LOCUS       sequence-438498-        8510 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of STING without transmembrane domain in mammalian cells 
            by retroviral transduction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8510)
  AUTHORS   Landau LM, Chaudhary N, Tien YC, Rogozinska M, Joshi S, Yao C, 
            Crowley J, Hullahalli K, Campbell IW, Waldor MK, Haigis M, Kagan JC
  TITLE     pLxIS-containing domains are biochemically flexible regulators of 
            interferons and metabolism.
  JOURNAL   Mol Cell. 2024 Jul 11;84(13):2436-2454.e10. doi: 
            10.1016/j.molcel.2024.05.030. Epub 2024 Jun 25.
  PUBMED    38925114
REFERENCE   2  (bases 1 to 8510)
  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.molcel.2024.05.030"; journalName: "Mol Cell"; date: 
            "2024-07-11- 11"; volume: "84"; issue: "13"; pages: "2436-2454.e10"
COMMENT     Sequence Label: pMSCV-FKBP-STING(FL)
FEATURES             Location/Qualifiers
     source          1..8510
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      7..16
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             16..81
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    2133..2699
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    2149..2701
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2293..2310)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2520..2539
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2713..2715,2716..2718,2719..3432)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2758..2779)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3019..3038)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3366..3387
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             3494..4008
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    4170..4186
                     /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(4194..4200,4201..4218,4219..4224))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4239..4260
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4377..4394)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4548..5136)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4628..4647)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5307..6098,6099..6167))
                     /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     5930..5949
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6168..6272)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6340..6358
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6396..6418)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6518..6537
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6731..6753
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6881..6900
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             7100..7616
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    7680..8021
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             8088..8504
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     8434..8456
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     8472..8488
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
ORIGIN
        1 agatctgcca ccatggacta caaggatcac gacggtgatt acaaggacca cgacatcgac
       61 tacaaggacg acgacgacaa gggcgttcaa gtagaaacaa tatctccagg agacgggcgt
      121 acatttccta agcgtggcca aacttgcgta gtccattata ccggcatgct ggaggacggt
      181 aagaaggtcg atagctctag agaccgtaac aaacctttca aatttatgct ggggaaacag
      241 gaggttatcc gtggttggga ggaaggcgtg gcccaaatgt ccgttgggca gcgtgctaag
      301 ttaaccatct cgccagatta cgcatatggg gctacaggcc atcctgggat cattcccccg
      361 catgcgacac ttgtgtttga tgtagagctg ttgaagttgg agacgcgggg agttcaagtc
      421 gaaacgattt caccaggtga tggacgcacc tttcctaaac ggggacaaac gtgcgtcgtg
      481 cattacaccg gtatgctgga agacggcaag aaagtcgatt caagccgcga tagaaataaa
      541 cccttcaaat tcatgcttgg aaaacaggaa gtaatacgcg gatgggaaga gggtgtggcg
      601 caaatgtctg tcgggcaaag agctaaactg acaataagtc ctgattacgc gtacggtgcg
      661 actgggcacc ctggtatcat tccgccgcac gcgacacttg tcttcgacgt cgagttactt
      721 aagcttgaga cgcggggggt gcaggtagag actatttcgc ccggtgacgg aagaactttc
      781 ccaaagcggg gacagacctg tgttgtccac tacactggga tgttggagga tggtaagaaa
      841 gttgactcaa gtcgtgatcg caacaaaccc tttaagttta tgcttgggaa gcaggaagtc
      901 atcagaggtt gggaagaagg cgtagctcag atgtcagtgg gacaacgggc caaattaacc
      961 atcagtccag actacgccta cggagccacc ggtcacccag ggatcatacc acctcacgca
