LOCUS       sequence-357255-        6488 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Generation of MS2 Virus-Like Particles packaged with the sequence 
            for the RSV P and M genes [GenBank: M74568.1].
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6488)
  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 6488)
  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_RSV
FEATURES             Location/Qualifiers
     source          1..6488
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(197..301)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     602..624
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     616..633
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     617..633
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        638..656
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    657..681
                     /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(690..695,696..713,714..719)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     RBS             739..761
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2026..2412
                     /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             2455..2472
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      2851..2898
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        2944..2962
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_RNA        4843..4861
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     terminator      4876..4923
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(5021..5038)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5131..5716)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5211..5230)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5890..6488,1..196))
                     /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"
ORIGIN
        1 gcgccgcttc atcctgcagc tcgttcagcg caccgctcag atcggttttc acaaacagca
       61 ccggacgacc ctgcgcgctc agacgaaaca ccgccgcatc agagcagcca atggtctgct
      121 gcgcccaatc atagccaaac agacgttcca cccacgctgc cgggctaccc gcatgcaggc
      181 catcctgttc aatcatactc ttcctttttc aatattattg aagcatttat cagggttatt
      241 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
      301 gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt taaaattcgc
      361 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
      421 ttataaatca aaagaataga ccgagatagg gttgagtggc cgctacaggg cgctcccatt
      481 cgccattcag gctgcgcaac tgttgggaag ggcgtttcgg tgcgggcctc ttcgctatta
      541 cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt
      601 tcccagtcac gacgttgtaa aacgacggcc agtgaggtaa tacgactcac tataggggaa
      661 ttgtgagcgg ataacaattc ccctctagat ttacacttta tgcttccggc tcgtatgttt
      721 ctagagaatt caaataattt tgtttaactt taagaaggag atttaaattg gctatcgctg
      781 taggtagccg gaattccatt cctaggaggt ttgacctgtg cgagctttta gtacccttga
      841 tagggagaac gaaaccttcg tcccctccgt tcgcgtttac gcggacggtg agactgaaga
      901 taactcattc tctttaaaat atcgttcgaa ctggactccc ggtcgtttta actcgactgg
      961 ggccaaaacg aaacagtggc actacccctc tccgtattca cggggggcgt taagtgtcac
     1021 atcgatagat caaggtgcct acaagcgaag tgggtcatcg tggggtcgcc cgtacgagga
     1081 gaaagccggt ttcggcttct ccctcgacgc acgctcctgc tacagcctct tccctgtaag
     1141 ccaaaacttg acttacatcg aagtgccgca gaacgttgcg aaccgggcgt cgaccgaagt
     1201 cctgcaaaag gtcacccagg gtaattttaa ccttggtgtt gctttagcag aggccaggtc
     1261 gacagcctca caactcgcga cgcaaaccat tgcgctcgtg aaggcgtaca ctgccgctcg
     1321 tcgcggtaat tggcgccagg cgctccgcta ccttgcccta aacgaagatc gaaagtttcg
     1381 atcaaaacac gtggccggca gatggttgga gttgcagttc ggttggttac cactaatgag
     1441 tgatatccag ggtgcatatg agatgcttac gaaggttcac cttcaagagt ttcttcctat
     1501 gagagccgta cgtcaggtcg gtactaacat caagttagat ggccgtctgt cgtatccagc
     1561 tgcaaacttc cagacaacgt gcaacatatc gcgacgtatc gtgatatggt tttacataaa
     1621 cgatgcacgt ttggcatggt tgtcgtccct aggtatcttg aacccactag gtatagtgtg
     1681 ggaaaaggtg cctttctcat tcgttgtcga ctggctccta cctgtaggta acatgctcga
     1741 gggccttacg gcccccgtgg gatgctccta catgtcagga acagttactg acgtaataac
     1801 gggtgagtcc atcataagcg ttgacgctcc ctacgggtgg actgtggaga gacagggcac
     1861 tgctaaggcc caaatctcag ccatgcatcg aggggtacaa tccgtatggc caacaactgg
     1921 cgcgtacgta aagtctcctt tctcgatggt ccatacctta gatgcgttag cattaatcag
     1981 gcaacggctc tctagataga gccctcaacc ggagtttgaa gcatggcttc taactttact
     2041 cagttcgttc tcgtcgacaa tggcggaact ggcgacgtga ctgtcgcccc aagcaacttc
     2101 gctaacgggg tcgctgaatg gatcagctct aactcgcgtt cacaggctta caaagtaacc
     2161 tgtagcgttc gtcagagctc tgcgcagaat cgcaaataca ccatcaaagt cgaggtgcct
     2221 aaagtggcaa cccagactgt tggtggtgta gagcttcctg tagccgcatg gcgttcgtac
     2281 ttaaatatgg aactaaccat tccaattttc gctacgaatt ccgactgcga gcttattgtt
     2341 aaggcaatgc aaggtcttct aaaagatgga aacccgattc cctcagcaat cgcagcaaac
     2401 tccggcatct acgctaactt tactcagttc gttctcgtcg acaatggcgg tacccatcac
