LOCUS       sequence-352513-        6261 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Generation of MS2 Virus-Like Particles packaged with the sequence 
            for the B.1.640 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  JOURNAL   Unpublished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 Apr 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            "Unpublished"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_B.1.640
FEATURES             Location/Qualifiers
     source          1..6261
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      30..77
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(175..192)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(285..870)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(365..384)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1044..1838)
                     /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(1839..1943)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2244..2266
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2258..2275
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2259..2275
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2280..2298
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2299..2323
                     /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(2332..2337,2338..2355,2356..2361)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     RBS             2381..2403
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3668..4054
                     /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             4097..4114
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      4493..4540
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        4586..4604
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_RNA        join(6258..6261,1..15)
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
ORIGIN
        1 gaggatcacc catgtccgct gagcaataac tagcataacc ccttggggcc tctaaacggg
       61 tcttgagggg ttttttgtaa tttcactgcc cgctttccag tcgggaaacc tgtcgtgcca
      121 gctgcattaa catggtcata gctgtttcct tgcgtattgg gcgctctccg cttcctcgct
      181 cactgactcg ctgcgctcgg tcgttcgggt aaagcctggg gtgcctaatg agcaaaaggc
      241 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
      301 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
      361 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
      421 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
      481 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
      541 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
      601 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
      661 gcgaggtatg taggcgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
      721 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
      781 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
      841 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
      901 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
      961 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     1021 gagtaaactt ggtctgacag ttattagaaa aattcatcca gcagacgata aaacgcaata
     1081 cgctggctat ccggtgccgc aatgccatac agcaccagaa aacgatccgc ccattcgccg
     1141 cccagttctt ccgcaatatc acgggtggcc agcgcaatat cctgataacg atccgccacg
     1201 cccagacggc cgcaatcaat aaagccgcta aaacggccat tttccaccat aatgttcggc
     1261 aggcacgcat caccatgggt caccaccaga tcttcgccat ccggcatgct cgctttcaga
     1321 cgcgcaaaca gctctgccgg tgccaggccc tgatgttctt catccagatc atcctgatcc
     1381 accaggcccg cttccatacg ggtacgcgca cgttcaatac gatgtttcgc ctgatgatca
     1441 aacggacagg tcgccgggtc cagggtatgc agacgacgca tggcatccgc cataatgctc
     1501 actttttctg ccggcgccag atggctagac agcagatcct gacccggcac ttcgcccagc
     1561 agcagccaat cacggcccgc ttcggtcacc acatccagca ccgccgcaca cggaacaccg
     1621 gtggtggcca gccagctcag acgcgccgct tcatcctgca gctcgttcag cgcaccgctc
     1681 agatcggttt tcacaaacag caccggacga ccctgcgcgc tcagacgaaa caccgccgca
     1741 tcagagcagc caatggtctg ctgcgcccaa tcatagccaa acagacgttc cacccacgct
     1801 gccgggctac ccgcatgcag gccatcctgt tcaatcatac tcttcctttt tcaatattat
     1861 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     1921 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctaa attgtaagcg
     1981 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     2041 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     2101 gccgctacag ggcgctccca ttcgccattc aggctgcgca actgttggga agggcgtttc
     2161 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt
     2221 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgaggt
     2281 aatacgactc actatagggg aattgtgagc ggataacaat tcccctctag atttacactt
     2341 tatgcttccg gctcgtatgt ttctagagaa ttcaaataat tttgtttaac tttaagaagg
     2401 agatttaaat tggctatcgc tgtaggtagc cggaattcca ttcctaggag gtttgacctg
