LOCUS       sequence-311685-        5481 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry vector for an inducible 3xFLAG-tagged MAVS mutant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5481)
  AUTHORS   Lopacinski AB, Sweatt AJ, Smolko CM, Gray-Gaillard E, Borgman CA, 
            Shah M, Janes KA
  TITLE     Modeling the complete kinetics of coxsackievirus B3 reveals human 
            determinants of host-cell feedback.
  JOURNAL   Cell Syst. 2021 Apr 21;12(4):304-323.e13. doi: 
            10.1016/j.cels.2021.02.004. Epub 2021 Mar 18.
  PUBMED    33740397
REFERENCE   2  (bases 1 to 5481)
  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.cels.2021.02.004"; journalName: "Cell Syst"; date: 
            "2021-04-21- 21"; volume: "12"; issue: "4"; pages: "304-323.e13"
COMMENT     Sequence Label: pEN_TT 3xFLAG-MAVS-Ala148
FEATURES             Location/Qualifiers
     source          1..5481
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      33..60
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     68..91
                     /label=pENTR-F
                     /note="pENTR vectors, forward primer"
     protein_bind    110..209
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        241..555
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    249..267
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    285..303
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    320..338
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    356..374
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    392..410
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    427..445
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    463..481
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     528..552
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             564..629
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     protein_bind    complement(2294..2393)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2505..2524)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(2583..2602)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     CDS             complement(3176..3709)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(3898..3903,3904..3920,3921..3926))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     rep_origin      4583..5171
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5072..5091
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      5336..5422
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 cccgccataa actgccaggc atcaaactaa gcagaaggcc atcctgacgg atggcctttt
       61 tgcgtttcta caaactcttc ctgttagtta gttacttaag ctcgggcccc aaataatgat
      121 tttattttga ctgatagtga cctgttcgtt gcaacaaatt gataagcaat gcttttttat
      181 aatgccaact ttgtacaaaa aagcaggctt taaaggaacc aattcagtcg actggatccc
      241 gagtttactc cctatcagtg atagagaacg tatgtcgagt ttactcccta tcagtgatag
      301 agaacgatgt cgagtttact ccctatcagt gatagagaac gtatgtcgag tttactccct
      361 atcagtgata gagaacgtat gtcgagttta ctccctatca gtgatagaga acgtatgtcg
      421 agtttatccc tatcagtgat agagaacgta tgtcgagttt actccctatc agtgatagag
      481 aacgtatgtc gaggtaggcg tgtacggtgg gaggcctata taagcagagc tcgtttagtg
      541 aaccgtcaga tcgccactag atggactaca aagaccatga cggtgattat aaagatcatg
      601 acatcgatta caaggatgac gatgacaaga ctagtatgcc gtttgctgaa gacaagacct
      661 ataagtatat ctgccgcaat ttcagcaatt tttgcaatgt ggatgttgta gagattctgc
      721 cttacctgcc ctgcctcaca gcaagagacc aggatcgact gcgggccacc tgcacactct
      781 cagggaaccg ggacaccctc tggcatctct tcaataccct tcagcggcgg cccggctggg
      841 tggagtactt cattgcggca ctgaggggct gtgagctagt tgatctcgcg gacgaagtgg
      901 cctctgtcta cgagagctac cagcctcgga cctcggaccg tcccccagac ccactggagc
      961 caccgtcact tcctgctgag aggccagggc cccccacacc tgctgcggcc cacagcatcc
     1021 cctacaacag ctgcagagag aaggagccaa gttaccccat gcctgtccag gagaccgcgg
     1081 cgccagagtc cccaggagag aattcagagc aagccctgca gacgctcagc cccagagcca
     1141 tcccaaggaa tccagatggt ggccccctgg agtcctcctc tgacctggca gccctcagcc
     1201 ctctgacctc cagcgggcat caggagaagg acacagaact gggcagtacc cacacagcag
     1261 gtgcgacctc cagcctcaca ccatcccgtg ggcctgtgtc tccatctgtc tccttccagc
     1321 ccctggcccg ttccaccccc agggcaagcc gcttgcctgg acccacaggg tcagttgtat
     1381 ctactggcac ctccttctcc tcctcatccc ctggcttggc ctctgcaggg gctgcagagg
     1441 gtaaacaggg tgcagagagt gaccaggccg agcctatcat ctgctccagt ggggcagagg
     1501 cacctgccaa ctctctgccc tccaaagtgc ctaccacctt gatgcctgtg aacacagtgg
     1561 ccctgaaagt gcctgccaac ccagcatctg tcagcacagt gccctccaag ttgccaacta
     1621 gctcaaagcc ccctggtgca gtgccttcta atgcgctcac caatccagca ccatccaaat
