LOCUS       sequence-271045-        6157 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Human Dnase1L3 with activity-reducing HUVS mutation R206C 
            in mammalian cells along with DsRed.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6157)
  AUTHORS   Shi G, Abbott KN, Wu W, Salter RD, Keyel PA
  TITLE     Dnase1L3 Regulates Inflammasome-Dependent Cytokine Secretion.
  JOURNAL   Front Immunol. 2017 May 8;8:522. doi: 10.3389/fimmu.2017.00522. 
            eCollection 2017.
  PUBMED    28533778
REFERENCE   2  (bases 1 to 6157)
  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: "Front 
            Immunol."; date: "2017-05-8"; volume: "8:522. doi"; pages: " 
            10.3389/fimmu.2017.00522. eCollection 2017"
COMMENT     Sequence Label: Dnase1L3 R206C pIRES-DsRed
FEATURES             Location/Qualifiers
     source          1..6157
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        111..414
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        415..618
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     569..589
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    join(1610..2184,2185..2187,2188..2196)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1777..1794)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2004..2023
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2197..2874
                     /codon_start=1
                     /product="improved tetrameric variant of DsRed fluorescent 
                     protein"
                     /label=DsRed2
                     /note="mammalian codon-optimized"
                     /translation="MASSENVITEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTVK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     ATVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGETHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDAKLDITSHNEDYTIVEQYERTEGRHH
                     LFL"
     primer_bind     complement(2377..2397)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     2789..2812
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    2995..3116
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3032..3051)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3086..3105
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3123..3578)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3260..3281)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3472..3491
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3605..3709
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3711..4068
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3711..3902
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3734..3754)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3919..4054
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4103..4897
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4103..4897
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4157..4176)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4767..4786
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5085..5104)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5129..5176
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5505..6093
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5994..6013
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat tagttattaa
       61 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
      121 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
      181 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
      241 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
      301 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
      361 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg
      421 cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt
      481 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca
      541 aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag
      601 gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta ccggactcag
      661 atctcgagct caagcttcga attcatgtca cgggagctgg ccccactgct gcttctcctc
      721 ctctccatcc acagcgccct ggccatgagg atctgctcct tcaacgtcag gtcctttggg
      781 gaaagcaagc aggaagacaa gaatgccatg gatgtcattg tgaaggtcat caaacgctgt
      841 gacatcatac tcgtgatgga aatcaaggac agcaacaaca ggatctgccc catactgatg
      901 gagaagctga acagaaattc aaggagaggc ataacgtaca actatgtgat tagctctcgg
      961 cttggaagaa acacatataa agaacaatat gcctttctct acaaggaaaa gctggtgtct
     1021 gtgaagagga gttatcacta ccatgactat caggatggag acgcagatgt gttttccagg
     1081 gagccctttg tggtctggtt ccaatctccc cacactgctg tcaaagactt cgtgattatc
     1141 cccctgcaca ccaccccaga gacatccgtt aaggagatcg atgagttggt tgaggtctac
     1201 acggacgtga aacaccgctg gaaggcggag aatttcattt tcatgggtga cttcaatgcc
     1261 ggctgcagct acgtccccaa gaaggcctgg aagaacatct gcttgaggac tgaccccagg
     1321 tttgtttggc tgatcgggga ccaagaggac accacggtga agaagagcac caactgtgca
     1381 tatgacagga ttgtgcttag aggacaagaa atcgtcagtt ctgttgttcc caagtcaaac
     1441 agtgtttttg acttccagaa agcttacaag ctgactgaag aggaggccct ggatgtcagc
     1501 gaccactttc cagttgaatt taaactacag tcttcaaggg ccttcaccaa cagcaaaaaa
     1561 tctgtcactc taaggaagaa aacaaagagc aaacgctcct agggatccgc ccctctccct
     1621 cccccccccc taacgttact ggccgaagcc gcttggaata aggccggtgt gcgtttgtct
     1681 atatgttatt ttccaccata ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc
     1741 ctgtcttctt gacgagcatt cctaggggtc tttcccctct cgccaaagga atgcaaggtc
     1801 tgttgaatgt cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa acaacgtctg
     1861 tagcgaccct ttgcaggcag cggaaccccc cacctggcga caggtgcctc tgcggccaaa
     1921 agccacgtgt ataagataca cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt
     1981 ggatagttgt ggaaagagtc aaatggctct cctcaagcgt attcaacaag gggctgaagg
     2041 atgcccagaa ggtaccccat tgtatgggat ctgatctggg gcctcggtac acatgcttta
     2101 catgtgttta gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg gacgtggttt
     2161 tcctttgaaa aacacgatga taatatggcc acaaccatgg cctcctccga gaacgtcatc
     2221 accgagttca tgcgcttcaa ggtgcgcatg gagggcaccg tgaacggcca cgagttcgag
     2281 atcgagggcg agggcgaggg ccgcccctac gagggccaca acaccgtgaa gctgaaggtg
     2341 accaagggcg gccccctgcc cttcgcctgg gacatcctgt ccccccagtt ccagtacggc
     2401 tccaaggtgt acgtgaagca ccccgccgac atccccgact acaagaagct gtccttcccc
     2461 gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggc gaccgtgacc
     2521 caggactcct ccctgcagga cggctgcttc atctacaagg tgaagttcat cggcgtgaac
     2581 ttcccctccg acggccccgt gatgcagaag aagaccatgg gctgggaggc ctccaccgag
     2641 cgcctgtacc cccgcgacgg cgtgctgaag ggcgagaccc acaaggccct gaagctgaag
     2701 gacggcggcc actacctggt ggagttcaag tctatctaca tggccaagaa gcccgtgcag
     2761 ctgcccggct actactacgt ggacgccaag ctggacatca cctcccacaa cgaggactac
     2821 accatcgtgg agcagtacga gcgcaccgag ggccgccacc acctgttcct gtagcggccg
     2881 cgactctaga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac
     2941 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
     3001 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     3061 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc
     3121 gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     3181 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     3241 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     3301 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     3361 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     3421 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     3481 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     3541 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa
     3601 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     3661 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga
     3721 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     3781 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     3841 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     3901 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     3961 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     4021 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga
     4081 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     4141 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     4201 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     4261 gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac
     4321 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     4381 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     4441 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     4501 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     4561 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     4621 gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     4681 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     4741 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     4801 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     4861 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
     4921 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     4981 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     5041 gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag
     5101 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt
     5161 gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc
     5221 ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc
     5281 ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata
     5341 gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     5401 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     5461 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     5521 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     5581 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     5641 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     5701 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     5761 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     5821 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     5881 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     5941 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     6001 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     6061 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     6121 ttcctggcct tttgctggcc ttttgctcac atgttct
//
