LOCUS       sequence-176520-        4690 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Luciferase reporter for CD69 enhancer (IGI-P0619) .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4690)
  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 4690)
  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-CD69 CaRE1 (minus strand)
FEATURES             Location/Qualifiers
     source          1..4690
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        484..515
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      542..551
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             548..2200
                     /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    2244..2365
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2255..2274)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2309..2328
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2613..2630)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2784..3372)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2864..2883)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3572..4363,4364..4432))
                     /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    4537..4585
                     /note="synthetic polyadenylation signal"
     misc_feature    4599..4690
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4639..4658
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gccggtacca taagtgtttg actaatgatc aaatggaaaa tagaatctgg
       61 tgaaggtgtc caaacgcaga aaggaaggtg tttacaggga ggacaatatg agagcccagc
      121 actttacctc ctgaccacac agtagcaggt gtttccttaa ggcctcacac caccactaga
      181 gagcaacagt tcacaagcga actgaaggtc tacgtgcaag agcccttgca ggaatgcaga
      241 agaacaggaa ggctgtgagg tgtaggcctt gtgtcagaaa ttggtctgag gagctcattt
      301 actccaaggc agttatttga aacacaaaat caagtgatgt gctttaattc gctggccctg
      361 tgccctctat tatttaaccc gattctttct tcgagtcata caaaacaatt actctaatta
      421 ggtacctgag ctcgctagcc tcgaggatat caagatctgg cctcggcggc caagcttaga
      481 cactagaggg tatataatgg aagctcgact tccagcttgg caatccggta ctgttggtaa
      541 agccaccatg gaagatgcca aaaacattaa gaagggccca gcgccattct acccactcga
      601 agacgggacc gccggcgagc agctgcacaa agccatgaag cgctacgccc tggtgcccgg
      661 caccatcgcc tttaccgacg cacatatcga ggtggacatt acctacgccg agtacttcga
      721 gatgagcgtt cggctggcag aagctatgaa gcgctatggg ctgaatacaa accatcggat
      781 cgtggtgtgc agcgagaata gcttgcagtt cttcatgccc gtgttgggtg ccctgttcat
      841 cggtgtggct gtggccccag ctaacgacat ctacaacgag cgcgagctgc tgaacagcat
      901 gggcatcagc cagcccaccg tcgtattcgt gagcaagaaa gggctgcaaa agatcctcaa
      961 cgtgcaaaag aagctaccga tcatacaaaa gatcatcatc atggatagca agaccgacta
     1021 ccagggcttc caaagcatgt acaccttcgt gacttcccat ttgccacccg gcttcaacga
     1081 gtacgacttc gtgcccgaga gcttcgaccg ggacaaaacc atcgccctga tcatgaacag
     1141 tagtggcagt accggattgc ccaagggcgt agccctaccg caccgcaccg cttgtgtccg
     1201 attcagtcat gcccgcgacc ccatcttcgg caaccagatc atccccgaca ccgctatcct
     1261 cagcgtggtg ccatttcacc acggcttcgg catgttcacc acgctgggct acttgatctg
     1321 cggctttcgg gtcgtgctca tgtaccgctt cgaggaggag ctattcttgc gcagcttgca
     1381 agactataag attcaatctg ccctgctggt gcccacacta tttagcttct tcgctaagag
     1441 cactctcatc gacaagtacg acctaagcaa cttgcacgag atcgccagcg gcggggcgcc
     1501 gctcagcaag gaggtaggtg aggccgtggc caaacgcttc cacctaccag gcatccgcca
     1561 gggctacggc ctgacagaaa caaccagcgc cattctgatc acccccgaag gggacgacaa
     1621 gcctggcgca gtaggcaagg tggtgccctt cttcgaggct aaggtggtgg acttggacac
     1681 cggtaagaca ctgggtgtga accagcgcgg cgagctgtgc gtccgtggcc ccatgatcat
     1741 gagcggctac gttaacaacc ccgaggctac aaacgctctc atcgacaagg acggctggct
     1801 gcacagcggc gacatcgcct actgggacga ggacgagcac ttcttcatcg tggaccggct
     1861 gaagagcctg atcaaataca agggctacca ggtagcccca gccgaactgg agagcatcct
     1921 gctgcaacac cccaacatct tcgacgccgg ggtcgccggc ctgcccgacg acgatgccgg
     1981 cgagctgccc gccgcagtcg tcgtgctgga acacggtaaa accatgaccg agaaggagat
     2041 cgtggactat gtggccagcc aggttacaac cgccaagaag ctgcgcggtg gtgttgtgtt
     2101 cgtggacgag gtgcctaaag gactgaccgg caagttggac gcccgcaaga tccgcgagat
     2161 tctcattaag gccaagaagg gcggcaagat cgccgtgtaa taattctaga gtcggggcgg
     2221 ccggccgctt cgagcagaca tgataagata cattgatgag tttggacaaa ccacaactag
     2281 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     2341 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     2401 tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtaaaat
     2461 cgataaggat ccgtcgaccg atgcccttga gagccttcaa cccagtcagc tccttccggt
     2521 gggcgcgggg catgactatc gtcgccgcac ttatgactgt cttctttatc atgcaactcg
     2581 taggacaggt gccggcagcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     2641 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     2701 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     2761 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     2821 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     2881 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     2941 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     3001 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     3061 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     3121 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     3181 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     3241 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     3301 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     3361 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     3421 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     3481 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     3541 agcggccgca aatgctaaac cactgcagtg gttaccagtg cttgatcagt gaggcaccga
     3601 tctcagcgat ctgcctattt cgttcgtcca tagtggcctg actccccgtc gtgtagatca
     3661 ctacgattcg tgagggctta ccatcaggcc ccagcgcagc aatgatgccg cgagagccgc
     3721 gttcaccggc ccccgatttg tcagcaatga accagccagc agggagggcc gagcgaagaa
     3781 gtggtcctgc tactttgtcc gcctccatcc agtctatgag ctgctgtcgt gatgctagag
     3841 taagaagttc gccagtgagt agtttccgaa gagttgtggc cattgctact ggcatcgtgg
     3901 tatcacgctc gtcgttcggt atggcttcgt tcaactctgg ttcccagcgg tcaagccggg
     3961 tcacatgatc acccatatta tgaagaaatg cagtcagctc cttagggcct ccgatcgttg
     4021 tcagaagtaa gttggccgcg gtgttgtcgc tcatggtaat ggcagcacta cacaattctc
     4081 ttaccgtcat gccatccgta agatgctttt ccgtgaccgg cgagtactca accaagtcgt
     4141 tttgtgagta gtgtatacgg cgaccaagct gctcttgccc ggcgtctata cgggacaaca
     4201 ccgcgccaca tagcagtact ttgaaagtgc tcatcatcgg gaatcgttct tcggggcgga
     4261 aagactcaag gatcttgccg ctattgagat ccagttcgat atagcccact cttgcaccca
     4321 gttgatcttc agcatctttt actttcacca gcgtttcggg gtgtgcaaaa acaggcaagc
     4381 aaaatgccgc aaagaaggga atgagtgcga cacgaaaatg ttggatgctc atactcgtcc
     4441 tttttcaata ttattgaagc atttatcagg gttactagta cgtctctcaa ggataagtaa
     4501 gtaatattaa ggtacgggag gtattggaca ggccgcaata aaatatcttt attttcatta
     4561 catctgtgtg ttggtttttt gtgtgaatcg atagtactaa catacgctct ccatcaaaac
     4621 aaaacgaaac aaaacaaact agcaaaatag gctgtcccca gtgcaagtgc aggtgccaga
     4681 acatttctct
//
