LOCUS       sequence-311820-        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-Gln93,Ala271
FEATURES             Location/Qualifiers
     source          1..5481
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        21..335
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    29..47
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    65..83
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    100..118
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    136..154
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    172..190
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    207..225
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    243..261
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     308..332
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             344..409
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     protein_bind    complement(2074..2173)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2285..2304)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(2363..2382)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     CDS             complement(2956..3489)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(3678..3683,3684..3700,3701..3706))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     rep_origin      4363..4951
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4852..4871
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      5116..5202
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5294..5321
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     5329..5352
                     /label=pENTR-F
                     /note="pENTR vectors, forward primer"
     protein_bind    5371..5470
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 aattcagtcg actggatccc gagtttactc cctatcagtg atagagaacg tatgtcgagt
       61 ttactcccta tcagtgatag agaacgatgt cgagtttact ccctatcagt gatagagaac
      121 gtatgtcgag tttactccct atcagtgata gagaacgtat gtcgagttta ctccctatca
      181 gtgatagaga acgtatgtcg agtttatccc tatcagtgat agagaacgta tgtcgagttt
      241 actccctatc agtgatagag aacgtatgtc gaggtaggcg tgtacggtgg gaggcctata
      301 taagcagagc tcgtttagtg aaccgtcaga tcgccactag atggactaca aagaccatga
      361 cggtgattat aaagatcatg acatcgatta caaggatgac gatgacaaga ctagtatgcc
      421 gtttgctgaa gacaagacct ataagtatat ctgccgcaat ttcagcaatt tttgcaatgt
      481 ggatgttgta gagattctgc cttacctgcc ctgcctcaca gcaagagacc aggatcgact
      541 gcgggccacc tgcacactct cagggaaccg ggacaccctc tggcatctct tcaataccct
      601 tcagcggcgg cccggctggg tggagtactt cattgcggca ctgaggggct gtgagctagt
      661 tgatctcgcg gacgaagtgg cctctgtcta ccagagctac cagcctcgga cctcggaccg
      721 tcccccagac ccactggagc caccgtcact tcctgctgag aggccagggc cccccacacc
      781 tgctgcggcc cacagcatcc cctacaacag ctgcagagag aaggagccaa gttaccccat
      841 gcctgtccag gagacccagg cgccagagtc cccaggagag aattcagagc aagccctgca
      901 gacgctcagc cccagagcca tcccaaggaa tccagatggt ggccccctgg agtcctcctc
      961 tgacctggca gccctcagcc ctctgacctc cagcgggcat caggagaagg acacagaact
     1021 gggcagtacc cacacagcag gtgcgacctc cagcctcaca ccatcccgtg ggcctgtgtc
     1081 tccatctgtc tccttccagc ccctggcccg ttccaccccc agggcaagcc gcttgcctgg
     1141 acccacaggg tcagttgtat ctactggcac ctccttctcc tcctcatccc ctggcttggc
     1201 ctctgcaggg gctgcagagg gtaaagcggg tgcagagagt gaccaggccg agcctatcat
     1261 ctgctccagt ggggcagagg cacctgccaa ctctctgccc tccaaagtgc ctaccacctt
     1321 gatgcctgtg aacacagtgg ccctgaaagt gcctgccaac ccagcatctg tcagcacagt
     1381 gccctccaag ttgccaacta gctcaaagcc ccctggtgca gtgccttcta atgcgctcac
     1441 caatccagca ccatccaaat tgcccatcaa ctcaacccgt gctggcatgg tgccatccaa
     1501 agtgcctact agcatggtgc tcaccaaggt gtctgccagc acagtcccca ctgacgggag
     1561 cagcagaaat gaggagaccc cagcagctcc aacacccgcc ggcgccactg gaggcagctc
     1621 agcctggcta gacagcagct ttgagaatag gggccttggg tcggagctga gtaagcctgg
     1681 cgtgctggca tcccaggtag acagcccgtt ctcgggctgc ttcgaggatc ttgccatcag
     1741 tgccagcacc tccttgggca tggggccctg ccatggccca gaggagaatg agtataagtc
     1801 cgagggcacc tttgggatcc acgtggctga gaaccccagc atccagctcc tggagggcaa
     1861 ccctgggcca cctgcggacc cggatggcgg ccccaggcca caagccgacc ggaagttcca
     1921 ggagagggag gtgccatgcc acaggccctc acctggggct ctgtggctcc aggtggctgt