     1021 accttagtgt tcgatgtgga acttttaaag ttagagacgc gcggtgtaca agtggaaaca
     1081 atcagtccag gggatgggag aacattcccc aaaagaggcc agacgtgtgt cgtgcactat
     1141 acggggatgt tagaagatgg gaaaaaggtc gattcgtcac gcgaccgcaa caaaccgttt
     1201 aaattcatgt tgggaaagca ggaggtgatt cgcggctggg aggaaggtgt ggcacagatg
     1261 agcgtgggcc agagagcaaa actgactatc tctcctgatt atgcctacgg cgcgacgggc
     1321 caccctggca taattccacc ccacgctacc ctggtattcg atgtcgagct tctgaagctt
     1381 gagggtgctg gggctggggc taatgttgcc cacgggctgg cctggtcata ctacattggg
     1441 tacttgcggt tgatcttacc agggctccag gcccggatcc gaatgttcaa tcagctacat
     1501 aacaacatgc tcagtggtgc agggagccga agactgtaca tcctctttcc attggactgt
     1561 ggggtgcctg acaacctgag tgtagttgat cccaacattc gattccgaga tatgctgccc
     1621 cagcaaaaca tcgaccgtgc tggcatcaag aatcgggttt attccaacag cgtctacgag
     1681 attctggaga acggacagcc agcaggcgtc tgtatcctgg agtacgccac ccccttgcag
     1741 accctgtttg ccatgtcaca ggatgccaaa gctggcttca gtcgggagga tcggcttgag
     1801 caggctaaac tcttctgccg gacacttgag gaaatcctgg aagatgtccc cgagtctcga
     1861 aataactgcc gcctcattgt ctaccaagaa cccacagacg gaaacagttt ctcactgtct
     1921 caggaggtgc tccggcacat tcgtcaggaa gaaaaggagg aggttacaat gaatgcgcct
     1981 atgactagtg tggccccgcc gccttcggtt ttatcacagg agccgcggct gttaattagc
     2041 ggtatggatc aaccgctgcc tcttcggacc gatttgatct gataagcggc cgcgtttaaa
     2101 cgtcgacggt atcgataagc ttgatatcga attccgcccc cccccctaac gttactggcc
     2161 gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat gttattttcc accatattgc
     2221 cgtcttttgg caatgtgagg gcccggaaac ctggccctgt cttcttgacg agcattccta
     2281 ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt gaatgtcgtg aaggaagcag
     2341 ttcctctgga agcttcttga agacaaacaa cgtctgtagc gaccctttgc aggcagcgga
     2401 accccccacc tggcgacagg tgcctctgcg gccaaaagcc acgtgtataa gatacacctg
     2461 caaaggcggc acaaccccag tgccacgttg tgagttggat agttgtggaa agagtcaaat
     2521 ggctctcctc aagcgtattc aacaaggggc tgaaggatgc ccagaaggta ccccattgta
     2581 tgggatctga tctggggcct cggtgcacat gctttacatg tgtttagtcg aggttaaaaa
     2641 aacgtctagg ccccccgaac cacggggacg tggttttcct ttgaaaaaca cgatgataat
     2701 atggccacaa ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
     2761 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtctggcga gggcgagggc
     2821 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
     2881 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
     2941 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
     3001 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     3061 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     3121 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     3181 aagcagaaga acggcatcaa ggcgaacttc aagatccgcc acaacatcga ggacggcagc
     3241 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     3301 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     3361 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     3421 ctgtacaagt aatgaattaa ttaagaattc gataaaataa aagattttat ttagtctcca
     3481 gaaaaagggg ggaatgaaag accccacctg taggtttggc aagctagctt aagtaacgcc
     3541 attttgcaag gcatggaaaa tacataactg agaatagaga agttcagatc aaggttagga
     3601 acagagagac agcagaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg
     3661 gctcagggcc aagaacagat ggtccccaga tgcggtcccg ccctcagcag tttctagaga
     3721 accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc ttatttgaac
     3781 taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa
     3841 agagcccaca acccctcact cggcgcgcca gtcctccgat agactgcgtc gcccgggtac
     3901 ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc tgttccttgg
     3961 gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcatgg gtaacagttt
     4021 cttgaagttg gagaacaaca ttctgagggt aggagtcgaa tattaagtaa tcctgactca
     4081 attagccact gttttgaatc cacatactcc aatactcctg aaatagttca ttatggacag
     4141 cgcagaaaga gctggggaga attgtgaaat tgttatccgc tcacaattcc acacaacata
