LOCUS       sequence-300138-        8961 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 S (surface 
            glycoprotein).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8961)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 8961)
  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"
COMMENT     Sequence Label: pGBW-m4164342
FEATURES             Location/Qualifiers
     source          1..8961
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        106..409
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          688..1698
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1622..1644
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     polyA_signal    5584..5705
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5595..5614)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5649..5668
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      5820..5866
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      5896..6032
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6077..6148
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(6374..6962)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6454..6473)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             7708..8502
                     /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"
     CDS             7708..8502
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(7762..7781)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     8372..8391
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      8515..8651
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    8796..8867
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(8872..8918)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    join(8936..8961,1..23)
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 cttcgatgga tagcgatttt attatctaac tgctgatcga gtgtagccag atctatagta
       61 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
      121 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
      181 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggactattt
      241 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
      301 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
      361 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgc gtcgaggtga
      421 gccccacgtt ctgcttcact ctccccatct cccccccctc cccaccccca attttgtatt
      481 tatttatttt ttaattattt tatgcagcga tgggggcggg gggggggggg gcgcgcgccr
      541 ggsggggsgg ggsggggsgr ggggsggggs ggggsgrggc ggagaggtgc ggcggcagcc
      601 aatcagagcg gcgcgctccg aaagtttcct tttatggcga ggcggcggcg gcggcggccc
      661 tataaaaagc gaagcgcgcg gcgggcggga gtcgctgcgt tgccttcgcc ccgtgccccg
      721 ctccgcgccg cctcgcgccg cccgccccgg ctctgactga ccgcgttact cccacaggtg
      781 agcgggcggg acggcccttc tccctccggg ctgtaattag cgcttggttt aatgacggct
      841 cgtttctttt ctgtggctgc gtgaaagcct taaagggctc cgggagggcc tttgtgcggg
      901 ggggagcggc tcggggggtg cgtgcgtgtg tgtgtgcgtg gggagcgccg cgtgcggccc
      961 gcgctgcccg gcggctgtga gcgctgcggg cgcggcgcgg ggctttgtgc gctccgcgtg
     1021 tgcgcgaggg gagcgcgggc cgggggcggt gccccgcggt gcgggggggc tgcgagggga
     1081 acaaaggctg cgtgcggggt gtgtgcgtgg gggggtgagc agggggtgtg ggcgcggcgg
     1141 tcgggctgta acccccccct ggcacccccc tccccgagtt gctgagcacg gcccggcttc
     1201 gggtgcgggg ctccgtgcgg ggcgtggcgc ggggctcgcc gtgccgggcg gggggkggcg
     1261 gcaggkgggg gtgccgggcg gggcggggcc gcctcgggcc ggggrgggct cgggggrggg
     1321 gcgcggcggc cccggagcgc cggcggctgt cgaggcgcgg cgagccgcag ccattgcctt
     1381 ttatggtaat cgtgcgagag ggcgcaggga cttcctttgt cccaaatctg gcggagccga
     1441 aatctgggag gcgccgccgc accccctcta gcgggcgcgg gcgaagcggt gcggcgccgg
     1501 caggaaggaa atgggcgggg agggccttcg tgcgtcgccg cgccgccgtc cccttctcca
     1561 tctccagcct cggggctgcc gcagggggac ggctgccttc gggggggacg gggcagggcg
     1621 gggttcggct tctggcgtgt gaccggcggc tttagagcct ctgctaacca tgttcatgcc
     1681 ttcttctttt tcctacagct cctgggcaac gtgctggttg ttgtgctgtc tcatcatttt
     1741 ggcaaagatc tttgtcgatc ctaccatcca ctcgacacac catgtttgtt ttcttggttc
     1801 ttttgccact ggtaagcagt cagtgcgtca acctgaccac ccggacacaa ctcccacccg
     1861 catacacgaa cagtttcact aggggggttt actaccctga caaagttttt agatctagtg
     1921 ttttgcattc cacgcaagat ctgttccttc cgttcttttc aaatgtcacg tggttccatg
