LOCUS       sequence-307294-        8418 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 8418)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 8418)
  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-m4134372
FEATURES             Location/Qualifiers
     source          1..8418
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(5..51)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    69..117
                     /note="synthetic polyadenylation signal"
     enhancer        198..501
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        503..705
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     656..676
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             4599..4619
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             4968..4985
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    5062..5183
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5073..5092)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5127..5146
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      5298..5344
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      5374..5510
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5555..5626
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(5852..6440)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5932..5951)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(7186..7254,7255..8046)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(7404..7423)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      8059..8195
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    8347..8418
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
ORIGIN
        1 aattcaaaaa acccctcaag acccgtttag aggccccaag gggttatgct atcaatcgtt
       61 gcgttacaca cacaaaaaac caacacacat ccatcttcga tggatagcga ttttattatc
      121 taactgctga tcgagtgtag ccagatctag taatcaatta cggggtcatt agttcatagc
      181 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
      241 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
      301 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
      361 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
      421 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
      481 ttagtcatcg ctattaccat gctgatgcgg ttttggcagt acatcaatgg gcgtggatag
      541 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt
      601 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa
      661 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt
      721 cagatcagat ctttgtcgat cctaccatcc actcgacaca cccgccagcg gccgccacca
      781 tgttcgtctt tcttgtgtta ctccccctgg tttcgtccca gtgcgtgaac ttgactacgc
      841 gaacccaact gccgccagca tatacaaata gtttcacccg tggagtatac taccctgata
      901 aagtgtttag atcaagcgta ctacactcta ctcaggacct gtttctcccc ttcttctcca
      961 atgtcacatg gtttcatgcg atccacgtta gcggcaccaa cgggacaaag cgctttgaca
     1021 accctgtgct gccattcaat gatggtgtgt attttgcttc taccgagaag agcaacataa
     1081 ttcggggctg gatcttcgga actaccctcg acagtaaaac tcagtcattg ctgattgtga
     1141 ataacgccac aaatgtcgtt atcaaggtct gtgaatttca attctgcaac gatccattcc
     1201 ttggggtgta ttaccataaa aataataagt cctggatgga gagcgaattt agggtatact
     1261 cgtctgccaa caattgtact ttcgagtatg tgtctcagcc cttcttaatg gatctggaag
     1321 gtaaacaggg caactttaag aacctccgcg agtttgtatt caaaaatata gacggatact
     1381 ttaagattta ttccaagcac acaccaatca atttggtgcg ggatctcccg caagggttca
     1441 gcgctctgga acctctagtg gacctcccca tcggcattaa catcacaaga ttccagacgc
     1501 tgttagccct gcatagatca taccttactc ccggtgacag tagttctggg tggaccgcag
     1561 gcgcggctgc atattacgtg ggatatttgc agcctcggac gtttctgctg aaatacaacg
     1621 agaacggcac cattacagat gccgtggatt gcgcccttga ccccctaagc gaaactaagt
     1681 gtacgcttaa aagcttcacc gtcgagaagg gaatttacca gacttccaac tttagggtcc
     1741 agccgacaga atcaatagtt aggttcccaa atatcaccaa tttgtgccct tttggggaag
     1801 tcttcaatgc tacacgattt gcttcggttt atgcctggaa caggaagcgt atctcaaatt
     1861 gcgtcgcgga ctacagcgtg ctctataata gtgcaagttt ctcgaccttt aaatgttatg