     1981 gacaggggtg ctggtagtca cactcctggt ggtgctgtac cggcggcgtc tgcactagca
     2041 attggaagac taatgcgttt tcgagatatc tagacccagc tttcttgtac aaagttggca
     2101 ttataagaaa gcattgctta tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata
     2161 aaatcattat ttgccatcca gctgcagctc tggcccgtgt ctcaaaatct ctgatgttac
     2221 attgcacaag ataaaaatat atcatcatga acaataaaac tgtctgctta cataaacagt
     2281 aatacaaggg gtgttatgag ccatattcaa cgggaaacgt cgaggccgcg attaaattcc
     2341 aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt
     2401 gcgacaatct atcgcttgta tgggaagccc gatgcgccag agttgtttct gaaacatggc
     2461 aaaggtagcg ttgccaatga tgttacagat gagatggtca gactaaactg gctgacggaa
     2521 tttatgcctc ttccgaccat caagcatttt atccgtactc ctgatgatgc atggttactc
     2581 accactgcga tccccggaaa aacagcattc caggtattag aagaatatcc tgattcaggt
     2641 gaaaatattg ttgatgcgct ggcagtgtcc ctgcgccggt tgcattcgat tcctgtttgt
     2701 aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat
     2761 aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc tgttgaacaa
     2821 gtctggaaag aaatgcataa acttttgcca ttctcaccgg attcagtcgt cactcatggt
     2881 gatttctcac ttgataacct tatttttgac gaggggaaat taggccggga agccgatctc
     2941 ggcttgaacg aattgttagg tggcggtact tgggtcgata tcaaagtgca tcacttcttc
     3001 ccgtatgccc aactttgtat agagagccac tgcgggatcg tcaccgtaat ctgcttgcac
     3061 gtagatcaca taagcaccaa gcgcgttggc ctcatgcttg aggagattga tgagcgcggt
     3121 ggcaatgccc tgcctccggt gctcgccgga gactgcgaga tcatagatat agatctcact
     3181 acgcggctgc tcaaacttgg gcagaacgta agccgcgaga gcgccaacaa ccgcttcttg
     3241 gtcgaaggca gcaagcgcga tgaatgtctt actacggagc aagttcccga ggtaatcgga
     3301 gtccggctga tgttgggagt aggtggctac gtctccgaac tcacgaccga aaagatcaag
     3361 agcagcccgc atggatttga cttggtcagg gccgagccta catgtgcgaa tgatgcccat
     3421 acttgagcca cctaactttg ttttagggcg actgccctgc tgcgtaacat cgttgctgct
     3481 gcgtaacatc gttgctgctc cataacatca aacatcgacc cacggcgtaa cgcgcttgct
     3541 gcttggatgc ccgaggcata gactgtacaa aaaaacagtc ataacaagcc atgaaaaccg
     3601 ccactgcgcc gttaccaccg ctgcgttcgg tcaaggttct ggaccagttg cgtgagcgca
     3661 tacgctactt gcattacagt ttacgaaccg aacaggctta tgtcaactgg gttcgtgcct
     3721 tcatccgttt ccacggtgtg cgtcacccgg caaccttggg cagcagcgaa gtcgaggcat
     3781 ttctgtcctg gctggcgaac gagcgcaagg tttcggtctc cacgcatcgt caggcattgg
     3841 cggccttgct gttcttctac ggcaaggtgc tgtgcacgga tctgccctgg cttcaggaga
     3901 tcggaagacc tcggccgtcg cggcgcttgc cggtggtgct gaccccggat gaagtggttc
     3961 gcatcctcgg ttttctggaa ggcgagcatc gtttgttcgc ccaggactct agctatagtt
     4021 ctagtggttg gctactaata ggttgtattg atgttagacg agtcggaatc gcagaccgat
     4081 accaggatct tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac
     4141 ggctttttca aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga
     4201 tgctcgatga gtttttctaa tcagaattgg ttaattggtt gtaacactgg cagagcatta
     4261 cgctgacttg acgggacggc gcaagctcat gaccaaaatc ccttaacgtg agttttcgtt
     4321 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     4381 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     4441 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     4501 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     4561 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     4621 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     4681 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     4741 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     4801 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     4861 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     4921 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     4981 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     5041 ggataaccgt attaccgcta gcatggatct cggggacgtc taactactaa gcgagagtag
     5101 ggaactgcca ggcatcaaat aaaacgaaag gctcagtcgg aagactgggc ctttcgtttt
     5161 atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa atccgccggg agcggatttg
     5221 aacgttgtga agcaacggcc cggagggtgg cgggcaggac gcccgccata aactgccagg
     5281 catcaaacta agcagaaggc catcctgacg gatggccttt ttgcgtttct acaaactctt
     5341 cctgttagtt agttacttaa gctcgggccc caaataatga ttttattttg actgatagtg
     5401 acctgttcgt tgcaacaaat tgataagcaa tgctttttta taatgccaac tttgtacaaa
     5461 aaagcaggct ttaaaggaac c
//
