LOCUS       sequence-343943-        6261 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Generation of MS2 Virus-Like Particles packaged with the sequence 
            for the WHO Lambda SARS-CoV-2 S gene (Amino Acid 319-869).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6261)
  TITLE     A library of SARS-CoV-2 Spike gene variant virus-like particles
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6261)
  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: 
            "Unpublished"
COMMENT     Sequence Label: MS2_SC2_S_Lambda
FEATURES             Location/Qualifiers
     source          1..6261
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             47..69
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1334..1720
                     /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             1763..1780
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      2159..2206
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        2252..2270
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_RNA        3924..3942
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     terminator      3957..4004
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(4102..4119)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4212..4797)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4292..4311)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4971..5765)
                     /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(5766..5870)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6171..6193
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6185..6202
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6186..6202
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6207..6225
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    6226..6250
                     /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(6259..6261,1..3,4..21,22..27)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
ORIGIN
        1 acactttatg cttccggctc gtatgtttct agagaattca aataattttg tttaacttta
       61 agaaggagat ttaaattggc tatcgctgta ggtagccgga attccattcc taggaggttt
      121 gacctgtgcg agcttttagt acccttgata gggagaacga aaccttcgtc ccctccgttc
      181 gcgtttacgc ggacggtgag actgaagata actcattctc tttaaaatat cgttcgaact
      241 ggactcccgg tcgttttaac tcgactgggg ccaaaacgaa acagtggcac tacccctctc
      301 cgtattcacg gggggcgtta agtgtcacat cgatagatca aggtgcctac aagcgaagtg
      361 ggtcatcgtg gggtcgcccg tacgaggaga aagccggttt cggcttctcc ctcgacgcac
      421 gctcctgcta cagcctcttc cctgtaagcc aaaacttgac ttacatcgaa gtgccgcaga
      481 acgttgcgaa ccgggcgtcg accgaagtcc tgcaaaaggt cacccagggt aattttaacc
      541 ttggtgttgc tttagcagag gccaggtcga cagcctcaca actcgcgacg caaaccattg
      601 cgctcgtgaa ggcgtacact gccgctcgtc gcggtaattg gcgccaggcg ctccgctacc
      661 ttgccctaaa cgaagatcga aagtttcgat caaaacacgt ggccggcaga tggttggagt
      721 tgcagttcgg ttggttacca ctaatgagtg atatccaggg tgcatatgag atgcttacga
      781 aggttcacct tcaagagttt cttcctatga gagccgtacg tcaggtcggt actaacatca
      841 agttagatgg ccgtctgtcg tatccagctg caaacttcca gacaacgtgc aacatatcgc
      901 gacgtatcgt gatatggttt tacataaacg atgcacgttt ggcatggttg tcgtccctag
      961 gtatcttgaa cccactaggt atagtgtggg aaaaggtgcc tttctcattc gttgtcgact
     1021 ggctcctacc tgtaggtaac atgctcgagg gccttacggc ccccgtggga tgctcctaca
     1081 tgtcaggaac agttactgac gtaataacgg gtgagtccat cataagcgtt gacgctccct
     1141 acgggtggac tgtggagaga cagggcactg ctaaggccca aatctcagcc atgcatcgag
     1201 gggtacaatc cgtatggcca acaactggcg cgtacgtaaa gtctcctttc tcgatggtcc
     1261 ataccttaga tgcgttagca ttaatcaggc aacggctctc tagatagagc cctcaaccgg
     1321 agtttgaagc atggcttcta actttactca gttcgttctc gtcgacaatg gcggaactgg
     1381 cgacgtgact gtcgccccaa gcaacttcgc taacggggtc gctgaatgga tcagctctaa
     1441 ctcgcgttca caggcttaca aagtaacctg tagcgttcgt cagagctctg cgcagaatcg
     1501 caaatacacc atcaaagtcg aggtgcctaa agtggcaacc cagactgttg gtggtgtaga
     1561 gcttcctgta gccgcatggc gttcgtactt aaatatggaa ctaaccattc caattttcgc
     1621 tacgaattcc gactgcgagc ttattgttaa ggcaatgcaa ggtcttctaa aagatggaaa
     1681 cccgattccc tcagcaatcg cagcaaactc cggcatctac gctaacttta ctcagttcgt
     1741 tctcgtcgac aatggcggta cccatcacca tcaccatcac ggtaccggcg acgtgactgt
     1801 cgccccaagc aacttcgcta acggggtcgc tgaatggatc agctctaact cgcgttcaca
     1861 ggcttacaaa gtaacctgta gcgttcgtca gagctctgcg cagaatcgca aatacaccat
     1921 caaagtcgag gtgcctaaag tggcaaccca gactgttggt ggtgtagagc ttcctgtagc
     1981 cgcatggcgt tcgtacttaa atatggaact aaccattcca attttcgcta cgaattccga
     2041 ctgcgagctt attgttaagg caatgcaagg tcttctaaaa gatggaaacc cgattccctc
     2101 agcaatcgca gcaaactccg gcatctacta atagacgccg gccaccgctg agcaataact
     2161 agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgtcga acagaaagta
     2221 atcgtattgt acacggccgc ataatcgaaa ttaatacgac tcactatagg agagtccaac
