LOCUS       sequence-389091-        7099 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of FLAG-tagged mouse STING1 C64S, C65S.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7099)
  AUTHORS   Li W, Li Y, Kang J, Jiang H, Gong W, Chen L, Wu C, Liu M, Wu X, Zhao
            Y, Ren J
  TITLE     4-octyl itaconate as a metabolite derivative inhibits inflammation 
            via alkylation of STING.
  JOURNAL   Cell Rep. 2023 Mar 28;42(3):112145. doi: 
            10.1016/j.celrep.2023.112145. Epub 2023 Feb 28.
  PUBMED    36862550
REFERENCE   2  (bases 1 to 7099)
  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: "Cell Rep.";
            date: "2023-03-28"; volume: "42(3):112145. doi"; pages: " 
            10.1016/j.celrep.2023.112145. Epub 2023 Feb 28"
COMMENT     Sequence Label: pcmv3-STING-C64S,C65S-FLAG
FEATURES             Location/Qualifiers
     source          1..7099
                     /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        253..632
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        633..836
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     787..807
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     833..857
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     1338..1357
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1386..1405
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1386..1404
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2555..2578
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2631..2648)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2637..2861
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2907..3335
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2994..3013)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3204..3225
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3344..3364)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3349..3678
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3529..3664
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3591..3610
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3727..4752
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             3727..4752
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     polyA_signal    4882..5003
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4919..4938)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4973..4992
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5052..5068)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5052..5068)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5065..5087)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5076..5092
                     /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(5100..5106,5107..5124,5125..5130))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5145..5166
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5283..5300)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5454..6042)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5534..5553)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6142..6957)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(6142..6957)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     6865..6884
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6949..6968
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
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 agtacattta tattggctca tgtccaatat
      241 gaccgccatg ttgacattga ttattgacta gttattaata gtaatcaatt acggggtcat
      301 tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg
      361 gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa
      421 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact
      481 tggcagtaca tcaagtgtat catatgccaa gtccgccccc tattgacgtc aatgacggta
      541 aatggcccgc ctggcattat gcccagtaca tgaccttacg ggactttcct acttggcagt
      601 acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag tacaccaatg
      661 ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg
      721 ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaat aaccccgccc
      781 cgttgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctcgtt
      841 tagtgaaccg tcagatcctc actctcttcc gcatcgctgt ctgcgagggc cagctgttgg
      901 gctcgcggtt gaggacaaac tcttcgcggt ctttccagta ctcttggatc ggaaacccgt
      961 cggcctccga acggtactcc gccaccgagg gacctgagcg agtccgcatc gaccggatcg
     1021 gaaaacctct cgagaaaggc gtctaaccag tcacagtcgc aaggtaggct gagcaccgtg
     1081 gcgggcggca gcgggtggcg gtcggggttg tttctggcgg aggtgctgct gatgatgtaa
     1141 ttaaagtagg cggtcttgag acggcggatg gtcgaggtga ggtgtggcag gcttgagatc
     1201 cagctgttgg ggtgagtact ccctctcaaa agcgggcatt acttctgcgc taagattgtc
     1261 agtttccaaa aacgaggagg atttgatatt cacctggccc gatctggcca tacacttgag
     1321 tgacaatgac atccactttg cctttctctc cacaggtgtc cactcccagg tccaagttta
     1381 aactttaata cgactcacta taggggccgc caccaagctt atgccatact ccaacctgca
     1441 tccagccatc ccacggccca gaggtcaccg ctccaaatat gtagccctca tctttctggt
     1501 ggccagcctg atgatccttt gggtggcaaa ggatccacca aatcacactc tgaagtacct
     1561 agcacttcac ctagcctcgc acgaacttgg actactgttg aaaaacctca gctccctggc
     1621 tgaagagctg tgccatgtcc agtccaggta ccagggcagc tactggaagg ctgtgcgcgc
     1681 ctgcctggga tgccccatcc actgtatggc tatgattcta ctatcgtctt atttctattt
     1741 cctccaaaac actgctgaca tatacctcag ttggatgttt ggccttctgg tcctctataa
     1801 gtccctaagc atgctcctgg gccttcagag cttgactcca gcggaagtct ctgcagtctg
     1861 tgaagaaaag aagttaaatg ttgcccacgg gctggcctgg tcatactaca ttgggtactt
     1921 gcggttgatc ttaccagggc tccaggcccg gatccgaatg ttcaatcagc tacataacaa
     1981 catgctcagt ggtgcaggga gccgaagact gtacatcctc tttccattgg actgtggggt
     2041 gcctgacaac ctgagtgtag ttgaccccaa cattcgattc cgagatatgc tgccccagca
     2101 aaacatcgac cgtgctggca tcaagaatcg ggtttattcc aacagcgtct acgagattct
