LOCUS       sequence-343941-        6261 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Generation of MS2 Virus-Like Particles packaged with the sequence 
            for the WHO Theta 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  JOURNAL   Unpublished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            "Unpublished"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_Theta
FEATURES             Location/Qualifiers
     source          1..6261
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(3..8,9..26,27..32)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     RBS             52..74
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1339..1725
                     /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             1768..1785
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      2164..2211
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        2257..2275
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_RNA        3929..3947
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     terminator      3962..4009
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(4107..4124)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4217..4802)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4297..4316)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4976..5770)
                     /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(5771..5875)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6176..6198
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6190..6207
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6191..6207
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6212..6230
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    6231..6255
                     /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)."
ORIGIN
        1 gatttacact ttatgcttcc ggctcgtatg tttctagaga attcaaataa ttttgtttaa
       61 ctttaagaag gagatttaaa ttggctatcg ctgtaggtag ccggaattcc attcctagga
      121 ggtttgacct gtgcgagctt ttagtaccct tgatagggag aacgaaacct tcgtcccctc
      181 cgttcgcgtt tacgcggacg gtgagactga agataactca ttctctttaa aatatcgttc
      241 gaactggact cccggtcgtt ttaactcgac tggggccaaa acgaaacagt ggcactaccc
      301 ctctccgtat tcacgggggg cgttaagtgt cacatcgata gatcaaggtg cctacaagcg
      361 aagtgggtca tcgtggggtc gcccgtacga ggagaaagcc ggtttcggct tctccctcga
      421 cgcacgctcc tgctacagcc tcttccctgt aagccaaaac ttgacttaca tcgaagtgcc
      481 gcagaacgtt gcgaaccggg cgtcgaccga agtcctgcaa aaggtcaccc agggtaattt
      541 taaccttggt gttgctttag cagaggccag gtcgacagcc tcacaactcg cgacgcaaac
      601 cattgcgctc gtgaaggcgt acactgccgc tcgtcgcggt aattggcgcc aggcgctccg
      661 ctaccttgcc ctaaacgaag atcgaaagtt tcgatcaaaa cacgtggccg gcagatggtt
      721 ggagttgcag ttcggttggt taccactaat gagtgatatc cagggtgcat atgagatgct
      781 tacgaaggtt caccttcaag agtttcttcc tatgagagcc gtacgtcagg tcggtactaa
      841 catcaagtta gatggccgtc tgtcgtatcc agctgcaaac ttccagacaa cgtgcaacat
      901 atcgcgacgt atcgtgatat ggttttacat aaacgatgca cgtttggcat ggttgtcgtc
      961 cctaggtatc ttgaacccac taggtatagt gtgggaaaag gtgcctttct cattcgttgt
     1021 cgactggctc ctacctgtag gtaacatgct cgagggcctt acggcccccg tgggatgctc
     1081 ctacatgtca ggaacagtta ctgacgtaat aacgggtgag tccatcataa gcgttgacgc
     1141 tccctacggg tggactgtgg agagacaggg cactgctaag gcccaaatct cagccatgca
     1201 tcgaggggta caatccgtat ggccaacaac tggcgcgtac gtaaagtctc ctttctcgat
     1261 ggtccatacc ttagatgcgt tagcattaat caggcaacgg ctctctagat agagccctca
     1321 accggagttt gaagcatggc ttctaacttt actcagttcg ttctcgtcga caatggcgga
     1381 actggcgacg tgactgtcgc cccaagcaac ttcgctaacg gggtcgctga atggatcagc
     1441 tctaactcgc gttcacaggc ttacaaagta acctgtagcg ttcgtcagag ctctgcgcag
     1501 aatcgcaaat acaccatcaa agtcgaggtg cctaaagtgg caacccagac tgttggtggt
     1561 gtagagcttc ctgtagccgc atggcgttcg tacttaaata tggaactaac cattccaatt
     1621 ttcgctacga attccgactg cgagcttatt gttaaggcaa tgcaaggtct tctaaaagat
     1681 ggaaacccga ttccctcagc aatcgcagca aactccggca tctacgctaa ctttactcag
     1741 ttcgttctcg tcgacaatgg cggtacccat caccatcacc atcacggtac cggcgacgtg
     1801 actgtcgccc caagcaactt cgctaacggg gtcgctgaat ggatcagctc taactcgcgt
     1861 tcacaggctt acaaagtaac ctgtagcgtt cgtcagagct ctgcgcagaa tcgcaaatac
     1921 accatcaaag tcgaggtgcc taaagtggca acccagactg ttggtggtgt agagcttcct
     1981 gtagccgcat ggcgttcgta cttaaatatg gaactaacca ttccaatttt cgctacgaat
     2041 tccgactgcg agcttattgt taaggcaatg caaggtcttc taaaagatgg aaacccgatt
     2101 ccctcagcaa tcgcagcaaa ctccggcatc tactaataga cgccggccac cgctgagcaa
     2161 taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gtcgaacaga
     2221 aagtaatcgt attgtacacg gccgcataat cgaaattaat acgactcact ataggagagt
     2281 ccaaccaaca gaatctattg ttagatttcc taatattaca aacttgtgcc cttttggtga
