LOCUS       sequence-448254-        4729 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For in vitro transcription of mouse Zc3h12a D141N mutant tagged with
            FLAG-HA at N-term.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4729)
  AUTHORS   Bataclan M, Leoni C, Moro SG, Pecoraro M, Wong EH, Heissmeyer V, 
            Monticelli S
  TITLE     Crosstalk between Regnase-1 and -3 shapes mast cell survival and 
            cytokine expression.
  JOURNAL   Life Sci Alliance. 2024 Jun 3;7(8):e202402784. doi: 
            10.26508/lsa.202402784. Print 2024 Aug.
  PUBMED    38830770
REFERENCE   2  (bases 1 to 4729)
  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"; journalName: "Life Sci 
            Alliance."; date: "2024-06-3"; volume: "7(8):e202402784. doi"; 
            pages: " 10.26508/lsa.202402784. Print 2024 Aug"
COMMENT     Sequence Label: pUC57 mini-ZsGreen-P2A-FLAG-HA-Regnase-1 D141N
FEATURES             Location/Qualifiers
     source          1..4729
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(371..390)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(372..390)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        430..534
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(535..603,604..1395)
                     /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     complement(753..772)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1566..2154
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2055..2074
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4189..4215)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YPYDVPDYA"
     CDS             complement(4189..4215)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(4228..4251)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      4251..4260
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(4276..4332)
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             complement(join(4351..4729,1..314))
                     /codon_start=1
                     /product="Zoanthus green fluorescent protein"
                     /label=ZsGreen1
                     /note="mammalian codon-optimized"
                     /translation="MAQSKHGLTKEMTMKYRMEGCVDGHKFVITGEGIGYPFKGKQAIN
                     LCVVEGGPLPFAEDILSAAFMYGNRVFTEYPQDIVDYFKNSCPAGYTWDRSFLFEDGAV
                     CICNADITVSVEENCMYHESKFYGVNFPADGPVMKKMTDNWEPSCEKIIPVPKQGILKG
                     DVSMYLLLKDGGRLRCQFDTVYKAKSVPRKMPDWHFIQHKLTREDRSDAKNQKWHLTEH
                     AIASGSALP"
ORIGIN
        1 cacacggcgc cgtcctcgaa caggaaggag cggtcccagg tgtagccggc ggggcaggag
       61 ttcttgaagt agtcgacgat gtcctggggg tactcggtga acacgcggtt gccgtacatg
      121 aaggcggcgg acaagatgtc ctcggcgaag ggcaaggggc cgccctccac cacgcacagg
      181 ttgatggcct gcttgccctt gaaggggtag ccgatgccct cgccggtgat cacgaacttg
      241 tggccgtcca cgcagccctc catgcggtac ttcatggtca tctccttggt caggccgtgc
      301 ttggactggg ccatggtgtc tgtttgaggt tgctagtgaa cacagttgtg tcagaagcaa
      361 atgtatcgat ccctatagtg agtcgtatta atcattggga cgtcaggtgg cacttttcgg
      421 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
      481 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
      541 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
      601 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
      661 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
      721 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
      781 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
      841 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
      901 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
