LOCUS       sequence-439016-        9095 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of IRF3 in mammalian cells by retroviral transduction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9095)
  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 9095)
  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-IRF3
FEATURES             Location/Qualifiers
     source          1..9095
                     /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    2718..3284
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    2734..3286
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2878..2895)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3105..3124
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(3298..3300,3301..3303,3304..4017)
                     /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(3343..3364)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3604..3623)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3951..3972
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             4079..4593
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    4755..4771
                     /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(4779..4785,4786..4803,4804..4809))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4824..4845
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4962..4979)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5133..5721)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5213..5232)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5892..6683,6684..6752))
                     /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     6515..6534
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6753..6857)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6925..6943
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6981..7003)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7103..7122
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7316..7338
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7466..7485
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             7685..8201
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    8265..8606
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             8673..9089
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     9019..9041
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     9057..9073
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
ORIGIN
        1 agatctgcca ccatggacta caaggatcac gacggtgatt acaaggacca cgacatcgac
       61 tacaaggacg acgacgacaa gggcgtccag gtagaaacaa tttctcctgg ggatggacgc
      121 acttttccca agaggggaca gacatgcgtt gtacactaca caggtatgct ggaggatgga
      181 aagaaggttg actcatctcg cgatcgcaac aaacctttca aatttatgct ggggaagcag
      241 gaagtaattc gcggatggga agaaggtgtc gcacagatga gcgtcgggca aagagctaaa
      301 ctgacaatca gtccagatta tgcatatggt gccacaggcc accccgggat aatccctcct
      361 cacgcaaccc tcgtctttga tgtcgagctg cttaagctgg agacacgggg agttcaagtt
      421 gaaactatca gtcctggaga cggccgcact ttcccaaaaa gggggcagac ctgtgtggtg
      481 cattataccg ggatgcttga ggatggtaaa aaggtggact cttctagaga taggaacaag
      541 ccctttaaat ttatgcttgg caaacaagag gtgattcggg ggtgggagga aggggttgct
      601 cagatgagcg tagggcaacg cgctaaattg acaatcagtc ctgattatgc atacggagcc
      661 actggacatc cagggattat tccaccccac gctactttgg tatttgatgt agaactgctg
      721 aagttggaga cccggggcgt acaggtagaa accatctccc ccggggatgg gaggaccttc
      781 cctaagcgag ggcagacttg cgtggtgcat tacaccggca tgctggagga cggtaagaag
      841 gttgactcta gtagagaccg caacaagccc ttcaaattta tgcttgggaa acaagaggtt
      901 atacgaggtt gggaagaggg tgtagctcag atgagtgtag gtcagcgagc aaagctcaca
      961 atcagcccag attatgctta tggcgctaca ggacatcccg ggattatacc cccccatgcc
     1021 acacttgtat ttgacgtcga acttttgaag cttgagaccc ggggggttca agtggagacc
     1081 atttcacccg gcgatggaag aaccttcccc aagcgggggc agacctgcgt cgtgcattac
     1141 actggtatgc tcgaagacgg gaagaaggta gattccagcc gggaccggaa caaaccattt
     1201 aagtttatgc tggggaagca agaagtcatt aggggttggg aggagggtgt tgcccagatg
     1261 tctgtaggcc agcgcgcaaa gcttactatt tcacccgact atgcatacgg cgccactgga
     1321 caccctggta ttataccacc acacgccact ctggtgttcg atgtcgagtt gctcaagttg
     1381 gaaggtgcag gggctggggc tatggagacc cccaagccta gaatattgcc ttggctcgta
     1441 agccagcttg acttggggca gttggaggga gtagcatggc tggacgagtc caggaccaga
     1501 ttcaggatac catggaaaca tggactgcgc caggacgcac agatggctga ttttggcata
