LOCUS       sequence-180044-        4766 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Luciferase reporter for IL2RA enhancer (IGI-P0347).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4766)
  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 4766)
  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 CaRE4 scramble (plus strand)
FEATURES             Location/Qualifiers
     source          1..4766
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        560..591
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      618..627
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             624..2276
                     /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    2320..2441
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2331..2350)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2385..2404
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2689..2706)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2860..3448)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2940..2959)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3648..4439,4440..4508))
                     /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    4613..4661
                     /note="synthetic polyadenylation signal"
     misc_feature    4675..4766
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4715..4734
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gccggtacct gagctcgcta gcctccctct tcctcctgta ccctcaaccc
       61 atcagctgcc tgtactgctg tagcaacaac ttctctctga aaaccttcta tttctggtgg
      121 ctacacacag gtgtctgctc cggcaggata tctcccagag gtgtttatag actccgtaaa
      181 gatcggcacg acaggagtca acactggtgc ggagaaacga aagcacgtca aggcggcaga
      241 gagtgctacg gtgcggctgg cgaggggaag cgtccttctt cagaattctg gctgttatcc
      301 tcttacttgg cacgagctcc gatggggtcc gtacaagtga ccggcctacg aggtcggcac
      361 gtaaggtgga gtgtccttta ctaataagcc cggtttgata acagaccgtg caggaccgag
      421 gactgcactg gatttcaccc taaagccccg tagcgcctta agccatcctg caggaggccg
      481 gctgctctag caggttgagg agtgccacgc ggctctgtgg gtaccgcctc acttggcctc
      541 ggcggccaag cttagacact agagggtata taatggaagc tcgacttcca gcttggcaat
      601 ccggtactgt tggtaaagcc accatggaag atgccaaaaa cattaagaag ggcccagcgc
      661 cattctaccc actcgaagac gggaccgccg gcgagcagct gcacaaagcc atgaagcgct
      721 acgccctggt gcccggcacc atcgccttta ccgacgcaca tatcgaggtg gacattacct
      781 acgccgagta cttcgagatg agcgttcggc tggcagaagc tatgaagcgc tatgggctga
      841 atacaaacca tcggatcgtg gtgtgcagcg agaatagctt gcagttcttc atgcccgtgt
      901 tgggtgccct gttcatcggt gtggctgtgg ccccagctaa cgacatctac aacgagcgcg
      961 agctgctgaa cagcatgggc atcagccagc ccaccgtcgt attcgtgagc aagaaagggc
     1021 tgcaaaagat cctcaacgtg caaaagaagc taccgatcat acaaaagatc atcatcatgg
     1081 atagcaagac cgactaccag ggcttccaaa gcatgtacac cttcgtgact tcccatttgc
     1141 cacccggctt caacgagtac gacttcgtgc ccgagagctt cgaccgggac aaaaccatcg
     1201 ccctgatcat gaacagtagt ggcagtaccg gattgcccaa gggcgtagcc ctaccgcacc
     1261 gcaccgcttg tgtccgattc agtcatgccc gcgaccccat cttcggcaac cagatcatcc
     1321 ccgacaccgc tatcctcagc gtggtgccat ttcaccacgg cttcggcatg ttcaccacgc
     1381 tgggctactt gatctgcggc tttcgggtcg tgctcatgta ccgcttcgag gaggagctat
     1441 tcttgcgcag cttgcaagac tataagattc aatctgccct gctggtgccc acactattta
     1501 gcttcttcgc taagagcact ctcatcgaca agtacgacct aagcaacttg cacgagatcg
     1561 ccagcggcgg ggcgccgctc agcaaggagg taggtgaggc cgtggccaaa cgcttccacc
     1621 taccaggcat ccgccagggc tacggcctga cagaaacaac cagcgccatt ctgatcaccc
     1681 ccgaagggga cgacaagcct ggcgcagtag gcaaggtggt gcccttcttc gaggctaagg
