LOCUS       sequence-424630-        6580 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transient expression of STING.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6580)
  AUTHORS   Wu J, Dobbs N, Yang K, Yan N
  TITLE     Interferon-Independent Activities of Mammalian STING Mediate 
            Antiviral Response and Tumor Immune Evasion.
  JOURNAL   Immunity. 2020 Jul 14;53(1):115-126.e5. doi: 
            10.1016/j.immuni.2020.06.009. Epub 2020 Jul 7.
  PUBMED    32640258
REFERENCE   2  (bases 1 to 6580)
  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"; doi: 
            "10.1016/j.immuni.2020.06.009"; journalName: "Immunity"; date: 
            "2020-07-14- 14"; volume: "53"; issue: "1"; pages: "115-126.e5"
COMMENT     Sequence Label: pcDNA3.1-hSTING (S366A)
FEATURES             Location/Qualifiers
     source          1..6580
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     82..101
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    234..458
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     447..464
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     primer_bind     complement(1753..1772)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1754..1772)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(1817..2020)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1846..1866)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2021..2400
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     2572..2591
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        2666..2770
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2771..2839,2840..3631)
                     /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     complement(2989..3008)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3802..4390
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4291..4310
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4544..4561
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4678..4699
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4714..4719,4720..4737,4738..4744)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4752..4768
                     /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)."
     primer_bind     4757..4779
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4776..4792
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4776..4792
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    4841..4962
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4852..4871)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4906..4925
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(5136..5930)
                     /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(5247..5266)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5857..5876
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(5997..6326)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6011..6146
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6065..6084)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     6311..6331
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(join(6340..6580,1..188))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6450..6471)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
       61 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
      121 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
      181 gggcgcgtgg ggataccccc tagagcccca gctggttctt tccgcctcag aagccataga
      241 gcccaccgca tccccagcat gcctgctatt gtcttcccaa tcctccccct tgctgtcctg
      301 ccccacccca ccccccagaa tagaatgaca cctactcaga caatgcgatg caatttcctc
      361 attttattag gaaaggacag tgggagtggc accttccagg gtcaaggaag gcacggggga
      421 ggggcaaaca acagatggct ggcaactaga aggcacagtc gaggctgatc agcgggttta
      481 aacgggccct ctagactcga gcggccgcca ctgtgctgga tatctgcaga attctcaaga
      541 gaaatccgtg cggagaggga ggggcttttc cattccagcg atgaggagct caggctcttg
      601 ggacatcgtg gaggtactgg gcaccgctga ggtcttcaag ctgcccacag taacctcttc
      661 cttttcctcc tgccgcaggt gccggagaac ctcctgggac agcgagaagc tgctgtcatc
      721 tgcaggttcc tggtaggcaa tgaggcggca gttgttctga gactcagggg catctgccag
      781 gatgtcctca agtgtccggc agaagagttt ggcctgctca agcctatcct cccggctaaa
      841 gccagcttga ctgtattgtg acatggcaaa caaagtctgc aagggggtgg cgtactccag
      901 gacacaggtg cccgcccgct gcccgttctc cagaagctca tagatgctgt tgctgtaaac
      961 ccgatccttg atgccagcac ggtcaccggt ctgctggggc agtttatcca ggaagcgaat
     1021 gttggggtca gccatactca ggttatcagg caccccacag tccaatggga ggagaatata
     1081 cagccgctgg ctcactgcac cccgtagcag gttgttgtaa tgctgattgt aagttcgaat
     1141 ccgggcctgg agctctggca ggatcagccg cagatatccg atgtaatatg accatgccag
     1201 cccatgggcc acgttgaaat tccctttttc acacactgca gagatctcag ctggggccag
     1261 gcccttgagg cccaggagga tgttcagtgc ctgcgagagg cccaggaggg caagcatcca
     1321 agtgaagggc gggccgaccg catttgggag ggagtagtag aaatagatgg acagcagcaa
     1381 cagggcccca cggcggaggg ggcagcccag gcaggcccgc acagtcctcc agtagctgcc
     1441 ccggtacctg gagtggatgt ggcgcagctc ctcagccagg ctgcagaccc cgtttaacag
     1501 cagtcccagc tgcagggagg ctaggtggag caccaggtac cggagagtgt gctctggtgg
     1561 ctctcctagc ccccaaaggg tcaccaggca ggcactcagc agaaccaagg ctgccttctg
     1621 ggccccgtga cccctgggac acgggatgga tggatgcagg ctggagtggg gcatggtgaa
     1681 ttccaccaca ctggactagt ggatccgagc tcggtaccaa gcttaagttt aaacgctagc
     1741 cagcttgggt ctccctatag tgagtcgtat taatttcgat aagccagtaa gcagtgggtt
     1801 ctctagttag ccagagagct ctgcttatat agacctccca ccgtacacgc ctaccgccca
