LOCUS       sequence-176524-        4989 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Luciferase reporter for IL2RA enhancer (IGI-P0623) .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4989)
  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 4989)
  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 CaRE2 (plus strand)
FEATURES             Location/Qualifiers
     source          1..4989
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        783..814
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      841..850
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             847..2499
                     /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    2543..2664
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2554..2573)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2608..2627
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2912..2929)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3083..3671)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3163..3182)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3871..4662,4663..4731))
                     /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    4836..4884
                     /note="synthetic polyadenylation signal"
     misc_feature    4898..4989
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4938..4957
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gccggtacct gctgtagagg tgctggacca acagctcatc tgagaggctg
       61 tgctccaaag tggagcagag ggtatggggc agcagcggga ggcagttgtg gggactgcaa
      121 ctgcgaggtt tcttttcaaa tagactttgt ttttagagca gttttagacg cacagcaaac
      181 tcgagtggaa gggagagaga gttcccctgt gcctcctgcc ctcacacccg cacagcgccc
      241 caactatcaa caccccgacc agcgtggcgc atttgttaga actgatgaaa ctacttcatt
      301 tcaaccgagc ggtgtacgat ctattcttct gggagaaagg ggggtgaaat tgcaacagtt
      361 agcccttcct gaaaataaat atgtgtgggg ggtatgttgg tatatttccg agcactgtag
      421 catattcacc agtcacctgc ctggtgcact tgggagtcat ccaccccaca gtcccctcca
      481 ccccacagtc ccctccaccc cacagtcccc tccaccccgc agtcccctcc accccccagt
      541 ctcctccacc ccacagtctc ctccaccccg cagtcccctc caccccccag tcccctccac
      601 cccacagtcc cctttgggcg tttttgcaca aagtattcag cgcacagctt agttgcatac
      661 actcaggtgt gaataggtga ctacctggga tgttgaaatc ctggacagct ggaatctgtg
      721 gtacctgagc tcgctagcct cgaggatatc aagatctggc ctcggcggcc aagcttagac
      781 actagagggt atataatgga agctcgactt ccagcttggc aatccggtac tgttggtaaa
      841 gccaccatgg aagatgccaa aaacattaag aagggcccag cgccattcta cccactcgaa
      901 gacgggaccg ccggcgagca gctgcacaaa gccatgaagc gctacgccct ggtgcccggc
      961 accatcgcct ttaccgacgc acatatcgag gtggacatta cctacgccga gtacttcgag
     1021 atgagcgttc ggctggcaga agctatgaag cgctatgggc tgaatacaaa ccatcggatc
     1081 gtggtgtgca gcgagaatag cttgcagttc ttcatgcccg tgttgggtgc cctgttcatc
     1141 ggtgtggctg tggccccagc taacgacatc tacaacgagc gcgagctgct gaacagcatg
     1201 ggcatcagcc agcccaccgt cgtattcgtg agcaagaaag ggctgcaaaa gatcctcaac
     1261 gtgcaaaaga agctaccgat catacaaaag atcatcatca tggatagcaa gaccgactac
     1321 cagggcttcc aaagcatgta caccttcgtg acttcccatt tgccacccgg cttcaacgag
     1381 tacgacttcg tgcccgagag cttcgaccgg gacaaaacca tcgccctgat catgaacagt
     1441 agtggcagta ccggattgcc caagggcgta gccctaccgc accgcaccgc ttgtgtccga
     1501 ttcagtcatg cccgcgaccc catcttcggc aaccagatca tccccgacac cgctatcctc
     1561 agcgtggtgc catttcacca cggcttcggc atgttcacca cgctgggcta cttgatctgc
     1621 ggctttcggg tcgtgctcat gtaccgcttc gaggaggagc tattcttgcg cagcttgcaa
     1681 gactataaga ttcaatctgc cctgctggtg cccacactat ttagcttctt cgctaagagc
     1741 actctcatcg acaagtacga cctaagcaac ttgcacgaga tcgccagcgg cggggcgccg
     1801 ctcagcaagg aggtaggtga ggccgtggcc aaacgcttcc acctaccagg catccgccag
