LOCUS       sequence-176515-        4739 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Luciferase reporter for IL2RA enhancer (IGI-P0613) .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4739)
  AUTHORS   Simeonov DR, Gowen BG, Boontanrart M, Roth TL, Gagnon JD, Mumbach 
            MR, Satpathy AT, Lee Y, Bray NL, Chan AY, Lituiev DS, Nguyen ML, 
            Gate RE, Subramaniam M, Li Z, Woo JM, Mitros T, Ray GJ, Curie GL, 
            Naddaf N, Chu JS, Ma H, Boyer E, Van Gool F, Huang H, Liu R, Tobin 
            VR, Schumann K, Daly MJ, Farh KK, Ansel KM, Ye CJ, Greenleaf WJ, 
            Anderson MS, Bluestone JA, Chang HY, Corn JE, Marson A
  TITLE     Discovery of stimulation-responsive immune enhancers with CRISPR 
            activation.
  JOURNAL   Nature. 2017 Sep 7;549(7670):111-115. doi: 10.1038/nature23875. Epub
            2017 Aug 30.
  PUBMED    28854172
REFERENCE   2  (bases 1 to 4739)
  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"; doi: 
            "10.1038/nature23875"; journalName: "Nature"; date: "2017-09-7- 7"; 
            volume: "549"; issue: "7670"; pages: "111-115"
COMMENT     Sequence Label: pGL4.23-IL2RA CaRE5 (minus strand)
FEATURES             Location/Qualifiers
     source          1..4739
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        533..564
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      591..600
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             597..2249
                     /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    2293..2414
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2304..2323)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2358..2377
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2662..2679)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2833..3421)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2913..2932)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3621..4412,4413..4481))
                     /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    4586..4634
                     /note="synthetic polyadenylation signal"
     misc_feature    4648..4739
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4688..4707
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gccggtacca atatgactcc atcccaagaa attggtttta aagatttgag
       61 gaaaaactgc ttttagattt cagagaaaga aaaagagtgt gtttctactt ccccctttta
      121 aatctgataa cttttatttt aatcaggttg acattctgtg gtgagcttgt ggttaactct
      181 ttcatgcccg gaagcctgtt gtcgaccgag aagactccca ggattagaac ataaccagcg
      241 cccaacattt catttgcaga agcatctgag gtcccaaaat gtgatggttt agcggcagct
      301 gcttgtaggg gtatttttgg tttcctgctt cttttttccc ttctaagtaa agacaaaata
      361 actagatccc tccaccccac caccccatca tttcgttgtt gtttaacaag tgttgagtaa
      421 ggcacagttg ttctccaaaa gggtcttacc ctgtgagtta ctcattttag gtacctgagc
      481 tcgctagcct cgaggatatc aagatctggc ctcggcggcc aagcttagac actagagggt
      541 atataatgga agctcgactt ccagcttggc aatccggtac tgttggtaaa gccaccatgg
      601 aagatgccaa aaacattaag aagggcccag cgccattcta cccactcgaa gacgggaccg
      661 ccggcgagca gctgcacaaa gccatgaagc gctacgccct ggtgcccggc accatcgcct
      721 ttaccgacgc acatatcgag gtggacatta cctacgccga gtacttcgag atgagcgttc
      781 ggctggcaga agctatgaag cgctatgggc tgaatacaaa ccatcggatc gtggtgtgca
      841 gcgagaatag cttgcagttc ttcatgcccg tgttgggtgc cctgttcatc ggtgtggctg
      901 tggccccagc taacgacatc tacaacgagc gcgagctgct gaacagcatg ggcatcagcc
      961 agcccaccgt cgtattcgtg agcaagaaag ggctgcaaaa gatcctcaac gtgcaaaaga
     1021 agctaccgat catacaaaag atcatcatca tggatagcaa gaccgactac cagggcttcc
     1081 aaagcatgta caccttcgtg acttcccatt tgccacccgg cttcaacgag tacgacttcg
     1141 tgcccgagag cttcgaccgg gacaaaacca tcgccctgat catgaacagt agtggcagta
     1201 ccggattgcc caagggcgta gccctaccgc accgcaccgc ttgtgtccga ttcagtcatg
     1261 cccgcgaccc catcttcggc aaccagatca tccccgacac cgctatcctc agcgtggtgc
     1321 catttcacca cggcttcggc atgttcacca cgctgggcta cttgatctgc ggctttcggg