     1561 ttccaggcct gggctgaagc ttccggggct tacactcccg gaaaagataa accagacgta
     1621 tcaacttgga aaaggaattt ccggtctgca ctcaatagaa aggaagtcct tagattggct
     1681 gccgataata gtaaggaccc ctacgatcca cataaggttt acgaatttgt tacacccggg
     1741 gctagagatt ttgtgcattt gggcgcatcc ccagacacaa atgggaagtc ctcactccca
     1801 cactctcaag aaaatctccc taagctcttc gatggattga tacttggccc actgaaggat
     1861 gaaggaagtt ctgatttggc aatagtcagt gacccaagcc aacagctgcc cagcccaaat
     1921 gttaataatt ttttgaaccc agctccccaa gagaacccct tgaagcaact tcttgcagag
     1981 gaacagtggg aatttgaagt caccgctttt tatcgaggac gccaggtatt tcaacaaaca
     2041 cttttctgcc caggaggtct ccggctcgtt ggctctaccg ccgacatgac tctgccatgg
     2101 cagccagtaa ctcttcccga tccagaggga ttcctgaccg ataaactcgt aaaagagtat
     2161 gttgggcaag tacttaaggg tctgggaaat ggcctcgcac tgtggcaagc cgggcagtgt
     2221 ctgtgggccc agaggcttgg gcatagtcat gctttctggg ccctgggaga ggagcttttg
     2281 ccagattctg gaaggggtcc tgatggagaa gtgcacaaag acaaagatgg agcagttttt
     2341 gatctgaggc ccttcgtagc tgatcttata gcattcatgg agggcagcgg ccattcaccc
     2401 cgatatacac tttggttctg catgggcgag atgtggcccc aggaccaacc ctgggtgaag
     2461 aggctggtaa tggttaaagt tgtccccact tgtcttaaag agcttcttga aatggctaga
     2521 gaaggaggtg ctagtagttt gaagaccgtc gatctgcata tttccaattc acaacctata
     2581 agtctgactt ctgaccagta caaagcctac cttcaggacc ttgttgagga catggatttc
     2641 caggccaccg gtaacatttg ataatgataa gcggccgcgt ttaaacgtcg acggtatcga
     2701 taagcttgat atcgaattcc gccccccccc ctaacgttac tggccgaagc cgcttggaat
     2761 aaggccggtg tgcgtttgtc tatatgttat tttccaccat attgccgtct tttggcaatg
     2821 tgagggcccg gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc
     2881 tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct ctggaagctt
     2941 cttgaagaca aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg
     3001 acaggtgcct ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac
     3061 cccagtgcca cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg
     3121 tattcaacaa ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg
     3181 ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc
     3241 cgaaccacgg ggacgtggtt ttcctttgaa aaacacgatg ataatatggc cacaaccatg
     3301 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
     3361 gacgtaaacg gccacaagtt cagcgtgtct ggcgagggcg agggcgatgc cacctacggc
     3421 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
     3481 gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag
     3541 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
     3601 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
     3661 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
     3721 ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc
     3781 atcaaggcga acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac
     3841 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     3901 ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg
     3961 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaatga
     4021 attaattaag aattcgataa aataaaagat tttatttagt ctccagaaaa aggggggaat
     4081 gaaagacccc acctgtaggt ttggcaagct agcttaagta acgccatttt gcaaggcatg
     4141 gaaaatacat aactgagaat agagaagttc agatcaaggt taggaacaga gagacagcag
     4201 aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa
     4261 cagatggtcc ccagatgcgg tcccgccctc agcagtttct agagaaccat cagatgtttc
     4321 cagggtgccc caaggacctg aaatgaccct gtgccttatt tgaactaacc aatcagttcg
     4381 cttctcgctt ctgttcgcgc gcttctgctc cccgagctca ataaaagagc ccacaacccc
     4441 tcactcggcg cgccagtcct ccgatagact gcgtcgcccg ggtacccgtg tatccaataa
     4501 accctcttgc agttgcatcc gacttgtggt ctcgctgttc cttgggaggg tctcctctga
     4561 gtgattgact acccgtcagc gggggtcttt catgggtaac agtttcttga agttggagaa
     4621 caacattctg agggtaggag tcgaatatta agtaatcctg actcaattag ccactgtttt
     4681 gaatccacat actccaatac tcctgaaata gttcattatg gacagcgcag aaagagctgg
