LOCUS       sequence-438541-        7832 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of MAVS truncated pLxIS domain in mammalian cells by 
            retroviral transduction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7832)
  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 7832)
  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-MAVS(pLxIS trunc)
FEATURES             Location/Qualifiers
     source          1..7832
                     /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..39
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    1455..2021
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1471..2023
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1615..1632)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1842..1861
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2035..2037,2038..2040,2041..2754)
                     /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(2080..2101)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2341..2360)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2688..2709
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             2816..3330
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    3492..3508
                     /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(3516..3522,3523..3540,3541..3546))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3561..3582
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3699..3716)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3870..4458)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3950..3969)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4629..5420,5421..5489))
                     /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     5252..5271
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5490..5594)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5662..5680
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5718..5740)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5840..5859
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6053..6075
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6203..6222
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             6422..6938
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    7002..7343
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             7410..7826
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     7756..7778
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
ORIGIN
        1 agatctgcca ccatggatta caaagacgat gatgataaga tgacttttgc agaagacaag
       61 acatacaagt atatcaggga taatcacagc aagttctgtt gtgtcgacgt cttggagatt
      121 ctgccctatc ttagttgcct caccgcatcc gatcaggaca ggcttcgggc ttcctatcgc
      181 caaataggca accgcgacac tttgtgggga ctgttcaata accttcaaag aaggccaggc
      241 tgggtagaag tatttattcg agctcttcaa atctgtgaat tgcccggact cgctgaccag
      301 gtcacacgag tttaccagtc atacctccca cccggcacaa gtttgagaag cctcgaacct
      361 ctccagcttc ctgacttccc agccgcagtt tctgggcctt ctgcatttgc acctggtcac
      421 aatatacccg atcacggctt gcgagaaaca ccttcatgtc caaagcccgt acaagacact
      481 caaccacccg aatccccagt tgagaatagc gagcagttgt tgcaaacaaa ttcaggtgcc
      541 gttgcaagaa tgtctggggg gtctctgatt ccaagcccca atcaacaagc cttgagtccc
      601 cagccctcac gagagcacca ggagcaggaa ccagaacttg gcggcgcaca tgccgccaat
      661 gtcgcctcag ttcctatcgc tacttatggg cccgtatccc ctactgtgtc attccagcct
      721 ctgcctagaa cagctcttag aacaaatctt ctgagtgggg taaccgtgag tgccttgagc
      781 gcagatacaa gtctgagtag tagtagcact ggttctgcct ttgctaaggg cgccggtgac
      841 caagcaaagg ccgctacttg cttctcaact acattgacaa actctgttac tacatcttca
      901 gtgccctccc cacgactcgt tcccgtcaag actatgtcct ccaagcttcc cctctcatcc
      961 aagtctactg ccgctatgac cagcacagta ctcactaata ccgccccctc taagctccct
     1021 agcaattcag tttacgccgg aaccgttcca agccgggtcc ctgcaagtgt cgccaaggct
     1081 cccgctaaca caatacctcc agagagaaat tccaagcaag caaaggagac acctgaggga
     1141 ccagcaacaa aagtaactac aggaggcaac caaacaggtc caaatagtag tatccgatca
     1201 ctgcactccg gtccccaagt tgatgagtct ccttccgcag acttgttggg ctctccagaa
     1261 ccactcgcaa cacaacaacc ccaggaggaa gaggagcact gcgccagttc tatgccttgg
     1321 gccaaatggt tgggagctac ttcagccctg ttggccgttt ttctggcagt gatgctttat
     1381 aggtctcggc ggctggcaca atgataagcg gccgcgttta aacgtcgacg gtatcgataa
     1441 gcttgatatc gaattccgcc ccccccccta acgttactgg ccgaagccgc ttggaataag
     1501 gccggtgtgc gtttgtctat atgttatttt ccaccatatt gccgtctttt ggcaatgtga
     1561 gggcccggaa acctggccct gtcttcttga cgagcattcc taggggtctt tcccctctcg
     1621 ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt
     1681 gaagacaaac aacgtctgta gcgacccttt gcaggcagcg gaacccccca cctggcgaca
     1741 ggtgcctctg cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc
     1801 agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa atggctctcc tcaagcgtat
     1861 tcaacaaggg gctgaaggat gcccagaagg taccccattg tatgggatct gatctggggc
     1921 ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga
     1981 accacgggga cgtggttttc ctttgaaaaa cacgatgata atatggccac aaccatggtg
     2041 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
     2101 gtaaacggcc acaagttcag cgtgtctggc gagggcgagg gcgatgccac ctacggcaag
     2161 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
     2221 accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac
     2281 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
     2341 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
     2401 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     2461 gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc
     2521 aaggcgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac
     2581 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     2641 agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
     2701 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaatgaatt
