LOCUS       sequence-197076-        5496 bp    DNA     circular SYN 15-APR-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 Apr 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"
     promoter        25..55
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      98..107
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             104..1756
                     /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"
     polyA_signal    1800..1921
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1811..1830)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1865..1884
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(3590..4178)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3670..3689)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4378..5169,5170..5238))
                     /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    5343..5391
                     /note="synthetic polyadenylation signal"
     misc_feature    5405..5496
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5445..5464
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gccggatctt cgagagaggg tatataatgg aagctcgact tccagggcta
       61 gcgacctaaa gcttggcaat ccggtactgt tggtaaagcc accatggaag atgccaaaaa
      121 cattaagaag ggcccagcgc cattctaccc actcgaagac gggaccgccg gcgagcagct
      181 gcacaaagcc atgaagcgct acgccctggt gcccggcacc atcgccttta ccgacgcaca
      241 tatcgaggtg gacattacct acgccgagta cttcgagatg agcgttcggc tggcagaagc
      301 tatgaagcgc tatgggctga atacaaacca tcggatcgtg gtgtgcagcg agaatagctt
      361 gcagttcttc atgcccgtgt tgggtgccct gttcatcggt gtggctgtgg ccccagctaa
      421 cgacatctac aacgagcgcg agctgctgaa cagcatgggc atcagccagc ccaccgtcgt
      481 attcgtgagc aagaaagggc tgcaaaagat cctcaacgtg caaaagaagc taccgatcat
      541 acaaaagatc atcatcatgg atagcaagac cgactaccag ggcttccaaa gcatgtacac
      601 cttcgtgact tcccatttgc cacccggctt caacgagtac gacttcgtgc ccgagagctt
      661 cgaccgggac aaaaccatcg ccctgatcat gaacagtagt ggcagtaccg gattgcccaa
      721 gggcgtagcc ctaccgcacc gcaccgcttg tgtccgattc agtcatgccc gcgaccccat
      781 cttcggcaac cagatcatcc ccgacaccgc tatcctcagc gtggtgccat ttcaccacgg
      841 cttcggcatg ttcaccacgc tgggctactt gatctgcggc tttcgggtcg tgctcatgta
      901 ccgcttcgag gaggagctat tcttgcgcag cttgcaagac tataagattc aatctgccct
      961 gctggtgccc acactattta gcttcttcgc taagagcact ctcatcgaca agtacgacct
     1021 aagcaacttg cacgagatcg ccagcggcgg ggcgccgctc agcaaggagg taggtgaggc
     1081 cgtggccaaa cgcttccacc taccaggcat ccgccagggc tacggcctga cagaaacaac
     1141 cagcgccatt ctgatcaccc ccgaagggga cgacaagcct ggcgcagtag gcaaggtggt
     1201 gcccttcttc gaggctaagg tggtggactt ggacaccggt aagacactgg gtgtgaacca
     1261 gcgcggcgag ctgtgcgtcc gtggccccat gatcatgagc ggctacgtta acaaccccga
     1321 ggctacaaac gctctcatcg acaaggacgg ctggctgcac agcggcgaca tcgcctactg
     1381 ggacgaggac gagcacttct tcatcgtgga ccggctgaag agcctgatca aatacaaggg
     1441 ctaccaggta gccccagccg aactggagag catcctgctg caacacccca acatcttcga
     1501 cgccggggtc gccggcctgc ccgacgacga tgccggcgag ctgcccgccg cagtcgtcgt
     1561 gctggaacac ggtaaaacca tgaccgagaa ggagatcgtg gactatgtgg ccagccaggt
     1621 tacaaccgcc aagaagctgc gcggtggtgt tgtgttcgtg gacgaggtgc ctaaaggact
     1681 gaccggcaag ttggacgccc gcaagatccg cgagattctc attaaggcca agaagggcgg
     1741 caagatcgcc gtgtaataat tctagagtcg gggcggccgg ccgcttcgag cagacatgat
     1801 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     1861 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     1921 taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt
     1981 ttaaagcaag taaaacctct acaaatgtgg taaaatcgat aaggatccca cccctgggcg
     2041 ggtttgtagt gcggcttgtg cgtctcccca gattgcagag gagaggaaat acatcccctt
     2101 caagatgaga ggacacagct gcaacgtctt attttaaaaa tatgtgtgac ttttactttc
     2161 tgcttcatac atgcatcacg tgtttgaata tactttctat tatttgtgag ttaacattgc
     2221 tgtttttctc caagtgtttg agtgaaattg attctccacg cagactctcc ttgctcaccc
