LOCUS       sequence-176523-        4849 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Luciferase reporter for CD69 enhancer (IGI-P0622) .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4849)
  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 4849)
  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 CaRE3 (minus strand)
FEATURES             Location/Qualifiers
     source          1..4849
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        643..674
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      701..710
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             707..2359
                     /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    2403..2524
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2414..2433)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2468..2487
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2772..2789)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2943..3531)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3023..3042)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3731..4522,4523..4591))
                     /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    4696..4744
                     /note="synthetic polyadenylation signal"
     misc_feature    4758..4849
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4798..4817
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gccggtacct gagagaggat gatgtattct aaattgaagt tttgattcac
       61 aagaagatta aaagccattc agaaacctaa ttcacccact gaaaggaaaa aaaaaaaaga
      121 gagatgagca gtttgtctcc ggaaattgtc ttaggtcgga agtctgtggt ccctgttcac
      181 atgtacccaa aagcatcctg ctgctgcagc tgtctgataa gcacagagta ccccaccttc
      241 tctgcacact ttgcatctag ctcatattac ctcatcttta cttcctttct gacgtctcac
      301 cctggattct acatataagg tcacacagga aggaaagctg cattgagttt tggtgtcctg
      361 aaagactttt gccaaccttg tccccgcact aatttctcta agcctcggct atactatttt
      421 ctcagctaca cgatgaaatg tgaatgataa tttctgccct aaaaatatca cttaattttt
      481 taacatatca tttatgaaag aagacacata aaatgtctcc ctaaaatgga aagttacata
      541 ttattgccat ctgtgtttta taaagaggtt gaaaggggtg gtacctgagc tcgctagcct
      601 cgaggatatc aagatctggc ctcggcggcc aagcttagac actagagggt atataatgga
      661 agctcgactt ccagcttggc aatccggtac tgttggtaaa gccaccatgg aagatgccaa
      721 aaacattaag aagggcccag cgccattcta cccactcgaa gacgggaccg ccggcgagca
      781 gctgcacaaa gccatgaagc gctacgccct ggtgcccggc accatcgcct ttaccgacgc
      841 acatatcgag gtggacatta cctacgccga gtacttcgag atgagcgttc ggctggcaga
      901 agctatgaag cgctatgggc tgaatacaaa ccatcggatc gtggtgtgca gcgagaatag
      961 cttgcagttc ttcatgcccg tgttgggtgc cctgttcatc ggtgtggctg tggccccagc
     1021 taacgacatc tacaacgagc gcgagctgct gaacagcatg ggcatcagcc agcccaccgt
     1081 cgtattcgtg agcaagaaag ggctgcaaaa gatcctcaac gtgcaaaaga agctaccgat
     1141 catacaaaag atcatcatca tggatagcaa gaccgactac cagggcttcc aaagcatgta
     1201 caccttcgtg acttcccatt tgccacccgg cttcaacgag tacgacttcg tgcccgagag
     1261 cttcgaccgg gacaaaacca tcgccctgat catgaacagt agtggcagta ccggattgcc
     1321 caagggcgta gccctaccgc accgcaccgc ttgtgtccga ttcagtcatg cccgcgaccc
     1381 catcttcggc aaccagatca tccccgacac cgctatcctc agcgtggtgc catttcacca
     1441 cggcttcggc atgttcacca cgctgggcta cttgatctgc ggctttcggg tcgtgctcat
     1501 gtaccgcttc gaggaggagc tattcttgcg cagcttgcaa gactataaga ttcaatctgc
     1561 cctgctggtg cccacactat ttagcttctt cgctaagagc actctcatcg acaagtacga
     1621 cctaagcaac ttgcacgaga tcgccagcgg cggggcgccg ctcagcaagg aggtaggtga
     1681 ggccgtggcc aaacgcttcc acctaccagg catccgccag ggctacggcc tgacagaaac
     1741 aaccagcgcc attctgatca cccccgaagg ggacgacaag cctggcgcag taggcaaggt
