LOCUS       sequence-438973-        6835 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Overexpression of MAVS pLxIS domain in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6835)
  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 6835)
  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: pET28a-FKBP-MAVS(pLxIS)
FEATURES             Location/Qualifiers
     source          1..6835
                     /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             1529..1546
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1599..1617)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1613..1660
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1697..2152
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1784..1803)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1994..2015
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2244..3059)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(2244..3059)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2967..2986
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3051..3070
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3181..3769
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3670..3689
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3923..3940
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3955..4094
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4020..4039)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4180..4202
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4196..4387)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5196..6278)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     6240..6259
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6279..6356)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6562..6581
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     6665..6684
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6665..6683
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    6684..6708
                     /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)."
     RBS             6723..6745
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             6764..6781
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             6791..6808
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             6815..6835
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
ORIGIN
        1 ggatccgcca ccatgggcgt tcaagtagaa acaatatctc caggagacgg gcgtacattt
       61 cctaagcgtg gccaaacttg cgtagtccat tataccggca tgctggagga cggtaagaag
      121 gtcgatagct ctagagaccg taacaaacct ttcaaattta tgctggggaa acaggaggtt
      181 atccgtggtt gggaggaagg cgtggcccaa atgtccgttg ggcagcgtgc taagttaacc
      241 atctcgccag attacgcata tggggctaca ggccatcctg ggatcattcc cccgcatgcg
      301 acacttgtgt ttgatgtaga gctgttgaag ttggagacgc ggggagttca agtcgaaacg
      361 atttcaccag gtgatggacg cacctttcct aaacggggac aaacgtgcgt cgtgcattac
      421 accggtatgc tggaagacgg caagaaagtc gattcaagcc gcgatagaaa taaacccttc
      481 aaattcatgc ttggaaaaca ggaagtaata cgcggatggg aagagggtgt ggcgcaaatg
      541 tctgtcgggc aaagagctaa actgacaata agtcctgatt acgcgtacgg tgcgactggg
      601 caccctggta tcattccgcc gcacgcgaca cttgtcttcg acgtcgagtt acttaagctt
      661 gagacgcggg gggtgcaggt agagactatt tcgcccggtg acggaagaac tttcccaaag
      721 cggggacaga cctgtgttgt ccactacact gggatgttgg aggatggtaa gaaagttgac
      781 tcaagtcgtg atcgcaacaa accctttaag tttatgcttg ggaagcagga agtcatcaga
      841 ggttgggaag aaggcgtagc tcagatgtca gtgggacaac gggccaaatt aaccatcagt
      901 ccagactacg cctacggagc caccggtcac ccagggatca taccacctca cgcaacctta
      961 gtgttcgatg tggaactttt aaagttagag acgcgcggtg tacaagtgga aacaatcagt
     1021 ccaggggatg ggagaacatt ccccaaaaga ggccagacgt gtgtcgtgca ctatacgggg
     1081 atgttagaag atgggaaaaa ggtcgattcg tcacgcgacc gcaacaaacc gtttaaattc
     1141 atgttgggaa agcaggaggt gattcgcggc tgggaggaag gtgtggcaca gatgagcgtg
     1201 ggccagagag caaaactgac tatctctcct gattatgcct acggcgcgac gggccaccct
     1261 ggcataattc caccccacgc taccctggta ttcgatgtcg agcttctgaa gcttgagggt
     1321 gctggggctg gggctgagat gagcaagcca ggtgtgctgg tatcccagtt ggacgagcca
     1381 ttctcagcct gctctgtgga ccttgccatt agccctagca gctccttggt ctcagaaccc
     1441 aaccatggtc cagaggagaa tgagtattcg tcctttagaa tctgataaga attcgagctc
     1501 cgtcgacaag cttgcggccg cactcgagca ccaccaccac caccactgag atccggctgc
     1561 taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactagcata
     1621 accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag gaactatatc
     1681 cggattggcg aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     1741 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     1801 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     1861 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     1921 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     1981 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     2041 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     2101 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat ttaggtggca
     2161 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     2221 tgtatccgct catgaattaa ttcttagaaa aactcatcga gcatcaaatg aaactgcaat
     2281 ttattcatat caggattatc aataccatat ttttgaaaaa gccgtttctg taatgaagga
     2341 gaaaactcac cgaggcagtt ccataggatg gcaagatcct ggtatcggtc tgcgattccg
     2401 actcgtccaa catcaataca acctattaat ttcccctcgt caaaaataag gttatcaagt
