LOCUS       sequence-352242-        5635 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Generation of MS2 Virus-Like Particles packaged with the sequence 
            for the Influenza B NSP 1 protein [B/Colorado/06/2017].
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5635)
  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 5635)
  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_FluB
FEATURES             Location/Qualifiers
     source          1..5635
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1258..1644
                     /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             1687..1704
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      2083..2130
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        2176..2194
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_RNA        3222..3240
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     terminator      3255..3302
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(3400..3417)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3510..4095)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3590..3609)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4269..5063)
                     /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(5064..5168)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5469..5491
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5483..5500
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5484..5500
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5505..5523
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5524..5548
                     /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(5557..5562,5563..5580,5581..5586)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     RBS             5606..5628
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
ORIGIN
        1 tggctatcgc tgtaggtagc cggaattcca ttcctaggag gtttgacctg tgcgagcttt
       61 tagtaccctt gatagggaga acgaaacctt cgtcccctcc gttcgcgttt acgcggacgg
      121 tgagactgaa gataactcat tctctttaaa atatcgttcg aactggactc ccggtcgttt
      181 taactcgact ggggccaaaa cgaaacagtg gcactacccc tctccgtatt cacggggggc
      241 gttaagtgtc acatcgatag atcaaggtgc ctacaagcga agtgggtcat cgtggggtcg
      301 cccgtacgag gagaaagccg gtttcggctt ctccctcgac gcacgctcct gctacagcct
      361 cttccctgta agccaaaact tgacttacat cgaagtgccg cagaacgttg cgaaccgggc
      421 gtcgaccgaa gtcctgcaaa aggtcaccca gggtaatttt aaccttggtg ttgctttagc
      481 agaggccagg tcgacagcct cacaactcgc gacgcaaacc attgcgctcg tgaaggcgta
      541 cactgccgct cgtcgcggta attggcgcca ggcgctccgc taccttgccc taaacgaaga
      601 tcgaaagttt cgatcaaaac acgtggccgg cagatggttg gagttgcagt tcggttggtt
      661 accactaatg agtgatatcc agggtgcata tgagatgctt acgaaggttc accttcaaga
      721 gtttcttcct atgagagccg tacgtcaggt cggtactaac atcaagttag atggccgtct
      781 gtcgtatcca gctgcaaact tccagacaac gtgcaacata tcgcgacgta tcgtgatatg
      841 gttttacata aacgatgcac gtttggcatg gttgtcgtcc ctaggtatct tgaacccact
      901 aggtatagtg tgggaaaagg tgcctttctc attcgttgtc gactggctcc tacctgtagg
      961 taacatgctc gagggcctta cggcccccgt gggatgctcc tacatgtcag gaacagttac
     1021 tgacgtaata acgggtgagt ccatcataag cgttgacgct ccctacgggt ggactgtgga
     1081 gagacagggc actgctaagg cccaaatctc agccatgcat cgaggggtac aatccgtatg
     1141 gccaacaact ggcgcgtacg taaagtctcc tttctcgatg gtccatacct tagatgcgtt
     1201 agcattaatc aggcaacggc tctctagata gagccctcaa ccggagtttg aagcatggct
     1261 tctaacttta ctcagttcgt tctcgtcgac aatggcggaa ctggcgacgt gactgtcgcc
     1321 ccaagcaact tcgctaacgg ggtcgctgaa tggatcagct ctaactcgcg ttcacaggct
     1381 tacaaagtaa cctgtagcgt tcgtcagagc tctgcgcaga atcgcaaata caccatcaaa
     1441 gtcgaggtgc ctaaagtggc aacccagact gttggtggtg tagagcttcc tgtagccgca
     1501 tggcgttcgt acttaaatat ggaactaacc attccaattt tcgctacgaa ttccgactgc
     1561 gagcttattg ttaaggcaat gcaaggtctt ctaaaagatg gaaacccgat tccctcagca
     1621 atcgcagcaa actccggcat ctacgctaac tttactcagt tcgttctcgt cgacaatggc
     1681 ggtacccatc accatcacca tcacggtacc ggcgacgtga ctgtcgcccc aagcaacttc
     1741 gctaacgggg tcgctgaatg gatcagctct aactcgcgtt cacaggctta caaagtaacc
     1801 tgtagcgttc gtcagagctc tgcgcagaat cgcaaataca ccatcaaagt cgaggtgcct
     1861 aaagtggcaa cccagactgt tggtggtgta gagcttcctg tagccgcatg gcgttcgtac
     1921 ttaaatatgg aactaaccat tccaattttc gctacgaatt ccgactgcga gcttattgtt
