LOCUS       sequence-176516-        4742 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Luciferase reporter for IL2RA enhancer (IGI-P0614) .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4742)
  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 4742)
  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 CaRE1 (plus strand)
FEATURES             Location/Qualifiers
     source          1..4742
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        536..567
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      594..603
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             600..2252
                     /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    2296..2417
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2307..2326)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2361..2380
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2665..2682)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2836..3424)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2916..2935)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3624..4415,4416..4484))
                     /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    4589..4637
                     /note="synthetic polyadenylation signal"
     misc_feature    4651..4742
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4691..4710
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gccggtacct gagataattt acactaaaac ccctggcaca cggttggcag
       61 tagttctccc cgactctcct aggaatgttt ttcctctgtg ctgagctgtt gtcactgctc
      121 tttgcaacac ttttgctcca cggaggggga ggatgtaact tgaaagggtt actaatccct
      181 cgaaaccctc agatcacaga taaactcatt tcccgatcat gccacgtaag ccaatgatct
      241 ggctttgata acctgatcac acgttgggga ccacattaag gttgccactc aaagctgtac
      301 tcctgttcaa agcgttggag caaggtttgc tgttctgccc ctgttctctg gggatagcac
      361 taaagggcac aacggaaagg ataacagtgc cccaaggagg catcactcac agatgaagct
      421 cttggcctca ctggctgaag agtgggaacc tcccagctgg gaaaggataa aaggtacctg
      481 agctcgctag cctcgaggat atcaagatct ggcctcggcg gccaagctta gacactagag
      541 ggtatataat ggaagctcga cttccagctt ggcaatccgg tactgttggt aaagccacca
      601 tggaagatgc caaaaacatt aagaagggcc cagcgccatt ctacccactc gaagacggga
      661 ccgccggcga gcagctgcac aaagccatga agcgctacgc cctggtgccc ggcaccatcg
      721 cctttaccga cgcacatatc gaggtggaca ttacctacgc cgagtacttc gagatgagcg
      781 ttcggctggc agaagctatg aagcgctatg ggctgaatac aaaccatcgg atcgtggtgt
      841 gcagcgagaa tagcttgcag ttcttcatgc ccgtgttggg tgccctgttc atcggtgtgg
      901 ctgtggcccc agctaacgac atctacaacg agcgcgagct gctgaacagc atgggcatca
      961 gccagcccac cgtcgtattc gtgagcaaga aagggctgca aaagatcctc aacgtgcaaa
     1021 agaagctacc gatcatacaa aagatcatca tcatggatag caagaccgac taccagggct
     1081 tccaaagcat gtacaccttc gtgacttccc atttgccacc cggcttcaac gagtacgact
     1141 tcgtgcccga gagcttcgac cgggacaaaa ccatcgccct gatcatgaac agtagtggca
     1201 gtaccggatt gcccaagggc gtagccctac cgcaccgcac cgcttgtgtc cgattcagtc
     1261 atgcccgcga ccccatcttc ggcaaccaga tcatccccga caccgctatc ctcagcgtgg
     1321 tgccatttca ccacggcttc ggcatgttca ccacgctggg ctacttgatc tgcggctttc
     1381 gggtcgtgct catgtaccgc ttcgaggagg agctattctt gcgcagcttg caagactata
     1441 agattcaatc tgccctgctg gtgcccacac tatttagctt cttcgctaag agcactctca
     1501 tcgacaagta cgacctaagc aacttgcacg agatcgccag cggcggggcg ccgctcagca
     1561 aggaggtagg tgaggccgtg gccaaacgct tccacctacc aggcatccgc cagggctacg
     1621 gcctgacaga aacaaccagc gccattctga tcacccccga aggggacgac aagcctggcg
     1681 cagtaggcaa ggtggtgccc ttcttcgagg ctaaggtggt ggacttggac accggtaaga