     2281 tgaggtttca gggctttgac ctgagggctg agaccacagg gcacagctcc tgacccacgt
     2341 ggctgaaggg cttggagcag gaggtaaggg ggctgccggc gtagctccaa ggtgatgaat
     2401 atttctactt gatatttctc ctggtatttc tgcactttaa tgactatttc tactaatact
     2461 tgtactaata atttttaatg gttcacattg agtaggtgac attttgatac attaatgagc
     2521 cttatcaatg acttttaaaa gttccttagt gtgccaggat actttgctgc agcccttgga
     2581 aggaagggga gagggtgtgg ctgtggcccc cattctaccc tctgagcatg agtcacaggt
     2641 ggcacgccta agagtaacat ccacggctga atcgtcaccg gcattgaatt acacttccaa
     2701 ggttagtcag ggaggctttc ctttaacata gcttgcccgt actaaaagct gtcagaatac
     2761 tggagaaatg taggccttga agatagaggg gtgttttcag gggtggccgc cttcactctg
     2821 gaatgctttg cgcctgctgg gatgttgcta gcatgcctgt ggggagaggc ccggagggct
     2881 gggctgcgag ggaggctgca ctggcttctg ggaagcgctg gtttctggag caggccagcc
     2941 tgcgggactc caggactgag gcttgcccag ccactgtggg tcctgacggt gcagtagggg
     3001 ctgcattttg acagacttac aacaagtatc ccagagaaag atgagagcag atggggatcc
     3061 tgaattttgg tttcctgggg acaaatgttt tctggcttgt tctcatgttt ctgggaaata
     3121 atgggattaa attcgagtat ctttcttggc tacaggaatt gatggaagag gaagtatttt
     3181 gaactcagtg aaatgagcat tcctctgcac taggccgacc cctttctgtg gcagcccact
     3241 ggaacataag caccccatgt tgggaatttc tttttttggt caccagtgta tcccagaccc
     3301 ctagagtagc tctcagtaaa tatttatgta atgaaaaatc tcaattctca caaccacctt
     3361 aggagggata atagcataat ccccatttca cagacagaat gaatgctcag agaagttgag
     3421 ccacttactc aatgtcactc agcctgggag tggtaaacta acacttatgc atcgtatgga
     3481 actgcccacg agtcacgttt gttctatgag aaggcagaaa atacctggac atgtgagcaa
     3541 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     3601 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     3661 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     3721 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     3781 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     3841 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     3901 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     3961 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     4021 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     4081 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     4141 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     4201 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     4261 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     4321 gtatatatga gtaaacttgg tctgacagcg gccgcaaatg ctaaaccact gcagtggtta
     4381 ccagtgcttg atcagtgagg caccgatctc agcgatctgc ctatttcgtt cgtccatagt
     4441 ggcctgactc cccgtcgtgt agatcactac gattcgtgag ggcttaccat caggccccag
     4501 cgcagcaatg atgccgcgag agccgcgttc accggccccc gatttgtcag caatgaacca
     4561 gccagcaggg agggccgagc gaagaagtgg tcctgctact ttgtccgcct ccatccagtc
     4621 tatgagctgc tgtcgtgatg ctagagtaag aagttcgcca gtgagtagtt tccgaagagt
     4681 tgtggccatt gctactggca tcgtggtatc acgctcgtcg ttcggtatgg cttcgttcaa
     4741 ctctggttcc cagcggtcaa gccgggtcac atgatcaccc atattatgaa gaaatgcagt
     4801 cagctcctta gggcctccga tcgttgtcag aagtaagttg gccgcggtgt tgtcgctcat
     4861 ggtaatggca gcactacaca attctcttac cgtcatgcca tccgtaagat gcttttccgt
     4921 gaccggcgag tactcaacca agtcgttttg tgagtagtgt atacggcgac caagctgctc
     4981 ttgcccggcg tctatacggg acaacaccgc gccacatagc agtactttga aagtgctcat
     5041 catcgggaat cgttcttcgg ggcggaaaga ctcaaggatc ttgccgctat tgagatccag
     5101 ttcgatatag cccactcttg cacccagttg atcttcagca tcttttactt tcaccagcgt
     5161 ttcggggtgt gcaaaaacag gcaagcaaaa tgccgcaaag aagggaatga gtgcgacacg
     5221 aaaatgttgg atgctcatac tcgtcctttt tcaatattat tgaagcattt atcagggtta
     5281 ctagtacgtc tctcaaggat aagtaagtaa tattaaggta cgggaggtat tggacaggcc
     5341 gcaataaaat atctttattt tcattacatc tgtgtgttgg ttttttgtgt gaatcgatag
     5401 tactaacata cgctctccat caaaacaaaa cgaaacaaaa caaactagca aaataggctg
     5461 tccccagtgc aagtgcaggt gccagaacat ttctct
//
