LOCUS       sequence-324879-        5481 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry vector for an inducible 3xFLAG-tagged MAVS.
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
FEATURES             Location/Qualifiers
     source          1..5481
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     25..48
                     /label=pENTR-F
                     /note="pENTR vectors, forward primer"
     protein_bind    67..166
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        198..512
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    206..224
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    242..260
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    277..295
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    313..331
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    349..367
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    384..402
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    420..438
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     485..509
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             521..586
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     protein_bind    complement(2251..2350)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2462..2481)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(2540..2559)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     CDS             complement(3133..3666)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(3855..3860,3861..3877,3878..3883))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     rep_origin      4540..5128
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5029..5048
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      5293..5379
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      join(5471..5481,1..17)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ctgacggatg gcctttttgc gtttctacaa actcttcctg ttagttagtt acttaagctc
       61 gggccccaaa taatgatttt attttgactg atagtgacct gttcgttgca acaaattgat
      121 aagcaatgct tttttataat gccaactttg tacaaaaaag caggctttaa aggaaccaat
      181 tcagtcgact ggatcccgag tttactccct atcagtgata gagaacgtat gtcgagttta
      241 ctccctatca gtgatagaga acgatgtcga gtttactccc tatcagtgat agagaacgta
      301 tgtcgagttt actccctatc agtgatagag aacgtatgtc gagtttactc cctatcagtg
      361 atagagaacg tatgtcgagt ttatccctat cagtgataga gaacgtatgt cgagtttact
      421 ccctatcagt gatagagaac gtatgtcgag gtaggcgtgt acggtgggag gcctatataa
      481 gcagagctcg tttagtgaac cgtcagatcg ccactagatg gactacaaag accatgacgg
      541 tgattataaa gatcatgaca tcgattacaa ggatgacgat gacaagacta gtatgccgtt
      601 tgctgaagac aagacctata agtatatctg ccgcaatttc agcaattttt gcaatgtgga
      661 tgttgtagag attctgcctt acctgccctg cctcacagca agagaccagg atcgactgcg
      721 ggccacctgc acactctcag ggaaccggga caccctctgg catctcttca atacccttca
      781 gcggcggccc ggctgggtgg agtacttcat tgcggcactg aggggctgtg agctagttga
      841 tctcgcggac gaagtggcct ctgtctacga gagctaccag cctcggacct cggaccgtcc
      901 cccagaccca ctggagccac cgtcacttcc tgctgagagg ccagggcccc ccacacctgc
      961 tgcggcccac agcatcccct acaacagctg cagagagaag gagccaagtt accccatgcc
     1021 tgtccaggag acccaggcgc cagagtcccc aggagagaat tcagagcaag ccctgcagac
     1081 gctcagcccc agagccatcc caaggaatcc agatggtggc cccctggagt cctcctctga
     1141 cctggcagcc ctcagccctc tgacctccag cgggcatcag gagaaggaca cagaactggg
     1201 cagtacccac acagcaggtg cgacctccag cctcacacca tcccgtgggc ctgtgtctcc
     1261 atctgtctcc ttccagcccc tggcccgttc cacccccagg gcaagccgct tgcctggacc
     1321 cacagggtca gttgtatcta ctggcacctc cttctcctcc tcatcccctg gcttggcctc
     1381 tgcaggggct gcagagggta aacagggtgc agagagtgac caggccgagc ctatcatctg
     1441 ctccagtggg gcagaggcac ctgccaactc tctgccctcc aaagtgccta ccaccttgat
     1501 gcctgtgaac acagtggccc tgaaagtgcc tgccaaccca gcatctgtca gcacagtgcc
     1561 ctccaagttg ccaactagct caaagccccc tggtgcagtg ccttctaatg cgctcaccaa
     1621 tccagcacca tccaaattgc ccatcaactc aacccgtgct ggcatggtgc catccaaagt
     1681 gcctactagc atggtgctca ccaaggtgtc tgccagcaca gtccccactg acgggagcag
     1741 cagaaatgag gagaccccag cagctccaac acccgccggc gccactggag gcagctcagc
     1801 ctggctagac agcagctttg agaatagggg ccttgggtcg gagctgagta agcctggcgt
     1861 gctggcatcc caggtagaca gcccgttctc gggctgcttc gaggatcttg ccatcagtgc
