LOCUS       sequence-347832-        6237 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Generation of MS2 Virus-Like Particles packaged with the sequence 
            for the SARS-CoV-2 E and N genes [CoV USA/WA-CDC-WA1/2020].
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6237)
  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 6237)
  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_EN_SC2
FEATURES             Location/Qualifiers
     source          1..6237
                     /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        3824..3842
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     terminator      3857..3904
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(4002..4019)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4112..4697)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4192..4211)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4871..5665)
                     /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(5666..5770)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6071..6093
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6085..6102
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6086..6102
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6107..6125
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    6126..6150
                     /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(6159..6164,6165..6182,6183..6188)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     RBS             6208..6230
                     /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 taggatgtac tcattcgttt cggaagagac
     2221 aggtacgtta atagttaata gcgtacttct ttttcttgct ttcgtggtat tcttgctagt
     2281 tacactagcc atccttactg cgcttcgatt gtgtgcgtac tgctgcaata ttgttaacgt
     2341 gagtcttgta aaaccttctt tttacgttta ctctcgtgtt aaaaatctga attcttctag
     2401 agttcctgat cttctggtct aaatgtctga taatggaccc caaaatcagc gaaatgcacc
     2461 ccgcattacg tttggtggac cctcagattc aactggcagt aaccagaatg gagaacgcag
     2521 tggggcgcga tcaaaacaac gtcggcccca aggtttaccc aataatactg cgtcttggtt
     2581 caccgctctc actcaacatg gcaaggaaga ccttaaattc cctcgaggac aaggcgttcc
     2641 aattaacacc aatagcagtc cagatgacca aattggctac taccgaagag ctaccagacg
     2701 aattcgtggt ggtgacggta aaatgaaaga tctcagtcca agatggtatt tctactacct
     2761 aggaactggg ccagaagctg gacttcccta tggtgctaac aaagacggca tcatatgggt
     2821 tgcaactgag ggagccttga atacaccaaa agatcacatt ggcacccgca atcctgctaa
     2881 caatgctgca atcgtgctac aacttcctca aggaacaaca ttgccaaaag gcttctacgc
     2941 agaagggagc agaggcggca gtcaagcctc ttctcgttcc tcatcacgta gtcgcaacag
     3001 ttcaagaaat tcaactccag gcagcagtag gggaacttct cctgctagaa tggctggcaa
     3061 tggcggtgat gctgctcttg ctttgctgct gcttgacaga ttgaaccagc ttgagagcaa
     3121 aatgtctggt aaaggccaac aacaacaagg ccaaactgtc actaagaaat ctgctgctga
     3181 ggcttctaag aagcctcggc aaaaacgtac tgccactaaa gcatacaatg taacacaagc
     3241 tttcggcaga cgtggtccag aacaaaccca aggaaatttt ggggaccagg aactaatcag
     3301 acaaggaact gattacaaac attggccgca aattgcacaa tttgccccca gcgcttcagc
     3361 gttcttcgga atgtcgcgca ttggcatgga agtcacacct tcgggaacgt ggttgaccta
     3421 cacaggtgcc atcaaattgg atgacaaaga tccaaatttc aaagatcaag tcattttgct
     3481 gaataagcat attgacgcat acaaaacatt cccaccaaca gagcctaaaa aggacaaaaa
     3541 gaagaaggct gatgaaactc aagccttacc gcagagacag aagaaacagc aaactgtgac
     3601 tcttcttcct gctgcagatt tggatgattt ctccaaacaa ttgcaacaat ccatgagcag
     3661 tgctgactca actcaggcct aaactcatgc agaccacaca aggcagatgg gctatataaa
     3721 cgttttcgct tttccgttta cgatatatag tctactcttg tgcagaatga attctcgtaa
     3781 ctacatagca caagtagatg tagttaactt taatctcaca tagacatgag gatcacccat
     3841 gtccgctgag caataactag cataacccct tggggcctct aaacgggtct tgaggggttt
     3901 tttgtaattt cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaacat
     3961 ggtcatagct gtttccttgc gtattgggcg ctctccgctt cctcgctcac tgactcgctg
     4021 cgctcggtcg ttcgggtaaa gcctggggtg cctaatgagc aaaaggccag caaaaggcca
     4081 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     4141 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     4201 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     4261 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     4321 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     4381 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     4441 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     4501 gcgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     4561 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     4621 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     4681 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     4741 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     4801 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     4861 ctgacagtta ttagaaaaat tcatccagca gacgataaaa cgcaatacgc tggctatccg
     4921 gtgccgcaat gccatacagc accagaaaac gatccgccca ttcgccgccc agttcttccg
     4981 caatatcacg ggtggccagc gcaatatcct gataacgatc cgccacgccc agacggccgc
     5041 aatcaataaa gccgctaaaa cggccatttt ccaccataat gttcggcagg cacgcatcac
     5101 catgggtcac caccagatct tcgccatccg gcatgctcgc tttcagacgc gcaaacagct
     5161 ctgccggtgc caggccctga tgttcttcat ccagatcatc ctgatccacc aggcccgctt
     5221 ccatacgggt acgcgcacgt tcaatacgat gtttcgcctg atgatcaaac ggacaggtcg
     5281 ccgggtccag ggtatgcaga cgacgcatgg catccgccat aatgctcact ttttctgccg
     5341 gcgccagatg gctagacagc agatcctgac ccggcacttc gcccagcagc agccaatcac
     5401 ggcccgcttc ggtcaccaca tccagcaccg ccgcacacgg aacaccggtg gtggccagcc
     5461 agctcagacg cgccgcttca tcctgcagct cgttcagcgc accgctcaga tcggttttca
     5521 caaacagcac cggacgaccc tgcgcgctca gacgaaacac cgccgcatca gagcagccaa
     5581 tggtctgctg cgcccaatca tagccaaaca gacgttccac ccacgctgcc gggctacccg
     5641 catgcaggcc atcctgttca atcatactct tcctttttca atattattga agcatttatc
     5701 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     5761 gggttccgcg cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt
     5821 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     5881 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtggcc gctacagggc
     5941 gctcccattc gccattcagg ctgcgcaact gttgggaagg gcgtttcggt gcgggcctct
     6001 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg
     6061 ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgaggtaat acgactcact
     6121 ataggggaat tgtgagcgga taacaattcc cctctagatt tacactttat gcttccggct
     6181 cgtatgtttc tagagaattc aaataatttt gtttaacttt aagaaggaga tttaaat
//
