LOCUS       sequence-179920-        5731 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Luciferase reporter for IL2RA enhancer (IGI-P0612).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5731)
  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 5731)
  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 CaRE3 (minus strand)
FEATURES             Location/Qualifiers
     source          1..5731
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1525..1556
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      1583..1592
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1589..3241
                     /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    3285..3406
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3296..3315)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3350..3369
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3654..3671)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3825..4413)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3905..3924)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4613..5404,5405..5473))
                     /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    5578..5626
                     /note="synthetic polyadenylation signal"
     misc_feature    5640..5731
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5680..5699
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gccggtacct tggtaactca tcagtgctgg cttgagagat aaactctatt
       61 atccatccct gaactaaaat catgacagaa gtggccaggg agctttgctg ctatcccccc
      121 aggaaacacg tcctccactc aaatggaaag aggaccctct gacaacatct gtgggaccca
      181 acagcactgg tcaccacaag ccacaaaatg ttaacaaagt cagttttcaa ttgttaggga
      241 cggaggactc agttcatgat tcatacaaac caactgttct ctcccagtgt tttctgggtg
      301 gcacaaccca caagtcaaca gtggcttggg aactagacat ttgagtagag ttgggttatt
      361 tgattcatag tggattttgg ttttccacgg gacccctgtg cccttgtcta gtagaatctg
      421 gtggaaatta caaactgcag aaattcaact cagtgccgca ataacaggat gcacctgtag
      481 atttcgtaga attagcagca gcattctttc aataccagtt tgagagaaat aaccctgttt
      541 gcatagtgcc aactggggca gaatctgaag tgtcttgcct gcctctcggc catgggaggc
      601 accgatctca gcaacatcta tccaaccgcc atccacttaa caagcaaaga tggaaggccc
      661 tcgtgctcca ggctatcctg caatggttca tttgcttttg atgggattat cactttgaca
      721 ggtgtgcttg caattaactg ggggctttct gcttccaatc caaattccca caggtggatc
      781 actggctctt ggggagcaaa aacccttctt tgtttgtttt tgcttctccc agagcctggg
      841 cagagctccc cattcctgga tcctacgttg atatgcactg aattgaatta tactaaaggg
      901 tgcacccaag agtgcatgag ctaagtaaaa tgtaatttgt gggacctcat gtggacattt
      961 caggtccatg tgtacaagat aaaggagaga aaatcctggg ggacaccaca ggcctgggaa
     1021 aatgacctcc tactaagcct aacaagaaca tccagtacat aaacagcagg ctctctgcaa
     1081 ccgcttcggt ccttccactg ccacgctccc agaaagcaaa gggttatctc ctgttcggat
     1141 tttggctctg agaccacagc tcagcacatc actcctctag agaaaagaaa gtctaagatg
     1201 agagagtgag gctgctttca tgtggtgcca gcgtatagtg cagggcaggc ctgattgctg
     1261 aacggatccc tgcagacgtg agcggtcact tccccttttg agggcccatt tcttttcctc
     1321 tttgaggaga ctgtgagcgt cctcagtgca gggctatgtt ttagcttctt ggctctcctc
     1381 acaggttcta gcataattct ttgattaatt gcagtcaacg tttattgatg gattatgtga
     1441 cagaggtgaa attctttgat cggtacctga gctcgctagc ctcgaggata tcaagatctg
     1501 gcctcggcgg ccaagcttag acactagagg gtatataatg gaagctcgac ttccagcttg
     1561 gcaatccggt actgttggta aagccaccat ggaagatgcc aaaaacatta agaagggccc
     1621 agcgccattc tacccactcg aagacgggac cgccggcgag cagctgcaca aagccatgaa
     1681 gcgctacgcc ctggtgcccg gcaccatcgc ctttaccgac gcacatatcg aggtggacat
     1741 tacctacgcc gagtacttcg agatgagcgt tcggctggca gaagctatga agcgctatgg
     1801 gctgaataca aaccatcgga tcgtggtgtg cagcgagaat agcttgcagt tcttcatgcc
     1861 cgtgttgggt gccctgttca tcggtgtggc tgtggcccca gctaacgaca tctacaacga
     1921 gcgcgagctg ctgaacagca tgggcatcag ccagcccacc gtcgtattcg tgagcaagaa
     1981 agggctgcaa aagatcctca acgtgcaaaa gaagctaccg atcatacaaa agatcatcat
     2041 catggatagc aagaccgact accagggctt ccaaagcatg tacaccttcg tgacttccca
     2101 tttgccaccc ggcttcaacg agtacgactt cgtgcccgag agcttcgacc gggacaaaac
     2161 catcgccctg atcatgaaca gtagtggcag taccggattg cccaagggcg tagccctacc
