LOCUS       sequence-297247-        8964 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 8964)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 8964)
  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-m4164344
FEATURES             Location/Qualifiers
     source          1..8964
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    3809..3930
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3820..3839)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3874..3893
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      4045..4091
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      4121..4257
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4302..4373
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(4599..5187)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4679..4698)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             5933..6727
                     /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             5933..6727
                     /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(5987..6006)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6597..6616
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      6740..6876
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7021..7092
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7097..7143)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7161..7209
                     /note="synthetic polyadenylation signal"
     enhancer        7292..7595
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7598..7873
                     /label=chicken beta-actin promoter
     intron          7874..8884
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     8808..8830
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
ORIGIN
        1 accatgtttg ttttcttggt tcttttgcca ctggtaagca gtcagtgcgt caacctgacc
       61 acccggacac aactcccacc cgcatacacg aacagtttca ctaggggggt ttactaccct
      121 gacaaagttt ttagatctag tgttttgcat tccacgcaag atctgttcct tccgttcttt
      181 tcaaatgtca cgtggttcca tgctattcac gtcagcggca ctaatggaac gaaacgattc
      241 gataaccccg tcttgccctt caacgatggc gtatatttcg cgtcaacaga gaaaagtaat
      301 ataattagag gctggatctt tggtacgact ctcgacagta agacgcagtc acttctgatt
      361 gtaaacaatg caactaatgt agtcatcaag gtgtgtgagt tccagttctg caatgatcct
      421 ttcttgggag tgtattatca taagaataac aaatcctgga tggagtctga gttccgggta
      481 tattcttcag caaacaattg cacgtttgaa tatgtaagcc aacctttcct tatggacctg
      541 gaagggaagc aaggtaattt caagaatctt agggaatttg tgtttaaaaa tatagatggc
      601 tacttcaaga tctactcaaa gcataccccg attaacctcg tcagagacct gccccaaggg
      661 ttcagtgcgc ttgaacccct ggtagatttg ccaatcggca taaatataac caggttccag
      721 acactgctcg ccctccaccg gagctacctt actccaggag attcatctag cgggtggaca
      781 gctggagcgg cagcttacta cgtcggttac cttcaaccga ggacattctt gcttaaatac
      841 aatgagaatg gcacgataac tgacgccgta gattgcgcgc ttgatccgct gtcagaaacg
      901 aagtgtacac ttaaaagttt tacggtcgaa aagggaatct accaaacatc aaattttagg
      961 gtacaaccta cggaatctat cgtacgcttc ccgaatatca ctaacttgtg tccgttcggc
     1021 gaggttttta atgcgaccag gtttgcttcc gtgtacgcct ggaacaggaa acggatctcc
     1081 aattgtgtcg ccgattactc cgtcttgtat aattcagcat ctttcagcac gtttaaatgt
     1141 tacggagttt cccccacaaa attgaatgac ctttgcttta cgaacgtcta cgcggattca
     1201 tttgtaatcc ggggggacga agttaggcaa attgcgccag ggcagactgg caagatagct
     1261 gactataatt ataaattgcc ggatgacttt acgggctgtg tgattgcttg gaactcaaat
     1321 aatctggact caaaggtagg gggaaattat aactaccttt acaggctgtt ccggaagagt
     1381 aatctgaagc cattcgaaag agatataagt acagagatct accaagctgg aagcaccccc
     1441 tgcaatggtg ttgaaggatt caattgttat ttcccattgc aatcctatgg ttttcaaccg
     1501 acgaatgggg tgggatacca accatatcga gttgtggttc tcagtttcga gttgcttcat
     1561 gctcctgcga cagtatgtgg accaaaaaaa tctactaatc tggtgaagaa taaatgcgtc
     1621 aattttaatt ttaatggtct gacaggtacc ggagtgctta cggaatctaa caagaagttt
     1681 cttccttttc aacagttcgg gagggacata gccgacacaa ctgacgcggt aagagatccg
     1741 cagacgttgg aaattctcga tataactcca tgctcatttg gaggagtgag tgttatcact
     1801 cctggtacga acaccagtaa tcaggttgca gttctttatc aggacgtgaa ttgtacagag
     1861 gtcccggttg cgatacacgc ggaccagctg acgcccacgt ggcgggttta ctcaactggg
     1921 tcaaatgtgt ttcaaaccag agccggctgc ctcataggag ccgagcatgt gaacaacagt
