LOCUS       sequence-300137-        9018 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 9018)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 9018)
  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-m4164341
FEATURES             Location/Qualifiers
     source          1..9018
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        93..396
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        399..674
                     /label=chicken beta-actin promoter
     intron          675..1685
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1609..1631
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     polyA_signal    5628..5749
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5639..5658)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5693..5712
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      5864..5910
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      5940..6076
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6121..6192
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(6418..7006)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6498..6517)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             7752..8546
                     /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             7752..8546
                     /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(7806..7825)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     8416..8435
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      8559..8695
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    8840..8911
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(8916..8962)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    join(8980..9018,1..10)
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 cgattttatt atctaactgc tgatcgagtg tagccagatc tatagtaatc aattacgggg
       61 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
      121 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
      181 gtaacgccaa tagggacttt ccattgacgt caatgggtgg actatttacg gtaaactgcc
      241 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
      301 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
      361 cagtacatct acgtattagt catcgctatt accatgcgtc gaggtgagcc ccacgttctg
      421 cttcactctc cccatctccc ccccctcccc acccccaatt ttgtatttat ttatttttta
      481 attattttat gcagcgatgg gggcgggggg ggggggggcg cgcgccaggc ggggsggggs
      541 ggggsgrggg gsggggsggg gsgrggcgga gaggtgcggc ggcagccaat cagagcggcg
      601 cgctccgaaa gtttcctttt atggcgaggc ggcggcggcg gcggccctat aaaaagcgaa
      661 gcgcgcggcg ggcgggagtc gctgcgttgc cttcgccccg tgccccgctc cgcgccgcct
      721 cgcgccgccc gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg
      781 gcccttctcc ctccgggctg taattagcgc ttggtttaat gacggctcgt ttcttttctg
      841 tggctgcgtg aaagccttaa agggctccgg gagggccttt gtgcgggggg gagcggctcg
      901 gggggtgcgt gcgtgtgtgt gtgcgtgggg agcgccgcgt gcggcccgcg ctgcccggcg
      961 gctgtgagcg ctgcgggcgc ggcgcggggc tttgtgcgct ccgcgtgtgc gcgaggggag
     1021 cgcgggccgg gggcggtgcc ccgcggtgcg ggggggctgc gaggggaaca aaggctgcgt
     1081 gcggggtgtg tgcgtggggg ggtgagcagg gggtgtgggc gcggcggtcg ggctgtaacc
     1141 cccccctggc acccccctcc ccgagttgct gagcacggcc cggcttcggg tgcggggctc
     1201 cgtgcggggc gtggcgcggg gctcgccgtg ccgggcgggg ggtggcggca ggkgggggtg
     1261 ccgggcgggg cggggccgcc tcgggccggg grgggctcgg gggaggggcg cggcggcccc
     1321 ggagcgccgg cggctgtcga ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt
     1381 gcgagagggc gcagggactt cctttgtccc aaatctggcg gagccgaaat ctgggaggcg
     1441 ccgccgcacc ccctctagcg ggcgcgggcg aagcggtgcg gcgccggcag gaaggaaatg
     1501 ggcggggagg gccttcgtgc gtcgccgcgc cgccgtcccc ttctccatct ccagcctcgg
     1561 ggctgccgca gggggacggc tgccttcggg ggggacgggg cagggcgggg ttcggcttct
     1621 ggcgtgtgac cggcggcttt agagcctctg ctaaccatgt tcatgccttc ttctttttcc
     1681 tacagctcct gggcaacgtg ctggttgttg tgctgtctca tcattttggc aaagatcttt
     1741 gtcgatccta ccatccactc gacacaccat gtttgttttc ttggttcttt tgccactggt
     1801 aagcagtcag tgcgtcaacc tgaccacccg gacacaactc ccacccgcat acacgaacag
     1861 tttcactagg ggggtttact accctgacaa agtttttaga tctagtgttt tgcattccac
     1921 gcaagatctg ttccttccgt tcttttcaaa tgtcacgtgg ttccatgcta ttcacgtcag
     1981 cggcactaat ggaacgaaac gattcgataa ccccgtcttg cccttcaacg atggcgtata
