LOCUS       sequence-190088-        6157 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Human Dnase1L3 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 pIRES-DsRed
FEATURES             Location/Qualifiers
     source          1..6157
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    join(1500..2074,2075..2077,2078..2086)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1667..1684)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1894..1913
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2087..2764
                     /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(2267..2287)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     2679..2702
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    2885..3006
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2922..2941)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2976..2995
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3013..3468)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3150..3171)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3362..3381
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3495..3599
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3601..3958
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3601..3792
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3624..3644)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3809..3944
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3993..4787
                     /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             3993..4787
                     /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(4047..4066)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4657..4676
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4975..4994)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5019..5066
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5395..5983
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5884..5903
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggactcag atctcgagct caagcttcga attcatgtca cgggagctgg ccccactgct
      601 gcttctcctc ctctccatcc acagcgccct ggccatgagg atctgctcct tcaacgtcag
      661 gtcctttggg gaaagcaagc aggaagacaa gaatgccatg gatgtcattg tgaaggtcat
      721 caaacgctgt gacatcatac tcgtgatgga aatcaaggac agcaacaaca ggatctgccc
      781 catactgatg gagaagctga acagaaattc aaggagaggc ataacgtaca actatgtgat
      841 tagctctcgg cttggaagaa acacatataa agaacaatat gcctttctct acaaggaaaa
      901 gctggtgtct gtgaagagga gttatcacta ccatgactat caggatggag acgcagatgt
      961 gttttccagg gagccctttg tggtctggtt ccaatctccc cacactgctg tcaaagactt
     1021 cgtgattatc cccctgcaca ccaccccaga gacatccgtt aaggagatcg atgagttggt
     1081 tgaggtctac acggacgtga aacaccgctg gaaggcggag aatttcattt tcatgggtga
     1141 cttcaatgcc ggctgcagct acgtccccaa gaaggcctgg aagaacatcc gcttgaggac
     1201 tgaccccagg tttgtttggc tgatcgggga ccaagaggac accacggtga agaagagcac
     1261 caactgtgca tatgacagga ttgtgcttag aggacaagaa atcgtcagtt ctgttgttcc
     1321 caagtcaaac agtgtttttg acttccagaa agcttacaag ctgactgaag aggaggccct
     1381 ggatgtcagc gaccactttc cagttgaatt taaactacag tcttcaaggg ccttcaccaa
     1441 cagcaaaaaa tctgtcactc taaggaagaa aacaaagagc aaacgctcct agggatccgc
     1501 ccctctccct cccccccccc taacgttact ggccgaagcc gcttggaata aggccggtgt
     1561 gcgtttgtct atatgttatt ttccaccata ttgccgtctt ttggcaatgt gagggcccgg
     1621 aaacctggcc ctgtcttctt gacgagcatt cctaggggtc tttcccctct cgccaaagga
     1681 atgcaaggtc tgttgaatgt cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa
     1741 acaacgtctg tagcgaccct ttgcaggcag cggaaccccc cacctggcga caggtgcctc
     1801 tgcggccaaa agccacgtgt ataagataca cctgcaaagg cggcacaacc ccagtgccac
     1861 gttgtgagtt ggatagttgt ggaaagagtc aaatggctct cctcaagcgt attcaacaag
     1921 gggctgaagg atgcccagaa ggtaccccat tgtatgggat ctgatctggg gcctcggtac
     1981 acatgcttta catgtgttta gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg
     2041 gacgtggttt tcctttgaaa aacacgatga taatatggcc acaaccatgg cctcctccga
     2101 gaacgtcatc accgagttca tgcgcttcaa ggtgcgcatg gagggcaccg tgaacggcca
     2161 cgagttcgag atcgagggcg agggcgaggg ccgcccctac gagggccaca acaccgtgaa
     2221 gctgaaggtg accaagggcg gccccctgcc cttcgcctgg gacatcctgt ccccccagtt
     2281 ccagtacggc tccaaggtgt acgtgaagca ccccgccgac atccccgact acaagaagct
     2341 gtccttcccc gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggc
     2401 gaccgtgacc caggactcct ccctgcagga cggctgcttc atctacaagg tgaagttcat
     2461 cggcgtgaac ttcccctccg acggccccgt gatgcagaag aagaccatgg gctgggaggc
     2521 ctccaccgag cgcctgtacc cccgcgacgg cgtgctgaag ggcgagaccc acaaggccct
     2581 gaagctgaag gacggcggcc actacctggt ggagttcaag tctatctaca tggccaagaa
     2641 gcccgtgcag ctgcccggct actactacgt ggacgccaag ctggacatca cctcccacaa
     2701 cgaggactac accatcgtgg agcagtacga gcgcaccgag ggccgccacc acctgttcct
     2761 gtagcggccg cgactctaga tcataatcag ccataccaca tttgtagagg ttttacttgc
     2821 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt
     2881 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     2941 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     3001 atcttaaggc gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
     3061 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     3121 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     3181 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     3241 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     3301 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     3361 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
     3421 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt
     3481 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     3541 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc
     3601 ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     3661 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     3721 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     3781 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     3841 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     3901 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga
     3961 tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     4021 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     4081 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     4141 agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc
     4201 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     4261 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     4321 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     4381 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     4441 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     4501 tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg
     4561 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     4621 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     4681 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     4741 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     4801 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     4861 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     4921 ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac
     4981 acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa
     5041 aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc
     5101 tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc
     5161 accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg
     5221 ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta
     5281 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     5341 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     5401 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     5461 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     5521 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     5581 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     5641 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     5701 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     5761 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     5821 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     5881 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     5941 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     6001 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     6061 ccctgattct gtggataacc gtattaccgc catgcattag ttattaatag taatcaatta
     6121 cggggtcatt agttcatagc ccatatatgg agttccg
//
