LOCUS       sequence-299986-        4839 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Generation of MS2 Virus Like Particles packaged with the sequence 
            for the SARS-CoV-2 Envelope Gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4839)
  AUTHORS   Crone MA, Priestman M, Ciechonska M, Jensen K, Sharp DJ, Anand A, 
            Randell P, Storch M, Freemont PS
  TITLE     A role for Biofoundries in rapid development and validation of 
            automated SARS-CoV-2 clinical diagnostics.
  JOURNAL   Nat Commun. 2020 Sep 8;11(1):4464. doi: 10.1038/s41467-020-18130-3.
  PUBMED    32900994
REFERENCE   2  (bases 1 to 4839)
  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: "Nat 
            Commun."; date: "2020-09-8"; volume: "11(1):4464. doi"; pages: " 
            10.1038/s41467-020-18130-3"
COMMENT     Sequence Label: pMC060-HisMS2_PLP_Env_pac
FEATURES             Location/Qualifiers
     source          1..4839
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(166..270)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     571..593
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     585..602
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     586..602
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        607..625
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    626..650
                     /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(659..664,665..682,683..688)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     RBS             708..730
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1995..2381
                     /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             2424..2441
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      2820..2867
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        2913..2931
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_RNA        3160..3178
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     terminator      3193..3240
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(3338..3355)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3448..4036)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3528..3547)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4210..4839,1..165))
                     /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 accgctcaga tcggttttca caaacagcac cggacgaccc tgcgcgctca gacgaaacac
       61 cgccgcatca gagcagccaa tggtctgctg cgcccaatca tagccaaaca gacgttccac
      121 ccacgctgcc gggctacccg catgcaggcc atcctgttca atcatactct tcctttttca
      181 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
      241 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctaaatt
      301 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
      361 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
      421 ttgagtggcc gctacagggc gctcccattc gccattcagg ctgcgcaact gttgggaagg
      481 gcgtttcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
      541 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
      601 gtgaggtaat acgactcact ataggggaat tgtgagcgga taacaattcc cctctagatt
      661 tacactttat gcttccggct cgtatgtttc tagagaattc aaataatttt gtttaacttt
      721 aagaaggaga tttaaattgg ctatcgctgt aggtagccgg aattccattc ctaggaggtt
      781 tgacctgtgc gagcttttag tacccttgat agggagaacg aaaccttcgt cccctccgtt
      841 cgcgtttacg cggacggtga gactgaagat aactcattct ctttaaaata tcgttcgaac
      901 tggactcccg gtcgttttaa ctcgactggg gccaaaacga aacagtggca ctacccctct
      961 ccgtattcac ggggggcgtt aagtgtcaca tcgatagatc aaggtgccta caagcgaagt
     1021 gggtcatcgt ggggtcgccc gtacgaggag aaagccggtt tcggcttctc cctcgacgca
     1081 cgctcctgct acagcctctt ccctgtaagc caaaacttga cttacatcga agtgccgcag
     1141 aacgttgcga accgggcgtc gaccgaagtc ctgcaaaagg tcacccaggg taattttaac
     1201 cttggtgttg ctttagcaga ggccaggtcg acagcctcac aactcgcgac gcaaaccatt
     1261 gcgctcgtga aggcgtacac tgccgctcgt cgcggtaatt ggcgccaggc gctccgctac
     1321 cttgccctaa acgaagatcg aaagtttcga tcaaaacacg tggccggcag atggttggag
     1381 ttgcagttcg gttggttacc actaatgagt gatatccagg gtgcatatga gatgcttacg