     2161 ggagaacgga cagccagcag gcgtctgtat cctggagtac gccaccccct tgcagaccct
     2221 gtttgccatg tcacaggatg ccaaagctgg cttcagtcgg gaggatcggc ttgagcaggc
     2281 taaactcttc tgccggacac ttgaggaaat cctggaagat gtccccgagt ctcgaaataa
     2341 ctgccgcctc attgtctacc aagaacccac agacggaaac agtttctcac tgtctcagga
     2401 ggtgctccgg cacattcgtc aggaagaaaa ggaggaggtt accatgaatg cccccatgac
     2461 ctcagtggca cctcctccct ccgtactgtc ccaagagcca agactcctca tcagtggtat
     2521 ggatcagcct ctcccactcc gcactgacct catcgattac aaggacgacg atgacaagtg
     2581 ataatctaga gcggccgccg aattcgggcc cgtttaaacc cgctgatcag cctcgactgt
     2641 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     2701 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     2761 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     2821 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac
     2881 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     2941 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     3001 cgctttcttc ccttcctttc tcgccacgtt cgcaggcttt ccccgtcaag ctctaaatcg
     3061 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     3121 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     3181 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     3241 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     3301 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
     3361 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     3421 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3481 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     3541 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     3601 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     3661 ggcctaggct tttgcaaaaa gctctcggga gcttgtatat ccattttcgg atctgatcag
     3721 cacgtgatga aaaagcctga actcaccgcg acgtctgtcg agaagtttct gatcgaaaag
     3781 ttcgacagcg tctccgacct gatgcagctc tcggagggcg aagaatctcg tgctttcagc
     3841 ttcgatgtag gagggcgtgg atatgtcctg cgggtaaata gctgcgccga tggtttctac
     3901 aaagatcgtt atgtttatcg gcactttgca tcggccgcgc tcccgattcc ggaagtgctt
     3961 gacattgggg aattcagcga gagcctgacc tattgcatct cccgccgtgc acagggtgtc
     4021 acgttgcaag acctgcctga aaccgaactg cccgctgttc tgcagccggt cgcggaggcc
     4081 atggatgcga tcgctgcggc cgatcttagc cagacgagcg ggttcggccc attcggaccg
     4141 caaggaatcg gtcaatacac tacatggcgt gatttcatat gcgcgattgc tgatccccat
     4201 gtgtatcact ggcaaactgt gatggacgac accgtcagtg cgtccgtcgc gcaggctctc
     4261 gatgagctga tgctttgggc cgaggactgc cccgaagtcc ggcacctcgt gcacgcggat
     4321 ttcggctcca acaatgtcct gacggacaat ggccgcataa cagcggtcat tgactggagc
     4381 gaggcgatgt tcggggattc ccaatacgag gtcgccaaca tcttcttctg gaggccgtgg
     4441 ttggcttgta tggagcagca gacgcgctac ttcgagcgga ggcatccgga gcttgcagga
     4501 tcgccgcggc tccgggcgta tatgctccgc attggtcttg accaactcta tcagagcttg
     4561 gttgacggca atttcgatga tgcagcttgg gcgcagggtc gatgcgacgc aatcgtccga
     4621 tccggagccg ggactgtcgg gcgtacacaa atcgcccgca gaagcgcggc cgtctggacc
     4681 gatggctgtg tagaagtact cgccgatagt ggaaaccgac gccccagcac tcgtccgagg
     4741 gcaaaggaat agcacgtgct acgagatttc gattccaccg ccgccttcta tgaaaggttg
     4801 ggcttcggaa tcgttttccg ggacgctggc tggatgatcc tccagcgcgg ggatctcatg
     4861 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc
     4921 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     4981 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg
     5041 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     5101 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     5161 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     5221 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     5281 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     5341 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     5401 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     5461 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     5521 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     5581 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     5641 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     5701 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     5761 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     5821 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     5881 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     5941 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     6001 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     6061 tctacggggt ctgagcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     6121 tatccgctca tgaattaatt cttagaaaaa ctcatcgagc atcaaatgaa actgcaattt
     6181 attcatatca ggattatcaa taccatattt ttgaaaaagc cgtttctgta atgaaggaga
     6241 aaactcaccg aggcagttcc ataggatggc aagatcctgg tatcggtctg cgattccgac
     6301 tcgtccaaca tcaatacaac ctattaattt cccctcgtca aaaataaggt tatcaagtga
     6361 gaaatcacca tgagtgacga ctgaatccgg tgagaatggc aaaagtttat gcatttcttt
     6421 ccagacttgt tcaacaggcc agccattacg ctcgtcatca aaatcactcg catcaaccaa
     6481 accgttattc attcgtgatt gcgcctgagc gagacgaaat acgcgatcgc tgttaaaagg
     6541 acaattacaa acaggaatcg aatgcaaccg gcgcaggaac actgccagcg catcaacaat
     6601 attttcacct gaatcaggat attcttctaa tacctggaat gctgttttcc cagggatcgc
     6661 agtggtgagt aaccatgcat catcaggagt acggataaaa tgcttgatgg tcggaagagg
     6721 cataaattcc gtcagccagt ttagtctgac catctcatct gtaacatcat tggcaacgct
     6781 acctttgcca tgtttcagaa acaactctgg cgcatcgggc ttcccataca atcgatagat
     6841 tgtcgcacct gattgcccga cattatcgcg agcccattta tacccatata aatcagcatc
     6901 catgttggaa tttaatcgcg gcctagagca agacgtttcc cgttgaatat ggctcataac
     6961 accccttgta ttactgttta tgtaagcaga cagttttatt gttcatgacc aaaatccctt
     7021 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aatccgcgca catttccccg
     7081 aaaagtgcca cctgacgtc
//