     4201 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     4261 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     4321 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     4381 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     4441 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     4501 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     4561 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     4621 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     4681 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     4741 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     4801 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     4861 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     4921 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     4981 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     5041 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     5101 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     5161 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     5221 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     5281 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     5341 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     5401 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     5461 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     5521 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     5581 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     5641 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     5701 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     5761 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     5821 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     5881 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     5941 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     6001 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     6061 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     6121 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     6181 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     6241 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     6301 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     6361 tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
     6421 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
     6481 gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg
     6541 agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc
     6601 aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
     6661 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg
     6721 ccagggtttt cccagtcacg acgttgtaaa acgacggcgc aaggaatggt gcatgcaagg
     6781 agatggcgcc caacagtccc ccggccacgg ggcctgccac catacccacg ccgaaacaag
     6841 cgctcatgag cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg gcgatatagg
     6901 cgccagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg gcgtagaggc
     6961 gattagtcca atttgttaaa gacaggatat cagtggtcca ggctctagtt ttgactcaac
     7021 aatatcacca gctgaagcct atagagtacg agccatagat aaaataaaag attttattta
     7081 gtctccagaa aaagggggga atgaaagacc ccacctgtag gtttggcaag ctagcttaag
     7141 taacgccatt ttgcaaggca tggaaaatac ataactgaga atagagaagt tcagatcaag
     7201 gttaggaaca gagagacagc agaatatggg ccaaacagga tatctgtggt aagcagttcc
     7261 tgccccggct cagggccaag aacagatggt ccccagatgc ggtcccgccc tcagcagttt
     7321 ctagagaacc atcagatgtt tccagggtgc cccaaggacc tgaaatgacc ctgtgcctta
     7381 tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct
     7441 caataaaaga gcccacaacc cctcactcgg cgcgccagtc ctccgataga ctgcgtcgcc
     7501 cgggtacccg tattcccaat aaagcctctt gctgtttgca tccgaatcgt ggactcgctg
     7561 atccttggga gggtctcctc agattgattg actgcccacc tcgggggtct ttcatttgga
     7621 ggttccaccg agatttggag acccctgccc agggaccacc gacccccccg ccgggaggta
     7681 agctggccag cggtcgtttc gtgtctgtct ctgtctttgt gcgtgtttgt gccggcatct
     7741 aatgtttgcg cctgcgtctg tactagttag ctaactagct ctgtatctgg cggacccgtg
     7801 gtggaactga cgagttctga acacccggcc gcaaccctgg gagacgtccc agggactttg
     7861 ggggccgttt ttgtggcccg acctgaggaa gggagtcgat gtggaatccg accccgtcag
     7921 gatatgtggt tctggtagga gacgagaacc taaaacagtt cccgcctccg tctgaatttt
     7981 tgctttcggt ttggaaccga agccgcgcgt cttgtctgct gcagcgctgc agcatcgttc
     8041 tgtgttgtct ctgtctgact gtgtttctgt atttgtctga aaattagggc cagactgtta
     8101 ccactccctt aagtttgacc ttaggtcact ggaaagatgt cgagcggatc gctcacaacc
     8161 agtcggtaga tgtcaagaag agacgttggg ttaccttctg ctctgcagaa tggccaacct
     8221 ttaacgtcgg atggccgcga gacggcacct ttaaccgaga cctcatcacc caggttaaga
     8281 tcaaggtctt ttcacctggc ccgcatggac acccagacca ggtcccctac atcgtgacct
     8341 gggaagcctt ggcttttgac ccccctccct gggtcaagcc ctttgtacac cctaagcctc
     8401 cgcctcctct tcctccatcc gccccgtctc tcccccttga acctcctcgt tcgaccccgc
     8461 ctcgatcctc cctttatcca gccctcactc cttctctagg cgccggaatt
//
