LOCUS       sequence-304890-        5447 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ZC3H12D promoter region cloned upstream of luciferase reporter gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5447)
  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 5447)
  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_PromoterWT
FEATURES             Location/Qualifiers
     source          1..5447
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    26..74
                     /note="synthetic polyadenylation signal"
     misc_feature    88..179
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     128..147
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     regulatory      897..906
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             903..2555
                     /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(969..987)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    2596..2717
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2607..2626)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2661..2680
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2965..2982)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3136..3724)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3216..3235)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3895..4686,4687..4755))
                     /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     4518..4537
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4756..4860)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4887..5342
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4974..4993)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5184..5205
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 tacgggaggt acttggagcg gccgcaataa aatatcttta ttttcattac atctgtgtgt
       61 tggttttttg tgtgaatcga tagtactaac atacgctctc catcaaaaca aaacgaaaca
      121 aaacaaacta gcaaaatagg ctgtccccag tgcaagtgca ggtgccagaa catttctcta
      181 tcgataggta ccgagctctt acgcgtgcta gcctgatcat cttctcagct ggaggcagga
      241 gaacttctgt tccaagttcc cacttggaac aggggttctc ccccttggac tctgaatgct
      301 gttctcctca tggagcctct cctggctttg ctgtgcagcc aatcccagcc gtcctcacgg
      361 agacttgctg ccctcacgga gggtttgttt tgcatgtggt tgctgaactt tgattcggtg
      421 actcatgctg aggtgcaaac tttaagtccc ccccgcccct ttaattgctt tcttcctttc
      481 cttttttttt tcttaaaaaa aaaaaaacca aaaaaaccca aagcataact acttttgcag
      541 ctgaacgtga ctgtggcgtg caggaagtgg agcattggca tgaagtggct cctagtggct
      601 gcttggcgca cgccagctgc cctcctctga ctccagtggc acctgggggc ctggcctcac
      661 tgacgggaga acattggcgt gaaggctgct ggcgactggg ccagcattca ttgtgaagac
      721 cggagggaca caccctgctg gtgagtgcat gtcccctggc tgtcagtcag tgtggccaac
      781 accacgcaga cttgagtcca ggggtggagg agagagactg gaaccccggg gcagaggcag
      841 gaggagctcg agatctgcga tctaagtaag cttggcattc cggtactgtt ggtaaagcca
      901 ccatggaaga cgccaaaaac ataaagaaag gcccggcgcc attctatccg ctggaagatg
      961 gaaccgctgg agagcaactg cataaggcta tgaagagata cgccctggtt cctggaacaa
     1021 ttgcttttac agatgcacat atcgaggtgg acatcactta cgctgagtac ttcgaaatgt
     1081 ccgttcggtt ggcagaagct atgaaacgat atgggctgaa tacaaatcac agaatcgtcg
     1141 tatgcagtga aaactctctt caattcttta tgccggtgtt gggcgcgtta tttatcggag
     1201 ttgcagttgc gcccgcgaac gacatttata atgaacgtga attgctcaac agtatgggca
     1261 tttcgcagcc taccgtggtg ttcgtttcca aaaaggggtt gcaaaaaatt ttgaacgtgc
     1321 aaaaaaagct cccaatcatc caaaaaatta ttatcatgga ttctaaaacg gattaccagg
     1381 gatttcagtc gatgtacacg ttcgtcacat ctcatctacc tcccggtttt aatgaatacg
     1441 attttgtgcc agagtccttc gatagggaca agacaattgc actgatcatg aactcctctg
     1501 gatctactgg tctgcctaaa ggtgtcgctc tgcctcatag aactgcctgc gtgagattct
     1561 cgcatgccag agatcctatt tttggcaatc aaatcattcc ggatactgcg attttaagtg
     1621 ttgttccatt ccatcacggt tttggaatgt ttactacact cggatatttg atatgtggat
     1681 ttcgagtcgt cttaatgtat agatttgaag aagagctgtt tctgaggagc cttcaggatt
     1741 acaagattca aagtgcgctg ctggtgccaa ccctattctc cttcttcgcc aaaagcactc
     1801 tgattgacaa atacgattta tctaatttac acgaaattgc ttctggtggc gctcccctct
     1861 ctaaggaagt cggggaagcg gttgccaaga ggttccatct gccaggtatc aggcaaggat
     1921 atgggctcac tgagactaca tcagctattc tgattacacc cgagggggat gataaaccgg
     1981 gcgcggtcgg taaagttgtt ccattttttg aagcgaaggt tgtggatctg gataccggga
     2041 aaacgctggg cgttaatcaa agaggcgaac tgtgtgtgag aggtcctatg attatgtccg
     2101 gttatgtaaa caatccggaa gcgaccaacg ccttgattga caaggatgga tggctacatt
     2161 ctggagacat agcttactgg gacgaagacg aacacttctt catcgttgac cgcctgaagt
     2221 ctctgattaa gtacaaaggc tatcaggtgg ctcccgctga attggaatcc atcttgctcc
     2281 aacaccccaa catcttcgac gcaggtgtcg caggtcttcc cgacgatgac gccggtgaac
     2341 ttcccgccgc cgttgttgtt ttggagcacg gaaagacgat gacggaaaaa gagatcgtgg
     2401 attacgtcgc cagtcaagta acaaccgcga aaaagttgcg cggaggagtt gtgtttgtgg
     2461 acgaagtacc gaaaggtctt accggaaaac tcgacgcaag aaaaatcaga gagatcctca
     2521 taaaggccaa gaagggcgga aagatcgccg tgtaattcta gagtcggggc ggccggccgc
     2581 ttcgagcaga catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt
     2641 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
     2701 gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg
     2761 aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtaaa atcgataagg
     2821 atccgtcgac cgatgccctt gagagccttc aacccagtca gctccttccg gtgggcgcgg
     2881 ggcatgacta tcgtcgccgc acttatgact gtcttcttta tcatgcaact cgtaggacag
     2941 gtgccggcag cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     3001 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     3061 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     3121 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     3181 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     3241 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     3301 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     3361 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     3421 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     3481 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     3541 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     3601 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     3661 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     3721 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     3781 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     3841 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     3901 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     3961 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     4021 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     4081 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     4141 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     4201 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     4261 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     4321 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     4381 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     4441 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     4501 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     4561 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     4621 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     4681 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     4741 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     4801 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     4861 cacatttccc cgaaaagtgc cacctgacgc gccctgtagc ggcgcattaa gcgcggcggg
     4921 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     4981 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     5041 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     5101 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     5161 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     5221 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     5281 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat
     5341 ttgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
     5401 gctattacgc cagcccaagc taccatgata agtaagtaat attaagg
//