     1801 ggtgcccttc ttcgaggcta aggtggtgga cttggacacc ggtaagacac tgggtgtgaa
     1861 ccagcgcggc gagctgtgcg tccgtggccc catgatcatg agcggctacg ttaacaaccc
     1921 cgaggctaca aacgctctca tcgacaagga cggctggctg cacagcggcg acatcgccta
     1981 ctgggacgag gacgagcact tcttcatcgt ggaccggctg aagagcctga tcaaatacaa
     2041 gggctaccag gtagccccag ccgaactgga gagcatcctg ctgcaacacc ccaacatctt
     2101 cgacgccggg gtcgccggcc tgcccgacga cgatgccggc gagctgcccg ccgcagtcgt
     2161 cgtgctggaa cacggtaaaa ccatgaccga gaaggagatc gtggactatg tggccagcca
     2221 ggttacaacc gccaagaagc tgcgcggtgg tgttgtgttc gtggacgagg tgcctaaagg
     2281 actgaccggc aagttggacg cccgcaagat ccgcgagatt ctcattaagg ccaagaaggg
     2341 cggcaagatc gccgtgtaat aattctagag tcggggcggc cggccgcttc gagcagacat
     2401 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     2461 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     2521 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt
     2581 tttttaaagc aagtaaaacc tctacaaatg tggtaaaatc gataaggatc cgtcgaccga
     2641 tgcccttgag agccttcaac ccagtcagct ccttccggtg ggcgcggggc atgactatcg
     2701 tcgccgcact tatgactgtc ttctttatca tgcaactcgt aggacaggtg ccggcagcgc
     2761 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     2821 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     2881 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     2941 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     3001 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     3061 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     3121 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     3181 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     3241 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     3301 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     3361 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     3421 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     3481 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     3541 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     3601 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     3661 aaatcaatct aaagtatata tgagtaaact tggtctgaca gcggccgcaa atgctaaacc
     3721 actgcagtgg ttaccagtgc ttgatcagtg aggcaccgat ctcagcgatc tgcctatttc
     3781 gttcgtccat agtggcctga ctccccgtcg tgtagatcac tacgattcgt gagggcttac
     3841 catcaggccc cagcgcagca atgatgccgc gagagccgcg ttcaccggcc cccgatttgt
     3901 cagcaatgaa ccagccagca gggagggccg agcgaagaag tggtcctgct actttgtccg
     3961 cctccatcca gtctatgagc tgctgtcgtg atgctagagt aagaagttcg ccagtgagta
     4021 gtttccgaag agttgtggcc attgctactg gcatcgtggt atcacgctcg tcgttcggta
     4081 tggcttcgtt caactctggt tcccagcggt caagccgggt cacatgatca cccatattat
     4141 gaagaaatgc agtcagctcc ttagggcctc cgatcgttgt cagaagtaag ttggccgcgg
     4201 tgttgtcgct catggtaatg gcagcactac acaattctct taccgtcatg ccatccgtaa
     4261 gatgcttttc cgtgaccggc gagtactcaa ccaagtcgtt ttgtgagtag tgtatacggc
     4321 gaccaagctg ctcttgcccg gcgtctatac gggacaacac cgcgccacat agcagtactt
     4381 tgaaagtgct catcatcggg aatcgttctt cggggcggaa agactcaagg atcttgccgc
     4441 tattgagatc cagttcgata tagcccactc ttgcacccag ttgatcttca gcatctttta
     4501 ctttcaccag cgtttcgggg tgtgcaaaaa caggcaagca aaatgccgca aagaagggaa
     4561 tgagtgcgac acgaaaatgt tggatgctca tactcgtcct ttttcaatat tattgaagca
     4621 tttatcaggg ttactagtac gtctctcaag gataagtaag taatattaag gtacgggagg
     4681 tattggacag gccgcaataa aatatcttta ttttcattac atctgtgtgt tggttttttg
     4741 tgtgaatcga tagtactaac atacgctctc catcaaaaca aaacgaaaca aaacaaacta
     4801 gcaaaatagg ctgtccccag tgcaagtgca ggtgccagaa catttctct
//
