LOCUS       sequence-300140-        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"
     terminator      complement(45..91)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    109..157
                     /note="synthetic polyadenylation signal"
     enhancer        240..543
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          822..1832
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1756..1778
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     polyA_signal    5721..5842
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5732..5751)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5786..5805
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      5957..6003
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      6033..6169
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6214..6285
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(6511..7099)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6591..6610)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             7845..8639
                     /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             7845..8639
                     /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(7899..7918)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     8509..8528
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      8652..8788
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    join(8933..8964,1..40)
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
ORIGIN
        1 ttccctctag tggtcataac agcagcttca gctacctctc aattcaaaaa acccctcaag
       61 acccgtttag aggccccaag gggttatgct atcaatcgtt gcgttacaca cacaaaaaac
      121 caacacacat ccatcttcga tggatagcga ttttattatc taactgctga tcgagtgtag
      181 ccagatctat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
      241 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
      301 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
      361 tgggtggact atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
      421 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
      481 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
      541 atgcgtcgag gtgagcccca cgttctgctt cactctcccc atctcccccc cctccccacc
      601 cccaattttg tatttattta ttttttaatt attttatgca gcgatggggg cggggggggg
      661 gggggcgcgc gccrggsggg gsggggsggg gsgrggggsg gggsggggsg rggcggagag
      721 gkgcggcggc agccaatcag agcggcgcgc tccgaaagtt tccttttatg gcgaggcggc
      781 ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc gggagtcgct gcgttgcctt
      841 cgccccgtgc cccgctccgc gccgcctcgc gccgcccgcc ccggctctga ctgaccgcgt
      901 tactcccaca ggtgagcggg cgggacggcc cttctccctc cgggctgtaa ttagcgcttg
      961 gtttaatgac ggctcgtttc ttttctgtgg ctgcgtgaaa gccttaaagg gctccgggag
     1021 ggcctttgtg cgggggggag cggctcgggg ggtgcgtgcg tgtgtgtgtg cgtggggagc
     1081 gccgcgtgcg gcccgcgctg tccggcggct gtgagcgctg cgggcgcggc gcggggcttt
     1141 gtgcgctccg cgtgtgcgcg aggggagcgc gggccggggg cggtgccccg cggtgcgggg
     1201 gggctgcgag gggaacaaag gctgcgtgcg gggtgtgtgc gtgggggggt gagcaggggg
     1261 tgtgggcgcg gcggtcgggc tgtaaccccc ccctggcacc cccctccccg agttgctgag
     1321 cacggcccgg cttcgggtgc ggggctccgt gcggggcgtg gcgcggggct cgccgtgccg
     1381 ggcggggggk ggcggcaggk gggggtgccg ggcggggcgg ggccgcckcg ggccggggrg
     1441 ggctcggggg aggggcgcgg cggccccgga gcgccggcgg ctgtcgaggc gcggcgagcc
     1501 gcagccattg ccttttatgg taatcgtgcg agagggcgca gggacttcct ttgtcccaaa
     1561 tctggcggag ccgaaatctg ggaggcgccg ccgcaccccc tctagcgggc gcgggcgaag
     1621 cggtgcggcg ccggcaggaa ggaaatgggc ggggagggcc ttcgtgcgtc gccgcgccgc
     1681 cgtccccttc tccatctcca gcctcggggc tgccgcaggg ggacggctgc cttcgggggg
     1741 gacggggcag ggcggggttc ggcttctggc gtgtgaccgg cggctttaga gcctctgcta
     1801 accatgttca tgccttcttc tttttcctac agctcctggg caacgtgctg gttgttgtgc
     1861 tgtctcatca ttttggcaaa gatctttgtc gatcctacca tccactcgac acaccatgtt
     1921 tgttttcttg gttcttttgc cactggtaag cagtcagtgc gtcaacctga ccacccggac
     1981 acaactccca cccgcataca cgaacagttt cactaggggg gtttactacc ctgacaaagt
     2041 ttttagatct agtgttttgc attccacgca agatctgttc cttccgttct tttcaaatgt
     2101 cacgtggttc catgctattc acgtcagcgg cactaatgga acgaaacgat tcgataaccc