     1921 cagcacctcc ttgggcatgg ggccctgcca tggcccagag gagaatgagt ataagtccga
     1981 gggcaccttt gggatccacg tggctgagaa ccccagcatc cagctcctgg agggcaaccc
     2041 tgggccacct gcggacccgg atggcggccc caggccacaa gccgaccgga agttccagga
     2101 gagggaggtg ccatgccaca ggccctcacc tggggctctg tggctccagg tggctgtgac
     2161 aggggtgctg gtagtcacac tcctggtggt gctgtaccgg cggcgtctgc actagcaatt
     2221 ggaagactaa tgcgttttcg agatatctag acccagcttt cttgtacaaa gttggcatta
     2281 taagaaagca ttgcttatca atttgttgca acgaacaggt cactatcagt caaaataaaa
     2341 tcattatttg ccatccagct gcagctctgg cccgtgtctc aaaatctctg atgttacatt
     2401 gcacaagata aaaatatatc atcatgaaca ataaaactgt ctgcttacat aaacagtaat
     2461 acaaggggtg ttatgagcca tattcaacgg gaaacgtcga ggccgcgatt aaattccaac
     2521 atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg
     2581 acaatctatc gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa acatggcaaa
     2641 ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct gacggaattt
     2701 atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg gttactcacc
     2761 actgcgatcc ccggaaaaac agcattccag gtattagaag aatatcctga ttcaggtgaa
     2821 aatattgttg atgcgctggc agtgtccctg cgccggttgc attcgattcc tgtttgtaat
     2881 tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac
     2941 ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc
     3001 tggaaagaaa tgcataaact tttgccattc tcaccggatt cagtcgtcac tcatggtgat
     3061 ttctcacttg ataaccttat ttttgacgag gggaaattag gccgggaagc cgatctcggc
     3121 ttgaacgaat tgttaggtgg cggtacttgg gtcgatatca aagtgcatca cttcttcccg
     3181 tatgcccaac tttgtataga gagccactgc gggatcgtca ccgtaatctg cttgcacgta
     3241 gatcacataa gcaccaagcg cgttggcctc atgcttgagg agattgatga gcgcggtggc
     3301 aatgccctgc ctccggtgct cgccggagac tgcgagatca tagatataga tctcactacg
     3361 cggctgctca aacttgggca gaacgtaagc cgcgagagcg ccaacaaccg cttcttggtc
     3421 gaaggcagca agcgcgatga atgtcttact acggagcaag ttcccgaggt aatcggagtc
     3481 cggctgatgt tgggagtagg tggctacgtc tccgaactca cgaccgaaaa gatcaagagc
     3541 agcccgcatg gatttgactt ggtcagggcc gagcctacat gtgcgaatga tgcccatact
     3601 tgagccacct aactttgttt tagggcgact gccctgctgc gtaacatcgt tgctgctgcg
     3661 taacatcgtt gctgctccat aacatcaaac atcgacccac ggcgtaacgc gcttgctgct
     3721 tggatgcccg aggcatagac tgtacaaaaa aacagtcata acaagccatg aaaaccgcca
     3781 ctgcgccgtt accaccgctg cgttcggtca aggttctgga ccagttgcgt gagcgcatac
     3841 gctacttgca ttacagttta cgaaccgaac aggcttatgt caactgggtt cgtgccttca
     3901 tccgtttcca cggtgtgcgt cacccggcaa ccttgggcag cagcgaagtc gaggcatttc
     3961 tgtcctggct ggcgaacgag cgcaaggttt cggtctccac gcatcgtcag gcattggcgg
     4021 ccttgctgtt cttctacggc aaggtgctgt gcacggatct gccctggctt caggagatcg
     4081 gaagacctcg gccgtcgcgg cgcttgccgg tggtgctgac cccggatgaa gtggttcgca
     4141 tcctcggttt tctggaaggc gagcatcgtt tgttcgccca ggactctagc tatagttcta
     4201 gtggttggct actaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc
     4261 aggatcttgc catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc
     4321 tttttcaaaa atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc
     4381 tcgatgagtt tttctaatca gaattggtta attggttgta acactggcag agcattacgc
     4441 tgacttgacg ggacggcgca agctcatgac caaaatccct taacgtgagt tttcgttcca
     4501 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     4561 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     4621 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     4681 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     4741 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     4801 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     4861 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     4921 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     4981 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     5041 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     5101 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     5161 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     5221 taaccgtatt accgctagca tggatctcgg ggacgtctaa ctactaagcg agagtaggga
     5281 actgccaggc atcaaataaa acgaaaggct cagtcggaag actgggcctt tcgttttatc
     5341 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac
     5401 gttgtgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggcat
     5461 caaactaagc agaaggccat c
//