     1861 tttgcgtcaa tggggcggag ttgttacgac attttggaaa gtcccgttga ttttggtgcc
     1921 aaaacaaact cccattgacg tcaatggggt ggagacttgg aaatccccgt gagtcaaacc
     1981 gctatccacg cccattgatg tactgccaaa accgcatcac catggtaata gcgatgacta
     2041 atacgtagat gtactgccaa gtaggaaagt cccataaggt catgtactgg gcataatgcc
     2101 aggcgggcca tttaccgtca ttgacgtcaa tagggggcgt acttggcata tgatacactt
     2161 gatgtactgc caagtgggca gtttaccgta aatactccac ccattgacgt caatggaaag
     2221 tccctattgg cgttactatg ggaacatacg tcattattga cgtcaatggg cgggggtcgt
     2281 tgggcggtca gccaggcggg ccatttaccg taagttatgt aacgcggaac tccatatatg
     2341 ggctatgaac taatgacccc gtaattgatt actattaata actagtcaat aatcaatgtc
     2401 aacgcgtata tctggcccgt acatcgcgaa gcagcgcaaa acgcctaacc ctaagcagat
     2461 tcttcatgca attgtcggtc aagccttgcc ttgttgtagc ttaaattttg ctcgcgcact
     2521 actcagcgac ctccaacaca caagcaggga gcagatactg gcttaactat gcggcatcag
     2581 agcagattgt actgagagtg caccataggg gatcgggaga tctcccgatc cgtcgacgtc
     2641 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     2701 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     2761 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     2821 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     2881 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     2941 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     3001 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     3061 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     3121 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     3181 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     3241 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     3301 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     3361 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     3421 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     3481 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     3541 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     3601 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     3661 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     3721 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     3781 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     3841 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     3901 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     3961 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     4021 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     4081 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     4141 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     4201 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     4261 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     4321 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     4381 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     4441 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     4501 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     4561 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     4621 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
     4681 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
     4741 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta
     4801 cgccaagctc tagctagagg tcgacggtat acagacatga taagatacat tgatgagttt
     4861 ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct
     4921 attgctttat ttgtaaccat tataagctgc aataaacaag ttggggtggg cgaagaactc
     4981 cagcatgaga tccccgcgct ggaggatcat ccagccggcg tcccggaaaa cgattccgaa
     5041 gcccaacctt tcatagaagg cggcggtgga atcgaaatct cgtgatggca ggttgggcgt
     5101 cgcttggtcg gtcatttcga accccagagt cccgctcaga agaactcgtc aagaaggcga
     5161 tagaaggcga tgcgctgcga atcgggagcg gcgataccgt aaagcacgag gaagcggtca
     5221 gcccattcgc cgccaagctc ttcagcaata tcacgggtag ccaacgctat gtcctgatag
     5281 cggtccgcca cacccagccg gccacagtcg atgaatccag aaaagcggcc attttccacc
     5341 atgatattcg gcaagcaggc atcgccatgg gtcacgacga gatcctcgcc gtcgggcatg
     5401 cgcgccttga gcctggcgaa cagttcggct ggcgcgagcc cctgatgctc ttcgtccaga
     5461 tcatcctgat cgacaagacc ggcttccatc cgagtacgtg ctcgctcgat gcgatgtttc
     5521 gcttggtggt cgaatgggca ggtagccgga tcaagcgtat gcagccgccg cattgcatca
     5581 gccatgatgg atactttctc ggcaggagca aggtgagatg acaggagatc ctgccccggc
     5641 acttcgccca atagcagcca gtcccttccc gcttcagtga caacgtcgag cacagctgcg
     5701 caaggaacgc ccgtcgtggc cagccacgat agccgcgctg cctcgtcctg cagttcattc
     5761 agggcaccgg acaggtcggt cttgacaaaa agaaccgggc gcccctgcgc tgacagccgg
     5821 aacacggcgg catcagagca gccgattgtc tgttgtgccc agtcatagcc gaatagcctc
     5881 tccacccaag cggccggaga acctgcgtgc aatccatctt gttcaatcat gcgaaacgat
     5941 cctcatcctg tctcttgatc agatccgaaa atggatatac aagctcccgg gagctttttg
     6001 caaaagccta ggcctccaaa aaagcctcct cactacttct ggaatagctc agaggcagag
     6061 gcggcctcgg cctctgcata aataaaaaaa attagtcagc catggggcgg agaatgggcg
     6121 gaactgggcg gagttagggg cgggatgggc ggagttaggg gcgggactat ggttgctgac
     6181 taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac
     6241 ctggttgctg actaattgag atgcatgctt tgcatacttc tgcctgctgg ggagcctggg
     6301 gactttccac accctaactg acacacattc cacagaatta attcgcgtta aatttttgtt
     6361 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     6421 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     6481 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     6541 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
//