     2161 cgtcttgccc ttcaacgatg gcgtatattt cgcgtcaaca gagaaaagta atataattag
     2221 aggctggatc tttggtacga ctctcgacag taagacgcag tcacttctga ttgtaaacaa
     2281 tgcaactaat gtagtcatca aggtgtgtga gttccagttc tgcaatgatc ctttcttggg
     2341 agtgtattat cataagaata acaaatcctg gatggagtct gagttccggg tatattcttc
     2401 agcaaacaat tgcacgtttg aatatgtaag ccaacctttc cttatggacc tggaagggaa
     2461 gcaaggtaat ttcaagaatc ttagggaatt tgtgtttaaa aatatagatg gctacttcaa
     2521 gatctactca aagcataccc cgattaacct cgtcagagac ctgccccaag ggttcagtgc
     2581 gcttgaaccc ctggtagatt tgccaatcgg cataaatata accaggttcc agacactgct
     2641 cgccctccac cggagctacc ttactccagg agattcatct agcgggtgga cagctggagc
     2701 ggcagcttac tacgtcggtt accttcaacc gaggacattc ttgcttaaat acaatgagaa
     2761 tggcacgata actgacgccg tagattgcgc gcttgatccg ctgtcagaaa cgaagtgtac
     2821 acttaaaagt tttacggtcg aaaagggaat ctaccaaaca tcaaatttta gggtacaacc
     2881 tacggaatct atcgtacgct tcccgaatat cactaacttg tgtccgttcg gcgaggtttt
     2941 taatgcgacc aggtttgctt ccgtgtacgc ctggaacagg aaacggatct ccaattgtgt
     3001 cgccgattac tccgtcttgt ataattcagc atctttcagc acgtttaaat gttacggagt
     3061 ttcccccaca aaattgaatg acctttgctt tacgaacgtc tacgcggatt catttgtaat
     3121 ccggggggac gaagttaggc aaattgcgcc agggcagact ggcaagatag ctgactataa
     3181 ttataaattg ccggatgact ttacgggctg tgtgattgct tggaactcaa ataatctgga
     3241 ctcaaaggta gggggaaatt ataactacct ttacaggctg ttccggaaga gtaatctgaa
     3301 gccattcgaa agagatataa gtacagagat ctaccaagct ggaagcaccc cctgcaatgg
     3361 tgttgaagga ttcaattgtt atttcccatt gcaatcctat ggttttcaac cgacgaatgg
     3421 ggtgggatac caaccatatc gagttgtggt tctcagtttc gagttgcttc atgctcctgc
     3481 gacagtatgt ggaccaaaaa aatctactaa tctggtgaag aataaatgcg tcaattttaa
     3541 ttttaatggt ctgacaggta ccggagtgct tacggaatct aacaagaagt ttcttccttt
     3601 tcaacagttc gggagggaca tagccgacac aactgacgcg gtaagagatc cgcagacgtt
     3661 ggaaattctc gatataactc catgctcatt tggaggagtg agtgttatca ctcctggtac
     3721 gaacaccagt aatcaggttg cagttcttta tcaggacgtg aattgtacag aggtcccggt
     3781 tgcgatacac gcggaccagc tgacgcccac gtggcgggtt tactcaactg ggtcaaatgt
     3841 gtttcaaacc agagccggct gcctcatagg agccgagcat gtgaacaaca gttatgaatg
     3901 tgacatacct atcggggcgg ggatctgcgc ttcatatcaa acgcaaacca attccccgcg
     3961 aagagcgcga agtgttgcaa gtcaaagtat tatagcgtat acgatgagct tgggtgctga
     4021 gaactccgtg gcatattcaa ataattccat cgcaattccc acgaatttta cgatttctgt
     4081 gacgacggag atattgccgg tttcaatgac taagacgagt gttgactgta cgatgtacat
     4141 ctgcggcgac tcaacggagt gttccaacct gctcctgcag tacggctcat tctgcaccca
     4201 actgaatcgg gcacttacgg gtattgctgt tgaacaagac aaaaatacgc aagaggtgtt
     4261 cgcccaggtc aagcagatct acaagacacc accgattaag gactttgggg ggtttaattt
     4321 ttctcagata cttcccgatc ccagtaaacc tagtaaaagg tcatttatag aggacctctt
     4381 gttcaacaag gtaacgctcg ctgatgcagg attcatcaag cagtatggcg attgtcttgg
     4441 ggacattgcg gcgcgcgacc tgatttgtgc ccagaagttc aacggactga cggtacttcc
     4501 gcctctcctg accgacgaaa tgatcgcaca atacacttct gcattgcttg ctggcaccat
     4561 aacctcaggt tggaccttcg gtgccggggc ggccttgcaa atcccctttg ctatgcaaat
     4621 ggcatatcga ttcaatggga taggagttac tcaaaatgtt ctgtatgaaa accaaaagct
     4681 gatagccaat caatttaata gtgctattgg gaaaattcag gacagcctga gtagtacggc
     4741 ttccgctctc ggcaaactcc aagatgtcgt taaccaaaat gcacaagctc tcaatacact
     4801 tgttaagcaa ctgagctcca acttcggcgc tatatcatca gttcttaacg atattctttc
     4861 ccgactcgac aaggtcgaag ccgaagtcca aatagacagg ctgataaccg ggaggctgca
     4921 aagtctccaa acgtatgtta cccaacaact catacgagcg gccgagatta gggcgtcagc
     4981 caacctcgcc gctactaaaa tgtctgaatg tgtactcggg cagtccaagc gcgtggactt
     5041 ttgtggcaag ggttatcatc tgatgagttt cccgcaatca gcgcctcacg gagtggtctt
     5101 cctccacgtg acgtatgtcc ctgcgcaaga aaaaaacttt acgacagccc ctgccatttg
     5161 tcatgacggg aaggcgcact ttccacgcga aggcgttttt gtatccaacg gcacccactg
     5221 gtttgtcaca cagcgaaatt tttatgagcc tcagatcata actactgaca atactttcgt
     5281 ctcagggaac tgcgatgtag tcatagggat cgtaaacaat acggtatacg atcccctcca
     5341 gcctgaactg gactcattta aagaggagct ggacaagtat ttcaaaaatc acacatcacc
     5401 ggacgtggac ctcggagata ttagtggtat taacgcaagc gtagtaaata ttcaaaagga