     1741 cactgggtgt gaaccagcgc ggcgagctgt gcgtccgtgg ccccatgatc atgagcggct
     1801 acgttaacaa ccccgaggct acaaacgctc tcatcgacaa ggacggctgg ctgcacagcg
     1861 gcgacatcgc ctactgggac gaggacgagc acttcttcat cgtggaccgg ctgaagagcc
     1921 tgatcaaata caagggctac caggtagccc cagccgaact ggagagcatc ctgctgcaac
     1981 accccaacat cttcgacgcc ggggtcgccg gcctgcccga cgacgatgcc ggcgagctgc
     2041 ccgccgcagt cgtcgtgctg gaacacggta aaaccatgac cgagaaggag atcgtggact
     2101 atgtggccag ccaggttaca accgccaaga agctgcgcgg tggtgttgtg ttcgtggacg
     2161 aggtgcctaa aggactgacc ggcaagttgg acgcccgcaa gatccgcgag attctcatta
     2221 aggccaagaa gggcggcaag atcgccgtgt aataattcta gagtcggggc ggccggccgc
     2281 ttcgagcaga catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt
     2341 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
     2401 gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg
     2461 aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtaaa atcgataagg
     2521 atccgtcgac cgatgccctt gagagccttc aacccagtca gctccttccg gtgggcgcgg
     2581 ggcatgacta tcgtcgccgc acttatgact gtcttcttta tcatgcaact cgtaggacag
     2641 gtgccggcag cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     2701 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     2761 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     2821 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     2881 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     2941 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     3001 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     3061 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     3121 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     3181 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     3241 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     3301 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     3361 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     3421 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     3481 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     3541 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagcggccg
     3601 caaatgctaa accactgcag tggttaccag tgcttgatca gtgaggcacc gatctcagcg
     3661 atctgcctat ttcgttcgtc catagtggcc tgactccccg tcgtgtagat cactacgatt
     3721 cgtgagggct taccatcagg ccccagcgca gcaatgatgc cgcgagagcc gcgttcaccg
     3781 gcccccgatt tgtcagcaat gaaccagcca gcagggaggg ccgagcgaag aagtggtcct
     3841 gctactttgt ccgcctccat ccagtctatg agctgctgtc gtgatgctag agtaagaagt
     3901 tcgccagtga gtagtttccg aagagttgtg gccattgcta ctggcatcgt ggtatcacgc
     3961 tcgtcgttcg gtatggcttc gttcaactct ggttcccagc ggtcaagccg ggtcacatga
     4021 tcacccatat tatgaagaaa tgcagtcagc tccttagggc ctccgatcgt tgtcagaagt
     4081 aagttggccg cggtgttgtc gctcatggta atggcagcac tacacaattc tcttaccgtc
     4141 atgccatccg taagatgctt ttccgtgacc ggcgagtact caaccaagtc gttttgtgag
     4201 tagtgtatac ggcgaccaag ctgctcttgc ccggcgtcta tacgggacaa caccgcgcca
     4261 catagcagta ctttgaaagt gctcatcatc gggaatcgtt cttcggggcg gaaagactca
     4321 aggatcttgc cgctattgag atccagttcg atatagccca ctcttgcacc cagttgatct
     4381 tcagcatctt ttactttcac cagcgtttcg gggtgtgcaa aaacaggcaa gcaaaatgcc
     4441 gcaaagaagg gaatgagtgc gacacgaaaa tgttggatgc tcatactcgt cctttttcaa
     4501 tattattgaa gcatttatca gggttactag tacgtctctc aaggataagt aagtaatatt
     4561 aaggtacggg aggtattgga caggccgcaa taaaatatct ttattttcat tacatctgtg
     4621 tgttggtttt ttgtgtgaat cgatagtact aacatacgct ctccatcaaa acaaaacgaa
     4681 acaaaacaaa ctagcaaaat aggctgtccc cagtgcaagt gcaggtgcca gaacatttct
     4741 ct
//
