atgctcagag aagttgagcc acttactcaa tgtcactcag cctgggagtg
     3061 gtaaactaac acttatgcat tgtatggaac tgcccacgag tcacgtttgt tctatgagaa
     3121 ggcagaaaat acctggacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     3181 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     3241 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     3301 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     3361 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     3421 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     3481 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     3541 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     3601 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     3661 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     3721 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     3781 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     3841 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     3901 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagcggc
     3961 cgcaaatgct aaaccactgc agtggttacc agtgcttgat cagtgaggca ccgatctcag
     4021 cgatctgcct atttcgttcg tccatagtgg cctgactccc cgtcgtgtag atcactacga
     4081 ttcgtgaggg cttaccatca ggccccagcg cagcaatgat gccgcgagag ccgcgttcac
     4141 cggcccccga tttgtcagca atgaaccagc cagcagggag ggccgagcga agaagtggtc
     4201 ctgctacttt gtccgcctcc atccagtcta tgagctgctg tcgtgatgct agagtaagaa
     4261 gttcgccagt gagtagtttc cgaagagttg tggccattgc tactggcatc gtggtatcac
     4321 gctcgtcgtt cggtatggct tcgttcaact ctggttccca gcggtcaagc cgggtcacat
     4381 gatcacccat attatgaaga aatgcagtca gctccttagg gcctccgatc gttgtcagaa
     4441 gtaagttggc cgcggtgttg tcgctcatgg taatggcagc actacacaat tctcttaccg
     4501 tcatgccatc cgtaagatgc ttttccgtga ccggcgagta ctcaaccaag tcgttttgtg
     4561 agtagtgtat acggcgacca agctgctctt gcccggcgtc tatacgggac aacaccgcgc
     4621 cacatagcag tactttgaaa gtgctcatca tcgggaatcg ttcttcgggg cggaaagact
     4681 caaggatctt gccgctattg agatccagtt cgatatagcc cactcttgca cccagttgat
     4741 cttcagcatc ttttactttc accagcgttt cggggtgtgc aaaaacaggc aagcaaaatg
     4801 ccgcaaagaa gggaatgagt gcgacacgaa aatgttggat gctcatactc gtcctttttc
     4861 aatattattg aagcatttat cagggttact agtacgtctc tcaaggataa gtaagtaata
     4921 ttaaggtacg ggaggtattg gacaggccgc aataaaatat ctttattttc attacatctg
     4981 tgtgttggtt ttttgtgtga atcgatagta ctaacatacg ctctccatca aaacaaaacg
     5041 aaacaaaaca aactagcaaa ataggctgtc cccagtgcaa gtgcaggtgc cagaacattt
     5101 ctctggccta actggccgga tctccctagg gtacttatat aagggggtgg gggcgcgttc
     5161 gtcctcagtc gcgatcgaac actcgagccg agcagacgtg cctacggacc gggctagcga
     5221 cctaaagctt ggcaatccgg tactgttggt aaagccacca tggaagatgc caaaaacatt
     5281 aagaagggcc cagcgccatt ctacccactc gaagacggga ccgccggcga gcagctgcac
     5341 aaagccatga agcgctacgc cctggtgccc ggcaccatcg cctttaccga cgcacatatc
     5401 gaggtggaca ttacctacgc cgagtacttc gagatgagcg ttcggctggc agaagctatg
     5461 aagcgctatg ggctgaatac aaaccatcgg atcgtggtgt gcagcgagaa tagcttgcag
     5521 ttcttcatgc ccgtgttggg tgccctgt
//
