LOCUS       sequence-304731-        5694 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 5694)
  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 5694)
  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_PromoterWT
FEATURES             Location/Qualifiers
     source          1..5694
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      690..699
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             696..2348
                     /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(762..780)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    2389..2510
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2400..2419)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2454..2473
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2832..2852
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     primer_bind     complement(3004..3021)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3175..3763)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3255..3274)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3934..4725,4726..4794))
                     /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     4557..4576
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4795..4899)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4926..5381
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5013..5032)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5223..5244
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5512..5560
                     /note="synthetic polyadenylation signal"
     misc_feature    5574..5665
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5614..5633
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 tagcctgatc atcttctcag ctggaggcag gagaacttct gttccaagtt cccacttgga
       61 acaggggttc tcccccttgg actctgaatg ctgttctcct catggagcct ctcctggctt
      121 tgctgtgcag ccaatcccag ccgtcctcac ggagacttgc tgccctcacg gagggtttgt
      181 tttgcatgtg gttgctgaac tttgattcgg tgactcatgc tgaggtgcaa actttaagtc
      241 ccccccgccc ctttaattgc tttcttcctt tccttttttt tttttcttaa aaaaaaaaaa
      301 accaaaaaaa cccaaagcat aactactttt gcagctgaac gtgactgtgg cgtgcaggaa
      361 gtggagcatt ggcatgaagt ggctcctagt ggctgcttgg cgcacgccag ctgccctcct
      421 ctgactccag tggcacctgg gggcctggcc tcactgacgg gagaacattg gcgtgaaggc
      481 tgctggcgac tgggccagca ttcattgtga agaccggagg gacacaccct gctggtgagt
      541 gcatgtcccc tggctgtcag tcagtgtggc caacaccacg cagacttgag tccaggggtg
      601 gaggagagag actggaaccc cggggcagag gcaggaggag ctcgagatct ggatctaagt
      661 aagcttggca ttccggtact gttggtaaag ccaccatgga agacgccaaa aacataaaga
      721 aaggcccggc gccattctat ccgctggaag atggaaccgc tggagagcaa ctgcataagg
      781 ctatgaagag atacgccctg gttcctggaa caattgcttt tacagatgca catatcgagg
      841 tggacatcac ttacgctgag tacttcgaaa tgtccgttcg gttggcagaa gctatgaaac
      901 gatatgggct gaatacaaat cacagaatcg tcgtatgcag tgaaaactct cttcaattct
      961 ttatgccggt gttgggcgcg ttatttatcg gagttgcagt tgcgcccgcg aacgacattt
     1021 ataatgaacg tgaattgctc aacagtatgg gcatttcgca gcctaccgtg gtgttcgttt
     1081 ccaaaaaggg gttgcaaaaa attttgaacg tgcaaaaaaa gctcccaatc atccaaaaaa
     1141 ttattatcat ggattctaaa acggattacc agggatttca gtcgatgtac acgttcgtca
     1201 catctcatct acctcccggt tttaatgaat acgattttgt gccagagtcc ttcgataggg
     1261 acaagacaat tgcactgatc atgaactcct ctggatctac tggtctgcct aaaggtgtcg
     1321 ctctgcctca tagaactgcc tgcgtgagat tctcgcatgc cagagatcct atttttggca
     1381 atcaaatcat tccggatact gcgattttaa gtgttgttcc attccatcac ggttttggaa
     1441 tgtttactac actcggatat ttgatatgtg gatttcgagt cgtcttaatg tatagatttg
     1501 aagaagagct gtttctgagg agccttcagg attacaagat tcaaagtgcg ctgctggtgc
     1561 caaccctatt ctccttcttc gccaaaagca ctctgattga caaatacgat ttatctaatt
     1621 tacacgaaat tgcttctggt ggcgctcccc tctctaagga agtcggggaa gcggttgcca
     1681 agaggttcca tctgccaggt atcaggcaag gatatgggct cactgagact acatcagcta
     1741 ttctgattac acccgagggg gatgataaac cgggcgcggt cggtaaagtt gttccatttt
     1801 ttgaagcgaa ggttgtggat ctggataccg ggaaaacgct gggcgttaat caaagaggcg
     1861 aactgtgtgt gagaggtcct atgattatgt ccggttatgt aaacaatccg gaagcgacca
     1921 acgccttgat tgacaaggat ggatggctac attctggaga catagcttac tgggacgaag
     1981 acgaacactt cttcatcgtt gaccgcctga agtctctgat taagtacaaa ggctatcagg
     2041 tggctcccgc tgaattggaa tccatcttgc tccaacaccc caacatcttc gacgcaggtg
     2101 tcgcaggtct tcccgacgat gacgccggtg aacttcccgc cgccgttgtt gttttggagc
     2161 acggaaagac gatgacggaa aaagagatcg tggattacgt cgccagtcaa gtaacaaccg
     2221 cgaaaaagtt gcgcggagga gttgtgtttg tggacgaagt accgaaaggt cttaccggaa
     2281 aactcgacgc aagaaaaatc agagagatcc tcataaaggc caagaagggc ggaaagatcg
     2341 ccgtgtaatt ctagagtcgg ggcggccggc cgcttcgagc agacatgata agatacattg
     2401 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
     2461 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
     2521 attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt
     2581 aaaacctcta caaatgtggt aaaatcgata aggatctgaa cgatggagcg gagaatgggc
     2641 ggaactgggc ggagttaggg gcgggatggg cggagttagg ggcgggacta tggttgctga
     2701 ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg actttccaca
     2761 cctggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg gggagcctgg
     2821 ggactttcca caccctaact gacacacatt ccacagcgga tccgtcgacc gatgcccttg
     2881 agagccttca acccagtcag ctccttccgg tgggcgcggg gcatgactat cgtcgccgca
     2941 cttatgactg tcttctttat catgcaactc gtaggacagg tgccggcagc gctcttccgc
     3001 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     3061 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     3121 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     3181 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     3241 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     3301 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     3361 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     3421 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     3481 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     3541 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     3601 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     3661 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     3721 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     3781 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     3841 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     3901 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     3961 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     4021 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     4081 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     4141 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     4201 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     4261 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     4321 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     4381 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     4441 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     4501 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     4561 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     4621 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     4681 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     4741 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     4801 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     4861 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     4921 acctgacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     4981 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     5041 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     5101 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     5161 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     5221 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     5281 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     5341 atttaacgcg aattttaaca aaatattaac gcttacaatt tgccattcgc cattcaggct
     5401 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agcccaagct
     5461 accatgataa gtaagtaata ttaaggtacg ggaggtactt ggagcggccg caataaaata
     5521 tctttatttt cattacatct gtgtgttggt tttttgtgtg aatcgatagt actaacatac
     5581 gctctccatc aaaacaaaac gaaacaaaac aaactagcaa aataggctgt ccccagtgca
     5641 agtgcaggtg ccagaacatt tctctatcga taggtaccga gctcttacgc gtgc
//
