LOCUS       sequence-438523-        7949 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of STING L373A with IRSp53 pLxIS (RLL361NLV) in mammalian
            cells by retroviral transduction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7949)
  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 7949)
  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 L373A with IRSp53 pLxIS
FEATURES             Location/Qualifiers
     source          1..7949
                     /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    1572..2138
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1588..2140
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1732..1749)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1959..1978
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2152..2154,2155..2157,2158..2871)
                     /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(2197..2218)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2458..2477)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2805..2826
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             2933..3447
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    3609..3625
                     /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(3633..3639,3640..3657,3658..3663))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3678..3699
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3816..3833)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3987..4575)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4067..4086)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4746..5537,5538..5606))
                     /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     5369..5388
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5607..5711)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5779..5797
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5835..5857)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5957..5976
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6170..6192
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6320..6339
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             6539..7055
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    7119..7460
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             7527..7943
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     7873..7895
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     7911..7927
                     /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 tgttacaatg aatgcgccta tgactagtgt ggccccgccg
     1441 ccttcggttt tatcacagga gccgaacctg gtaattagcg gtatggatca accgctgcct
     1501 gcccggaccg atttgatctg ataagcggcc gcgtttaaac gtcgacggta tcgataagct
     1561 tgatatcgaa ttccgccccc ccccctaacg ttactggccg aagccgcttg gaataaggcc
     1621 ggtgtgcgtt tgtctatatg ttattttcca ccatattgcc gtcttttggc aatgtgaggg
     1681 cccggaaacc tggccctgtc ttcttgacga gcattcctag gggtctttcc cctctcgcca
     1741 aaggaatgca aggtctgttg aatgtcgtga aggaagcagt tcctctggaa gcttcttgaa
     1801 gacaaacaac gtctgtagcg accctttgca ggcagcggaa ccccccacct ggcgacaggt
     1861 gcctctgcgg ccaaaagcca cgtgtataag atacacctgc aaaggcggca caaccccagt
     1921 gccacgttgt gagttggata gttgtggaaa gagtcaaatg gctctcctca agcgtattca
     1981 acaaggggct gaaggatgcc cagaaggtac cccattgtat gggatctgat ctggggcctc
     2041 ggtgcacatg ctttacatgt gtttagtcga ggttaaaaaa acgtctaggc cccccgaacc
     2101 acggggacgt ggttttcctt tgaaaaacac gatgataata tggccacaac catggtgagc
     2161 aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta
     2221 aacggccaca agttcagcgt gtctggcgag ggcgagggcg atgccaccta cggcaagctg
     2281 accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc
     2341 accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac
     2401 ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac
     2461 gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc
     2521 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag
     2581 tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag
     2641 gcgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac
     2701 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc
     2761 acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag
     2821 ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta atgaattaat
     2881 taagaattcg ataaaataaa agattttatt tagtctccag aaaaaggggg gaatgaaaga
     2941 ccccacctgt aggtttggca agctagctta agtaacgcca ttttgcaagg catggaaaat
     3001 acataactga gaatagagaa gttcagatca aggttaggaa cagagagaca gcagaatatg
     3061 ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcagggcca agaacagatg
     3121 gtccccagat gcggtcccgc cctcagcagt ttctagagaa ccatcagatg tttccagggt
     3181 gccccaagga cctgaaatga ccctgtgcct tatttgaact aaccaatcag ttcgcttctc
     3241 gcttctgttc gcgcgcttct gctccccgag ctcaataaaa gagcccacaa cccctcactc
     3301 ggcgcgccag tcctccgata gactgcgtcg cccgggtacc cgtgtatcca ataaaccctc
     3361 ttgcagttgc atccgacttg tggtctcgct gttccttggg agggtctcct ctgagtgatt
     3421 gactacccgt cagcgggggt ctttcatggg taacagtttc ttgaagttgg agaacaacat
     3481 tctgagggta ggagtcgaat attaagtaat cctgactcaa ttagccactg ttttgaatcc
     3541 acatactcca atactcctga aatagttcat tatggacagc gcagaaagag ctggggagaa
     3601 ttgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     3661 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     3721 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     3781 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     3841 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     3901 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     3961 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     4021 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     4081 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     4141 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     4201 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     4261 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     4321 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     4381 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     4441 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     4501 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     4561 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     4621 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     4681 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     4741 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     4801 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     4861 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     4921 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     4981 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     5041 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     5101 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     5161 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     5221 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     5281 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     5341 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     5401 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     5461 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     5521 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     5581 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     5641 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     5701 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc
     5761 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt
     5821 gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa
     5881 gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg
     5941 ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt
     6001 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc gccattcagg
     6061 ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg
     6121 aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga
     6181 cgttgtaaaa cgacggcgca aggaatggtg catgcaagga gatggcgccc aacagtcccc
     6241 cggccacggg gcctgccacc atacccacgc cgaaacaagc gctcatgagc ccgaagtggc
     6301 gagcccgatc ttccccatcg gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg
     6361 cgccggtgat gccggccacg atgcgtccgg cgtagaggcg attagtccaa tttgttaaag
     6421 acaggatatc agtggtccag gctctagttt tgactcaaca atatcaccag ctgaagccta
     6481 tagagtacga gccatagata aaataaaaga ttttatttag tctccagaaa aaggggggaa
     6541 tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat
     6601 ggaaaataca taactgagaa tagagaagtt cagatcaagg ttaggaacag agagacagca
     6661 gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga
     6721 acagatggtc cccagatgcg gtcccgccct cagcagtttc tagagaacca tcagatgttt
     6781 ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc
     6841 gcttctcgct tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc
     6901 ctcactcggc gcgccagtcc tccgatagac tgcgtcgccc gggtacccgt attcccaata
     6961 aagcctcttg ctgtttgcat ccgaatcgtg gactcgctga tccttgggag ggtctcctca
     7021 gattgattga ctgcccacct cgggggtctt tcatttggag gttccaccga gatttggaga
     7081 cccctgccca gggaccaccg acccccccgc cgggaggtaa gctggccagc ggtcgtttcg
     7141 tgtctgtctc tgtctttgtg cgtgtttgtg ccggcatcta atgtttgcgc ctgcgtctgt
     7201 actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac gagttctgaa
     7261 cacccggccg caaccctggg agacgtccca gggactttgg gggccgtttt tgtggcccga
     7321 cctgaggaag ggagtcgatg tggaatccga ccccgtcagg atatgtggtt ctggtaggag
     7381 acgagaacct aaaacagttc ccgcctccgt ctgaattttt gctttcggtt tggaaccgaa
     7441 gccgcgcgtc ttgtctgctg cagcgctgca gcatcgttct gtgttgtctc tgtctgactg
     7501 tgtttctgta tttgtctgaa aattagggcc agactgttac cactccctta agtttgacct
     7561 taggtcactg gaaagatgtc gagcggatcg ctcacaacca gtcggtagat gtcaagaaga
     7621 gacgttgggt taccttctgc tctgcagaat ggccaacctt taacgtcgga tggccgcgag
     7681 acggcacctt taaccgagac ctcatcaccc aggttaagat caaggtcttt tcacctggcc
     7741 cgcatggaca cccagaccag gtcccctaca tcgtgacctg ggaagccttg gcttttgacc
     7801 cccctccctg ggtcaagccc tttgtacacc ctaagcctcc gcctcctctt cctccatccg
     7861 ccccgtctct cccccttgaa cctcctcgtt cgaccccgcc tcgatcctcc ctttatccag
     7921 ccctcactcc ttctctaggc gccggaatt
//