     1381 tcgtgctcat gtaccgcttc gaggaggagc tattcttgcg cagcttgcaa gactataaga
     1441 ttcaatctgc cctgctggtg cccacactat ttagcttctt cgctaagagc actctcatcg
     1501 acaagtacga cctaagcaac ttgcacgaga tcgccagcgg cggggcgccg ctcagcaagg
     1561 aggtaggtga ggccgtggcc aaacgcttcc acctaccagg catccgccag ggctacggcc
     1621 tgacagaaac aaccagcgcc attctgatca cccccgaagg ggacgacaag cctggcgcag
     1681 taggcaaggt ggtgcccttc ttcgaggcta aggtggtgga cttggacacc ggtaagacac
     1741 tgggtgtgaa ccagcgcggc gagctgtgcg tccgtggccc catgatcatg agcggctacg
     1801 ttaacaaccc cgaggctaca aacgctctca tcgacaagga cggctggctg cacagcggcg
     1861 acatcgccta ctgggacgag gacgagcact tcttcatcgt ggaccggctg aagagcctga
     1921 tcaaatacaa gggctaccag gtagccccag ccgaactgga gagcatcctg ctgcaacacc
     1981 ccaacatctt cgacgccggg gtcgccggcc tgcccgacga cgatgccggc gagctgcccg
     2041 ccgcagtcgt cgtgctggaa cacggtaaaa ccatgaccga gaaggagatc gtggactatg
     2101 tggccagcca ggttacaacc gccaagaagc tgcgcggtgg tgttgtgttc gtggacgagg
     2161 tgcctaaagg actgaccggc aagttggacg cccgcaagat ccgcgagatt ctcattaagg
     2221 ccaagaaggg cggcaagatc gccgtgtaat aattctagag tcggggcggc cggccgcttc
     2281 gagcagacat gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa
     2341 aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct
     2401 gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg
     2461 tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtaaaatc gataaggatc
     2521 cgtcgaccga tgcccttgag agccttcaac ccagtcagct ccttccggtg ggcgcggggc
     2581 atgactatcg tcgccgcact tatgactgtc ttctttatca tgcaactcgt aggacaggtg
     2641 ccggcagcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     2701 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     2761 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     2821 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     2881 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     2941 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     3001 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     3061 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     3121 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     3181 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     3241 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     3301 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     3361 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     3421 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     3481 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     3541 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gcggccgcaa
     3601 atgctaaacc actgcagtgg ttaccagtgc ttgatcagtg aggcaccgat ctcagcgatc
     3661 tgcctatttc gttcgtccat agtggcctga ctccccgtcg tgtagatcac tacgattcgt
     3721 gagggcttac catcaggccc cagcgcagca atgatgccgc gagagccgcg ttcaccggcc
     3781 cccgatttgt cagcaatgaa ccagccagca gggagggccg agcgaagaag tggtcctgct
     3841 actttgtccg cctccatcca gtctatgagc tgctgtcgtg atgctagagt aagaagttcg
     3901 ccagtgagta gtttccgaag agttgtggcc attgctactg gcatcgtggt atcacgctcg
     3961 tcgttcggta tggcttcgtt caactctggt tcccagcggt caagccgggt cacatgatca
     4021 cccatattat gaagaaatgc agtcagctcc ttagggcctc cgatcgttgt cagaagtaag
     4081 ttggccgcgg tgttgtcgct catggtaatg gcagcactac acaattctct taccgtcatg
     4141 ccatccgtaa gatgcttttc cgtgaccggc gagtactcaa ccaagtcgtt ttgtgagtag
     4201 tgtatacggc gaccaagctg ctcttgcccg gcgtctatac gggacaacac cgcgccacat
     4261 agcagtactt tgaaagtgct catcatcggg aatcgttctt cggggcggaa agactcaagg
     4321 atcttgccgc tattgagatc cagttcgata tagcccactc ttgcacccag ttgatcttca
     4381 gcatctttta ctttcaccag cgtttcgggg tgtgcaaaaa caggcaagca aaatgccgca
     4441 aagaagggaa tgagtgcgac acgaaaatgt tggatgctca tactcgtcct ttttcaatat
     4501 tattgaagca tttatcaggg ttactagtac gtctctcaag gataagtaag taatattaag
     4561 gtacgggagg tattggacag gccgcaataa aatatcttta ttttcattac atctgtgtgt
     4621 tggttttttg tgtgaatcga tagtactaac atacgctctc catcaaaaca aaacgaaaca
     4681 aaacaaacta gcaaaatagg ctgtccccag tgcaagtgca ggtgccagaa catttctct
//