     2221 gcaccgcacc gcttgtgtcc gattcagtca tgcccgcgac cccatcttcg gcaaccagat
     2281 catccccgac accgctatcc tcagcgtggt gccatttcac cacggcttcg gcatgttcac
     2341 cacgctgggc tacttgatct gcggctttcg ggtcgtgctc atgtaccgct tcgaggagga
     2401 gctattcttg cgcagcttgc aagactataa gattcaatct gccctgctgg tgcccacact
     2461 atttagcttc ttcgctaaga gcactctcat cgacaagtac gacctaagca acttgcacga
     2521 gatcgccagc ggcggggcgc cgctcagcaa ggaggtaggt gaggccgtgg ccaaacgctt
     2581 ccacctacca ggcatccgcc agggctacgg cctgacagaa acaaccagcg ccattctgat
     2641 cacccccgaa ggggacgaca agcctggcgc agtaggcaag gtggtgccct tcttcgaggc
     2701 taaggtggtg gacttggaca ccggtaagac actgggtgtg aaccagcgcg gcgagctgtg
     2761 cgtccgtggc cccatgatca tgagcggcta cgttaacaac cccgaggcta caaacgctct
     2821 catcgacaag gacggctggc tgcacagcgg cgacatcgcc tactgggacg aggacgagca
     2881 cttcttcatc gtggaccggc tgaagagcct gatcaaatac aagggctacc aggtagcccc
     2941 agccgaactg gagagcatcc tgctgcaaca ccccaacatc ttcgacgccg gggtcgccgg
     3001 cctgcccgac gacgatgccg gcgagctgcc cgccgcagtc gtcgtgctgg aacacggtaa
     3061 aaccatgacc gagaaggaga tcgtggacta tgtggccagc caggttacaa ccgccaagaa
     3121 gctgcgcggt ggtgttgtgt tcgtggacga ggtgcctaaa ggactgaccg gcaagttgga
     3181 cgcccgcaag atccgcgaga ttctcattaa ggccaagaag ggcggcaaga tcgccgtgta
     3241 ataattctag agtcggggcg gccggccgct tcgagcagac atgataagat acattgatga
     3301 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     3361 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     3421 cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa
     3481 cctctacaaa tgtggtaaaa tcgataagga tccgtcgacc gatgcccttg agagccttca
     3541 acccagtcag ctccttccgg tgggcgcggg gcatgactat cgtcgccgca cttatgactg
     3601 tcttctttat catgcaactc gtaggacagg tgccggcagc gctcttccgc ttcctcgctc
     3661 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     3721 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     3781 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     3841 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     3901 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     3961 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     4021 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     4081 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     4141 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     4201 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     4261 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     4321 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     4381 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     4441 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     4501 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     4561 tatgagtaaa cttggtctga cagcggccgc aaatgctaaa ccactgcagt ggttaccagt
     4621 gcttgatcag tgaggcaccg atctcagcga tctgcctatt tcgttcgtcc atagtggcct
     4681 gactccccgt cgtgtagatc actacgattc gtgagggctt accatcaggc cccagcgcag
     4741 caatgatgcc gcgagagccg cgttcaccgg cccccgattt gtcagcaatg aaccagccag
     4801 cagggagggc cgagcgaaga agtggtcctg ctactttgtc cgcctccatc cagtctatga
     4861 gctgctgtcg tgatgctaga gtaagaagtt cgccagtgag tagtttccga agagttgtgg
     4921 ccattgctac tggcatcgtg gtatcacgct cgtcgttcgg tatggcttcg ttcaactctg
     4981 gttcccagcg gtcaagccgg gtcacatgat cacccatatt atgaagaaat gcagtcagct
     5041 ccttagggcc tccgatcgtt gtcagaagta agttggccgc ggtgttgtcg ctcatggtaa
     5101 tggcagcact acacaattct cttaccgtca tgccatccgt aagatgcttt tccgtgaccg
     5161 gcgagtactc aaccaagtcg ttttgtgagt agtgtatacg gcgaccaagc tgctcttgcc
     5221 cggcgtctat acgggacaac accgcgccac atagcagtac tttgaaagtg ctcatcatcg
     5281 ggaatcgttc ttcggggcgg aaagactcaa ggatcttgcc gctattgaga tccagttcga
     5341 tatagcccac tcttgcaccc agttgatctt cagcatcttt tactttcacc agcgtttcgg
     5401 ggtgtgcaaa aacaggcaag caaaatgccg caaagaaggg aatgagtgcg acacgaaaat
     5461 gttggatgct catactcgtc ctttttcaat attattgaag catttatcag ggttactagt
     5521 acgtctctca aggataagta agtaatatta aggtacggga ggtattggac aggccgcaat
     5581 aaaatatctt tattttcatt acatctgtgt gttggttttt tgtgtgaatc gatagtacta
     5641 acatacgctc tccatcaaaa caaaacgaaa caaaacaaac tagcaaaata ggctgtcccc
     5701 agtgcaagtg caggtgccag aacatttctc t
//