     2761 aattaagaat tcgataaaat aaaagatttt atttagtctc cagaaaaagg ggggaatgaa
     2821 agaccccacc tgtaggtttg gcaagctagc ttaagtaacg ccattttgca aggcatggaa
     2881 aatacataac tgagaataga gaagttcaga tcaaggttag gaacagagag acagcagaat
     2941 atgggccaaa caggatatct gtggtaagca gttcctgccc cggctcaggg ccaagaacag
     3001 atggtcccca gatgcggtcc cgccctcagc agtttctaga gaaccatcag atgtttccag
     3061 ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga actaaccaat cagttcgctt
     3121 ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata aaagagccca caacccctca
     3181 ctcggcgcgc cagtcctccg atagactgcg tcgcccgggt acccgtgtat ccaataaacc
     3241 ctcttgcagt tgcatccgac ttgtggtctc gctgttcctt gggagggtct cctctgagtg
     3301 attgactacc cgtcagcggg ggtctttcat gggtaacagt ttcttgaagt tggagaacaa
     3361 cattctgagg gtaggagtcg aatattaagt aatcctgact caattagcca ctgttttgaa
     3421 tccacatact ccaatactcc tgaaatagtt cattatggac agcgcagaaa gagctgggga
     3481 gaattgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     3541 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     3601 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     3661 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     3721 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     3781 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     3841 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     3901 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     3961 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     4021 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     4081 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     4141 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     4201 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     4261 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     4321 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     4381 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     4441 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     4501 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     4561 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     4621 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     4681 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     4741 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     4801 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     4861 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     4921 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     4981 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     5041 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     5101 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     5161 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     5221 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     5281 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     5341 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     5401 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     5461 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     5521 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     5581 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt
     5641 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc
     5701 ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg
     5761 taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt
     5821 cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg
     5881 tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca ttcgccattc
     5941 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
     6001 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
     6061 cgacgttgta aaacgacggc gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc
     6121 ccccggccac ggggcctgcc accataccca cgccgaaaca agcgctcatg agcccgaagt
     6181 ggcgagcccg atcttcccca tcggtgatgt cggcgatata ggcgccagca accgcacctg
     6241 tggcgccggt gatgccggcc acgatgcgtc cggcgtagag gcgattagtc caatttgtta
     6301 aagacaggat atcagtggtc caggctctag ttttgactca acaatatcac cagctgaagc
     6361 ctatagagta cgagccatag ataaaataaa agattttatt tagtctccag aaaaaggggg
     6421 gaatgaaaga ccccacctgt aggtttggca agctagctta agtaacgcca ttttgcaagg
     6481 catggaaaat acataactga gaatagagaa gttcagatca aggttaggaa cagagagaca
     6541 gcagaatatg ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcagggcca
     6601 agaacagatg gtccccagat gcggtcccgc cctcagcagt ttctagagaa ccatcagatg
     6661 tttccagggt gccccaagga cctgaaatga ccctgtgcct tatttgaact aaccaatcag
     6721 ttcgcttctc gcttctgttc gcgcgcttct gctccccgag ctcaataaaa gagcccacaa
     6781 cccctcactc ggcgcgccag tcctccgata gactgcgtcg cccgggtacc cgtattccca
     6841 ataaagcctc ttgctgtttg catccgaatc gtggactcgc tgatccttgg gagggtctcc
     6901 tcagattgat tgactgccca cctcgggggt ctttcatttg gaggttccac cgagatttgg
     6961 agacccctgc ccagggacca ccgacccccc cgccgggagg taagctggcc agcggtcgtt
     7021 tcgtgtctgt ctctgtcttt gtgcgtgttt gtgccggcat ctaatgtttg cgcctgcgtc
     7081 tgtactagtt agctaactag ctctgtatct ggcggacccg tggtggaact gacgagttct
     7141 gaacacccgg ccgcaaccct gggagacgtc ccagggactt tgggggccgt ttttgtggcc
     7201 cgacctgagg aagggagtcg atgtggaatc cgaccccgtc aggatatgtg gttctggtag
     7261 gagacgagaa cctaaaacag ttcccgcctc cgtctgaatt tttgctttcg gtttggaacc
     7321 gaagccgcgc gtcttgtctg ctgcagcgct gcagcatcgt tctgtgttgt ctctgtctga
     7381 ctgtgtttct gtatttgtct gaaaattagg gccagactgt taccactccc ttaagtttga
     7441 ccttaggtca ctggaaagat gtcgagcgga tcgctcacaa ccagtcggta gatgtcaaga
     7501 agagacgttg ggttaccttc tgctctgcag aatggccaac ctttaacgtc ggatggccgc
     7561 gagacggcac ctttaaccga gacctcatca cccaggttaa gatcaaggtc ttttcacctg
     7621 gcccgcatgg acacccagac caggtcccct acatcgtgac ctgggaagcc ttggcttttg
     7681 acccccctcc ctgggtcaag ccctttgtac accctaagcc tccgcctcct cttcctccat
     7741 ccgccccgtc tctccccctt gaacctcctc gttcgacccc gcctcgatcc tccctttatc
     7801 cagccctcac tccttctcta ggcgccggaa tt
//