     1981 tatgaatgtg acatacctat cggggcgggg atctgcgctt catatcaaac gcaaaccaat
     2041 tccccgcgaa gagcgcgaag tgttgcaagt caaagtatta tagcgtatac gatgagcttg
     2101 ggtgctgaga actccgtggc atattcaaat aattccatcg caattcccac gaattttacg
     2161 atttctgtga cgacggagat attgccggtt tcaatgacta agacgagtgt tgactgtacg
     2221 atgtacatct gcggcgactc aacggagtgt tccaacctgc tcctgcagta cggctcattc
     2281 tgcacccaac tgaatcgggc acttacgggt attgctgttg aacaagacaa aaatacgcaa
     2341 gaggtgttcg cccaggtcaa gcagatctac aagacaccac cgattaagga ctttgggggg
     2401 tttaattttt ctcagatact tcccgatccc agtaaaccta gtaaaaggtc atttatagag
     2461 gacctcttgt tcaacaaggt aacgctcgct gatgcaggat tcatcaagca gtatggcgat
     2521 tgtcttgggg acattgcggc gcgcgacctg atttgtgccc agaagttcaa cggactgacg
     2581 gtacttccgc ctctcctgac cgacgaaatg atcgcacaat acacttctgc attgcttgct
     2641 ggcaccataa cctcaggttg gaccttcggt gccggggcgg ccttgcaaat cccctttgct
     2701 atgcaaatgg catatcgatt caatgggata ggagttactc aaaatgttct gtatgaaaac
     2761 caaaagctga tagccaatca atttaatagt gctattggga aaattcagga cagcctgagt
     2821 agtacggctt ccgctctcgg caaactccaa gatgtcgtta accaaaatgc acaagctctc
     2881 aatacacttg ttaagcaact gagctccaac ttcggcgcta tatcatcagt tcttaacgat
     2941 attctttccc gactcgacaa ggtcgaagcc gaagtccaaa tagacaggct gataaccggg
     3001 aggctgcaaa gtctccaaac gtatgttacc caacaactca tacgagcggc cgagattagg
     3061 gcgtcagcca acctcgccgc tactaaaatg tctgaatgtg tactcgggca gtccaagcgc
     3121 gtggactttt gtggcaaggg ttatcatctg atgagtttcc cgcaatcagc gcctcacgga
     3181 gtggtcttcc tccacgtgac gtatgtccct gcgcaagaaa aaaactttac gacagcccct
     3241 gccatttgtc atgacgggaa ggcgcacttt ccacgcgaag gcgtttttgt atccaacggc
     3301 acccactggt ttgtcacaca gcgaaatttt tatgagcctc agatcataac tactgacaat
     3361 actttcgtct cagggaactg cgatgtagtc atagggatcg taaacaatac ggtatacgat
     3421 cccctccagc ctgaactgga ctcatttaaa gaggagctgg acaagtattt caaaaatcac
     3481 acatcaccgg acgtggacct cggagatatt agtggtatta acgcaagcgt agtaaatatt
     3541 caaaaggaaa tcgaccgctt gaatgaagtc gcgaaaaacc tgaacgagtc tcttattgat
     3601 cttcaagaac tcggtaagta tgaacaatac atcaagtggc cttggtacat atggcttgga
     3661 ttcattgcgg ggttgatcgc aatcgtaatg gttaccatta tgttgtgttg catgacaagc
     3721 tgttgcagct gcctcaaagg ttgttgttct tgcggtagtt gctgcaagta agactgagat
     3781 acagcgtacc ttcagctcac agacatgata agatacattg atgagtttgg acaaaccaca
     3841 actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt
     3901 gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt
     3961 caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt
     4021 attggcccat ctctatcggt atcgtagcat aaccccttgg ggcctctaaa cgggtcttga
     4081 ggggtttttt gtgcccctcg ggccggattg ctatctaccg gcattggcgc agaaaaaaat
     4141 gcctgatgcg acgctgcgcg tcttatactc ccacatatgc cagattcagc aacggatacg
     4201 gcttccccaa cttgcccact tccatacgtg tcctccttac cagaaattta tccttaaggt
     4261 cgtcagctat cctgcaggcg atctctcgat ttcgatcaag acattccttt aatggtcttt
     4321 tctggacacc actaggggtc agaagtagtt catcaaactt tcttccctcc ctaatctcat
     4381 tggttacctt gggctatcga aacttaatta accagtcaag tcagctactt ggcgagatcg
     4441 acttgtctgg gtttcgacta cgctcagaat tgcgtcagtc aagttcgatc tggtccttgc
     4501 tattgcaccc gttctccgat tacgagtttc atttaaatca tgtgagcaaa aggccagcaa
     4561 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     4621 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     4681 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     4741 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     4801 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     4861 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     4921 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     4981 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     5041 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     5101 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     5161 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     5221 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     5281 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     5341 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     5401 ctatttcgtt catccatagt tgcatttaaa tttccgaact ctccaaggcc ctcgtcggaa
     5461 aatcttcaaa cctttcgtcc gatccatctt gcaggctacc tctcgaacga actatcgcaa