     2461 gagaaatcac catgagtgac gactgaatcc ggtgagaatg gcaaaagttt atgcatttct
     2521 ttccagactt gttcaacagg ccagccatta cgctcgtcat caaaatcact cgcatcaacc
     2581 aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa atacgcgatc gctgttaaaa
     2641 ggacaattac aaacaggaat cgaatgcaac cggcgcagga acactgccag cgcatcaaca
     2701 atattttcac ctgaatcagg atattcttct aatacctgga atgctgtttt cccggggatc
     2761 gcagtggtga gtaaccatgc atcatcagga gtacggataa aatgcttgat ggtcggaaga
     2821 ggcataaatt ccgtcagcca gtttagtctg accatctcat ctgtaacatc attggcaacg
     2881 ctacctttgc catgtttcag aaacaactct ggcgcatcgg gcttcccata caatcgatag
     2941 attgtcgcac ctgattgccc gacattatcg cgagcccatt tatacccata taaatcagca
     3001 tccatgttgg aatttaatcg cggcctagag caagacgttt cccgttgaat atggctcata
     3061 acaccccttg tattactgtt tatgtaagca gacagtttta ttgttcatga ccaaaatccc
     3121 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     3181 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     3241 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     3301 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
     3361 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     3421 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     3481 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     3541 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     3601 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     3661 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     3721 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     3781 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     3841 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     3901 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat
     3961 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcaatggtg cactctcagt
     4021 acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg ctacgtgact
     4081 gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     4141 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     4201 ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt
     4261 cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc cagctcgttg agtttctcca
     4321 gaagcgttaa tgtctggctt ctgataaagc gggccatgtt aagggcggtt ttttcctgtt
     4381 tggtcactga tgcctccgtg taagggggat ttctgttcat gggggtaatg ataccgatga
     4441 aacgagagag gatgctcacg atacgggtta ctgatgatga acatgcccgg ttactggaac
     4501 gttgtgaggg taaacaactg gcggtatgga tgcggcggga ccagagaaaa atcactcagg
     4561 gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc acagggtagc cagcagcatc
     4621 ctgcgatgca gatccggaac ataatggtgc agggcgctga cttccgcgtt tccagacttt
     4681 acgaaacacg gaaaccgaag accattcatg ttgttgctca ggtcgcagac gttttgcagc
     4741 agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac
     4801 cccgccagcc tagccgggtc ctcaacgaca ggagcacgat catgcgcacc cgtggggccg
     4861 ccatgccggc gataatggcc tgcttctcgc cgaaacgttt ggtggcggga ccagtgacga
     4921 aggcttgagc gagggcgtgc aagattccga ataccgcaag cgacaggccg atcatcgtcg
     4981 cgctccagcg aaagcggtcc tcgccgaaaa tgacccagag cgctgccggc acctgtccta
     5041 cgagttgcat gataaagaag acagtcataa gtgcggcgac gatagtcatg ccccgcgccc
     5101 accggaagga gctgactggg ttgaaggctc tcaagggcat cggtcgagat cccggtgcct
     5161 aatgagtgag ctaacttaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     5221 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     5281 ttgggcgcca gggtggtttt tcttttcacc agtgagacgg gcaacagctg attgcccttc
     5341 accgcctggc cctgagagag ttgcagcaag cggtccacgc tggtttgccc cagcaggcga
     5401 aaatcctgtt tgatggtggt taacggcggg atataacatg agctgtcttc ggtatcgtcg
     5461 tatcccacta ccgagatatc cgcaccaacg cgcagcccgg actcggtaat ggcgcgcatt
     5521 gcgcccagcg ccatctgatc gttggcaacc agcatcgcag tgggaacgat gccctcattc
     5581 agcatttgca tggtttgttg aaaaccggac atggcactcc agtcgccttc ccgttccgct
     5641 atcggctgaa tttgattgcg agtgagatat ttatgccagc cagccagacg cagacgcgcc
     5701 gagacagaac ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga
     5761 tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa taatactgtt gatgggtgtc
     5821 tggtcagaga catcaagaaa taacgccgga acattagtgc aggcagcttc cacagcaatg
     5881 gcatcctggt catccagcgg atagttaatg atcagcccac tgacgcgttg cgcgagaaga
     5941 ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga caccaccacg
     6001 ctggcaccca gttgatcggc gcgagattta atcgccgcga caatttgcga cggcgcgtgc
     6061 agggccagac tggaggtggc aacgccaatc agcaacgact gtttgcccgc cagttgttgt
     6121 gccacgcggt tgggaatgta attcagctcc gccatcgccg cttccacttt ttcccgcgtt
     6181 ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa cggtctgata agagacaccg
     6241 gcatactctg cgacatcgta taacgttact ggtttcacat tcaccaccct gaattgactc
     6301 tcttccgggc gctatcatgc cataccgcga aaggttttgc gccattcgat ggtgtccggg
     6361 atctcgacgc tctcccttat gcgactcctg cattaggaag cagcccagta gtaggttgag
     6421 gccgttgagc accgccgccg caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc
     6481 cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgctcatga gcccgaagtg
     6541 gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt
     6601 ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg atcgagatct cgatcccgcg
     6661 aaattaatac gactcactat aggggaattg tgagcggata acaattcccc tctagaaata
     6721 attttgttta actttaagaa ggagatatac catgggcagc agccatcatc atcatcatca
     6781 cagcagcggc ctggtgccgc gcggcagcca tatggaaaac ctgtattttc agggc
//