      961 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     1021 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     1081 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     1141 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     1201 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     1261 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     1321 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     1381 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     1441 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     1501 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     1561 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     1621 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     1681 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     1741 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     1801 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     1861 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     1921 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     1981 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     2041 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     2101 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     2161 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     2221 ccgatactag tttgcaatga aaataaatgt tttttattag gcagaatcca gatgctcaag
     2281 gcccttcata atatccccca gtttagtagt tggacttagg gaacaaagga acctttaata
     2341 gaaattggac agcaagaaag cgagcctcga ggcggccgct tactcactga ggtgctggga
     2401 cttgtaagac aggatctctg cggccagctg ctgcggatcc agcagctgtg ggaagcgtct
     2461 cattacagct tctaccaggt gtggggggaa gacaccacac agcttggtat atacaccagc
     2521 cctttctttg gccaggtcac ccaccctgcc ccaggggccc ccaccagccc cggggctggg
     2581 tctctggggc tcctgaagga caggagtggg tgggggtaat gggtacggct cagaccagta
     2641 ctctctggat gggtaggtgg gtggcggggg cacatagtcg gtggggacac tgaagtggcc
     2701 agcacccatg gctggtctaa aaggagaaaa ggaaggtgct gctgggacct tggatccata
     2761 gctgtgataa cctgcatagg ggccccgagt ggggcccgac tccccagggc tgcctcctcg
     2821 cacgccccat aattctgaca tctggctctc cagggaacca atgcctgaat caaggcactc
     2881 ctgggaggtg tatgggagtg agtcctgggt gtgtgcaagc cactctgtgg gcccaaagct
     2941 gcccccacta acagggaggc tcctgccagc tggagcacag gtctgcggtg agccttctcg
     3001 gtggggaccc ggagatgacc tggcacccag ctttttccca tccaggctgc ctggttcagc
     3061 ctctagggac acagagctgg actgagaggc aggggaaggc ctctggctac ttttgtcctt
     3121 gactggagtc ctgggaggtg agaggagagc gttggcacgg agctcatcgg ccacagagcg
     3181 ctggggacgg cttggccgct cagggtggaa aaatcggcac ttgattccat acgtacattt
     3241 cttcccatag gggcatggct gcttcctgtg ctcagaaggc agtggtttct tacgaaggaa
     3301 gttgtccagg ctaggcccat gccgtcctaa agggtcgtca gggggcatga acttgtcatt
     3361 gacgaaggag tacatgagca gccgctcctc gatgaagcgt ttccactcct gcctctcgcc
     3421 ttggaggtct cggtacgtgt cattggagac caccactccg tcggattcga aggccagctt
     3481 cacaatgaag cggtcatcat agcacaccac gcgcttgccg ccgacccgcc tggatggcgt
     3541 gaacaccaag atcttctttt tctctagttc ccgaaggatg tgctggtctg tgataggcac
     3601 atctggtcga ggctgttcct tcctccaaga tggcacaaac acggtaatat ctgtgtggcc
     3661 tcgctccaga aaccagttca cagccagcag aatgccccgg caagagaaga cttccttgtt
     3721 cccatggctc atggccacat tgcttccgtt gatgaccaca ggtctcaggt cgctgccctc
     3781 tctgtcctcc tctgggagtg agggttctag agtggaaggc ttgggggtgc ttccaccccg
     3841 gggcaccaga gggggctggg ggctgggggc cgtcagggcc tggcattctc gttcagtagc
     3901 tgagccatgc ttcaccaatt cccctagcac cgtgttggtg tctgcttgga ctcccagctt
     3961 ctgcaggaca ctgtggatct cagaggacga gtaccccagt ttacggaaaa agtccacctt
     4021 catctgcagc tcggaagtag gggcctcttt agccacagga tcctggtctg gctgaggtcg
     4081 accctggctg ctgtgtctgt cctcaagact ccacagactc atggtggggt tggattcttg
     4141 gacaggcttc gttccacaag ggtcactcat tgtacagccg gaggagccgg cgtagtcggg
     4201 cacgtcgtag gggtatcctc catcgagctt gtcatcgtcg tccttgtagt ccatggtggc
     4261 acgcgtctgc agcctaggac cggggttttc ttccacgtct cctgcttgct ttaacagaga
     4321 gaagttcgtg gctccggaac cggtttcgaa gggcaaggcg gagccggagg cgatggcgtg
     4381 ctcggtcagg tgccacttct ggttcttggc gtcgctgcgg tcctcgcggg tcagcttgtg
     4441 ctggatgaag tgccagtcgg gcatcttgcg gggcacggac ttggccttgt acacggtgtc
     4501 gaactggcag cgcaagcggc caccgtcctt cagcagcagg tacatgctca cgtcgccctt
     4561 caagatgccc tgcttgggca cggggatgat cttctcgcag gagggctccc agttgtcggt
     4621 catcttcttc atcacggggc cgtcggcggg gaagttcacg ccgtagaact tggactcgtg
     4681 gtacatgcag ttctcctcca cgctcacggt gatgtcggcg ttgcagatg
//