     5461 aatcgaccgc ttgaatgaag tcgcgaaaaa cctgaacgag tctcttattg atcttcaaga
     5521 actcggtaag tatgaacaat acatcaagtg gccttggtac atatggcttg gattcattgc
     5581 ggggttgatc gcaatcgtaa tggttaccat tatgttgtgt tgcatgacaa gctgttgcag
     5641 ctgcctcaaa ggttgttgtt cttgcggtag ttgctgcaag taagactgag atacagcgta
     5701 ccttcagctc acagacatga taagatacat tgatgagttt ggacaaacca caactagaat
     5761 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     5821 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     5881 gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtattggccc
     5941 atctctatcg gtatcgtagc ataacccctt ggggcctcta aacgggtctt gaggggtttt
     6001 ttgtgcccct cgggccggat tgctatctac cggcattggc gcagaaaaaa atgcctgatg
     6061 cgacgctgcg cgtcttatac tcccacatat gccagattca gcaacggata cggcttcccc
     6121 aacttgccca cttccatacg tgtcctcctt accagaaatt tatccttaag gtcgtcagct
     6181 atcctgcagg cgatctctcg atttcgatca agacattcct ttaatggtct tttctggaca
     6241 ccactagggg tcagaagtag ttcatcaaac tttcttccct ccctaatctc attggttacc
     6301 ttgggctatc gaaacttaat taaccagtca agtcagctac ttggcgagat cgacttgtct
     6361 gggtttcgac tacgctcaga attgcgtcag tcaagttcga tctggtcctt gctattgcac
     6421 ccgttctccg attacgagtt tcatttaaat catgtgagca aaaggccagc aaaaggccat
     6481 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     6541 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     6601 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     6661 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     6721 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     6781 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     6841 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     6901 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     6961 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     7021 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     7081 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     7141 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     7201 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     7261 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     7321 ttcatccata gttgcattta aatttccgaa ctctccaagg ccctcgtcgg aaaatcttca
     7381 aacctttcgt ccgatccatc ttgcaggcta cctctcgaac gaactatcgc aagtctcttg
     7441 gccggccttg cgccttggct attgcttggc agcgcctatc gccaggtatt actccaatcc
     7501 cgaatatccg agatcgggat cacccgagag aagttcaacc tacatcctca atcccgatct
     7561 atccgagatc cgaggaatat cgaaatcggg gcgcgcctgg tgtaccgaga acgatcctct
     7621 cagtgcgagt ctcgacgatc catatcgttg cttggcagtc agccagtcgg aatccagctt
     7681 gggacccagg aagtccaatc gtcagatatt gtactcaagc ctggtcacgg cagcgtaccg
     7741 atctgtttaa acctagatat tgatagtctg atcggtcaac gtataatcga gtcctagctt
     7801 ttgcaaacat ctatcaagag acaggatcag caggaggctt tcgcatgatt gaacaagatg
     7861 gattgcacgc aggttctccg gcggcttggg tggagaggct attcggctat gactgggcac
     7921 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgtccgg
     7981 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc
     8041 ggctatcgtg gctggcgacg acgggcgttc cttgcgcggc tgtgctcgac gttgtcactg
     8101 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     8161 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     8221 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     8281 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     8341 cgccagccga actgttcgcc aggctcaagg cgtctatgcc cgacggcgag gatctcgtcg
     8401 tgacccacgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     8461 tcatcgactg tggccgtctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     8521 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttccttgtg ctttacggta
     8581 tcgccgcgcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgac
     8641 cgattctagg tgcattggcg cagaaaaaaa tgcctgatgc gacgctgcgc gtcttatact
     8701 cccacatatg ccagattcag caacggatac ggcttcccca acttgcccac ttccatacgt
     8761 gtcctcctta ccagaaattt atccttaagg tcgtttaaac tcgactctgg ctctatcgaa
     8821 tctccgtcgt ttcgagctta cgcgaacagc cgtggcgctc atttgctcgt cgggcatcga
     8881 atctcgtcag ctatcgtcag cttacctttt tggcagcgat cgcggctccc gacatcttgg
     8941 accattagct ccacaggtat cttc
//
