LOCUS       sequence-176521-        4522 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Luciferase reporter for CD69 enhancer (IGI-P0620) .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4522)
  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 4522)
  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 CaRE2 (minus strand)
FEATURES             Location/Qualifiers
     source          1..4522
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        316..347
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      374..383
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             380..2032
                     /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    2076..2197
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2087..2106)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2141..2160
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2445..2462)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2616..3204)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2696..2715)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3404..4195,4196..4264))
                     /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    4369..4417
                     /note="synthetic polyadenylation signal"
     misc_feature    4431..4522
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4471..4490
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gccggtacct atgcattagt agcagtcaat tttcaaatca cattccttta
       61 tgatttctag cctgggtctc tacaacttgc aaccagaata tatatctctt aggccttctc
      121 catttgaagc tgttgctact gatgtgtgct ggttctattt tgtttccttt ttatctgatt
      181 ctgctattcc ccacccactt ctctaaactt tacttcatag aaaacctgtc tatggtgttt
      241 ggacaatagc acggtacctg agctcgctag cctcgaggat atcaagatct ggcctcggcg
      301 gccaagctta gacactagag ggtatataat ggaagctcga cttccagctt ggcaatccgg
      361 tactgttggt aaagccacca tggaagatgc caaaaacatt aagaagggcc cagcgccatt
      421 ctacccactc gaagacggga ccgccggcga gcagctgcac aaagccatga agcgctacgc
      481 cctggtgccc ggcaccatcg cctttaccga cgcacatatc gaggtggaca ttacctacgc
      541 cgagtacttc gagatgagcg ttcggctggc agaagctatg aagcgctatg ggctgaatac
      601 aaaccatcgg atcgtggtgt gcagcgagaa tagcttgcag ttcttcatgc ccgtgttggg
      661 tgccctgttc atcggtgtgg ctgtggcccc agctaacgac atctacaacg agcgcgagct
      721 gctgaacagc atgggcatca gccagcccac cgtcgtattc gtgagcaaga aagggctgca
      781 aaagatcctc aacgtgcaaa agaagctacc gatcatacaa aagatcatca tcatggatag
      841 caagaccgac taccagggct tccaaagcat gtacaccttc gtgacttccc atttgccacc
      901 cggcttcaac gagtacgact tcgtgcccga gagcttcgac cgggacaaaa ccatcgccct
      961 gatcatgaac agtagtggca gtaccggatt gcccaagggc gtagccctac cgcaccgcac
     1021 cgcttgtgtc cgattcagtc atgcccgcga ccccatcttc ggcaaccaga tcatccccga
     1081 caccgctatc ctcagcgtgg tgccatttca ccacggcttc ggcatgttca ccacgctggg
     1141 ctacttgatc tgcggctttc gggtcgtgct catgtaccgc ttcgaggagg agctattctt
     1201 gcgcagcttg caagactata agattcaatc tgccctgctg gtgcccacac tatttagctt
     1261 cttcgctaag agcactctca tcgacaagta cgacctaagc aacttgcacg agatcgccag
     1321 cggcggggcg ccgctcagca aggaggtagg tgaggccgtg gccaaacgct tccacctacc
     1381 aggcatccgc cagggctacg gcctgacaga aacaaccagc gccattctga tcacccccga
     1441 aggggacgac aagcctggcg cagtaggcaa ggtggtgccc ttcttcgagg ctaaggtggt
     1501 ggacttggac accggtaaga cactgggtgt gaaccagcgc ggcgagctgt gcgtccgtgg
     1561 ccccatgatc atgagcggct acgttaacaa ccccgaggct acaaacgctc tcatcgacaa
     1621 ggacggctgg ctgcacagcg gcgacatcgc ctactgggac gaggacgagc acttcttcat
     1681 cgtggaccgg ctgaagagcc tgatcaaata caagggctac caggtagccc cagccgaact
     1741 ggagagcatc ctgctgcaac accccaacat cttcgacgcc ggggtcgccg gcctgcccga
     1801 cgacgatgcc ggcgagctgc ccgccgcagt cgtcgtgctg gaacacggta aaaccatgac
     1861 cgagaaggag atcgtggact atgtggccag ccaggttaca accgccaaga agctgcgcgg
     1921 tggtgttgtg ttcgtggacg aggtgcctaa aggactgacc ggcaagttgg acgcccgcaa
     1981 gatccgcgag attctcatta aggccaagaa gggcggcaag atcgccgtgt aataattcta
     2041 gagtcggggc ggccggccgc ttcgagcaga catgataaga tacattgatg agtttggaca
     2101 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     2161 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
     2221 tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa
     2281 atgtggtaaa atcgataagg atccgtcgac cgatgccctt gagagccttc aacccagtca
     2341 gctccttccg gtgggcgcgg ggcatgacta tcgtcgccgc acttatgact gtcttcttta
     2401 tcatgcaact cgtaggacag gtgccggcag cgctcttccg cttcctcgct cactgactcg
     2461 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     2521 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     2581 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     2641 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     2701 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     2761 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     2821 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     2881 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     2941 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     3001 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     3061 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     3121 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     3181 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     3241 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     3301 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     3361 acttggtctg acagcggccg caaatgctaa accactgcag tggttaccag tgcttgatca
     3421 gtgaggcacc gatctcagcg atctgcctat ttcgttcgtc catagtggcc tgactccccg
     3481 tcgtgtagat cactacgatt cgtgagggct taccatcagg ccccagcgca gcaatgatgc
     3541 cgcgagagcc gcgttcaccg gcccccgatt tgtcagcaat gaaccagcca gcagggaggg
     3601 ccgagcgaag aagtggtcct gctactttgt ccgcctccat ccagtctatg agctgctgtc
     3661 gtgatgctag agtaagaagt tcgccagtga gtagtttccg aagagttgtg gccattgcta
     3721 ctggcatcgt ggtatcacgc tcgtcgttcg gtatggcttc gttcaactct ggttcccagc
     3781 ggtcaagccg ggtcacatga tcacccatat tatgaagaaa tgcagtcagc tccttagggc
     3841 ctccgatcgt tgtcagaagt aagttggccg cggtgttgtc gctcatggta atggcagcac
     3901 tacacaattc tcttaccgtc atgccatccg taagatgctt ttccgtgacc ggcgagtact
     3961 caaccaagtc gttttgtgag tagtgtatac ggcgaccaag ctgctcttgc ccggcgtcta
     4021 tacgggacaa caccgcgcca catagcagta ctttgaaagt gctcatcatc gggaatcgtt
     4081 cttcggggcg gaaagactca aggatcttgc cgctattgag atccagttcg atatagccca
     4141 ctcttgcacc cagttgatct tcagcatctt ttactttcac cagcgtttcg gggtgtgcaa
     4201 aaacaggcaa gcaaaatgcc gcaaagaagg gaatgagtgc gacacgaaaa tgttggatgc
     4261 tcatactcgt cctttttcaa tattattgaa gcatttatca gggttactag tacgtctctc
     4321 aaggataagt aagtaatatt aaggtacggg aggtattgga caggccgcaa taaaatatct
     4381 ttattttcat tacatctgtg tgttggtttt ttgtgtgaat cgatagtact aacatacgct
     4441 ctccatcaaa acaaaacgaa acaaaacaaa ctagcaaaat aggctgtccc cagtgcaagt
     4501 gcaggtgcca gaacatttct ct
//
