LOCUS       sequence-347829-        5590 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Generation of MS2 Virus-Like Particles packaged with the sequence 
            for the Influenza A Matrix Protein [A/Illinois/20/2018(H1N1)].
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5590)
  AUTHORS   Unruh LAH, Crone MA, Freemont PS, Chindelevitch L
  TITLE     A standardised, high-throughput approach to diagnostic group testing
            method validation
  JOURNAL   medRxiv 2024.04.17.24305965
REFERENCE   2  (bases 1 to 5590)
  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"; journalName: "medRxiv 
            2024.04.17.24305965"
COMMENT     Sequence Label: HisMS2_VLP_FluA
FEATURES             Location/Qualifiers
     source          1..5590
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1251..1637
                     /codon_start=1
                     /product="bacteriophage MS2 coat protein"
                     /label=MS2
                     /note="binds to a specific stem-loop structure in the viral
                     RNA (Peabody, 1993)"
                     /translation="ASNFTQFVLVDNGGTGDVTVAPSNFANGVAEWISSNSRSQAYKVT
                     CSVRQSSAQNRKYTIKVEVPKVATQTVGGVELPVAAWRSYLNMELTIPIFATNSDCELI
                     VKAMQGLLKDGNPIPSAIAANSGIY"
     CDS             1680..1697
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      2076..2123
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        2169..2187
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_RNA        3170..3188
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     terminator      3203..3250
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      complement(3458..4043)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3538..3557)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4217..5011)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     promoter        complement(5012..5116)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5417..5439
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5431..5448
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5432..5448
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5453..5471
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5472..5496
                     /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)."
     promoter        join(5505..5510,5511..5528,5529..5534)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     RBS             5554..5576
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
ORIGIN
        1 cgctgtaggt agccggaatt ccattcctag gaggtttgac ctgtgcgagc ttttagtacc
       61 cttgataggg agaacgaaac cttcgtcccc tccgttcgcg tttacgcgga cggtgagact
      121 gaagataact cattctcttt aaaatatcgt tcgaactgga ctcccggtcg ttttaactcg
      181 actggggcca aaacgaaaca gtggcactac ccctctccgt attcacgggg ggcgttaagt
      241 gtcacatcga tagatcaagg tgcctacaag cgaagtgggt catcgtgggg tcgcccgtac
      301 gaggagaaag ccggtttcgg cttctccctc gacgcacgct cctgctacag cctcttccct
      361 gtaagccaaa acttgactta catcgaagtg ccgcagaacg ttgcgaaccg ggcgtcgacc
      421 gaagtcctgc aaaaggtcac ccagggtaat tttaaccttg gtgttgcttt agcagaggcc
      481 aggtcgacag cctcacaact cgcgacgcaa accattgcgc tcgtgaaggc gtacactgcc
      541 gctcgtcgcg gtaattggcg ccaggcgctc cgctaccttg ccctaaacga agatcgaaag
      601 tttcgatcaa aacacgtggc cggcagatgg ttggagttgc agttcggttg gttaccacta
      661 atgagtgata tccagggtgc atatgagatg cttacgaagg ttcaccttca agagtttctt
      721 cctatgagag ccgtacgtca ggtcggtact aacatcaagt tagatggccg tctgtcgtat
      781 ccagctgcaa acttccagac aacgtgcaac atatcgcgac gtatcgtgat atggttttac
      841 ataaacgatg cacgtttggc atggttgtcg tccctaggta tcttgaaccc actaggtata
      901 gtgtgggaaa aggtgccttt ctcattcgtt gtcgactggc tcctacctgt aggtaacatg
      961 ctcgagggcc ttacggcccc cgtgggatgc tcctacatgt caggaacagt tactgacgta
     1021 ataacgggtg agtccatcat aagcgttgac gctccctacg ggtggactgt ggagagacag
     1081 ggcactgcta aggcccaaat ctcagccatg catcgagggg tacaatccgt atggccaaca
     1141 actggcgcgt acgtaaagtc tcctttctcg atggtccata ccttagatgc gttagcatta
     1201 atcaggcaac ggctctctag atagagccct caaccggagt ttgaagcatg gcttctaact
     1261 ttactcagtt cgttctcgtc gacaatggcg gaactggcga cgtgactgtc gccccaagca
     1321 acttcgctaa cggggtcgct gaatggatca gctctaactc gcgttcacag gcttacaaag
     1381 taacctgtag cgttcgtcag agctctgcgc agaatcgcaa atacaccatc aaagtcgagg
     1441 tgcctaaagt ggcaacccag actgttggtg gtgtagagct tcctgtagcc gcatggcgtt
     1501 cgtacttaaa tatggaacta accattccaa ttttcgctac gaattccgac tgcgagctta
     1561 ttgttaaggc aatgcaaggt cttctaaaag atggaaaccc gattccctca gcaatcgcag
     1621 caaactccgg catctacgct aactttactc agttcgttct cgtcgacaat ggcggtaccc
     1681 atcaccatca ccatcacggt accggcgacg tgactgtcgc cccaagcaac ttcgctaacg
