LOCUS       sequence-176518-        5319 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Luciferase reporter for IL2RA enhancer (IGI-P0616) .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5319)
  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 5319)
  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 CaRE6 (plus strand)
FEATURES             Location/Qualifiers
     source          1..5319
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1113..1144
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      1171..1180
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1177..2829
                     /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    2873..2994
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2884..2903)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2938..2957
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3242..3259)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3413..4001)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3493..3512)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4201..4992,4993..5061))
                     /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    5166..5214
                     /note="synthetic polyadenylation signal"
     misc_feature    5228..5319
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5268..5287
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gccggtacca aagagatgtg ggggacagtc attttagaga gcggacacca
       61 ctgctgaaga agaaggatat cccggttggc tgctctagag ccggcaggac ggatgctgag
      121 gttgttcgca gcagcgtgtg tgcagagccc ccgggatctt gcagactggg atttgtcagg
      181 gcaggtgtgg acattgaggc tatttctaac aagctcctac atgatgctgg ggctgcagag
      241 ccccaggccg cacttggagt atgagagttt ccagggagtg aagccccggc tgctctctct
      301 gccccctctc catctgcccc cttggtgagg acaggactcc catccctcca gtgcacagtg
      361 agtgctgctc aatctgcatc cacagcggtt tgtgcccaga tgcttcctca ctcagcgtct
      421 aacagggtgc agtggaaacg tcattgactg gaggaaaggt gtgtgggtga gaaacgtttg
      481 tcggttgagt aactttttgt cgttattttt tttctctgat gttcaaagta agagaaaact
      541 catctggccc ctggagccca tgtgagcaac atgaaagata aagggaaaag ggggctgata
      601 aagatgtgga ccgagtctga gataagattc tttgcccctt gggtgaatcc aggtgaagtt
      661 gttttactct tagcccagcc agccctggat aaaggcttca tccctcttct ctttcctgtc
      721 catatatcag aactttaagc tctacttgta cccagtgaag attagagtaa cgcacggagc
      781 acaggaaatg tctaggtgat gatttcactg agctttacaa gggacacagg tacgcatcaa
      841 gagcttgggc cactgggtaa agagggatct ggttcttgct ctaacgcggt tcatcttctg
      901 tgtcagaatc agatagattc tgggtgtcag agacccgaat cttcctcttg ttccgctcac
      961 tgcccccctg tggccccttc tcatttcaaa tgctctgtct gtaatatgaa atgaaaagaa
     1021 ttcctcccag acccctagcc aaatttcacg gtacctgagc tcgctagcct cgaggatatc
     1081 aagatctggc ctcggcggcc aagcttagac actagagggt atataatgga agctcgactt
     1141 ccagcttggc aatccggtac tgttggtaaa gccaccatgg aagatgccaa aaacattaag
     1201 aagggcccag cgccattcta cccactcgaa gacgggaccg ccggcgagca gctgcacaaa
     1261 gccatgaagc gctacgccct ggtgcccggc accatcgcct ttaccgacgc acatatcgag
     1321 gtggacatta cctacgccga gtacttcgag atgagcgttc ggctggcaga agctatgaag
     1381 cgctatgggc tgaatacaaa ccatcggatc gtggtgtgca gcgagaatag cttgcagttc
     1441 ttcatgcccg tgttgggtgc cctgttcatc ggtgtggctg tggccccagc taacgacatc
     1501 tacaacgagc gcgagctgct gaacagcatg ggcatcagcc agcccaccgt cgtattcgtg
     1561 agcaagaaag ggctgcaaaa gatcctcaac gtgcaaaaga agctaccgat catacaaaag
     1621 atcatcatca tggatagcaa gaccgactac cagggcttcc aaagcatgta caccttcgtg
     1681 acttcccatt tgccacccgg cttcaacgag tacgacttcg tgcccgagag cttcgaccgg
     1741 gacaaaacca tcgccctgat catgaacagt agtggcagta ccggattgcc caagggcgta
     1801 gccctaccgc accgcaccgc ttgtgtccga ttcagtcatg cccgcgaccc catcttcggc
     1861 aaccagatca tccccgacac cgctatcctc agcgtggtgc catttcacca cggcttcggc
     1921 atgttcacca cgctgggcta cttgatctgc ggctttcggg tcgtgctcat gtaccgcttc
     1981 gaggaggagc tattcttgcg cagcttgcaa gactataaga ttcaatctgc cctgctggtg