     4741 ggagaattgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     4801 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     4861 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     4921 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     4981 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     5041 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     5101 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     5161 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     5221 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     5281 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     5341 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     5401 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     5461 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     5521 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     5581 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     5641 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     5701 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     5761 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     5821 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     5881 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     5941 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     6001 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     6061 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     6121 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     6181 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     6241 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     6301 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     6361 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     6421 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     6481 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     6541 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     6601 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     6661 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     6721 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     6781 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     6841 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     6901 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt
     6961 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt
     7021 ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg
     7081 tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg
     7141 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca
     7201 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
     7261 ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag
     7321 tcacgacgtt gtaaaacgac ggcgcaagga atggtgcatg caaggagatg gcgcccaaca
     7381 gtcccccggc cacggggcct gccaccatac ccacgccgaa acaagcgctc atgagcccga
     7441 agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac
     7501 ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta gaggcgatta gtccaatttg
     7561 ttaaagacag gatatcagtg gtccaggctc tagttttgac tcaacaatat caccagctga
     7621 agcctataga gtacgagcca tagataaaat aaaagatttt atttagtctc cagaaaaagg
     7681 ggggaatgaa agaccccacc tgtaggtttg gcaagctagc ttaagtaacg ccattttgca
     7741 aggcatggaa aatacataac tgagaataga gaagttcaga tcaaggttag gaacagagag
     7801 acagcagaat atgggccaaa caggatatct gtggtaagca gttcctgccc cggctcaggg
     7861 ccaagaacag atggtcccca gatgcggtcc cgccctcagc agtttctaga gaaccatcag
     7921 atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga actaaccaat
     7981 cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata aaagagccca
     8041 caacccctca ctcggcgcgc cagtcctccg atagactgcg tcgcccgggt acccgtattc
     8101 ccaataaagc ctcttgctgt ttgcatccga atcgtggact cgctgatcct tgggagggtc
     8161 tcctcagatt gattgactgc ccacctcggg ggtctttcat ttggaggttc caccgagatt
     8221 tggagacccc tgcccaggga ccaccgaccc ccccgccggg aggtaagctg gccagcggtc
     8281 gtttcgtgtc tgtctctgtc tttgtgcgtg tttgtgccgg catctaatgt ttgcgcctgc
     8341 gtctgtacta gttagctaac tagctctgta tctggcggac ccgtggtgga actgacgagt
     8401 tctgaacacc cggccgcaac cctgggagac gtcccaggga ctttgggggc cgtttttgtg
     8461 gcccgacctg aggaagggag tcgatgtgga atccgacccc gtcaggatat gtggttctgg
     8521 taggagacga gaacctaaaa cagttcccgc ctccgtctga atttttgctt tcggtttgga
     8581 accgaagccg cgcgtcttgt ctgctgcagc gctgcagcat cgttctgtgt tgtctctgtc
     8641 tgactgtgtt tctgtatttg tctgaaaatt agggccagac tgttaccact cccttaagtt
     8701 tgaccttagg tcactggaaa gatgtcgagc ggatcgctca caaccagtcg gtagatgtca
     8761 agaagagacg ttgggttacc ttctgctctg cagaatggcc aacctttaac gtcggatggc
     8821 cgcgagacgg cacctttaac cgagacctca tcacccaggt taagatcaag gtcttttcac
     8881 ctggcccgca tggacaccca gaccaggtcc cctacatcgt gacctgggaa gccttggctt
     8941 ttgacccccc tccctgggtc aagccctttg tacaccctaa gcctccgcct cctcttcctc
     9001 catccgcccc gtctctcccc cttgaacctc ctcgttcgac cccgcctcga tcctcccttt
     9061 atccagccct cactccttct ctaggcgccg gaatt
//