     1861 ggctacggcc tgacagaaac aaccagcgcc attctgatca cccccgaagg ggacgacaag
     1921 cctggcgcag taggcaaggt ggtgcccttc ttcgaggcta aggtggtgga cttggacacc
     1981 ggtaagacac tgggtgtgaa ccagcgcggc gagctgtgcg tccgtggccc catgatcatg
     2041 agcggctacg ttaacaaccc cgaggctaca aacgctctca tcgacaagga cggctggctg
     2101 cacagcggcg acatcgccta ctgggacgag gacgagcact tcttcatcgt ggaccggctg
     2161 aagagcctga tcaaatacaa gggctaccag gtagccccag ccgaactgga gagcatcctg
     2221 ctgcaacacc ccaacatctt cgacgccggg gtcgccggcc tgcccgacga cgatgccggc
     2281 gagctgcccg ccgcagtcgt cgtgctggaa cacggtaaaa ccatgaccga gaaggagatc
     2341 gtggactatg tggccagcca ggttacaacc gccaagaagc tgcgcggtgg tgttgtgttc
     2401 gtggacgagg tgcctaaagg actgaccggc aagttggacg cccgcaagat ccgcgagatt
     2461 ctcattaagg ccaagaaggg cggcaagatc gccgtgtaat aattctagag tcggggcggc
     2521 cggccgcttc gagcagacat gataagatac attgatgagt ttggacaaac cacaactaga
     2581 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
     2641 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt
     2701 cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtaaaatc
     2761 gataaggatc cgtcgaccga tgcccttgag agccttcaac ccagtcagct ccttccggtg
     2821 ggcgcggggc atgactatcg tcgccgcact tatgactgtc ttctttatca tgcaactcgt
     2881 aggacaggtg ccggcagcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     2941 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     3001 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     3061 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     3121 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     3181 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     3241 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     3301 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     3361 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     3421 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     3481 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     3541 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     3601 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     3661 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     3721 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     3781 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     3841 gcggccgcaa atgctaaacc actgcagtgg ttaccagtgc ttgatcagtg aggcaccgat
     3901 ctcagcgatc tgcctatttc gttcgtccat agtggcctga ctccccgtcg tgtagatcac
     3961 tacgattcgt gagggcttac catcaggccc cagcgcagca atgatgccgc gagagccgcg
     4021 ttcaccggcc cccgatttgt cagcaatgaa ccagccagca gggagggccg agcgaagaag
     4081 tggtcctgct actttgtccg cctccatcca gtctatgagc tgctgtcgtg atgctagagt
     4141 aagaagttcg ccagtgagta gtttccgaag agttgtggcc attgctactg gcatcgtggt
     4201 atcacgctcg tcgttcggta tggcttcgtt caactctggt tcccagcggt caagccgggt
     4261 cacatgatca cccatattat gaagaaatgc agtcagctcc ttagggcctc cgatcgttgt
     4321 cagaagtaag ttggccgcgg tgttgtcgct catggtaatg gcagcactac acaattctct
     4381 taccgtcatg ccatccgtaa gatgcttttc cgtgaccggc gagtactcaa ccaagtcgtt
     4441 ttgtgagtag tgtatacggc gaccaagctg ctcttgcccg gcgtctatac gggacaacac
     4501 cgcgccacat agcagtactt tgaaagtgct catcatcggg aatcgttctt cggggcggaa
     4561 agactcaagg atcttgccgc tattgagatc cagttcgata tagcccactc ttgcacccag
     4621 ttgatcttca gcatctttta ctttcaccag cgtttcgggg tgtgcaaaaa caggcaagca
     4681 aaatgccgca aagaagggaa tgagtgcgac acgaaaatgt tggatgctca tactcgtcct
     4741 ttttcaatat tattgaagca tttatcaggg ttactagtac gtctctcaag gataagtaag
     4801 taatattaag gtacgggagg tattggacag gccgcaataa aatatcttta ttttcattac
     4861 atctgtgtgt tggttttttg tgtgaatcga tagtactaac atacgctctc catcaaaaca
     4921 aaacgaaaca aaacaaacta gcaaaatagg ctgtccccag tgcaagtgca ggtgccagaa
     4981 catttctct
//
