LOCUS       sequence-324888-        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-Ala271
FEATURES             Location/Qualifiers
     source          1..5481
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(join(173..178,179..195,196..201))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     rep_origin      858..1446
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1347..1366
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      1611..1697
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1789..1816
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     1824..1847
                     /label=pENTR-F
                     /note="pENTR vectors, forward primer"
     protein_bind    1866..1965
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        1997..2311
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    2005..2023
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2041..2059
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2076..2094
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2112..2130
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2148..2166
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2183..2201
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2219..2237
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2284..2308
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2320..2385
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     protein_bind    complement(4050..4149)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4261..4280)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(4339..4358)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     CDS             complement(4932..5465)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
ORIGIN
        1 catcaaacat cgacccacgg cgtaacgcgc ttgctgcttg gatgcccgag gcatagactg
       61 tacaaaaaaa cagtcataac aagccatgaa aaccgccact gcgccgttac caccgctgcg
      121 ttcggtcaag gttctggacc agttgcgtga gcgcatacgc tacttgcatt acagtttacg
      181 aaccgaacag gcttatgtca actgggttcg tgccttcatc cgtttccacg gtgtgcgtca
      241 cccggcaacc ttgggcagca gcgaagtcga ggcatttctg tcctggctgg cgaacgagcg
      301 caaggtttcg gtctccacgc atcgtcaggc attggcggcc ttgctgttct tctacggcaa
      361 ggtgctgtgc acggatctgc cctggcttca ggagatcgga agacctcggc cgtcgcggcg
      421 cttgccggtg gtgctgaccc cggatgaagt ggttcgcatc ctcggttttc tggaaggcga
      481 gcatcgtttg ttcgcccagg actctagcta tagttctagt ggttggctac taataggttg
      541 tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa
      601 ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga
      661 taatcctgat atgaataaat tgcagtttca tttgatgctc gatgagtttt tctaatcaga
      721 attggttaat tggttgtaac actggcagag cattacgctg acttgacggg acggcgcaag
      781 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
      841 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
      901 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
      961 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     1021 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     1081 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     1141 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     1201 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     1261 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     1321 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     1381 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     1441 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     1501 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgctagcatg
     1561 gatctcgggg acgtctaact actaagcgag agtagggaac tgccaggcat caaataaaac
     1621 gaaaggctca gtcggaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc
     1681 tcctgagtag gacaaatccg ccgggagcgg atttgaacgt tgtgaagcaa cggcccggag
     1741 ggtggcgggc aggacgcccg ccataaactg ccaggcatca aactaagcag aaggccatcc
     1801 tgacggatgg cctttttgcg tttctacaaa ctcttcctgt tagttagtta cttaagctcg
     1861 ggccccaaat aatgatttta ttttgactga tagtgacctg ttcgttgcaa caaattgata
     1921 agcaatgctt ttttataatg ccaactttgt acaaaaaagc aggctttaaa ggaaccaatt
     1981 cagtcgactg gatcccgagt ttactcccta tcagtgatag agaacgtatg tcgagtttac
     2041 tccctatcag tgatagagaa cgatgtcgag tttactccct atcagtgata gagaacgtat