     2341 agtttttaac gccaccagat ttgcatctgt ttatgcttgg aacaggaaga gaatcagcaa
     2401 ctgtgttgct gattattctg tcctatataa ttccgcatca ttttccactt ttaagtgtta
     2461 tggagtgtct cctactaaat taaatgatct ctgctttact aatgtctatg cagattcatt
     2521 tgtaattaga ggtgatgaag tcagacaaat cgctccaggg caaactggaa agattgctga
     2581 ttataattat aaattaccag atgattttac aggctgcgtt atagcttgga attctaacaa
     2641 tcttgattct aaggttggtg gtaattataa ttacctgtat agattgttta ggaagtctaa
     2701 tctcaaacct tttgagagag atatttcaac tgaaatctat caggccggta gcacaccttg
     2761 taatggtgtt aaaggtttta attgttactt tcctttacaa tcatatggtt tccaacccac
     2821 ttatggtgtt ggttaccaac catacagagt agtagtactt tcttttgaac ttctacatgc
     2881 accagcaact gtttgtggac ctaaaaagtc tactaatttg gttaaaaaca aatgtgtcaa
     2941 tttcaacttc aatggtttaa caggcacagg tgttcttact gagtctaaca aaaagtttct
     3001 gcctttccaa caatttggca gagacattgc tgacactact gatgctgtcc gtgatccaca
     3061 gacacttgag attcttgaca ttacaccatg ttcttttggt ggtgtcagtg ttataacacc
     3121 aggaacaaat acttctaacc aggttgctgt tctttatcag gatgttaact gcacagaagt
     3181 ccctgttgct attcatgcag atcaacttac tcctacttgg cgtgtttatt ctacaggttc
     3241 taatgttttt caaacacgtg caggctgttt aataggggct gaacatgtca acaactcata
     3301 tgagtgtgac atacccattg gtgcaggtat atgcgctagt tatcagactc agactaattc
     3361 tcatcggcgg gcacgtagtg tagctagtca atccatcatt gcctacacta tgtcacttgg
     3421 tgcagaaaat tcagttgctt actctaataa ctctattgcc atacccacaa attttactat
     3481 tagtgttacc acagaaattc taccagtgtc tatgaccaag acatcagtag attgtacaat
     3541 gtacatttgt ggtgattcaa ctgaatgcag caatcttttg ttgcaatatg gcagtttttg
     3601 tacacaatta aaccgtgctt taactggaat agctgttgaa caagacaaaa acacccaaga
     3661 agtttttgca caagtcaaac aaatttacaa aacaccacca attaaagatt ttggtggttt
     3721 taatttttca caaatattac cagatccatc aaaaccaagc aagaggtcat ttattgaaga
     3781 tctacttttc aacaaagtga cacttgcaga tgctggcttc atcaaacaat atggtgattg
     3841 ccttggtgat attgctgcta gagacctcat ttgtgcacaa aagtttaacg gccttactgt
     3901 tttgccacct ttgctcacag atgaaatgac atgaggatca cccatgtccg ctgagcaata
     3961 actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgt aatttcactg
     4021 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aacatggtca tagctgtttc
     4081 cttgcgtatt gggcgctctc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     4141 gtaaagcctg gggtgcctaa tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     4201 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     4261 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     4321 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     4381 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     4441 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     4501 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     4561 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcgct acagagttct
     4621 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     4681 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     4741 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     4801 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     4861 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     4921 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttattaga
     4981 aaaattcatc cagcagacga taaaacgcaa tacgctggct atccggtgcc gcaatgccat
     5041 acagcaccag aaaacgatcc gcccattcgc cgcccagttc ttccgcaata tcacgggtgg
     5101 ccagcgcaat atcctgataa cgatccgcca cgcccagacg gccgcaatca ataaagccgc
     5161 taaaacggcc attttccacc ataatgttcg gcaggcacgc atcaccatgg gtcaccacca
     5221 gatcttcgcc atccggcatg ctcgctttca gacgcgcaaa cagctctgcc ggtgccaggc
     5281 cctgatgttc ttcatccaga tcatcctgat ccaccaggcc cgcttccata cgggtacgcg
     5341 cacgttcaat acgatgtttc gcctgatgat caaacggaca ggtcgccggg tccagggtat
     5401 gcagacgacg catggcatcc gccataatgc tcactttttc tgccggcgcc agatggctag
     5461 acagcagatc ctgacccggc acttcgccca gcagcagcca atcacggccc gcttcggtca
     5521 ccacatccag caccgccgca cacggaacac cggtggtggc cagccagctc agacgcgccg
     5581 cttcatcctg cagctcgttc agcgcaccgc tcagatcggt tttcacaaac agcaccggac
     5641 gaccctgcgc gctcagacga aacaccgccg catcagagca gccaatggtc tgctgcgccc
     5701 aatcatagcc aaacagacgt tccacccacg ctgccgggct acccgcatgc aggccatcct
     5761 gttcaatcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     5821 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     5881 ttccccgaaa agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa
     5941 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
     6001 atcaaaagaa tagaccgaga tagggttgag tggccgctac agggcgctcc cattcgccat
     6061 tcaggctgcg caactgttgg gaagggcgtt tcggtgcggg cctcttcgct attacgccag
     6121 ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag
     6181 tcacgacgtt gtaaaacgac ggccagtgag gtaatacgac tcactatagg ggaattgtga
     6241 gcggataaca attcccctct a
//