     1741 tggtggactt ggacaccggt aagacactgg gtgtgaacca gcgcggcgag ctgtgcgtcc
     1801 gtggccccat gatcatgagc ggctacgtta acaaccccga ggctacaaac gctctcatcg
     1861 acaaggacgg ctggctgcac agcggcgaca tcgcctactg ggacgaggac gagcacttct
     1921 tcatcgtgga ccggctgaag agcctgatca aatacaaggg ctaccaggta gccccagccg
     1981 aactggagag catcctgctg caacacccca acatcttcga cgccggggtc gccggcctgc
     2041 ccgacgacga tgccggcgag ctgcccgccg cagtcgtcgt gctggaacac ggtaaaacca
     2101 tgaccgagaa ggagatcgtg gactatgtgg ccagccaggt tacaaccgcc aagaagctgc
     2161 gcggtggtgt tgtgttcgtg gacgaggtgc ctaaaggact gaccggcaag ttggacgccc
     2221 gcaagatccg cgagattctc attaaggcca agaagggcgg caagatcgcc gtgtaataat
     2281 tctagagtcg gggcggccgg ccgcttcgag cagacatgat aagatacatt gatgagtttg
     2341 gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta
     2401 ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc
     2461 attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct
     2521 acaaatgtgg taaaatcgat aaggatccgt cgaccgatgc ccttgagagc cttcaaccca
     2581 gtcagctcct tccggtgggc gcggggcatg actatcgtcg ccgcacttat gactgtcttc
     2641 tttatcatgc aactcgtagg acaggtgccg gcagcgctct tccgcttcct cgctcactga
     2701 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     2761 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     2821 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     2881 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     2941 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     3001 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     3061 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     3121 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     3181 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     3241 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     3301 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     3361 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     3421 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     3481 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     3541 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     3601 gtaaacttgg tctgacagcg gccgcaaatg ctaaaccact gcagtggtta ccagtgcttg
     3661 atcagtgagg caccgatctc agcgatctgc ctatttcgtt cgtccatagt ggcctgactc
     3721 cccgtcgtgt agatcactac gattcgtgag ggcttaccat caggccccag cgcagcaatg
     3781 atgccgcgag agccgcgttc accggccccc gatttgtcag caatgaacca gccagcaggg
     3841 agggccgagc gaagaagtgg tcctgctact ttgtccgcct ccatccagtc tatgagctgc
     3901 tgtcgtgatg ctagagtaag aagttcgcca gtgagtagtt tccgaagagt tgtggccatt
     3961 gctactggca tcgtggtatc acgctcgtcg ttcggtatgg cttcgttcaa ctctggttcc
     4021 cagcggtcaa gccgggtcac atgatcaccc atattatgaa gaaatgcagt cagctcctta
     4081 gggcctccga tcgttgtcag aagtaagttg gccgcggtgt tgtcgctcat ggtaatggca
     4141 gcactacaca attctcttac cgtcatgcca tccgtaagat gcttttccgt gaccggcgag
     4201 tactcaacca agtcgttttg tgagtagtgt atacggcgac caagctgctc ttgcccggcg
     4261 tctatacggg acaacaccgc gccacatagc agtactttga aagtgctcat catcgggaat
     4321 cgttcttcgg ggcggaaaga ctcaaggatc ttgccgctat tgagatccag ttcgatatag
     4381 cccactcttg cacccagttg atcttcagca tcttttactt tcaccagcgt ttcggggtgt
     4441 gcaaaaacag gcaagcaaaa tgccgcaaag aagggaatga gtgcgacacg aaaatgttgg
     4501 atgctcatac tcgtcctttt tcaatattat tgaagcattt atcagggtta ctagtacgtc
     4561 tctcaaggat aagtaagtaa tattaaggta cgggaggtat tggacaggcc gcaataaaat
     4621 atctttattt tcattacatc tgtgtgttgg ttttttgtgt gaatcgatag tactaacata
     4681 cgctctccat caaaacaaaa cgaaacaaaa caaactagca aaataggctg tccccagtgc
     4741 aagtgcaggt gccagaacat ttctct
//
