LOCUS       sequence-304643-        5448 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ZC3H12D promoter region carrying mutations in BHLHE40 binding sites,
            cloned upstream of luciferase reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5448)
  AUTHORS   Emming S, Bianchi N, Polletti S, Balestrieri C, Leoni C, Montagner 
            S, Chirichella M, Delaleu N, Natoli G, Monticelli S
  TITLE     A molecular network regulating the proinflammatory phenotype of 
            human memory T lymphocytes.
  JOURNAL   Nat Immunol. 2020 Apr;21(4):388-399. doi: 10.1038/s41590-020-0622-8.
            Epub 2020 Mar 16.
  PUBMED    32205878
REFERENCE   2  (bases 1 to 5448)
  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/s41590-020-0622-8"; journalName: "Nat Immunol"; date: 
            "2020-04"; volume: "21"; issue: "4"; pages: "388-399"
COMMENT     Sequence Label: pGL3-Basic-ZC3H12D_PromoterMUT
FEATURES             Location/Qualifiers
     source          1..5448
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      526..535
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             532..2184
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(598..616)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    2225..2346
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2236..2255)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2290..2309
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2594..2611)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2765..3353)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2845..2864)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3524..4315,4316..4384))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4147..4166
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4385..4489)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4516..4971
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4603..4622)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4813..4834
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5102..5150
                     /note="synthetic polyadenylation signal"
     misc_feature    5164..5255
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5204..5223
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ctcacggagg gtttgttttg gatccggttg ctgaactttg attcggtgac tcatgctgag
       61 gtgcaaactt taagtccccc ccgccccttt aattgctttc ttcctttcct tttttttttc
      121 ttaaaaaaaa aaaaaaccaa aaaaacccaa agcataacta cttttgcagc tgaattcgac
      181 tgtggcgtgc aggaagtgga gcattggcat gaagtggctc ctagtggctg cttggcgcac
      241 gccagctgcc ctcctctgac tccagtggga tatcggggcc tggcctcact gacgggagaa
      301 cattggcgtg aaggctgctg gcgactgggc cagcattcat tgtgaagacc ggagggacac
      361 accctgctgg tgagtgcatg tcccctggct gtcagtcagt gtggccaaca ccacgcagac
      421 ttgagtccag gggtggagga gagagactgg aaccccgggg cagaggcagg aggagctcga
      481 gatctgcgat ctaagtaagc ttggcattcc ggtactgttg gtaaagccac catggaagac
      541 gccaaaaaca taaagaaagg cccggcgcca ttctatccgc tggaagatgg aaccgctgga
      601 gagcaactgc ataaggctat gaagagatac gccctggttc ctggaacaat tgcttttaca
      661 gatgcacata tcgaggtgga catcacttac gctgagtact tcgaaatgtc cgttcggttg
      721 gcagaagcta tgaaacgata tgggctgaat acaaatcaca gaatcgtcgt atgcagtgaa
      781 aactctcttc aattctttat gccggtgttg ggcgcgttat ttatcggagt tgcagttgcg
      841 cccgcgaacg acatttataa tgaacgtgaa ttgctcaaca gtatgggcat ttcgcagcct
      901 accgtggtgt tcgtttccaa aaaggggttg caaaaaattt tgaacgtgca aaaaaagctc
      961 ccaatcatcc aaaaaattat tatcatggat tctaaaacgg attaccaggg atttcagtcg
     1021 atgtacacgt tcgtcacatc tcatctacct cccggtttta atgaatacga ttttgtgcca
     1081 gagtccttcg atagggacaa gacaattgca ctgatcatga actcctctgg atctactggt
     1141 ctgcctaaag gtgtcgctct gcctcataga actgcctgcg tgagattctc gcatgccaga
     1201 gatcctattt ttggcaatca aatcattccg gatactgcga ttttaagtgt tgttccattc
     1261 catcacggtt ttggaatgtt tactacactc ggatatttga tatgtggatt tcgagtcgtc
     1321 ttaatgtata gatttgaaga agagctgttt ctgaggagcc ttcaggatta caagattcaa
     1381 agtgcgctgc tggtgccaac cctattctcc ttcttcgcca aaagcactct gattgacaaa
     1441 tacgatttat ctaatttaca cgaaattgct tctggtggcg ctcccctctc taaggaagtc
     1501 ggggaagcgg ttgccaagag gttccatctg ccaggtatca ggcaaggata tgggctcact
     1561 gagactacat cagctattct gattacaccc gagggggatg ataaaccggg cgcggtcggt
     1621 aaagttgttc cattttttga agcgaaggtt gtggatctgg ataccgggaa aacgctgggc
     1681 gttaatcaaa gaggcgaact gtgtgtgaga ggtcctatga ttatgtccgg ttatgtaaac
     1741 aatccggaag cgaccaacgc cttgattgac aaggatggat ggctacattc tggagacata
     1801 gcttactggg acgaagacga acacttcttc atcgttgacc gcctgaagtc tctgattaag
     1861 tacaaaggct atcaggtggc tcccgctgaa ttggaatcca tcttgctcca acaccccaac
     1921 atcttcgacg caggtgtcgc aggtcttccc gacgatgacg ccggtgaact tcccgccgcc
     1981 gttgttgttt tggagcacgg aaagacgatg acggaaaaag agatcgtgga ttacgtcgcc
     2041 agtcaagtaa caaccgcgaa aaagttgcgc ggaggagttg tgtttgtgga cgaagtaccg
     2101 aaaggtctta ccggaaaact cgacgcaaga aaaatcagag agatcctcat aaaggccaag
     2161 aagggcggaa agatcgccgt gtaattctag agtcggggcg gccggccgct tcgagcagac
     2221 atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc
     2281 tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa
     2341 caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag
     2401 gttttttaaa gcaagtaaaa cctctacaaa tgtggtaaaa tcgataagga tccgtcgacc
     2461 gatgcccttg agagccttca acccagtcag ctccttccgg tgggcgcggg gcatgactat
     2521 cgtcgccgca cttatgactg tcttctttat catgcaactc gtaggacagg tgccggcagc
     2581 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     2641 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     2701 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     2761 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     2821 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     2881 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     2941 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     3001 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     3061 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     3121 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     3181 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     3241 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     3301 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     3361 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     3421 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     3481 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     3541 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     3601 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     3661 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     3721 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     3781 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     3841 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     3901 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     3961 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     4021 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     4081 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     4141 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     4201 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     4261 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     4321 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     4381 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     4441 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     4501 gaaaagtgcc acctgacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     4561 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     4621 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     4681 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     4741 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     4801 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     4861 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     4921 tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt tgccattcgc
     4981 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
     5041 agcccaagct accatgataa gtaagtaata ttaaggtacg ggaggtactt ggagcggccg
     5101 caataaaata tctttatttt cattacatct gtgtgttggt tttttgtgtg aatcgatagt
     5161 actaacatac gctctccatc aaaacaaaac gaaacaaaac aaactagcaa aataggctgt
     5221 ccccagtgca agtgcaggtg ccagaacatt tctctatcga taggtaccga gctcttacgc
     5281 gtgctagcct gatcatcttc tcagctggag gcaggagaac ttctgttcca agttcccact
     5341 tggaacaggg gttctccccc ttggactctg aatgctgttc tcctcatgga gcctctcctg
     5401 gctttgctgt gcagccaatc ccagccgtcc tcacggagac ttgctgcc
//
