LOCUS       sequence-449846-        8086 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector to express mouse Regnase-3 WT.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8086)
  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 8086)
  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: pCDNA3-Regnase-3 WT
FEATURES             Location/Qualifiers
     source          1..8086
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      944..953
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3639..3657)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3640..3657)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3677..3694)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3683..3907
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3953..4381
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4040..4059)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4250..4271
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4390..4410)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4395..4724
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4575..4710
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4637..4656
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4791..5585
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4845..4864)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5455..5474
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5759..5880
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5796..5815)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5850..5869
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5929..5945)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5929..5945)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5942..5964)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5953..5969
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5977..5983,5984..6001,6002..6007))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6022..6043
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6160..6177)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6331..6919)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6411..6430)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7090..7881,7882..7950))
                     /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     7713..7732
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7951..8055)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
      901 gagctcggat ccactagtaa cggccgccag tgtgctggaa ttcgccacca tggctgcgga
      961 gaagacattg caggaatacg gggtgctttg cattcaagaa tacaggaaaa gcagcaaagt
     1021 ggagtcaagt gcacggaaca gcttcatggg cctgaaggat cacctggggc atgatcttgg
     1081 ccatctttat atggagagca ctgacccaca gatgagtgca gctgtgcctt ggcctatggt
     1141 agagaaacca acaatggata cagtcaattc ggggaaggaa ggaaaagggg tgtcagaaga
     1201 gaatgtgagc tccggcgact ctgagggaag cacgagttct gatcatgagt cagaacaact
     1261 gagtagcctt tcggtagagc catgctcatt aaccaagact cacagacagc tatgtcgctc
     1321 tccctgctta gagcctcgcc tactgaaaca cagtgacatt ttgcaagact ttaaaccaga
     1381 agagtcccag actccatcta aggaagtgaa gaaaccccct gatgtggttc gagaatacca
     1441 aacaaaactg gagtttgcac ttaagttagg ttattctgaa gaacaggtcc agcttgtgct
     1501 aaacaaactt ggtactgatg ctttaatcaa tgatatattg ggagaacttg tcaaacttgg
     1561 aaataaaagt gaagccgatc aaacagttag tactattaac agcgtaatgc gagaaacctc
     1621 atccctggag tctcagaggt ctgaatctcc aatgcaagag gttgtagtag atgacggtga
     1681 gaatctgaga ccagtagtta ttgatggcag caatgtggca atgagccatg gaaacaaaga
     1741 agtattctcg tgccgaggaa taaaattggc ggtggattgg tttttggaaa gaggccataa
     1801 agatatcacg gtctttgttc ctgcttggag aaaagaacag tcccgccctg acgccctcat
     1861 cacagatcag gaaattttac gtaaactaga gaaggaaaaa atcctggtat tcacaccatc
     1921 acggcgtgtc cagggcaggc gagtggtgtg ctatgatgac agattcatcg tgaagctggc
     1981 ttttgagtcc gatggtataa ttgtgtccaa tgataactac agagatttgg caaatgagaa
     2041 accagaatgg aagaagttca tagatgagcg gttattgatg tattcctttg tcaatgacaa
     2101 gttcatgccc ccagatgacc ctcttggcag acatggccca agtctggata attttctgag
     2161 gaagaaaccc attgttcctg agcacaaaaa gcagccttgt ccctatggaa agaagtgtac
     2221 gtatggacac aagtgcaaat actaccaccc agaaaggggc agtcagcctc agcggtcagt
     2281 ggctgatgaa cttcgtgcca tgtctagaaa cacagcagcc aaaactacaa atgaaggggg
     2341 actggtgaaa agcaacagtg ttccttgcag caccaaggca gacagcactt cggatgttaa
     2401 acgcggtgct ccaaagaggc agtcagaccc aagcataagg actcacgtct accaagacat
     2461 agaggagaag cttcccacca aaaacaagtt ggaaaccagg tctgtacctt ccttagttag
     2521 tatcccggcc acttctactg caaaacccca aagcactaca cctttaagca atggccttcc
     2581 atctggagtt catttcccac ctcaggatca aagaccacag ggacaatacc ctccaatgat
     2641 gatggcaacc aaaaatcatg gaacgccaat gccttatgaa cagtatccaa aatgtgactc
     2701 ccctgttgac gttggatatt attccatgtt gaatgcatat tctaatctga gtatctcagg
     2761 cccccgaagt cctgaaaggc gtttctcact agacactgat tacagagtaa actctgtggc
     2821 ttcagactgc agcagtgaag ggagcatgag ttgtgggagc agcgactcct acgtggggta
     2881 caatgaccgc tcctatgtca gctctcctga cccacagctg gaagagagtc tcaagtgtca
     2941 gcacatgcac ccccacagcc gccttaattc ccagcccttc ctgcagaatt tccacgatcc
     3001 cttaaccaga gtccagagtt acagtcacga agaaccaaag ttccatccca agcgtcctct
     3061 tccacacttg gctatgcatc tgcagcaccc tgccgtgggt gcccgctcca gctgtcctgg
     3121 cgattacccc tccccaccaa gttcagcaca ttcgaaggca ccacacctag ggaggtcctt
     3181 agtggccacg agaatagaca gcatttctga ttcccgactc tatgatagtt ctccttcaag
     3241 acaaagaaag ccttattctc gccaggaagg cctaggaagc tggggtaggc caagctatgg
     3301 gctggaagca tatggctatc ggcagacgta ctccttaccc gataattcca cgcccccatg
     3361 ctatgagtcg atcaccttcc agagcctgcc tgagcagcag gaacccacct ggagaatccc
     3421 ctactgtgga atgccacacg atcctccaag gtaccaggac aaccgagaga agatttttat
     3481 caatttgtgt aacatcttcc cccctgacct tgtgaggctc gtcatgaaaa ggaatcctca
     3541 tatgacggat gcccagcagc tcgccgcagc catcttagtg gagaaatcgc agctgggtta
     3601 ctgagcggcc gctcgagcat gcatctagag ggccctattc tatagtgtca cctaaatgct
     3661 agagctcgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     3721 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     3781 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     3841 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
     3901 tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc
     3961 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     4021 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     4081 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     4141 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     4201 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     4261 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     4321 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     4381 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     4441 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     4501 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     4561 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     4621 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc
     4681 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt
     4741 gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac
     4801 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     4861 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     4921 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg
     4981 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     5041 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     5101 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     5161 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     5221 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     5281 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc
     5341 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     5401 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     5461 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     5521 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     5581 tctgagcggg actctggggt tcgaaatgac cgaccaagtg acgcccaacc tgccatcacg
     5641 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     5701 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
     5761 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     5821 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     5881 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     5941 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     6001 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     6061 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     6121 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     6181 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     6241 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     6301 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     6361 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     6421 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     6481 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     6541 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     6601 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     6661 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     6721 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     6781 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     6841 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     6901 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     6961 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     7021 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     7081 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     7141 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     7201 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     7261 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     7321 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     7381 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     7441 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     7501 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     7561 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     7621 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     7681 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     7741 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     7801 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     7861 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     7921 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     7981 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     8041 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
