LOCUS       sequence-438540-        7979 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of MAVS in mammalian cells by retroviral transduction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7979)
  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 7979)
  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
FEATURES             Location/Qualifiers
     source          1..7979
                     /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    1602..2168
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1618..2170
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1762..1779)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1989..2008
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2182..2184,2185..2187,2188..2901)
                     /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(2227..2248)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2488..2507)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2835..2856
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             2963..3477
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    3639..3655
                     /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(3663..3669,3670..3687,3688..3693))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3708..3729
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3846..3863)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4017..4605)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4097..4116)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4776..5567,5568..5636))
                     /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     5399..5418
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5637..5741)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5809..5827
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5865..5887)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5987..6006
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6200..6222
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6350..6369
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             6569..7085
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    7149..7490
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             7557..7973
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     7903..7925
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     7941..7957
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
ORIGIN
        1 agatctgcca ccatggacta caaagacgat gacgacaaga tgacctttgc agaggacaag
       61 acatataagt acatccgaga taatcatagt aagttctgtt gcgtcgacgt gcttgagata
      121 ctcccatact tgagctgcct tactgcttcc gatcaagaca gactgcgcgc tagttatagg
      181 caaattggta atagagatac actgtggggg ttgtttaaca atctgcaacg acgacctggc
      241 tgggtagagg ttttcattag agctctgcaa atatgtgaac ttcctggtct tgccgaccaa
      301 gttactcgcg tatatcaatc ttacctgcct ccagggacat cactccgcag cctcgaacca
      361 ctccagctgc ccgatttccc agctgctgtg agtgggccca gtgctttcgc ccccggtcat
      421 aatatacccg atcatggcct gcgagagaca ccatcttgtc ccaagcctgt tcaggataca
      481 caacctcctg aatctcctgt cgaaaactcc gagcaactcc tgcaaacaaa ttccggggca
      541 gtagcaagaa tgtccggcgg ctcactcatc ccctccccca accaacaagc attgtcccca
      601 caaccctcta gagagcacca ggagcaagaa cccgagctcg gcggtgcaca tgccgctaac
      661 gtggcttctg tcccaatcgc cacctacggc ccagtcagcc caactgtctc ttttcagcct
      721 ctgccaagga ctgcactgcg aacaaacctt ctttctggcg taacagtcag cgccttgtcc
      781 gccgatacct ctttgagcag ctctagcacc ggatcagcat tcgccaaggg cgctggggac
      841 caggctaaag ctgccacatg cttctccaca acactcacaa actccgtgac aactagctct
      901 gtgccctccc ctcgccttgt tccagttaag accatgtcta gcaagctccc tttgtcctcc
      961 aaatcaaccg ctgcaatgac aagtactgtg cttaccaaca ccgctccatc caaattgcct
     1021 agtaacagtg tgtacgccgg tacagtacca tcacgagttc cagcttccgt cgcaaaggct
     1081 cccgccaata caattcctcc agagcgcaac agcaaacaag ccaaggaaac cccagagggt
     1141 cctgccacta aggtgactac aggggggaac cagacaggac ctaatagttc catccgctcc
     1201 ctgcacagtg ggccagagat gagtaagcct ggtgtcttgg tctctcagct cgatgagccc
     1261 ttttccgcat gttcagttga ccttgccata agccctagca gcagtcttgt tagcgagcca
     1321 aatcatggcc ccgaggaaaa tgaatattct tcttttcgga tccaagttga cgagagtcct
     1381 agcgccgacc ttcttggttc ccccgaaccc cttgccacac aacaaccaca ggaggaggag
     1441 gaacactgcg catccagtat gccatgggcc aagtggctcg gtgcaacatc tgccttgctc
     1501 gctgtcttcc tggcagtgat gctttaccga tccagacgct tggcacaatg ataagcggcc
     1561 gcgtttaaac gtcgacggta tcgataagct tgatatcgaa ttccgccccc ccccctaacg
     1621 ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca
     1681 ccatattgcc gtcttttggc aatgtgaggg cccggaaacc tggccctgtc ttcttgacga
     1741 gcattcctag gggtctttcc cctctcgcca aaggaatgca aggtctgttg aatgtcgtga
     1801 aggaagcagt tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg accctttgca
     1861 ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag
     1921 atacacctgc aaaggcggca caaccccagt gccacgttgt gagttggata gttgtggaaa
     1981 gagtcaaatg gctctcctca agcgtattca acaaggggct gaaggatgcc cagaaggtac
     2041 cccattgtat gggatctgat ctggggcctc ggtgcacatg ctttacatgt gtttagtcga
     2101 ggttaaaaaa acgtctaggc cccccgaacc acggggacgt ggttttcctt tgaaaaacac
     2161 gatgataata tggccacaac catggtgagc aagggcgagg agctgttcac cggggtggtg
     2221 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtctggcgag
     2281 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     2341 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc
     2401 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     2461 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     2521 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     2581 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
     2641 atggccgaca agcagaagaa cggcatcaag gcgaacttca agatccgcca caacatcgag
     2701 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     2761 gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac
     2821 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     2881 atggacgagc tgtacaagta atgaattaat taagaattcg ataaaataaa agattttatt
     2941 tagtctccag aaaaaggggg gaatgaaaga ccccacctgt aggtttggca agctagctta
     3001 agtaacgcca ttttgcaagg catggaaaat acataactga gaatagagaa gttcagatca
     3061 aggttaggaa cagagagaca gcagaatatg ggccaaacag gatatctgtg gtaagcagtt
     3121 cctgccccgg ctcagggcca agaacagatg gtccccagat gcggtcccgc cctcagcagt
     3181 ttctagagaa ccatcagatg tttccagggt gccccaagga cctgaaatga ccctgtgcct
     3241 tatttgaact aaccaatcag ttcgcttctc gcttctgttc gcgcgcttct gctccccgag
     3301 ctcaataaaa gagcccacaa cccctcactc ggcgcgccag tcctccgata gactgcgtcg
     3361 cccgggtacc cgtgtatcca ataaaccctc ttgcagttgc atccgacttg tggtctcgct
     3421 gttccttggg agggtctcct ctgagtgatt gactacccgt cagcgggggt ctttcatggg
     3481 taacagtttc ttgaagttgg agaacaacat tctgagggta ggagtcgaat attaagtaat
     3541 cctgactcaa ttagccactg ttttgaatcc acatactcca atactcctga aatagttcat
     3601 tatggacagc gcagaaagag ctggggagaa ttgtgaaatt gttatccgct cacaattcca
     3661 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     3721 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     3781 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     3841 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     3901 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     3961 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     4021 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     4081 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     4141 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     4201 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     4261 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     4321 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     4381 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     4441 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     4501 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     4561 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     4621 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     4681 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     4741 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     4801 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     4861 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     4921 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     4981 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     5041 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     5101 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     5161 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     5221 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     5281 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     5341 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     5401 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     5461 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     5521 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     5581 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     5641 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     5701 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     5761 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     5821 acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
     5881 ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag
     5941 ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag
     6001 attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa
     6061 taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
     6121 cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt
     6181 tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggcgca aggaatggtg
     6241 catgcaagga gatggcgccc aacagtcccc cggccacggg gcctgccacc atacccacgc
     6301 cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg
     6361 cgatataggc gccagcaacc gcacctgtgg cgccggtgat gccggccacg atgcgtccgg
     6421 cgtagaggcg attagtccaa tttgttaaag acaggatatc agtggtccag gctctagttt
     6481 tgactcaaca atatcaccag ctgaagccta tagagtacga gccatagata aaataaaaga
     6541 ttttatttag tctccagaaa aaggggggaa tgaaagaccc cacctgtagg tttggcaagc
     6601 tagcttaagt aacgccattt tgcaaggcat ggaaaataca taactgagaa tagagaagtt
     6661 cagatcaagg ttaggaacag agagacagca gaatatgggc caaacaggat atctgtggta
     6721 agcagttcct gccccggctc agggccaaga acagatggtc cccagatgcg gtcccgccct
     6781 cagcagtttc tagagaacca tcagatgttt ccagggtgcc ccaaggacct gaaatgaccc
     6841 tgtgccttat ttgaactaac caatcagttc gcttctcgct tctgttcgcg cgcttctgct
     6901 ccccgagctc aataaaagag cccacaaccc ctcactcggc gcgccagtcc tccgatagac
     6961 tgcgtcgccc gggtacccgt attcccaata aagcctcttg ctgtttgcat ccgaatcgtg
     7021 gactcgctga tccttgggag ggtctcctca gattgattga ctgcccacct cgggggtctt
     7081 tcatttggag gttccaccga gatttggaga cccctgccca gggaccaccg acccccccgc
     7141 cgggaggtaa gctggccagc ggtcgtttcg tgtctgtctc tgtctttgtg cgtgtttgtg
     7201 ccggcatcta atgtttgcgc ctgcgtctgt actagttagc taactagctc tgtatctggc
     7261 ggacccgtgg tggaactgac gagttctgaa cacccggccg caaccctggg agacgtccca
     7321 gggactttgg gggccgtttt tgtggcccga cctgaggaag ggagtcgatg tggaatccga
     7381 ccccgtcagg atatgtggtt ctggtaggag acgagaacct aaaacagttc ccgcctccgt
     7441 ctgaattttt gctttcggtt tggaaccgaa gccgcgcgtc ttgtctgctg cagcgctgca
     7501 gcatcgttct gtgttgtctc tgtctgactg tgtttctgta tttgtctgaa aattagggcc
     7561 agactgttac cactccctta agtttgacct taggtcactg gaaagatgtc gagcggatcg
     7621 ctcacaacca gtcggtagat gtcaagaaga gacgttgggt taccttctgc tctgcagaat
     7681 ggccaacctt taacgtcgga tggccgcgag acggcacctt taaccgagac ctcatcaccc
     7741 aggttaagat caaggtcttt tcacctggcc cgcatggaca cccagaccag gtcccctaca
     7801 tcgtgacctg ggaagccttg gcttttgacc cccctccctg ggtcaagccc tttgtacacc
     7861 ctaagcctcc gcctcctctt cctccatccg ccccgtctct cccccttgaa cctcctcgtt
     7921 cgaccccgcc tcgatcctcc ctttatccag ccctcactcc ttctctaggc gccggaatt
//