     2461 tgcgagcttt tagtaccctt gatagggaga acgaaacctt cgtcccctcc gttcgcgttt
     2521 acgcggacgg tgagactgaa gataactcat tctctttaaa atatcgttcg aactggactc
     2581 ccggtcgttt taactcgact ggggccaaaa cgaaacagtg gcactacccc tctccgtatt
     2641 cacggggggc gttaagtgtc acatcgatag atcaaggtgc ctacaagcga agtgggtcat
     2701 cgtggggtcg cccgtacgag gagaaagccg gtttcggctt ctccctcgac gcacgctcct
     2761 gctacagcct cttccctgta agccaaaact tgacttacat cgaagtgccg cagaacgttg
     2821 cgaaccgggc gtcgaccgaa gtcctgcaaa aggtcaccca gggtaatttt aaccttggtg
     2881 ttgctttagc agaggccagg tcgacagcct cacaactcgc gacgcaaacc attgcgctcg
     2941 tgaaggcgta cactgccgct cgtcgcggta attggcgcca ggcgctccgc taccttgccc
     3001 taaacgaaga tcgaaagttt cgatcaaaac acgtggccgg cagatggttg gagttgcagt
     3061 tcggttggtt accactaatg agtgatatcc agggtgcata tgagatgctt acgaaggttc
     3121 accttcaaga gtttcttcct atgagagccg tacgtcaggt cggtactaac atcaagttag
     3181 atggccgtct gtcgtatcca gctgcaaact tccagacaac gtgcaacata tcgcgacgta
     3241 tcgtgatatg gttttacata aacgatgcac gtttggcatg gttgtcgtcc ctaggtatct
     3301 tgaacccact aggtatagtg tgggaaaagg tgcctttctc attcgttgtc gactggctcc
     3361 tacctgtagg taacatgctc gagggcctta cggcccccgt gggatgctcc tacatgtcag
     3421 gaacagttac tgacgtaata acgggtgagt ccatcataag cgttgacgct ccctacgggt
     3481 ggactgtgga gagacagggc actgctaagg cccaaatctc agccatgcat cgaggggtac
     3541 aatccgtatg gccaacaact ggcgcgtacg taaagtctcc tttctcgatg gtccatacct
     3601 tagatgcgtt agcattaatc aggcaacggc tctctagata gagccctcaa ccggagtttg
     3661 aagcatggct tctaacttta ctcagttcgt tctcgtcgac aatggcggaa ctggcgacgt
     3721 gactgtcgcc ccaagcaact tcgctaacgg ggtcgctgaa tggatcagct ctaactcgcg
     3781 ttcacaggct tacaaagtaa cctgtagcgt tcgtcagagc tctgcgcaga atcgcaaata
     3841 caccatcaaa gtcgaggtgc ctaaagtggc aacccagact gttggtggtg tagagcttcc
     3901 tgtagccgca tggcgttcgt acttaaatat ggaactaacc attccaattt tcgctacgaa
     3961 ttccgactgc gagcttattg ttaaggcaat gcaaggtctt ctaaaagatg gaaacccgat
     4021 tccctcagca atcgcagcaa actccggcat ctacgctaac tttactcagt tcgttctcgt
     4081 cgacaatggc ggtacccatc accatcacca tcacggtacc ggcgacgtga ctgtcgcccc
     4141 aagcaacttc gctaacgggg tcgctgaatg gatcagctct aactcgcgtt cacaggctta
     4201 caaagtaacc tgtagcgttc gtcagagctc tgcgcagaat cgcaaataca ccatcaaagt
     4261 cgaggtgcct aaagtggcaa cccagactgt tggtggtgta gagcttcctg tagccgcatg
     4321 gcgttcgtac ttaaatatgg aactaaccat tccaattttc gctacgaatt ccgactgcga
     4381 gcttattgtt aaggcaatgc aaggtcttct aaaagatgga aacccgattc cctcagcaat
     4441 cgcagcaaac tccggcatct actaatagac gccggccacc gctgagcaat aactagcata
     4501 accccttggg gcctctaaac gggtcttgag gggttttttg tcgaacagaa agtaatcgta
     4561 ttgtacacgg ccgcataatc gaaattaata cgactcacta taggagagtc caaccaacag
     4621 aatctattgt tagatttcct aatattacaa acttgtgccc ttttggtgaa gtttttaacg
     4681 ccaccagctt tgcatctgtt tatgcttgga acaggaagag aatcagcaac tgtgttgctg
     4741 attattctgt cctatataat tccgcatcat tttccacttt taagtgttat ggagtgtctc
     4801 ctactaaatt aaatgatctc tgctttacta gtgtctatgc agattcattt gtaattagag
     4861 gtgatgaagt cagacaaatc gctccagggc aaactggaaa gattgctgat tataattata
     4921 aattaccaga tgattttaca ggctgcgtta tagcttggaa ttctaacaat cttgattcta
     4981 aggttggtgg taataataat tacctgtata gattgtttag gaagtctaat ctcaaacctt
     5041 ttgagagaga tatttcaact gaaatctatc aggccggtag cacaccttgt aatggtgttg
     5101 aaggttttaa ttgttaccgt cctttacaat catatggttt ccaacccact tatggtgttg
     5161 gttaccaacc atacagagta gtagtacttt cttttgaact tctacatgca ccagcaactg
     5221 tttgtggacc taaaaagtct actaatttgg ttaaaaacaa atgtgtcaat ttcaacttca
     5281 atggtttaac aggcacaggt gttcttactg agtctaacaa aaagtttctg cctttccaac
     5341 aatttggcag agacattgct gacactactg atgctgtccg tgatccacag acacttgaga
     5401 ttcttgacat tacaccatgt tcttttggtg gtgtcagtgt tataacacca ggaacaaata
     5461 cttctaacca ggttgctgtt ctttatcagg gtgttaactg cacagaagtc cctgttgcta
     5521 ttcatgcaga tcaacttact cctacttggc gtgtttattc tacaggttct aatgtttttc
     5581 aaacacgtgc aggctgttta ataggggctg aacatgtcaa caactcatat gagtgtgaca
     5641 tacccattgg tgcaggtata tgcgctagtt atcagactca gactaattct catcggcggg
     5701 cacgtagtgt agctagtcaa tccatcattg cctacactat gtcacttggt gcagaaaatt
     5761 cagttgctta ctctaataac tctattgcca tacccacaaa ttttactatt agtgttacca
     5821 cagaaattct accagtgtct atgaccaaga catcagtaga ttgtacaatg tacatttgtg
     5881 gtgattcaac tgaatgcagc aatcttttgt tgcaatatgg cagtttttgt acacaattaa
     5941 accgtgcttt aactggaata gctgttgaac aagacaaaaa cacccaagaa gtttttgcac
     6001 aagtcaaaca aatttacaaa acaccaccaa ttaaagattt tggtggtttt aatttttcac
     6061 aaatattacc agatccatca aaaccaagca agaggtcatt tattgaagat ctacttttca
     6121 acaaagtgac acttgcagat gctggcttca tcaaacaata tggtgattgc cttggtgata
     6181 ttgctgctag agacctcatt tgtgcacaaa agtttaacgg ccttaatgtt ttgccacctt
     6241 tgctcacaga tgaaatgaca t
//
