LOCUS       sequence-297246-        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"
     polyA_signal    3806..3927
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3817..3836)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3871..3890
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      4042..4088
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      4118..4254
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4299..4370
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(4596..5184)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4676..4695)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             5930..6724
                     /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             5930..6724
                     /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(5984..6003)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6594..6613
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      6737..6873
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7018..7089
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7094..7140)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7158..7206
                     /note="synthetic polyadenylation signal"
     enhancer        7289..7592
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7595..7870
                     /label=chicken beta-actin promoter
     intron          7871..8881
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     8805..8827
                     /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 gctgctaaga ctgagataca
     3781 gcgtaccttc agctcacaga catgataaga tacattgatg agtttggaca aaccacaact
     3841 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta
     3901 accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag
     3961 gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtatt
     4021 ggcccatctc tatcggtatc gtagcataac cccttggggc ctctaaacgg gtcttgaggg
     4081 gttttttgtg cccctcgggc cggattgcta tctaccggca ttggcgcaga aaaaaatgcc
     4141 tgatgcgacg ctgcgcgtct tatactccca catatgccag attcagcaac ggatacggct
     4201 tccccaactt gcccacttcc atacgtgtcc tccttaccag aaatttatcc ttaaggtcgt
     4261 cagctatcct gcaggcgatc tctcgatttc gatcaagaca ttcctttaat ggtcttttct
     4321 ggacaccact aggggtcaga agtagttcat caaactttct tccctcccta atctcattgg
     4381 ttaccttggg ctatcgaaac ttaattaacc agtcaagtca gctacttggc gagatcgact
     4441 tgtctgggtt tcgactacgc tcagaattgc gtcagtcaag ttcgatctgg tccttgctat
     4501 tgcacccgtt ctccgattac gagtttcatt taaatcatgt gagcaaaagg ccagcaaaag
     4561 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     4621 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     4681 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     4741 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     4801 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     4861 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     4921 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     4981 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     5041 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     5101 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     5161 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     5221 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     5281 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     5341 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     5401 tttcgttcat ccatagttgc atttaaattt ccgaactctc caaggccctc gtcggaaaat
     5461 cttcaaacct ttcgtccgat ccatcttgca ggctacctct cgaacgaact atcgcaagtc
     5521 tcttggccgg ccttgcgcct tggctattgc ttggcagcgc ctatcgccag gtattactcc
     5581 aatcccgaat atccgagatc gggatcaccc gagagaagtt caacctacat cctcaatccc
     5641 gatctatccg agatccgagg aatatcgaaa tcggggcgcg cctggtgtac cgagaacgat
     5701 cctctcagtg cgagtctcga cgatccatat cgttgcttgg cagtcagcca gtcggaatcc
     5761 agcttgggac ccaggaagtc caatcgtcag atattgtact caagcctggt cacggcagcg
     5821 taccgatctg tttaaaccta gatattgata gtctgatcgg tcaacgtata atcgagtcct
     5881 agcttttgca aacatctatc aagagacagg atcagcagga ggctttcgca tgattgaaca
     5941 agatggattg cacgcaggtt ctccggcggc ttgggtggag aggctattcg gctatgactg
     6001 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     6061 tccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc
     6121 agcgcggcta tcgtggctgg cgacgacggg cgttccttgc gcggctgtgc tcgacgttgt
     6181 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     6241 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     6301 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     6361 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     6421 gctcgcgcca gccgaactgt tcgccaggct caaggcgtct atgcccgacg gcgaggatct
     6481 cgtcgtgacc cacggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     6541 tggattcatc gactgtggcc gtctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     6601 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc ttgtgcttta
     6661 cggtatcgcc gcgcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     6721 ctgaccgatt ctaggtgcat tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt
     6781 atactcccac atatgccaga ttcagcaacg gatacggctt ccccaacttg cccacttcca
     6841 tacgtgtcct ccttaccaga aatttatcct taaggtcgtt taaactcgac tctggctcta
     6901 tcgaatctcc gtcgtttcga gcttacgcga acagccgtgg cgctcatttg ctcgtcgggc
     6961 atcgaatctc gtcagctatc gtcagcttac ctttttggca gcgatcgcgg ctcccgacat
     7021 cttggaccat tagctccaca ggtatcttct tccctctagt ggtcataaca gcagcttcag
     7081 ctacctctca attcaaaaaa cccctcaaga cccgtttaga ggccccaagg ggttatgcta
     7141 tcaatcgttg cgttacacac acaaaaaacc aacacacatc catcttcgat ggatagcgat
     7201 tttattatct aactgctgat cgagtgtagc cagatctata gtaatcaatt acggggtcat
     7261 tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg
     7321 gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa
     7381 cgccaatagg gactttccat tgacgtcaat gggtggacta tttacggtaa actgcccact
     7441 tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta
     7501 aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt
     7561 acatctacgt attagtcatc gctattacca tgcgtcgagg tgagccccac gttctgcttc
     7621 actctcccca tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta
     7681 ttttatgcag cgatgggggc gggggggggg ggggcgcgcg ccaggcgggg cggggcgggg
     7741 cgaggggcgg ggcggggcga ggcggagagg tgcggcggca gccaatcaga gcggcgcgct
     7801 ccgaaagttt ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa agcgaagcgc
     7861 gcggcgggcg ggagtcgctg cgttgccttc gccccgtgcc ccgctccgcg ccgcctcgcg
     7921 ccgcccgccc cggctctgac tgaccgcgtt actcccacag gtgagcgggc gggacggccc
     7981 ttctccctcc gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc
     8041 tgcgtgaaag ccttaaaggg ctccgggagg gcctttgtgc gggggggagc ggctcggggg
     8101 gtgcgtgcgt gtgtgtgtgc gtggggagcg ccgcgtgcgg cccgcgctgc ccggcggctg
     8161 tgagcgctgc gggcgcggcg cggggctttg tgcgctccgc gtgtgcgcga ggggagcgcg
     8221 ggccgggggc ggtgccccgc ggtgcggggg ggctgcgagg ggaacaaagg ctgcgtgcgg
     8281 ggtgtgtgcg tgggggggtg agcagggggt gtgggcgcgg cggtcgggct gtaacccccc
     8341 cctggcaccc ccctccccga gttgctgagc acggcccggc ttcgggtgcg gggctccgtg
     8401 cggggcgtgg cgcggggctc gccgtgccgg gcggggggtg gcggcaggtg ggggtgccgg
     8461 gcggggcggg gccgcctcgg gccggggagg gctcggggga ggggcgcggc ggccccggag
     8521 cgccggcggc tgtcgaggcg cggcgagccg cagccattgc cttttatggt aatcgtgcga
     8581 gagggcgcag ggacttcctt tgtcccaaat ctggcggagc cgaaatctgg gaggcgccgc
     8641 cgcaccccct ctagcgggcg cgggcgaagc ggtgcggcgc cggcaggaag gaaatgggcg
     8701 gggagggcct tcgtgcgtcg ccgcgccgcc gtccccttct ccatctccag cctcggggct
     8761 gccgcagggg gacggctgcc ttcggggggg acggggcagg gcggggttcg gcttctggcg
     8821 tgtgaccggc ggctttagag cctctgctaa ccatgttcat gccttcttct ttttcctaca
     8881 gctcctgggc aacgtgctgg ttgttgtgct gtctcatcat tttggcaaag atctttgtcg
     8941 atcctaccat ccactcgaca c
//