     2101 gtcgagttta ctccctatca gtgatagaga acgtatgtcg agtttactcc ctatcagtga
     2161 tagagaacgt atgtcgagtt tatccctatc agtgatagag aacgtatgtc gagtttactc
     2221 cctatcagtg atagagaacg tatgtcgagg taggcgtgta cggtgggagg cctatataag
     2281 cagagctcgt ttagtgaacc gtcagatcgc cactagatgg actacaaaga ccatgacggt
     2341 gattataaag atcatgacat cgattacaag gatgacgatg acaagactag tatgccgttt
     2401 gctgaagaca agacctataa gtatatctgc cgcaatttca gcaatttttg caatgtggat
     2461 gttgtagaga ttctgcctta cctgccctgc ctcacagcaa gagaccagga tcgactgcgg
     2521 gccacctgca cactctcagg gaaccgggac accctctggc atctcttcaa tacccttcag
     2581 cggcggcccg gctgggtgga gtacttcatt gcggcactga ggggctgtga gctagttgat
     2641 ctcgcggacg aagtggcctc tgtctacgag agctaccagc ctcggacctc ggaccgtccc
     2701 ccagacccac tggagccacc gtcacttcct gctgagaggc cagggccccc cacacctgct
     2761 gcggcccaca gcatccccta caacagctgc agagagaagg agccaagtta ccccatgcct
     2821 gtccaggaga cccaggcgcc agagtcccca ggagagaatt cagagcaagc cctgcagacg
     2881 ctcagcccca gagccatccc aaggaatcca gatggtggcc ccctggagtc ctcctctgac
     2941 ctggcagccc tcagccctct gacctccagc gggcatcagg agaaggacac agaactgggc
     3001 agtacccaca cagcaggtgc gacctccagc ctcacaccat cccgtgggcc tgtgtctcca
     3061 tctgtctcct tccagcccct ggcccgttcc acccccaggg caagccgctt gcctggaccc
     3121 acagggtcag ttgtatctac tggcacctcc ttctcctcct catcccctgg cttggcctct
     3181 gcaggggctg cagagggtaa agcgggtgca gagagtgacc aggccgagcc tatcatctgc
     3241 tccagtgggg cagaggcacc tgccaactct ctgccctcca aagtgcctac caccttgatg
     3301 cctgtgaaca cagtggccct gaaagtgcct gccaacccag catctgtcag cacagtgccc
     3361 tccaagttgc caactagctc aaagccccct ggtgcagtgc cttctaatgc gctcaccaat
     3421 ccagcaccat ccaaattgcc catcaactca acccgtgctg gcatggtgcc atccaaagtg
     3481 cctactagca tggtgctcac caaggtgtct gccagcacag tccccactga cgggagcagc
     3541 agaaatgagg agaccccagc agctccaaca cccgccggcg ccactggagg cagctcagcc
     3601 tggctagaca gcagctttga gaataggggc cttgggtcgg agctgagtaa gcctagcgtg
     3661 ctggcatccc aggtagacag cccgttctcg ggctgcttcg aggatcttgc catcagtgcc
     3721 agcacctcct tgggcatggg gccctgccat ggcccagagg agaatgagta taagtccgag
     3781 ggcacctttg ggatccacgt ggctgagaac cccagcatcc agctcctgga gggcaaccct
     3841 gggccacctg cggacccgga tggcggcccc aggccacaag ccgaccggaa gttccaggag
     3901 agggaggtgc catgccacag gccctcacct ggggctctgt ggctccaggt ggctgtgaca
     3961 ggggtgctgg tagtcacact cctggtggtg ctgtaccggc ggcgtctgca ctagcaattg
     4021 gaagactaat gcgttttcga gatatctaga cccagctttc ttgtacaaag ttggcattat
     4081 aagaaagcat tgcttatcaa tttgttgcaa cgaacaggtc actatcagtc aaaataaaat
     4141 cattatttgc catccagctg cagctctggc ccgtgtctca aaatctctga tgttacattg
     4201 cacaagataa aaatatatca tcatgaacaa taaaactgtc tgcttacata aacagtaata
     4261 caaggggtgt tatgagccat attcaacggg aaacgtcgag gccgcgatta aattccaaca
     4321 tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga
     4381 caatctatcg cttgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag
     4441 gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta
     4501 tgcctcttcc gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca
     4561 ctgcgatccc cggaaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa
     4621 atattgttga tgcgctggca gtgtccctgc gccggttgca ttcgattcct gtttgtaatt
     4681 gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga atgaataacg
     4741 gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct
     4801 ggaaagaaat gcataaactt ttgccattct caccggattc agtcgtcact catggtgatt
     4861 tctcacttga taaccttatt tttgacgagg ggaaattagg ccgggaagcc gatctcggct
     4921 tgaacgaatt gttaggtggc ggtacttggg tcgatatcaa agtgcatcac ttcttcccgt
     4981 atgcccaact ttgtatagag agccactgcg ggatcgtcac cgtaatctgc ttgcacgtag
     5041 atcacataag caccaagcgc gttggcctca tgcttgagga gattgatgag cgcggtggca
     5101 atgccctgcc tccggtgctc gccggagact gcgagatcat agatatagat ctcactacgc
     5161 ggctgctcaa acttgggcag aacgtaagcc gcgagagcgc caacaaccgc ttcttggtcg
     5221 aaggcagcaa gcgcgatgaa tgtcttacta cggagcaagt tcccgaggta atcggagtcc
     5281 ggctgatgtt gggagtaggt ggctacgtct ccgaactcac gaccgaaaag atcaagagca
     5341 gcccgcatgg atttgacttg gtcagggccg agcctacatg tgcgaatgat gcccatactt
     5401 gagccaccta actttgtttt agggcgactg ccctgctgcg taacatcgtt gctgctgcgt
     5461 aacatcgttg ctgctccata a
//
