LOCUS       sequence-347830-        6009 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Generation of MS2 Virus-Like Particles packaged with the sequence 
            for the SARS-CoV-2 N gene [CoV USA/WA-CDC-WA1/2020].
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6009)
  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 6009)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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_SC2
FEATURES             Location/Qualifiers
     source          1..6009
                     /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        3596..3614
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     terminator      3629..3676
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(3774..3791)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3884..4469)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3964..3983)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4643..5437)
                     /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(5438..5542)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5843..5865
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5857..5874
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5858..5874
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5879..5897
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5898..5922
                     /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(5931..5936,5937..5954,5955..5960)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     RBS             5980..6002
                     /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 taggatgtct gataatggac cccaaaatca
     2221 gcgaaatgca ccccgcatta cgtttggtgg accctcagat tcaactggca gtaaccagaa
     2281 tggagaacgc agtggggcgc gatcaaaaca acgtcggccc caaggtttac ccaataatac
     2341 tgcgtcttgg ttcaccgctc tcactcaaca tggcaaggaa gaccttaaat tccctcgagg
     2401 acaaggcgtt ccaattaaca ccaatagcag tccagatgac caaattggct actaccgaag
     2461 agctaccaga cgaattcgtg gtggtgacgg taaaatgaaa gatctcagtc caagatggta
     2521 tttctactac ctaggaactg ggccagaagc tggacttccc tatggtgcta acaaagacgg
     2581 catcatatgg gttgcaactg agggagcctt gaatacacca aaagatcaca ttggcacccg
     2641 caatcctgct aacaatgctg caatcgtgct acaacttcct caaggaacaa cattgccaaa
     2701 aggcttctac gcagaaggga gcagaggcgg cagtcaagcc tcttctcgtt cctcatcacg
     2761 tagtcgcaac agttcaagaa attcaactcc aggcagcagt aggggaactt ctcctgctag
     2821 aatggctggc aatggcggtg atgctgctct tgctttgctg ctgcttgaca gattgaacca
     2881 gcttgagagc aaaatgtctg gtaaaggcca acaacaacaa ggccaaactg tcactaagaa
     2941 atctgctgct gaggcttcta agaagcctcg gcaaaaacgt actgccacta aagcatacaa
     3001 tgtaacacaa gctttcggca gacgtggtcc agaacaaacc caaggaaatt ttggggacca
     3061 ggaactaatc agacaaggaa ctgattacaa acattggccg caaattgcac aatttgcccc
     3121 cagcgcttca gcgttcttcg gaatgtcgcg cattggcatg gaagtcacac cttcgggaac
     3181 gtggttgacc tacacaggtg ccatcaaatt ggatgacaaa gatccaaatt tcaaagatca
     3241 agtcattttg ctgaataagc atattgacgc atacaaaaca ttcccaccaa cagagcctaa
     3301 aaaggacaaa aagaagaagg ctgatgaaac tcaagcctta ccgcagagac agaagaaaca
     3361 gcaaactgtg actcttcttc ctgctgcaga tttggatgat ttctccaaac aattgcaaca
     3421 atccatgagc agtgctgact caactcaggc ctaaactcat gcagaccaca caaggcagat
     3481 gggctatata aacgttttcg cttttccgtt tacgatatat agtctactct tgtgcagaat
     3541 gaattctcgt aactacatag cacaagtaga tgtagttaac tttaatctca catagacatg
     3601 aggatcaccc atgtccgctg agcaataact agcataaccc cttggggcct ctaaacgggt
     3661 cttgaggggt tttttgtaat ttcactgccc gctttccagt cgggaaacct gtcgtgccag
     3721 ctgcattaac atggtcatag ctgtttcctt gcgtattggg cgctctccgc ttcctcgctc
     3781 actgactcgc tgcgctcggt cgttcgggta aagcctgggg tgcctaatga gcaaaaggcc
     3841 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     3901 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     3961 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     4021 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     4081 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     4141 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     4201 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     4261 cgaggtatgt aggcgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     4321 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     4381 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     4441 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     4501 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     4561 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     4621 agtaaacttg gtctgacagt tattagaaaa attcatccag cagacgataa aacgcaatac
     4681 gctggctatc cggtgccgca atgccataca gcaccagaaa acgatccgcc cattcgccgc
     4741 ccagttcttc cgcaatatca cgggtggcca gcgcaatatc ctgataacga tccgccacgc
     4801 ccagacggcc gcaatcaata aagccgctaa aacggccatt ttccaccata atgttcggca
     4861 ggcacgcatc accatgggtc accaccagat cttcgccatc cggcatgctc gctttcagac
     4921 gcgcaaacag ctctgccggt gccaggccct gatgttcttc atccagatca tcctgatcca
     4981 ccaggcccgc ttccatacgg gtacgcgcac gttcaatacg atgtttcgcc tgatgatcaa
     5041 acggacaggt cgccgggtcc agggtatgca gacgacgcat ggcatccgcc ataatgctca
     5101 ctttttctgc cggcgccaga tggctagaca gcagatcctg acccggcact tcgcccagca
     5161 gcagccaatc acggcccgct tcggtcacca catccagcac cgccgcacac ggaacaccgg
     5221 tggtggccag ccagctcaga cgcgccgctt catcctgcag ctcgttcagc gcaccgctca
     5281 gatcggtttt cacaaacagc accggacgac cctgcgcgct cagacgaaac accgccgcat
     5341 cagagcagcc aatggtctgc tgcgcccaat catagccaaa cagacgttcc acccacgctg
     5401 ccgggctacc cgcatgcagg ccatcctgtt caatcatact cttccttttt caatattatt
     5461 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     5521 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctaaa ttgtaagcgt
     5581 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
     5641 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgg
     5701 ccgctacagg gcgctcccat tcgccattca ggctgcgcaa ctgttgggaa gggcgtttcg
     5761 gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta
     5821 agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgaggta
     5881 atacgactca ctatagggga attgtgagcg gataacaatt cccctctaga tttacacttt
     5941 atgcttccgg ctcgtatgtt tctagagaat tcaaataatt ttgtttaact ttaagaagga
     6001 gatttaaat
//
