LOCUS       sequence-304889-        5693 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 5693)
  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 5693)
  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-Enhancer-ZC3H12D_PromoterMUT
FEATURES             Location/Qualifiers
     source          1..5693
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..92
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     41..60
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     regulatory      810..819
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             816..2468
                     /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(882..900)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    2509..2630
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2520..2539)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2574..2593
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2952..2972
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     primer_bind     complement(3124..3141)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3295..3883)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3375..3394)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4054..4845,4846..4914))
                     /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     4677..4696
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4915..5019)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5046..5501
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5133..5152)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5343..5364
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5632..5680
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 aacatacgct ctccatcaaa acaaaacgaa acaaaacaaa ctagcaaaat aggctgtccc
       61 cagtgcaagt gcaggtgcca gaacatttct ctatcgatag gtaccgagct cttacgcgtg
      121 ctagcctgat catcttctca gctggaggca ggagaacttc tgttccaagt tcccacttgg
      181 aacaggggtt ctcccccttg gactctgaat gctgttctcc tcatggagcc tctcctggct
      241 ttgctgtgca gccaatccca gccgtcctca cggagacttg ctgccctcac ggagggtttg
      301 ttttggatcc ggttgctgac tttgattcgg tgactcatgc tgaggtgcaa actttaagtc
      361 ccccccgccc ctttaattgc tttcttcctt tccttttttt tttttcttaa aaaaaaaaaa
      421 accaaaaaaa cccaaagcat aactactttt gcagctgaat tcgactgtgg cgtgcaggaa
      481 gtggagcatt ggcatgaagt ggctcctagt ggctgcttgg cgcacgccag ctgccctcct
      541 ctgactccag tgggatatcg gggcctggcc tcactgacgg gagaacattg gcgtgaaggc
      601 tgctggcgac tgggccagca ttcattgtga agaccggagg gacacaccct gctggtgagt
      661 gcatgtcccc tggctgtcag tcagtgtggc caacaccacg cagacttgag tccaggggtg
      721 gaggagagag actggaaccc cggggcagag gcaggaggag ctcgagatct ggatctaagt
      781 aagcttggca ttccggtact gttggtaaag ccaccatgga agacgccaaa aacataaaga
      841 aaggcccggc gccattctat ccgctggaag atggaaccgc tggagagcaa ctgcataagg
      901 ctatgaagag atacgccctg gttcctggaa caattgcttt tacagatgca catatcgagg
      961 tggacatcac ttacgctgag tacttcgaaa tgtccgttcg gttggcagaa gctatgaaac
     1021 gatatgggct gaatacaaat cacagaatcg tcgtatgcag tgaaaactct cttcaattct
     1081 ttatgccggt gttgggcgcg ttatttatcg gagttgcagt tgcgcccgcg aacgacattt
     1141 ataatgaacg tgaattgctc aacagtatgg gcatttcgca gcctaccgtg gtgttcgttt
     1201 ccaaaaaggg gttgcaaaaa attttgaacg tgcaaaaaaa gctcccaatc atccaaaaaa
     1261 ttattatcat ggattctaaa acggattacc agggatttca gtcgatgtac acgttcgtca
     1321 catctcatct acctcccggt tttaatgaat acgattttgt gccagagtcc ttcgataggg
     1381 acaagacaat tgcactgatc atgaactcct ctggatctac tggtctgcct aaaggtgtcg
     1441 ctctgcctca tagaactgcc tgcgtgagat tctcgcatgc cagagatcct atttttggca
     1501 atcaaatcat tccggatact gcgattttaa gtgttgttcc attccatcac ggttttggaa
     1561 tgtttactac actcggatat ttgatatgtg gatttcgagt cgtcttaatg tatagatttg
     1621 aagaagagct gtttctgagg agccttcagg attacaagat tcaaagtgcg ctgctggtgc
     1681 caaccctatt ctccttcttc gccaaaagca ctctgattga caaatacgat ttatctaatt
     1741 tacacgaaat tgcttctggt ggcgctcccc tctctaagga agtcggggaa gcggttgcca
     1801 agaggttcca tctgccaggt atcaggcaag gatatgggct cactgagact acatcagcta
     1861 ttctgattac acccgagggg gatgataaac cgggcgcggt cggtaaagtt gttccatttt
     1921 ttgaagcgaa ggttgtggat ctggataccg ggaaaacgct gggcgttaat caaagaggcg
     1981 aactgtgtgt gagaggtcct atgattatgt ccggttatgt aaacaatccg gaagcgacca
     2041 acgccttgat tgacaaggat ggatggctac attctggaga catagcttac tgggacgaag
     2101 acgaacactt cttcatcgtt gaccgcctga agtctctgat taagtacaaa ggctatcagg
     2161 tggctcccgc tgaattggaa tccatcttgc tccaacaccc caacatcttc gacgcaggtg
     2221 tcgcaggtct tcccgacgat gacgccggtg aacttcccgc cgccgttgtt gttttggagc
     2281 acggaaagac gatgacggaa aaagagatcg tggattacgt cgccagtcaa gtaacaaccg
     2341 cgaaaaagtt gcgcggagga gttgtgtttg tggacgaagt accgaaaggt cttaccggaa
     2401 aactcgacgc aagaaaaatc agagagatcc tcataaaggc caagaagggc ggaaagatcg
     2461 ccgtgtaatt ctagagtcgg ggcggccggc cgcttcgagc agacatgata agatacattg
     2521 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
     2581 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
     2641 attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt
     2701 aaaacctcta caaatgtggt aaaatcgata aggatctgaa cgatggagcg gagaatgggc
     2761 ggaactgggc ggagttaggg gcgggatggg cggagttagg ggcgggacta tggttgctga
     2821 ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg actttccaca
     2881 cctggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg gggagcctgg
     2941 ggactttcca caccctaact gacacacatt ccacagcgga tccgtcgacc gatgcccttg
     3001 agagccttca acccagtcag ctccttccgg tgggcgcggg gcatgactat cgtcgccgca
     3061 cttatgactg tcttctttat catgcaactc gtaggacagg tgccggcagc gctcttccgc
     3121 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     3181 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     3241 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     3301 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     3361 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     3421 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     3481 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     3541 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     3601 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     3661 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     3721 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     3781 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     3841 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     3901 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     3961 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     4021 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     4081 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     4141 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     4201 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     4261 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     4321 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     4381 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     4441 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     4501 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     4561 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     4621 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     4681 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     4741 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     4801 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     4861 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     4921 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     4981 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     5041 acctgacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     5101 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     5161 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     5221 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     5281 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     5341 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     5401 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     5461 atttaacgcg aattttaaca aaatattaac gcttacaatt tgccattcgc cattcaggct
     5521 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agcccaagct
     5581 accatgataa gtaagtaata ttaaggtacg ggaggtactt ggagcggccg caataaaata
     5641 tctttatttt cattacatct gtgtgttggt tttttgtgtg aatcgatagt act
//