     1921 gcgtgtcccc gaccaaacta aatgacctct gttttacaaa cgtttacgcc gattcttttg
     1981 tgattcgcgg tgatgaggta cgccaaatcg caccgggcca gactggcaag attgccgatt
     2041 acaactacaa gttgcctgac gacttcaccg gatgtgttat agcctggaat agcaataatc
     2101 tggattctaa ggtcggaggt aactataact acctgtatag actgttccgt aagtcaaacc
     2161 tgaaaccctt tgagcgggac atctccacag agatttacca agcagggtct accccctgca
     2221 atggggttga aggattcaat tgctactttc cattacagag ctacgggttc cagcctacga
     2281 acggggttgg ttaccagcct tatagggtag tggtgctgtc gttcgaactg ttgcacgctc
     2341 cagctacagt gtgcgggcct aaaaaatcaa ctaacctggt taaaaataag tgtgttaatt
     2401 ttaacttcaa tggcttaacc ggaactggag tgctcacgga gtcaaacaag aaatttctac
     2461 cgtttcagca attcgggagg gatatcgcag acactacgga cgcggtcaga gatccacaga
     2521 cccttgagat ccttgacata acaccctgct ctttcggagg tgtctccgta attacaccag
     2581 gcactaatac ctctaatcag gtggccgtgc tttatcagga tgttaactgt acagaggtcc
     2641 cggtggctat ccatgccgac caattgacac ctacgtggcg cgtctattct accggctcca
     2701 acgtcttcca gacccgggct ggatgcttga taggcgcaga acacgtaaac aactcttacg
     2761 aatgtgatat tcccattgga gccgggatct gcgcaagcta ccagacccaa acaaacagtc
     2821 ccagacgggc tcgtagcgtg gcctctcaga gcataatagc ctataccatg tcccttggcg
     2881 cggagaatag tgtggcatac tcaaataata gcatcgctat ccccactaac tttactattt
     2941 ccgtgacgac tgagattctg ccagtgtcta tgactaagac atccgtggat tgcacaatgt
     3001 atatttgtgg ggattccaca gagtgtagca acttgttgct gcaatatgga tcattctgta
     3061 cgcagctgaa tcgcgccctc accggcatcg ctgtggaaca ggacaagaac actcaagagg
     3121 tctttgctca ggtgaagcaa atctacaaga ccccacctat aaaagacttc gggggtttta
     3181 acttctcgca gattctcccc gaccccagca agccttcaaa aaggagcttt attgaggacc
     3241 tgttgttcaa caaggttact ttagccgatg ccggttttat caaacagtac ggcgactgtc
     3301 ttggcgatat agcagcccgg gatctgattt gcgcacaaaa attcaatggc cttaccgtac
     3361 tgccacccct tctcaccgac gaaatgatcg cgcagtacac ctccgcactg ctggctggga
     3421 ccatcacttc gggatggacc tttggggctg gtgctgcttt gcaaattccc ttcgccatgc
     3481 agatggccta tcgattcaat ggcatagggg taacacagaa cgtattgtat gaaaatcaaa
     3541 agctaatcgc taaccagttt aattcagcca ttggtaagat ccaggattca ctctccagta
     3601 cagcctctgc cctgggaaag ctccaggacg tggtcaacca gaacgcgcag gcattgaata
     3661 cactcgtcaa acagctgagt agcaatttcg gcgcaatcag tagtgtctta aacgatatac
     3721 tgagtaggct tgataaagta gaggccgagg tccagattga cagactaatc acggggcgtc
     3781 ttcagagttt acagacctat gtgacccaac agctgatccg agcagcggaa attcgagcaa
     3841 gcgcgaacct ggccgctact aaaatgagtg agtgcgtgct gggccaaagt aaacgagtcg
     3901 acttttgcgg caaagggtat catctcatgt ctttccctca gtccgcccct cacggtgttg
     3961 ttttcctcca cgtgacatac gttcccgcgc aggaaaagaa ttttactacg gcacccgcta
     4021 tttgccatga cggaaaggct cattttccac gggagggagt gttcgtttca aacgggacac
     4081 actggtttgt aactcagcgg aacttttatg agcctcaaat catcacgaca gacaatacct
     4141 tcgtgagtgg caactgcgat gtagtcatag gcattgtgaa taacactgtt tatgatccac
     4201 ttcagccgga actcgacagc ttcaaagagg aattggataa gtactttaaa aatcatacca
     4261 gtccagacgt agatctcggc gatatatcgg gcattaacgc cagtgtggtc aatatacaaa
     4321 aggagattga cagactgaat gaagtagcta agaacctcaa cgaatctctg attgacctgc
     4381 aagagctagg gaagtacgag cagtatatca aatggccatg gtacatatgg ctgggtttta
     4441 tcgcgggtct tattgccata gtcatggtga cgattatgtt gtgctgtatg acctcctgct
     4501 gttcttgttt gaaaggctgc tgctcctgtg gtagctgttg caaattcgat gaagatgact
     4561 ctgaacccgt gttgaaaggc gttaagctac actacacaga aaacctttac tttcagtcga
     4621 tgagtgacaa aatcattcat ctaaccgatg actcctttga tacagacgtg ttgaaggccg
     4681 atggtgcgat actggtagat ttctgggctg aatggtgtgg cccgtgcaag atgatcgcac
     4741 ccattctgga cgagatcgcc gacgagtatc aaggaaaatt aactgtcgct aagcttaata
     4801 tagatcagaa cccagggacg gcacctaaat acggcatcag gggaatacct accctcctgc
     4861 tctttaagaa tggggaagtt gccgcgacaa aagtgggagc cctgtctaag ggccagctaa
     4921 aagagttctt ggacgctaac ctggcaggtt caggctcggg gcacatgcac catcaccatc
     4981 accatagcag cgggctcgtg ccacgcggct aagctagctt gactgactga gatacagcgt