     1981 ctattcacgt cagcggcact aatggaacga aacgattcga taaccccgtc ttgcccttca
     2041 acgatggcgt atatttcgcg tcaacagaga aaagtaatat aattagaggc tggatctttg
     2101 gtacgactct cgacagtaag acgcagtcac ttctgattgt aaacaatgca actaatgtag
     2161 tcatcaaggt gtgtgagttc cagttctgca atgatccttt cttgggagtg tattatcata
     2221 agaataacaa atcctggatg gagtctgagt tccgggtata ttcttcagca aacaattgca
     2281 cgtttgaata tgtaagccaa cctttcctta tggacctgga agggaagcaa ggtaatttca
     2341 agaatcttag ggaatttgtg tttaaaaata tagatggcta cttcaagatc tactcaaagc
     2401 ataccccgat taacctcgtc agagacctgc cccaagggtt cagtgcgctt gaacccctgg
     2461 tagatttgcc aatcggcata aatataacca ggttccagac actgctcgcc ctccaccgga
     2521 gctaccttac tccaggagat tcatctagcg ggtggacagc tggagcggca gcttactacg
     2581 tcggttacct tcaaccgagg acattcttgc ttaaatacaa tgagaatggc acgataactg
     2641 acgccgtaga ttgcgcgctt gatccgctgt cagaaacgaa gtgtacactt aaaagtttta
     2701 cggtcgaaaa gggaatctac caaacatcaa attttagggt acaacctacg gaatctatcg
     2761 tacgcttccc gaatatcact aacttgtgtc cgttcggcga ggtttttaat gcgaccaggt
     2821 ttgcttccgt gtacgcctgg aacaggaaac ggatctccaa ttgtgtcgcc gattactccg
     2881 tcttgtataa ttcagcatct ttcagcacgt ttaaatgtta cggagtttcc cccacaaaat
     2941 tgaatgacct ttgctttacg aacgtctacg cggattcatt tgtaatccgg ggggacgaag
     3001 ttaggcaaat tgcgccaggg cagactggca agatagctga ctataattat aaattgccgg
     3061 atgactttac gggctgtgtg attgcttgga actcaaataa tctggactca aaggtagggg
     3121 gaaattataa ctacctttac aggctgttcc ggaagagtaa tctgaagcca ttcgaaagag
     3181 atataagtac agagatctac caagctggaa gcaccccctg caatggtgtt gaaggattca
     3241 attgttattt cccattgcaa tcctatggtt ttcaaccgac gaatggggtg ggataccaac
     3301 catatcgagt tgtggttctc agtttcgagt tgcttcatgc tcctgcgaca gtatgtggac
     3361 caaaaaaatc tactaatctg gtgaagaata aatgcgtcaa ttttaatttt aatggtctga
     3421 caggtaccgg agtgcttacg gaatctaaca agaagtttct tccttttcaa cagttcggga
     3481 gggacatagc cgacacaact gacgcggtaa gagatccgca gacgttggaa attctcgata
     3541 taactccatg ctcatttgga ggagtgagtg ttatcactcc tggtacgaac accagtaatc
     3601 aggttgcagt tctttatcag gacgtgaatt gtacagaggt cccggttgcg atacacgcgg
     3661 accagctgac gcccacgtgg cgggtttact caactgggtc aaatgtgttt caaaccagag
     3721 ccggctgcct cataggagcc gagcatgtga acaacagtta tgaatgtgac atacctatcg
     3781 gggcggggat ctgcgcttca tatcaaacgc aaaccaattc cccgcgaaga gcgcgaagtg
     3841 ttgcaagtca aagtattata gcgtatacga tgagcttggg tgctgagaac tccgtggcat
     3901 attcaaataa ttccatcgca attcccacga attttacgat ttctgtgacg acggagatat
     3961 tgccggtttc aatgactaag acgagtgttg actgtacgat gtacatctgc ggcgactcaa
     4021 cggagtgttc caacctgctc ctgcagtacg gctcattctg cacccaactg aatcgggcac
     4081 ttacgggtat tgctgttgaa caagacaaaa atacgcaaga ggtgttcgcc caggtcaagc
     4141 agatctacaa gacaccaccg attaaggact ttggggggtt taatttttct cagatacttc
     4201 ccgatcccag taaacctagt aaaaggtcat ttatagagga cctcttgttc aacaaggtaa
     4261 cgctcgctga tgcaggattc atcaagcagt atggcgattg tcttggggac attgcggcgc
     4321 gcgacctgat ttgtgcccag aagttcaacg gactgacggt acttccgcct ctcctgaccg
     4381 acgaaatgat cgcacaatac acttctgcat tgcttgctgg caccataacc tcaggttgga
     4441 ccttcggtgc cggggcggcc ttgcaaatcc cctttgctat gcaaatggca tatcgattca
     4501 atgggatagg agttactcaa aatgttctgt atgaaaacca aaagctgata gccaatcaat
     4561 ttaatagtgc tattgggaaa attcaggaca gcctgagtag tacggcttcc gctctcggca
     4621 aactccaaga tgtcgttaac caaaatgcac aagctctcaa tacacttgtt aagcaactga
     4681 gctccaactt cggcgctata tcatcagttc ttaacgatat tctttcccga ctcgacaagg
     4741 tcgaagccga agtccaaata gacaggctga taaccgggag gctgcaaagt ctccaaacgt
     4801 atgttaccca acaactcata cgagcggccg agattagggc gtcagccaac ctcgccgcta
     4861 ctaaaatgtc tgaatgtgta ctcgggcagt ccaagcgcgt ggacttttgt ggcaagggtt
     4921 atcatctgat gagtttcccg caatcagcgc ctcacggagt ggtcttcctc cacgtgacgt
     4981 atgtccctgc gcaagaaaaa aactttacga cagcccctgc catttgtcat gacgggaagg
     5041 cgcactttcc acgcgaaggc gtttttgtat ccaacggcac ccactggttt gtcacacagc
     5101 gaaattttta tgagcctcag atcataacta ctgacaatac tttcgtctca gggaactgcg
     5161 atgtagtcat agggatcgta aacaatacgg tatacgatcc cctccagcct gaactggact
     5221 catttaaaga ggagctggac aagtatttca aaaatcacac atcaccggac gtggacctcg
     5281 gagatattag tggtattaac gcaagcgtag taaatattca aaaggaaatc gaccgcttga
     5341 atgaagtcgc gaaaaacctg aacgagtctc ttattgatct tcaagaactc ggtaagtatg