     1981 aaggcaatgc aaggtcttct aaaagatgga aacccgattc cctcagcaat cgcagcaaac
     2041 tccggcatct actaatagac gccggccacc gctgagcaat aactagcata accccttggg
     2101 gcctctaaac gggtcttgag gggttttttg tcgaacagaa agtaatcgta ttgtacacgg
     2161 ccgcataatc gaaattaata cgactcacta taggatggcg aacaacaaca tgaccacaac
     2221 acaaattgag gtgggtccgg gagcaaccaa tgccaccata aactttgaag caggaattct
     2281 ggagtgctat gaaaggcttt catggcaaag agcccttgac taccccggtc aagaccgcct
     2341 aaacagacta aaaagaaaat tagagtcaag aataaagact cacaacaaaa gtgagcctga
     2401 aagtaaaagg atgtcccttg aagagagaaa agcaattgga gtaaaaatga tgaaagtact
     2461 cctatttatg aatccgtctg ctggaattga agggtttgag ccatactgta tgaacagttc
     2521 ctcaaatagc aactgtacga aatacaattg gaccgattac ccttcaacac cagagaggtg
     2581 ccttgatgac atagaggaag aaccagagga tgttgatggc ccaactgaaa tagtattaag
     2641 ggacatgaac aacaaagatg caaggcaaaa gataaaggag gaagtaaaca ctcagaaaga
     2701 agggaagttc cgtttgacaa taaaaaggga tatgcgtaat gtattgtcct tgagagtgtt
     2761 ggtaaatgga acattcctca aacaccccaa tggatacaag tccttatcaa ctctgcatag
     2821 attgaatgca tatgaccaga gtggaaggct tgttgctaaa cttgttgcca ctgatgatct
     2881 tacagtggag gatgaagaag atggccatcg gatcctcaac tcactcttcg agcgtctcaa
     2941 tgaaggacat tcaaagccaa ttcgagcagc tgaaactgcg gtgggagtct tatcccaatt
     3001 tggtcaagag caccgattat caccagaaga gggagacaat tagattggtc acggaagaac
     3061 tttatctttt aagtaaaaga attgatgata acatactatt ccacaaaaca gtgatagcta
     3121 acagctccat aatagctgac atggttgtat cattatcatt attagaaaca ttgtatgaaa
     3181 tgaaggatgt ggttgaagtg tacagcaggc agtgcttgtg aacatgagga tcacccatgt
     3241 ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt
     3301 tgtaatttca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaacatgg
     3361 tcatagctgt ttccttgcgt attgggcgct ctccgcttcc tcgctcactg actcgctgcg
     3421 ctcggtcgtt cgggtaaagc ctggggtgcc taatgagcaa aaggccagca aaaggccagg
     3481 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     3541 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     3601 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     3661 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     3721 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     3781 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     3841 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     3901 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     3961 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     4021 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     4081 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     4141 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     4201 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     4261 gacagttatt agaaaaattc atccagcaga cgataaaacg caatacgctg gctatccggt
     4321 gccgcaatgc catacagcac cagaaaacga tccgcccatt cgccgcccag ttcttccgca
     4381 atatcacggg tggccagcgc aatatcctga taacgatccg ccacgcccag acggccgcaa
     4441 tcaataaagc cgctaaaacg gccattttcc accataatgt tcggcaggca cgcatcacca
     4501 tgggtcacca ccagatcttc gccatccggc atgctcgctt tcagacgcgc aaacagctct
     4561 gccggtgcca ggccctgatg ttcttcatcc agatcatcct gatccaccag gcccgcttcc
     4621 atacgggtac gcgcacgttc aatacgatgt ttcgcctgat gatcaaacgg acaggtcgcc
     4681 gggtccaggg tatgcagacg acgcatggca tccgccataa tgctcacttt ttctgccggc
     4741 gccagatggc tagacagcag atcctgaccc ggcacttcgc ccagcagcag ccaatcacgg
     4801 cccgcttcgg tcaccacatc cagcaccgcc gcacacggaa caccggtggt ggccagccag
     4861 ctcagacgcg ccgcttcatc ctgcagctcg ttcagcgcac cgctcagatc ggttttcaca
     4921 aacagcaccg gacgaccctg cgcgctcaga cgaaacaccg ccgcatcaga gcagccaatg
     4981 gtctgctgcg cccaatcata gccaaacaga cgttccaccc acgctgccgg gctacccgca
     5041 tgcaggccat cctgttcaat catactcttc ctttttcaat attattgaag catttatcag
     5101 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     5161 gttccgcgca catttccccg aaaagtgcca cctaaattgt aagcgttaat attttgttaa
     5221 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
     5281 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtggccgc tacagggcgc
     5341 tcccattcgc cattcaggct gcgcaactgt tgggaagggc gtttcggtgc gggcctcttc
     5401 gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc
     5461 agggttttcc cagtcacgac gttgtaaaac gacggccagt gaggtaatac gactcactat
     5521 aggggaattg tgagcggata acaattcccc tctagattta cactttatgc ttccggctcg
     5581 tatgtttcta gagaattcaa ataattttgt ttaactttaa gaaggagatt taaat
//