     5041 accttcagct cacagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa
     5101 tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
     5161 ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc
     5221 agggggaggt gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtattggcc
     5281 catctctatc ggtatcgtag cataacccct tggggcctct aaacgggtct tgaggggttt
     5341 tttgtgcccc tcgggccgga ttgctatcta ccggcattgg cgcagaaaaa aatgcctgat
     5401 gcgacgctgc gcgtcttata ctcccacata tgccagattc agcaacggat acggcttccc
     5461 caacttgccc acttccatac gtgtcctcct taccagaaat ttatccttaa ggtcgtcagc
     5521 tatcctgcag gcgatctctc gatttcgatc aagacattcc tttaatggtc ttttctggac
     5581 accactaggg gtcagaagta gttcatcaaa ctttcttccc tccctaatct cattggttac
     5641 cttgggctat cgaaacttaa ttaaccagtc aagtcagcta cttggcgaga tcgacttgtc
     5701 tgggtttcga ctacgctcag aattgcgtca gtcaagttcg atctggtcct tgctattgca
     5761 cccgttctcc gattacgagt ttcatttaaa tcatgtgagc aaaaggccag caaaaggcca
     5821 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     5881 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     5941 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     6001 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     6061 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     6121 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     6181 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     6241 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     6301 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     6361 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     6421 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     6481 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     6541 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     6601 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     6661 gttcatccat agttgcattt aaatttccga actctccaag gccctcgtcg gaaaatcttc
     6721 aaacctttcg tccgatccat cttgcaggct acctctcgaa cgaactatcg caagtctctt
     6781 ggccggcctt gcgccttggc tattgcttgg cagcgcctat cgccaggtat tactccaatc
     6841 ccgaatatcc gagatcggga tcacccgaga gaagttcaac ctacatcctc aatcccgatc
     6901 tatccgagat ccgaggaata tcgaaatcgg ggcgcgcctg gtgtaccgag aacgatcctc
     6961 tcagtgcgag tctcgacgat ccatatcgtt gcttggcagt cagccagtcg gaatccagct
     7021 tgggacccag gaagtccaat cgtcagatat tgtactcaag cctggtcacg gcagcgtacc
     7081 gatctgttta aacctagata ttgatagtct gatcggtcaa cgtataatcg agtcctagct
     7141 tttgcaaaca tctatcaaga gacaggatca gcaggaggct ttcgcatgag tattcaacat
     7201 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     7261 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcgcgagt gggttacatc
     7321 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgctttcca
     7381 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     7441 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtattcacca
     7501 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     7561 accatgagtg ataacactgc ggccaactta cttctgacaa cgattggagg accgaaggag
     7621 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     7681 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     7741 acaaccttgc gtaaactatt aactggcgaa ctacttactc tagcttcccg gcaacagttg
     7801 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     7861 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     7921 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     7981 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     8041 tggtaaccga ttctaggtgc attggcgcag aaaaaaatgc ctgatgcgac gctgcgcgtc
     8101 ttatactccc acatatgcca gattcagcaa cggatacggc ttccccaact tgcccacttc
     8161 catacgtgtc ctccttacca gaaatttatc cttaagatcc cgaatcgttt aaactcgact
     8221 ctggctctat cgaatctccg tcgtttcgag cttacgcgaa cagccgtggc gctcatttgc
     8281 tcgtcgggca tcgaatctcg tcagctatcg tcagcttacc tttttggcag cgatcgcggc
     8341 tcccgacatc ttggaccatt agctccacag gtatcttctt ccctctagtg gtcataacag
     8401 cagcttcagc tacctctc
//