     5401 aacaatacat caagtggcct tggtacatat ggcttggatt cattgcgggg ttgatcgcaa
     5461 tcgtaatggt taccattatg ttgtgttgca tgacaagctg ttgcagctgc ctcaaaggtt
     5521 gttgttcttg cggtagttgc tgctaagact gagatacagc gtaccttcag ctcacagaca
     5581 tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct
     5641 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac
     5701 aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg
     5761 ttttttaaag caagtaaaac ctctacaaat gtggtattgg cccatctcta tcggtatcgt
     5821 agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgtgcc cctcgggccg
     5881 gattgctatc taccggcatt ggcgcagaaa aaaatgcctg atgcgacgct gcgcgtctta
     5941 tactcccaca tatgccagat tcagcaacgg atacggcttc cccaacttgc ccacttccat
     6001 acgtgtcctc cttaccagaa atttatcctt aaggtcgtca gctatcctgc aggcgatctc
     6061 tcgatttcga tcaagacatt cctttaatgg tcttttctgg acaccactag gggtcagaag
     6121 tagttcatca aactttcttc cctccctaat ctcattggtt accttgggct atcgaaactt
     6181 aattaaccag tcaagtcagc tacttggcga gatcgacttg tctgggtttc gactacgctc
     6241 agaattgcgt cagtcaagtt cgatctggtc cttgctattg cacccgttct ccgattacga
     6301 gtttcattta aatcatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     6361 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     6421 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     6481 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     6541 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     6601 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     6661 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     6721 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     6781 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     6841 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     6901 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     6961 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     7021 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     7081 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     7141 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcat
     7201 ttaaatttcc gaactctcca aggccctcgt cggaaaatct tcaaaccttt cgtccgatcc
     7261 atcttgcagg ctacctctcg aacgaactat cgcaagtctc ttggccggcc ttgcgccttg
     7321 gctattgctt ggcagcgcct atcgccaggt attactccaa tcccgaatat ccgagatcgg
     7381 gatcacccga gagaagttca acctacatcc tcaatcccga tctatccgag atccgaggaa
     7441 tatcgaaatc ggggcgcgcc tggtgtaccg agaacgatcc tctcagtgcg agtctcgacg
     7501 atccatatcg ttgcttggca gtcagccagt cggaatccag cttgggaccc aggaagtcca
     7561 atcgtcagat attgtactca agcctggtca cggcagcgta ccgatctgtt taaacctaga
     7621 tattgatagt ctgatcggtc aacgtataat cgagtcctag cttttgcaaa catctatcaa
     7681 gagacaggat cagcaggagg ctttcgcatg attgaacaag atggattgca cgcaggttct
     7741 ccggcggctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     7801 tctgatgccg ccgtgttccg gctgtcagcg caggggcgtc cggttctttt tgtcaagacc
     7861 gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc gtggctggcg
     7921 acgacgggcg ttccttgcgc ggctgtgctc gacgttgtca ctgaagcggg aagggactgg
     7981 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     8041 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     8101 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     8161 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     8221 gccaggctca aggcgtctat gcccgacggc gaggatctcg tcgtgaccca cggcgatgcc
     8281 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgt
     8341 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     8401 cttggcggcg aatgggctga ccgcttcctt gtgctttacg gtatcgccgc gcccgattcg
     8461 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gaccgattct aggtgcattg
     8521 gcgcagaaaa aaatgcctga tgcgacgctg cgcgtcttat actcccacat atgccagatt
     8581 cagcaacgga tacggcttcc ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa
     8641 tttatcctta aggtcgttta aactcgactc tggctctatc gaatctccgt cgtttcgagc
     8701 ttacgcgaac agccgtggcg ctcatttgct cgtcgggcat cgaatctcgt cagctatcgt
     8761 cagcttacct ttttggcagc gatcgcggct cccgacatct tggaccatta gctccacagg
     8821 tatcttcttc cctctagtgg tcataacagc agcttcagct acctctcaat tcaaaaaacc
     8881 cctcaagacc cgtttagagg ccccaagggg ttatgctatc aatcgttgcg ttacacacac
     8941 aaaaaaccaa cacacatcca t
//