     5521 gtctcttggc cggccttgcg ccttggctat tgcttggcag cgcctatcgc caggtattac
     5581 tccaatcccg aatatccgag atcgggatca cccgagagaa gttcaaccta catcctcaat
     5641 cccgatctat ccgagatccg aggaatatcg aaatcggggc gcgcctggtg taccgagaac
     5701 gatcctctca gtgcgagtct cgacgatcca tatcgttgct tggcagtcag ccagtcggaa
     5761 tccagcttgg gacccaggaa gtccaatcgt cagatattgt actcaagcct ggtcacggca
     5821 gcgtaccgat ctgtttaaac ctagatattg atagtctgat cggtcaacgt ataatcgagt
     5881 cctagctttt gcaaacatct atcaagagac aggatcagca ggaggctttc gcatgattga
     5941 acaagatgga ttgcacgcag gttctccggc ggcttgggtg gagaggctat tcggctatga
     6001 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     6061 gcgtccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga
     6121 ggcagcgcgg ctatcgtggc tggcgacgac gggcgttcct tgcgcggctg tgctcgacgt
     6181 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     6241 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     6301 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     6361 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     6421 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg tctatgcccg acggcgagga
     6481 tctcgtcgtg acccacggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     6541 ttctggattc atcgactgtg gccgtctggg tgtggcggac cgctatcagg acatagcgtt
     6601 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tccttgtgct
     6661 ttacggtatc gccgcgcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     6721 cttctgaccg attctaggtg cattggcgca gaaaaaaatg cctgatgcga cgctgcgcgt
     6781 cttatactcc cacatatgcc agattcagca acggatacgg cttccccaac ttgcccactt
     6841 ccatacgtgt cctccttacc agaaatttat ccttaaggtc gtttaaactc gactctggct
     6901 ctatcgaatc tccgtcgttt cgagcttacg cgaacagccg tggcgctcat ttgctcgtcg
     6961 ggcatcgaat ctcgtcagct atcgtcagct tacctttttg gcagcgatcg cggctcccga
     7021 catcttggac cattagctcc acaggtatct tcttccctct agtggtcata acagcagctt
     7081 cagctacctc tcaattcaaa aaacccctca agacccgttt agaggcccca aggggttatg
     7141 ctatcaatcg ttgcgttaca cacacaaaaa accaacacac atccatcttc gatggatagc
     7201 gattttatta tctaactgct gatcgagtgt agccagatct atagtaatca attacggggt
     7261 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc
     7321 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
     7381 taacgccaat agggactttc cattgacgtc aatgggtgga ctatttacgg taaactgccc
     7441 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg
     7501 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
     7561 agtacatcta cgtattagtc atcgctatta ccatgcgtcg aggtgagccc cacgttctgc
     7621 ttcactctcc ccatctcccc cccctcccca cccccaattt tgtatttatt tattttttaa
     7681 ttattttatg cagcgatggg ggcggggggg gggggggcgc gcgccaggcg gggcggggcg
     7741 gggcgagggg cggggcgggg cgaggcggag aggtgcggcg gcagccaatc agagcggcgc
     7801 gctccgaaag tttcctttta tggcgaggcg gcggcggcgg cggccctata aaaagcgaag
     7861 cgcgcggcgg gcgggagtcg ctgcgttgcc ttcgccccgt gccccgctcc gcgccgcctc
     7921 gcgccgcccg ccccggctct gactgaccgc gttactccca caggtgagcg ggcgggacgg
     7981 cccttctccc tccgggctgt aattagcgct tggtttaatg acggctcgtt tcttttctgt
     8041 ggctgcgtga aagccttaaa gggctccggg agggcctttg tgcggggggg agcggctcgg
     8101 ggggtgcgtg cgtgtgtgtg tgcgtgggga gcgccgcgtg cggcccgcgc tgcccggcgg
     8161 ctgtgagcgc tgcgggcgcg gcgcggggct ttgtgcgctc cgcgtgtgcg cgaggggagc
     8221 gcgggccggg ggcggtgccc cgcggtgcgg gggggctgcg aggggaacaa aggctgcgtg
     8281 cggggtgtgt gcgtgggggg gtgagcaggg ggtgtgggcg cggcggtcgg gctgtaaccc
     8341 ccccctggca cccccctccc cgagttgctg agcacggccc ggcttcgggt gcggggctcc
     8401 gtgcggggcg tggcgcgggg ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc
     8461 cgggcggggc ggggccgcct cgggccgggg agggctcggg ggaggggcgc ggcggccccg
     8521 gagcgccggc ggctgtcgag gcgcggcgag ccgcagccat tgccttttat ggtaatcgtg
     8581 cgagagggcg cagggacttc ctttgtccca aatctggcgg agccgaaatc tgggaggcgc
     8641 cgccgcaccc cctctagcgg gcgcgggcga agcggtgcgg cgccggcagg aaggaaatgg
     8701 gcggggaggg ccttcgtgcg tcgccgcgcc gccgtcccct tctccatctc cagcctcggg
     8761 gctgccgcag ggggacggct gccttcgggg gggacggggc agggcggggt tcggcttctg
     8821 gcgtgtgacc ggcggcttta gagcctctgc taaccatgtt catgccttct tctttttcct
     8881 acagctcctg ggcaacgtgc tggttgttgt gctgtctcat cattttggca aagatctttg
     8941 tcgatcctac catccactcg acac
//