     1741 gggtcgctga atggatcagc tctaactcgc gttcacaggc ttacaaagta acctgtagcg
     1801 ttcgtcagag ctctgcgcag aatcgcaaat acaccatcaa agtcgaggtg cctaaagtgg
     1861 caacccagac tgttggtggt gtagagcttc ctgtagccgc atggcgttcg tacttaaata
     1921 tggaactaac cattccaatt ttcgctacga attccgactg cgagcttatt gttaaggcaa
     1981 tgcaaggtct tctaaaagat ggaaacccga ttccctcagc aatcgcagca aactccggca
     2041 tctactaata gacgccggcc accgctgagc aataactagc ataacccctt ggggcctcta
     2101 aacgggtctt gaggggtttt ttgtcgaaca gaaagtaatc gtattgtaca cggccgcata
     2161 atcgaaatta atacgactca ctataggatg agtcttctaa ccgaggtcga aacgtacgtt
     2221 ctttctatca tcccgtcagg ccccctcaaa gccgagatcg cacagagact ggaaagtgtc
     2281 tttgcaggaa agaacacaga tcttgaggct ctcatggaat ggctaaagac aagaccaatc
     2341 ttgtcacctc tgactaaggg aattttagga tttgtgttca cgctcaccgt gcccagtgag
     2401 cgaggactgc agcgtagacg ctttatccaa aatgccctaa atggaaatgg ggacccgaac
     2461 aacatggata gagcagttaa actatacaag aagctcaaaa gagaaataac attccatggg
     2521 gccaaggagg tgtcactaag ctattcaact ggtgcacttg caagttgcat gggcctcata
     2581 tacaacagaa tgggaacagt gaccacagaa gctgctttcg gtctagtttg tgccacttgt
     2641 gaacagattg ctgattcaca gcatcggtct cacagacaaa tggctactac cacaaatcca
     2701 ctaatcaggc atgaaaacag aatggtgctg gctagcacta cggcaaaggc tatggaacag
     2761 gtggctggat cgagtgaaca ggcagcagag gccatggagg ttgctaataa gactaggcag
     2821 atggtacatg caatgagaac tattgggact catcctagct ccagtgctgg tctgagagat
     2881 gaccttcttg aaaatttgca ggcataccag aagcgaatgg gagtgcagat gcagcggttc
     2941 aagtgatcct ctcgtcattg cagcaaacat cattgggatc ttgcacctga tattgtggat
     3001 tactgatcgt ctttttttca aatgcattta tcgtcgcttt aaatacggtt tgaaaagagg
     3061 gccttctacg gaaggagtgc ctgagtccat gagggaagaa tatcaacagg agcagcagag
     3121 tgctgtggat gttgacgatg gtcattttgt caacatagag ctagagtaaa catgaggatc
     3181 acccatgtcc gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag
     3241 gggttttttg taatttcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     3301 taacatggtc atagctgttt ccttgcgtat tgggcgctct ccgcttcctc gctcactgac
     3361 tcgctgcgct cggtcgttcg ggtaaagcct ggggtgccta atgagcaaaa ggccagcaaa
     3421 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     3481 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     3541 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     3601 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     3661 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     3721 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     3781 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     3841 atgtaggcgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     3901 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     3961 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     4021 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     4081 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     4141 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     4201 cttggtctga cagttattag aaaaattcat ccagcagacg ataaaacgca atacgctggc
     4261 tatccggtgc cgcaatgcca tacagcacca gaaaacgatc cgcccattcg ccgcccagtt
     4321 cttccgcaat atcacgggtg gccagcgcaa tatcctgata acgatccgcc acgcccagac
     4381 ggccgcaatc aataaagccg ctaaaacggc cattttccac cataatgttc ggcaggcacg
     4441 catcaccatg ggtcaccacc agatcttcgc catccggcat gctcgctttc agacgcgcaa
     4501 acagctctgc cggtgccagg ccctgatgtt cttcatccag atcatcctga tccaccaggc
     4561 ccgcttccat acgggtacgc gcacgttcaa tacgatgttt cgcctgatga tcaaacggac
     4621 aggtcgccgg gtccagggta tgcagacgac gcatggcatc cgccataatg ctcacttttt
     4681 ctgccggcgc cagatggcta gacagcagat cctgacccgg cacttcgccc agcagcagcc
     4741 aatcacggcc cgcttcggtc accacatcca gcaccgccgc acacggaaca ccggtggtgg
     4801 ccagccagct cagacgcgcc gcttcatcct gcagctcgtt cagcgcaccg ctcagatcgg
     4861 ttttcacaaa cagcaccgga cgaccctgcg cgctcagacg aaacaccgcc gcatcagagc
     4921 agccaatggt ctgctgcgcc caatcatagc caaacagacg ttccacccac gctgccgggc
     4981 tacccgcatg caggccatcc tgttcaatca tactcttcct ttttcaatat tattgaagca
     5041 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     5101 aaataggggt tccgcgcaca tttccccgaa aagtgccacc taaattgtaa gcgttaatat
     5161 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     5221 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtggccgcta
     5281 cagggcgctc ccattcgcca ttcaggctgc gcaactgttg ggaagggcgt ttcggtgcgg
     5341 gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg
     5401 gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga ggtaatacga
     5461 ctcactatag gggaattgtg agcggataac aattcccctc tagatttaca ctttatgctt
     5521 ccggctcgta tgtttctaga gaattcaaat aattttgttt aactttaaga aggagattta
     5581 aattggctat
//