     1441 aaggttcacc ttcaagagtt tcttcctatg agagccgtac gtcaggtcgg tactaacatc
     1501 aagttagatg gccgtctgtc gtatccagct gcaaacttcc agacaacgtg caacatatcg
     1561 cgacgtatcg tgatatggtt ttacataaac gatgcacgtt tggcatggtt gtcgtcccta
     1621 ggtatcttga acccactagg tatagtgtgg gaaaaggtgc ctttctcatt cgttgtcgac
     1681 tggctcctac ctgtaggtaa catgctcgag ggccttacgg cccccgtggg atgctcctac
     1741 atgtcaggaa cagttactga cgtaataacg ggtgagtcca tcataagcgt tgacgctccc
     1801 tacgggtgga ctgtggagag acagggcact gctaaggccc aaatctcagc catgcatcga
     1861 ggggtacaat ccgtatggcc aacaactggc gcgtacgtaa agtctccttt ctcgatggtc
     1921 cataccttag atgcgttagc attaatcagg caacggctct ctagatagag ccctcaaccg
     1981 gagtttgaag catggcttct aactttactc agttcgttct cgtcgacaat ggcggaactg
     2041 gcgacgtgac tgtcgcccca agcaacttcg ctaacggggt cgctgaatgg atcagctcta
     2101 actcgcgttc acaggcttac aaagtaacct gtagcgttcg tcagagctct gcgcagaatc
     2161 gcaaatacac catcaaagtc gaggtgccta aagtggcaac ccagactgtt ggtggtgtag
     2221 agcttcctgt agccgcatgg cgttcgtact taaatatgga actaaccatt ccaattttcg
     2281 ctacgaattc cgactgcgag cttattgtta aggcaatgca aggtcttcta aaagatggaa
     2341 acccgattcc ctcagcaatc gcagcaaact ccggcatcta cgctaacttt actcagttcg
     2401 ttctcgtcga caatggcggt acccatcacc atcaccatca cggtaccggc gacgtgactg
     2461 tcgccccaag caacttcgct aacggggtcg ctgaatggat cagctctaac tcgcgttcac
     2521 aggcttacaa agtaacctgt agcgttcgtc agagctctgc gcagaatcgc aaatacacca
     2581 tcaaagtcga ggtgcctaaa gtggcaaccc agactgttgg tggtgtagag cttcctgtag
     2641 ccgcatggcg ttcgtactta aatatggaac taaccattcc aattttcgct acgaattccg
     2701 actgcgagct tattgttaag gcaatgcaag gtcttctaaa agatggaaac ccgattccct
     2761 cagcaatcgc agcaaactcc ggcatctact aatagacgcc ggccaccgct gagcaataac
     2821 tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgtcg aacagaaagt
     2881 aatcgtattg tacacggccg cataatcgaa attaatacga ctcactatag gatgtactca
     2941 ttcgtttcgg aagagacagg tacgttaata gttaatagcg tacttctttt tcttgctttc
     3001 gtggtattct tgctagttac actagccatc cttactgcgc ttcgattgtg tgcgtactgc
     3061 tgcaatattg ttaacgtgag tcttgtaaaa ccttcttttt acgtttactc tcgtgttaaa
     3121 aatctgaatt cttctagagt tcctgatctt ctggtctaaa catgaggatc acccatgtcc
     3181 gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg
     3241 taatttcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taacatggtc
     3301 atagctgttt ccttgcgtat tgggcgctct ccgcttcctc gctcactgac tcgctgcgct
     3361 cggtcgttcg ggtaaagcct ggggtgccta atgagcaaaa ggccagcaaa aggccaggaa
     3421 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     3481 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     3541 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     3601 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     3661 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     3721 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     3781 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     3841 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     3901 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     3961 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     4021 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     4081 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     4141 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     4201 tgacagttat tagaaaaatt catccagcag acgataaaac gcaatacgct ggctatccgg
     4261 tgccgcaatg ccatacagca ccagaaaacg atccgcccat tcgccgccca gttcttccgc
     4321 aatatcacgg gtggccagcg caatatcctg ataacgatcc gccacgccca gacggccgca
     4381 atcaataaag ccgctaaaac ggccattttc caccataatg ttcggcaggc acgcatcacc
     4441 atgggtcacc accagatctt cgccatccgg catgctcgct ttcagacgcg caaacagctc
     4501 tgccggtgcc aggccctgat gttcttcatc cagatcatcc tgatccacca ggcccgcttc
     4561 catacgggta cgcgcacgtt caatacgatg tttcgcctga tgatcaaacg gacaggtcgc
     4621 cgggtccagg gtatgcagac gacgcatggc atccgccata atgctcactt tttctgccgg
     4681 cgccagatgg ctagacagca gatcctgacc cggcacttcg cccagcagca gccaatcacg
     4741 gcccgcttcg gtcaccacat ccagcaccgc cgcacacgga acaccggtgg tggccagcca
     4801 gctcagacgc gccgcttcat cctgcagctc gttcagcgc
//