     2041 tttcgcgtca acagagaaaa gtaatataat tagaggctgg atctttggta cgactctcga
     2101 cagtaagacg cagtcacttc tgattgtaaa caatgcaact aatgtagtca tcaaggtgtg
     2161 tgagttccag ttctgcaatg atcctttctt gggagtgtat tatcataaga ataacaaatc
     2221 ctggatggag tctgagttcc gggtatattc ttcagcaaac aattgcacgt ttgaatatgt
     2281 aagccaacct ttccttatgg acctggaagg gaagcaaggt aatttcaaga atcttaggga
     2341 atttgtgttt aaaaatatag atggctactt caagatctac tcaaagcata ccccgattaa
     2401 cctcgtcaga gacctgcccc aagggttcag tgcgcttgaa cccctggtag atttgccaat
     2461 cggcataaat ataaccaggt tccagacact gctcgccctc caccggagct accttactcc
     2521 aggagattca tctagcgggt ggacagctgg agcggcagct tactacgtcg gttaccttca
     2581 accgaggaca ttcttgctta aatacaatga gaatggcacg ataactgacg ccgtagattg
     2641 cgcgcttgat ccgctgtcag aaacgaagtg tacacttaaa agttttacgg tcgaaaaggg
     2701 aatctaccaa acatcaaatt ttagggtaca acctacggaa tctatcgtac gcttcccgaa
     2761 tatcactaac ttgtgtccgt tcggcgaggt ttttaatgcg accaggtttg cttccgtgta
     2821 cgcctggaac aggaaacgga tctccaattg tgtcgccgat tactccgtct tgtataattc
     2881 agcatctttc agcacgttta aatgttacgg agtttccccc acaaaattga atgacctttg
     2941 ctttacgaac gtctacgcgg attcatttgt aatccggggg gacgaagtta ggcaaattgc
     3001 gccagggcag actggcaaga tagctgacta taattataaa ttgccggatg actttacggg
     3061 ctgtgtgatt gcttggaact caaataatct ggactcaaag gtagggggaa attataacta
     3121 cctttacagg ctgttccgga agagtaatct gaagccattc gaaagagata taagtacaga
     3181 gatctaccaa gctggaagca ccccctgcaa tggtgttgaa ggattcaatt gttatttccc
     3241 attgcaatcc tatggttttc aaccgacgaa tggggtggga taccaaccat atcgagttgt
     3301 ggttctcagt ttcgagttgc ttcatgctcc tgcgacagta tgtggaccaa aaaaatctac
     3361 taatctggtg aagaataaat gcgtcaattt taattttaat ggtctgacag gtaccggagt
     3421 gcttacggaa tctaacaaga agtttcttcc ttttcaacag ttcgggaggg acatagccga
     3481 cacaactgac gcggtaagag atccgcagac gttggaaatt ctcgatataa ctccatgctc
     3541 atttggagga gtgagtgtta tcactcctgg tacgaacacc agtaatcagg ttgcagttct
     3601 ttatcaggac gtgaattgta cagaggtccc ggttgcgata cacgcggacc agctgacgcc
     3661 cacgtggcgg gtttactcaa ctgggtcaaa tgtgtttcaa accagagccg gctgcctcat
     3721 aggagccgag catgtgaaca acagttatga atgtgacata cctatcgggg cggggatctg
     3781 cgcttcatat caaacgcaaa ccaattcccc gcgaagagcg cgaagtgttg caagtcaaag
     3841 tattatagcg tatacgatga gcttgggtgc tgagaactcc gtggcatatt caaataattc
     3901 catcgcaatt cccacgaatt ttacgatttc tgtgacgacg gagatattgc cggtttcaat
     3961 gactaagacg agtgttgact gtacgatgta catctgcggc gactcaacgg agtgttccaa
     4021 cctgctcctg cagtacggct cattctgcac ccaactgaat cgggcactta cgggtattgc
     4081 tgttgaacaa gacaaaaata cgcaagaggt gttcgcccag gtcaagcaga tctacaagac
     4141 accaccgatt aaggactttg gggggtttaa tttttctcag atacttcccg atcccagtaa
     4201 acctagtaaa aggtcattta tagaggacct cttgttcaac aaggtaacgc tcgctgatgc
     4261 aggattcatc aagcagtatg gcgattgtct tggggacatt gcggcgcgcg acctgatttg
     4321 tgcccagaag ttcaacggac tgacggtact tccgcctctc ctgaccgacg aaatgatcgc
     4381 acaatacact tctgcattgc ttgctggcac cataacctca ggttggacct tcggtgccgg
     4441 ggcggccttg caaatcccct ttgctatgca aatggcatat cgattcaatg ggataggagt
     4501 tactcaaaat gttctgtatg aaaaccaaaa gctgatagcc aatcaattta atagtgctat
     4561 tgggaaaatt caggacagcc tgagtagtac ggcttccgct ctcggcaaac tccaagatgt
     4621 cgttaaccaa aatgcacaag ctctcaatac acttgttaag caactgagct ccaacttcgg
     4681 cgctatatca tcagttctta acgatattct ttcccgactc gacaaggtcg aagccgaagt
     4741 ccaaatagac aggctgataa ccgggaggct gcaaagtctc caaacgtatg ttacccaaca
     4801 actcatacga gcggccgaga ttagggcgtc agccaacctc gccgctacta aaatgtctga
     4861 atgtgtactc gggcagtcca agcgcgtgga cttttgtggc aagggttatc atctgatgag
     4921 tttcccgcaa tcagcgcctc acggagtggt cttcctccac gtgacgtatg tccctgcgca
     4981 agaaaaaaac tttacgacag cccctgccat ttgtcatgac gggaaggcgc actttccacg
     5041 cgaaggcgtt tttgtatcca acggcaccca ctggtttgtc acacagcgaa atttttatga
     5101 gcctcagatc ataactactg acaatacttt cgtctcaggg aactgcgatg tagtcatagg
     5161 gatcgtaaac aatacggtat acgatcccct ccagcctgaa ctggactcat ttaaagagga
     5221 gctggacaag tatttcaaaa atcacacatc accggacgtg gacctcggag atattagtgg
     5281 tattaacgca agcgtagtaa atattcaaaa ggaaatcgac cgcttgaatg aagtcgcgaa
     5341 aaacctgaac gagtctctta ttgatcttca agaactcggt aagtatgaac aatacatcaa
