LOCUS       sequence-448258-        7222 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector to express mouse Regnase-1 D141N mutant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7222)
  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 7222)
  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-1 D141N
FEATURES             Location/Qualifiers
     source          1..7222
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(464..1255,1256..1324))
                     /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     1087..1106
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1325..1429)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1504..1523)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1695..2074
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2075..2278
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2229..2249
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2323..2342
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2323..2341
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(4235..4253)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4236..4253)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(4273..4290)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4279..4503
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4549..4977
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4636..4655)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4846..4867
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4986..5006)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4991..5320
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5171..5306
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5233..5252
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5387..6181
                     /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(5441..5460)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6051..6070
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6355..6476
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6392..6411)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6446..6465
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6525..6541)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6525..6541)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6538..6560)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6549..6565
                     /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(6573..6579,6580..6597,6598..6603))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6618..6639
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6756..6773)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(6927..7222,1..293))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7007..7026)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
       61 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
      121 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
      181 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
      241 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
      301 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
      361 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
      421 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
      481 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
      541 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
      601 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
      661 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
      721 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
      781 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
      841 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
      901 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
      961 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     1021 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     1081 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     1141 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     1201 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     1261 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     1321 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     1381 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     1441 gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat ggtgcactct
     1501 cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt
     1561 ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga
     1621 caattgcatg aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc
     1681 cagatatacg cgttgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc
     1741 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
     1801 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
     1861 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
     1921 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
     1981 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
     2041 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa
     2101 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
     2161 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc
     2221 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctct
     2281 ctggctaact agagaaccca ctgcttactg gcttatcgaa attaatacga ctcactatag
     2341 ggagacccaa gcttggtacc gagctcggat ccactagtaa cggccgccag tgtgctggaa
     2401 ttcgccacca tgagtgaccc ttgtggaacg aagcctgtcc aagaatccaa ccccaccatg
     2461 agtctgtgga gtcttgagga cagacacagc agccagggtc gacctcagcc agaccaggat
     2521 cctgtggcta aagaggcccc tacttccgag ctgcagatga aggtggactt tttccgtaaa
     2581 ctggggtact cgtcctctga gatccacagt gtcctgcaga agctgggagt ccaagcagac
     2641 accaacacgg tgctagggga attggtgaag catggctcag ctactgaacg agaatgccag
     2701 gccctgacgg cccccagccc ccagccccct ctggtgcccc ggggtggaag cacccccaag
     2761 ccttccactc tagaaccctc actcccagag gaggacagag agggcagcga cctgagacct
     2821 gtggtcatca acggaagcaa tgtggccatg agccatggga acaaggaagt cttctcttgc
     2881 cggggcattc tgctggctgt gaactggttt ctggagcgag gccacacaga tattaccgtg
     2941 tttgtgccat cttggaggaa ggaacagcct cgaccagatg tgcctatcac agaccagcac
     3001 atccttcggg aactagagaa aaagaagatc ttggtgttca cgccatccag gcgggtcggc
     3061 ggcaagcgcg tggtgtgcta tgatgaccgc ttcattgtga agctggcctt cgaatccgac
     3121 ggagtggtgg tctccaatga cacgtaccga gacctccaag gcgagaggca ggagtggaaa
     3181 cgcttcatcg aggagcggct gctcatgtac tccttcgtca atgacaagtt catgccccct
     3241 gacgaccctt taggacggca tgggcctagc ctggacaact tccttcgtaa gaaaccactg
     3301 ccttctgagc acaggaagca gccatgcccc tatgggaaga aatgtacgta tggaatcaag
     3361 tgccgatttt tccaccctga gcggccaagc cgtccccagc gctctgtggc cgatgagctc
     3421 cgtgccaacg ctctcctctc acctcccagg actccagtca aggacaaaag tagccagagg
     3481 ccttcccctg cctctcagtc cagctctgtg tccctagagg ctgaaccagg cagcctggat
     3541 gggaaaaagc tgggtgccag gtcatctccg ggtccccacc gagaaggctc accgcagacc
     3601 tgtgctccag ctggcaggag cctccctgtt agtgggggca gctttgggcc cacagagtgg
     3661 cttgcacaca cccaggactc actcccatac acctcccagg agtgccttga ttcaggcatt
     3721 ggttccctgg agagccagat gtcagaatta tggggcgtgc gaggaggcag ccctggggag
     3781 tcgggcccca ctcggggccc ctatgcaggt tatcacagct atggatccaa ggtcccagca
     3841 gcaccttcct tttctccttt tagaccagcc atgggtgctg gccacttcag tgtccccacc
     3901 gactatgtgc ccccgccacc cacctaccca tccagagagt actggtctga gccgtaccca
     3961 ttacccccac ccactcctgt ccttcaggag ccccagagac ccagccccgg ggctggtggg
     4021 ggcccctggg gcagggtggg tgacctggcc aaagaaaggg ctggtgtata taccaagctg
     4081 tgtggtgtct tccccccaca cctggtagaa gctgtaatga gacgcttccc acagctgctg
     4141 gatccgcagc agctggccgc agagatcctg tcttacaagt cccagcacct cagtgagtaa
     4201 gcggccgctc gagcatgcat ctagagggcc ctattctata gtgtcaccta aatgctagag
     4261 ctcgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     4321 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     4381 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     4441 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta
     4501 tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta
     4561 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     4621 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     4681 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     4741 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     4801 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     4861 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     4921 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat
     4981 tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     5041 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     5101 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     5161 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     5221 actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa
     5281 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat
     5341 atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     5401 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     5461 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     5521 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc
     5581 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     5641 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     5701 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     5761 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     5821 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     5881 cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt
     5941 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     6001 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     6061 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     6121 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     6181 agcgggactc tggggttcga aatgaccgac caagtgacgc ccaacctgcc atcacgagat
     6241 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     6301 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg
     6361 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     6421 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     6481 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
     6541 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     6601 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     6661 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     6721 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     6781 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     6841 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     6901 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     6961 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     7021 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     7081 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     7141 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     7201 cccgaccgct gcgccttatc cg
//