     1681 tgcccatcaa ctcaacccgt gctggcatgg tgccatccaa agtgcctact agcatggtgc
     1741 tcaccaaggt gtctgccagc acagtcccca ctgacgggag cagcagaaat gaggagaccc
     1801 cagcagctcc aacacccgcc ggcgccactg gaggcagctc agcctggcta gacagcagct
     1861 ttgagaatag gggccttggg tcggagctga gtaagcctgg cgtgctggca tcccaggtag
     1921 acagcccgtt ctcgggctgc ttcgaggatc ttgccatcag tgccagcacc tccttgggca
     1981 tggggccctg ccatggccca gaggagaatg agtataagtc cgagggcacc tttgggatcc
     2041 acgtggctga gaaccccagc atccagctcc tggagggcaa ccctgggcca cctgcggacc
     2101 cggatggcgg ccccaggcca caagccgacc ggaagttcca ggagagggag gtgccatgcc
     2161 acaggccctc acctggggct ctgtggctcc aggtggctgt gacaggggtg ctggtagtca
     2221 cactcctggt ggtgctgtac cggcggcgtc tgcactagca attggaagac taatgcgttt
     2281 tcgagatatc tagacccagc tttcttgtac aaagttggca ttataagaaa gcattgctta
     2341 tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata aaatcattat ttgccatcca
     2401 gctgcagctc tggcccgtgt ctcaaaatct ctgatgttac attgcacaag ataaaaatat
     2461 atcatcatga acaataaaac tgtctgctta cataaacagt aatacaaggg gtgttatgag
     2521 ccatattcaa cgggaaacgt cgaggccgcg attaaattcc aacatggatg ctgatttata
     2581 tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct atcgcttgta
     2641 tgggaagccc gatgcgccag agttgtttct gaaacatggc aaaggtagcg ttgccaatga
     2701 tgttacagat gagatggtca gactaaactg gctgacggaa tttatgcctc ttccgaccat
     2761 caagcatttt atccgtactc ctgatgatgc atggttactc accactgcga tccccggaaa
     2821 aacagcattc caggtattag aagaatatcc tgattcaggt gaaaatattg ttgatgcgct
     2881 ggcagtgtcc ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt ttaacagcga
     2941 tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag
     3001 tgattttgat gacgagcgta atggctggcc tgttgaacaa gtctggaaag aaatgcataa
     3061 acttttgcca ttctcaccgg attcagtcgt cactcatggt gatttctcac ttgataacct
     3121 tatttttgac gaggggaaat taggccggga agccgatctc ggcttgaacg aattgttagg
     3181 tggcggtact tgggtcgata tcaaagtgca tcacttcttc ccgtatgccc aactttgtat
     3241 agagagccac tgcgggatcg tcaccgtaat ctgcttgcac gtagatcaca taagcaccaa
     3301 gcgcgttggc ctcatgcttg aggagattga tgagcgcggt ggcaatgccc tgcctccggt
     3361 gctcgccgga gactgcgaga tcatagatat agatctcact acgcggctgc tcaaacttgg
     3421 gcagaacgta agccgcgaga gcgccaacaa ccgcttcttg gtcgaaggca gcaagcgcga
     3481 tgaatgtctt actacggagc aagttcccga ggtaatcgga gtccggctga tgttgggagt
     3541 aggtggctac gtctccgaac tcacgaccga aaagatcaag agcagcccgc atggatttga
     3601 cttggtcagg gccgagccta catgtgcgaa tgatgcccat acttgagcca cctaactttg
     3661 ttttagggcg actgccctgc tgcgtaacat cgttgctgct gcgtaacatc gttgctgctc
     3721 cataacatca aacatcgacc cacggcgtaa cgcgcttgct gcttggatgc ccgaggcata
     3781 gactgtacaa aaaaacagtc ataacaagcc atgaaaaccg ccactgcgcc gttaccaccg
     3841 ctgcgttcgg tcaaggttct ggaccagttg cgtgagcgca tacgctactt gcattacagt
     3901 ttacgaaccg aacaggctta tgtcaactgg gttcgtgcct tcatccgttt ccacggtgtg
     3961 cgtcacccgg caaccttggg cagcagcgaa gtcgaggcat ttctgtcctg gctggcgaac
     4021 gagcgcaagg tttcggtctc cacgcatcgt caggcattgg cggccttgct gttcttctac
     4081 ggcaaggtgc tgtgcacgga tctgccctgg cttcaggaga tcggaagacc tcggccgtcg
     4141 cggcgcttgc cggtggtgct gaccccggat gaagtggttc gcatcctcgg ttttctggaa
     4201 ggcgagcatc gtttgttcgc ccaggactct agctatagtt ctagtggttg gctactaata
     4261 ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct tgccatccta
     4321 tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca aaaatatggt
     4381 attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga gtttttctaa
     4441 tcagaattgg ttaattggtt gtaacactgg cagagcatta cgctgacttg acgggacggc
     4501 gcaagctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     4561 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     4621 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     4681 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     4741 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     4801 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     4861 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     4921 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     4981 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     5041 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     5101 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     5161 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     5221 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcta
     5281 gcatggatct cggggacgtc taactactaa gcgagagtag ggaactgcca ggcatcaaat
     5341 aaaacgaaag gctcagtcgg aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa
     5401 cgctctcctg agtaggacaa atccgccggg agcggatttg aacgttgtga agcaacggcc
     5461 cggagggtgg cgggcaggac g
//