     2461 catcaccatc acggtaccgg cgacgtgact gtcgccccaa gcaacttcgc taacggggtc
     2521 gctgaatgga tcagctctaa ctcgcgttca caggcttaca aagtaacctg tagcgttcgt
     2581 cagagctctg cgcagaatcg caaatacacc atcaaagtcg aggtgcctaa agtggcaacc
     2641 cagactgttg gtggtgtaga gcttcctgta gccgcatggc gttcgtactt aaatatggaa
     2701 ctaaccattc caattttcgc tacgaattcc gactgcgagc ttattgttaa ggcaatgcaa
     2761 ggtcttctaa aagatggaaa cccgattccc tcagcaatcg cagcaaactc cggcatctac
     2821 taatagacgc cggccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg
     2881 gtcttgaggg gttttttgtc gaacagaaag taatcgtatt gtacacggcc gcataatcga
     2941 aattaatacg actcactata ggggggcaaa taaatcatca tggaaaagtt tgctcctgaa
     3001 ttccatggag aagatgcaaa caacagggct actaaattcc tagaatcaat aaagggcaaa
     3061 ttcacatcac ccaaagatcc caagaaaaaa gatagtatca tatctgtcaa ctcaatagat
     3121 atagaagtaa ccaaagaaag ccctataaca tcaaattcaa ctattatcaa cccaacaaat
     3181 gagacagatg atactgcagg gaacaagccc aattatcaaa gaaaacctct agtaagtttc
     3241 aaagaagacc ctacaccaag tgataatccc ttttctaaac tatacaaaga aaccatagaa
     3301 acatttgata acaatgaaga agaatccagc tattcatacg aagaaataaa tgatcagaca
     3361 aacgataata taacagcaag attagatagg attgatgaaa aattaagtga aatactagga
     3421 atgcttcaca cattagtagt ggcaagtgca ggacctacat ctgctcggga tggtataaga
     3481 gatgccatga ttggtttaag agaagaaatg atagaaaaaa tcagaactga agcattaatg
     3541 accaatgaca gattagaagc tatggcaaga ctcaggaatg aggaaagtga aaagatggca
     3601 aaagacacat cagatgaagt gtctctcaat ccaacatcag agaaattgaa caacctattg
     3661 gaagggaatg atagtgacaa tgatctatca cttgaagatt tctgattagt taccactctt
     3721 cacatcaaca cacaatacca acagaagacc aacaaactaa ccaacccaat catccaacca
     3781 aacatccatc cgccaatcag ccaaacagcc aacaaaacaa ccagccaatc caaaactaac
     3841 cacccggaaa aaatctataa tatagttaca aaaaaaggaa agggtggggc aaatatggaa
     3901 acatacgtga acaagcttca cgaaggctcc acatacacag ctgctgttca atacaatgtc
     3961 ttagaaaaag acgatgaccc tgcatcactt acaatatggg tgcccatgtt ccaatcatct
     4021 atgccagcag atttacttat aaaagaacta gctaatgtca acatactagt gaaacaaata
     4081 tccacaccca agggaccttc actaagagtc atgataaact caagaagtgc agtgctagca
     4141 caaatgccca gcaaatttac catatgcgct aatgtgtcct tggatgaaag aagcaaacta
     4201 gcatatgatg taaccacacc ctgtgaaatc aaggcatgta gtctaacatg cctaaaatca
     4261 aaaaatatgt tgactacagt taaagatctc actatgaaga cactcaaccc tacacatgat
     4321 attattgctt tatgtgaatt tgaaaacata gtaacatcaa aaaaagtcat aataccaaca
     4381 tacctaagat ccatcagtgt cagaaataaa gatctgaaca cacttgaaaa tataacaacc
     4441 actgaattca aaaatgctat cacaaatgca aaaatcatcc cttactcagg attactatta
     4501 gtcatcacag tgactgacaa caaaggagca ttcaaataca taaagccaca aagtcaattc
     4561 atagtagatc ttggagctta cctagaaaaa gaaagtatat attatgttac cacaaattgg
     4621 aaacacacag ctacacgatt tgcaatcaaa cccatggaag attaaccttt ttcctctaca
     4681 tcagtgtgtt aattcataca aactttctac ctacattctt cacttcacca tcacaatcac
     4741 aaacactctg tggttcaacc aatcaaacaa aacttatctg aagtcccaga tcatcccaag
     4801 tcattgttta tcagatctag tactcaaata agttaataaa aaacatgagg atcacccatg
     4861 tccgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt
     4921 ttgtaatttc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaacatg
     4981 gtcatagctg tttccttgcg tattgggcgc tctccgcttc ctcgctcact gactcgctgc
     5041 gctcggtcgt tcgggtaaag cctggggtgc ctaatgagca aaaggccagc aaaaggccag
     5101 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     5161 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     5221 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     5281 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     5341 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     5401 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     5461 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     5521 cgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     5581 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     5641 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     5701 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     5761 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     5821 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     5881 tgacagttat tagaaaaatt catccagcag acgataaaac gcaatacgct ggctatccgg
     5941 tgccgcaatg ccatacagca ccagaaaacg atccgcccat tcgccgccca gttcttccgc
     6001 aatatcacgg gtggccagcg caatatcctg ataacgatcc gccacgccca gacggccgca
     6061 atcaataaag ccgctaaaac ggccattttc caccataatg ttcggcaggc acgcatcacc
     6121 atgggtcacc accagatctt cgccatccgg catgctcgct ttcagacgcg caaacagctc
     6181 tgccggtgcc aggccctgat gttcttcatc cagatcatcc tgatccacca ggcccgcttc
     6241 catacgggta cgcgcacgtt caatacgatg tttcgcctga tgatcaaacg gacaggtcgc
     6301 cgggtccagg gtatgcagac gacgcatggc atccgccata atgctcactt tttctgccgg
     6361 cgccagatgg ctagacagca gatcctgacc cggcacttcg cccagcagca gccaatcacg
     6421 gcccgcttcg gtcaccacat ccagcaccgc cgcacacgga acaccggtgg tggccagcca
     6481 gctcagac
//