     5401 gtggccttgg tacatatggc ttggattcat tgcggggttg atcgcaatcg taatggttac
     5461 cattatgttg tgttgcatga caagctgttg cagctgcctc aaaggttgtt gttcttgcgg
     5521 tagttgctgc aagttcgacg aagatgattc tgaacctgtt cttaaaggcg taaaattgca
     5581 ctacacgtaa gactgagata cagcgtacct tcagctcaca gacatgataa gatacattga
     5641 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
     5701 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
     5761 ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta
     5821 aaacctctac aaatgtggta ttggcccatc tctatcggta tcgtagcata accccttggg
     5881 gcctctaaac gggtcttgag gggttttttg tgcccctcgg gccggattgc tatctaccgg
     5941 cattggcgca gaaaaaaatg cctgatgcga cgctgcgcgt cttatactcc cacatatgcc
     6001 agattcagca acggatacgg cttccccaac ttgcccactt ccatacgtgt cctccttacc
     6061 agaaatttat ccttaaggtc gtcagctatc ctgcaggcga tctctcgatt tcgatcaaga
     6121 cattccttta atggtctttt ctggacacca ctaggggtca gaagtagttc atcaaacttt
     6181 cttccctccc taatctcatt ggttaccttg ggctatcgaa acttaattaa ccagtcaagt
     6241 cagctacttg gcgagatcga cttgtctggg tttcgactac gctcagaatt gcgtcagtca
     6301 agttcgatct ggtccttgct attgcacccg ttctccgatt acgagtttca tttaaatcat
     6361 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     6421 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     6481 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     6541 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     6601 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     6661 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     6721 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     6781 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     6841 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     6901 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     6961 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     7021 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     7081 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     7141 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     7201 acctatctca gcgatctgtc tatttcgttc atccatagtt gcatttaaat ttccgaactc
     7261 tccaaggccc tcgtcggaaa atcttcaaac ctttcgtccg atccatcttg caggctacct
     7321 ctcgaacgaa ctatcgcaag tctcttggcc ggccttgcgc cttggctatt gcttggcagc
     7381 gcctatcgcc aggtattact ccaatcccga atatccgaga tcgggatcac ccgagagaag
     7441 ttcaacctac atcctcaatc ccgatctatc cgagatccga ggaatatcga aatcggggcg
     7501 cgcctggtgt accgagaacg atcctctcag tgcgagtctc gacgatccat atcgttgctt
     7561 ggcagtcagc cagtcggaat ccagcttggg acccaggaag tccaatcgtc agatattgta
     7621 ctcaagcctg gtcacggcag cgtaccgatc tgtttaaacc tagatattga tagtctgatc
     7681 ggtcaacgta taatcgagtc ctagcttttg caaacatcta tcaagagaca ggatcagcag
     7741 gaggctttcg catgattgaa caagatggat tgcacgcagg ttctccggcg gcttgggtgg
     7801 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     7861 tccggctgtc agcgcagggg cgtccggttc tttttgtcaa gaccgacctg tccggtgccc
     7921 tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggcgacgacg ggcgttcctt
     7981 gcgcggctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     8041 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     8101 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     8161 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     8221 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgt
     8281 ctatgcccga cggcgaggat ctcgtcgtga cccacggcga tgcctgcttg ccgaatatca
     8341 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccgtctgggt gtggcggacc
     8401 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     8461 ctgaccgctt ccttgtgctt tacggtatcg ccgcgcccga ttcgcagcgc atcgccttct
     8521 atcgccttct tgacgagttc ttctgaccga ttctaggtgc attggcgcag aaaaaaatgc
     8581 ctgatgcgac gctgcgcgtc ttatactccc acatatgcca gattcagcaa cggatacggc
     8641 ttccccaact tgcccacttc catacgtgtc ctccttacca gaaatttatc cttaaggtcg
     8701 tttaaactcg actctggctc tatcgaatct ccgtcgtttc gagcttacgc gaacagccgt
     8761 ggcgctcatt tgctcgtcgg gcatcgaatc tcgtcagcta tcgtcagctt acctttttgg
     8821 cagcgatcgc ggctcccgac atcttggacc attagctcca caggtatctt cttccctcta
     8881 gtggtcataa cagcagcttc agctacctct caattcaaaa aacccctcaa gacccgttta
     8941 gaggccccaa ggggttatgc tatcaatcgt tgcgttacac acacaaaaaa ccaacacaca
     9001 tccatcttcg atggatag
//