     2281 caacagaatc tattgttaga tttcctaata ttacaaactt gtgccctttt ggtgaagttt
     2341 ttaacgccac cagatttgca tctgtttatg cttggaacag gaagagaatc agcaactgtg
     2401 ttgctgatta ttctgtccta tataattccg catcattttc cacttttaag tgttatggag
     2461 tgtctcctac taaattaaat gatctctgct ttactaatgt ctatgcagat tcatttgtaa
     2521 ttagaggtga tgaagtcaga caaatcgctc cagggcaaac tggaaagatt gctgattata
     2581 attataaatt accagatgat tttacaggct gcgttatagc ttggaattct aacaatcttg
     2641 attctaaggt tggtggtaat tataattacc agtatagatt gtttaggaag tctaatctca
     2701 aaccttttga gagagatatt tcaactgaaa tctatcaggc cggtagcaca ccttgtaatg
     2761 gtgttgaagg ttttaattgt tactctcctt tacaatcata tggtttccaa cccactaatg
     2821 gtgttggtta ccaaccatac agagtagtag tactttcttt tgaacttcta catgcaccag
     2881 caactgtttg tggacctaaa aagtctacta atttggttaa aaacaaatgt gtcaatttca
     2941 acttcaatgg tttaacaggc acaggtgttc ttactgagtc taacaaaaag tttctgcctt
     3001 tccaacaatt tggcagagac attgctgaca ctactgatgc tgtccgtgat ccacagacac
     3061 ttgagattct tgacattaca ccatgttctt ttggtggtgt cagtgttata acaccaggaa
     3121 caaatacttc taaccaggtt gctgttcttt atcaggatgt taactgcaca gaagtccctg
     3181 ttgctattca tgcagatcaa cttactccta cttggcgtgt ttattctaca ggttctaatg
     3241 tttttcaaac acgtgcaggc tgtttaatag gggctgaaca tgtcaacaac tcatatgagt
     3301 gtgacatacc cattggtgca ggtatatgcg ctagttatca gactcagact aattctcctc
     3361 ggcgggcacg tagtgtagct agtcaatcca tcattgccta cactatgtca cttggtgcag
     3421 aaaattcagt tgcttactct aataactcta ttgccatacc cacaaatttt actattagtg
     3481 ttaccacaga aattctacca gtgtctatga ccaagacatc agtagattgt acaatgtaca
     3541 tttgtggtga ttcaactgaa tgcagcaatc ttttgttgca atatggcagt ttttgtacac
     3601 aattaaaccg tgctttaact ggaatagctg ttgaacaaga caaaaacacc caagaagttt
     3661 ttgcacaagt caaacaaatt tacaaaacac caccaattaa agattttggt ggttttaatt
     3721 tttcacaaat attaccagat ccatcaaaac caagcaagag gtcatttatt gaagatctac
     3781 ttttcaacaa agtgacactt gcagatgctg gcttcatcaa acaatatggt gattgccttg
     3841 gtgatattgc tgctagagac ctcatttgtg cacaaaagtt taacggcctt aatgttttgc
     3901 cacctttgct cacagatgaa atgacatgag gatcacccat gtccgctgag caataactag
     3961 cataacccct tggggcctct aaacgggtct tgaggggttt tttgtaattt cactgcccgc
     4021 tttccagtcg ggaaacctgt cgtgccagct gcattaacat ggtcatagct gtttccttgc
     4081 gtattgggcg ctctccgctt cctcgctcac tgactcgctg cgctcggtcg ttcgggtaaa
     4141 gcctggggtg cctaatgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     4201 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     4261 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     4321 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     4381 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     4441 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     4501 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     4561 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcgctacaga gttcttgaag
     4621 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     4681 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     4741 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     4801 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     4861 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     4921 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ttagaaaaat
     4981 tcatccagca gacgataaaa cgcaatacgc tggctatccg gtgccgcaat gccatacagc
     5041 accagaaaac gatccgccca ttcgccgccc agttcttccg caatatcacg ggtggccagc
     5101 gcaatatcct gataacgatc cgccacgccc agacggccgc aatcaataaa gccgctaaaa
     5161 cggccatttt ccaccataat gttcggcagg cacgcatcac catgggtcac caccagatct
     5221 tcgccatccg gcatgctcgc tttcagacgc gcaaacagct ctgccggtgc caggccctga
     5281 tgttcttcat ccagatcatc ctgatccacc aggcccgctt ccatacgggt acgcgcacgt
     5341 tcaatacgat gtttcgcctg atgatcaaac ggacaggtcg ccgggtccag ggtatgcaga
     5401 cgacgcatgg catccgccat aatgctcact ttttctgccg gcgccagatg gctagacagc
     5461 agatcctgac ccggcacttc gcccagcagc agccaatcac ggcccgcttc ggtcaccaca
     5521 tccagcaccg ccgcacacgg aacaccggtg gtggccagcc agctcagacg cgccgcttca
     5581 tcctgcagct cgttcagcgc accgctcaga tcggttttca caaacagcac cggacgaccc
     5641 tgcgcgctca gacgaaacac cgccgcatca gagcagccaa tggtctgctg cgcccaatca
     5701 tagccaaaca gacgttccac ccacgctgcc gggctacccg catgcaggcc atcctgttca
     5761 atcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     5821 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     5881 cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     5941 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     6001 aagaatagac cgagataggg ttgagtggcc gctacagggc gctcccattc gccattcagg
     6061 ctgcgcaact gttgggaagg gcgtttcggt gcgggcctct tcgctattac gccagctggc
     6121 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
     6181 acgttgtaaa acgacggcca gtgaggtaat acgactcact ataggggaat tgtgagcgga
     6241 taacaattcc cctctagatt t
//