     2041 cccacactat ttagcttctt cgctaagagc actctcatcg acaagtacga cctaagcaac
     2101 ttgcacgaga tcgccagcgg cggggcgccg ctcagcaagg aggtaggtga ggccgtggcc
     2161 aaacgcttcc acctaccagg catccgccag ggctacggcc tgacagaaac aaccagcgcc
     2221 attctgatca cccccgaagg ggacgacaag cctggcgcag taggcaaggt ggtgcccttc
     2281 ttcgaggcta aggtggtgga cttggacacc ggtaagacac tgggtgtgaa ccagcgcggc
     2341 gagctgtgcg tccgtggccc catgatcatg agcggctacg ttaacaaccc cgaggctaca
     2401 aacgctctca tcgacaagga cggctggctg cacagcggcg acatcgccta ctgggacgag
     2461 gacgagcact tcttcatcgt ggaccggctg aagagcctga tcaaatacaa gggctaccag
     2521 gtagccccag ccgaactgga gagcatcctg ctgcaacacc ccaacatctt cgacgccggg
     2581 gtcgccggcc tgcccgacga cgatgccggc gagctgcccg ccgcagtcgt cgtgctggaa
     2641 cacggtaaaa ccatgaccga gaaggagatc gtggactatg tggccagcca ggttacaacc
     2701 gccaagaagc tgcgcggtgg tgttgtgttc gtggacgagg tgcctaaagg actgaccggc
     2761 aagttggacg cccgcaagat ccgcgagatt ctcattaagg ccaagaaggg cggcaagatc
     2821 gccgtgtaat aattctagag tcggggcggc cggccgcttc gagcagacat gataagatac
     2881 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     2941 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     3001 aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc
     3061 aagtaaaacc tctacaaatg tggtaaaatc gataaggatc cgtcgaccga tgcccttgag
     3121 agccttcaac ccagtcagct ccttccggtg ggcgcggggc atgactatcg tcgccgcact
     3181 tatgactgtc ttctttatca tgcaactcgt aggacaggtg ccggcagcgc tcttccgctt
     3241 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     3301 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     3361 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     3421 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     3481 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     3541 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     3601 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     3661 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     3721 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     3781 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     3841 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     3901 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     3961 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     4021 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     4081 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     4141 aaagtatata tgagtaaact tggtctgaca gcggccgcaa atgctaaacc actgcagtgg
     4201 ttaccagtgc ttgatcagtg aggcaccgat ctcagcgatc tgcctatttc gttcgtccat
     4261 agtggcctga ctccccgtcg tgtagatcac tacgattcgt gagggcttac catcaggccc
     4321 cagcgcagca atgatgccgc gagagccgcg ttcaccggcc cccgatttgt cagcaatgaa
     4381 ccagccagca gggagggccg agcgaagaag tggtcctgct actttgtccg cctccatcca
     4441 gtctatgagc tgctgtcgtg atgctagagt aagaagttcg ccagtgagta gtttccgaag
     4501 agttgtggcc attgctactg gcatcgtggt atcacgctcg tcgttcggta tggcttcgtt
     4561 caactctggt tcccagcggt caagccgggt cacatgatca cccatattat gaagaaatgc
     4621 agtcagctcc ttagggcctc cgatcgttgt cagaagtaag ttggccgcgg tgttgtcgct
     4681 catggtaatg gcagcactac acaattctct taccgtcatg ccatccgtaa gatgcttttc
     4741 cgtgaccggc gagtactcaa ccaagtcgtt ttgtgagtag tgtatacggc gaccaagctg
     4801 ctcttgcccg gcgtctatac gggacaacac cgcgccacat agcagtactt tgaaagtgct
     4861 catcatcggg aatcgttctt cggggcggaa agactcaagg atcttgccgc tattgagatc
     4921 cagttcgata tagcccactc ttgcacccag ttgatcttca gcatctttta ctttcaccag
     4981 cgtttcgggg tgtgcaaaaa caggcaagca aaatgccgca aagaagggaa tgagtgcgac
     5041 acgaaaatgt tggatgctca tactcgtcct ttttcaatat tattgaagca tttatcaggg
     5101 ttactagtac gtctctcaag gataagtaag taatattaag gtacgggagg tattggacag
     5161 gccgcaataa aatatcttta ttttcattac atctgtgtgt tggttttttg tgtgaatcga
     5221 tagtactaac atacgctctc catcaaaaca aaacgaaaca aaacaaacta gcaaaatagg
     5281 ctgtccccag tgcaagtgca ggtgccagaa catttctct
//
