LOCUS       sequence-307282-       10004 bp    DNA     circular SYN 15-APR-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 10004)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 10004)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pGBW-m4134207
FEATURES             Location/Qualifiers
     source          1..10004
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    60..131
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(136..182)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    200..248
                     /note="synthetic polyadenylation signal"
     enhancer        329..632
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        634..836
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     787..807
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             4730..4750
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             4754..4819
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    4872..4993
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4883..4902)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4937..4956
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      5108..5154
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      5184..5320
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5365..5436
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(5662..6250)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5742..5761)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        6750..7959
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     7796..7815
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             7963..8562
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(7963..7982)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     8459..8479
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    8589..8637
                     /note="synthetic polyadenylation signal"
     CDS             join(8903..8971,8972..9763)
                     /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(9121..9140)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      9776..9912
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
ORIGIN
        1 gcatcgaatc tcgtcagcta tcgtcagctt acctttttgg cagcgatcgc ggctcccgac
       61 atcttggacc attagctcca caggtatctt cttccctcta gtggtcataa cagcagcttc
      121 agctacctct caattcaaaa aacccctcaa gacccgttta gaggccccaa ggggttatgc
      181 tatcaatcgt tgcgttacac acacaaaaaa ccaacacaca tccatcttcg atggatagcg
      241 attttattat ctaactgctg atcgagtgta gccagatcta gtaatcaatt acggggtcat
      301 tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg
      361 gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa
      421 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact
      481 tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta
      541 aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt
      601 acatctacgt attagtcatc gctattacca tgctgatgcg gttttggcag tacatcaatg
      661 ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg
      721 ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc
      781 cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctggtt
      841 tagtgaaccg tcagatcaga tctttgtcga tcctaccatc cactcgacac acccgccagc
      901 ggccgccacc atgttcgtct ttcttgtgtt actccccctg gtttcgtccc agtgcgtgaa
      961 cttgactacg cgaacccaac tgccgccagc atatacaaat agtttcaccc gtggagtata
     1021 ctaccctgat aaagtgttta gatcaagcgt actacactct actcaggacc tgtttctccc
     1081 cttcttctcc aatgtcacat ggtttcatgc gatccacgtt agcggcacca acgggacaaa
     1141 gcgctttgac aaccctgtgc tgccattcaa tgatggtgtg tattttgctt ctaccgagaa
     1201 gagcaacata attcggggct ggatcttcgg aactaccctc gacagtaaaa ctcagtcatt
     1261 gctgattgtg aataacgcca caaatgtcgt tatcaaggtc tgtgaatttc aattctgcaa
     1321 cgatccattc cttggggtgt attaccataa aaataataag tcctggatgg agagcgaatt
     1381 tagggtatac tcgtctgcca acaattgtac tttcgagtat gtgtctcagc ccttcttaat
     1441 ggatctggaa ggtaaacagg gcaactttaa gaacctccgc gagtttgtat tcaaaaatat
     1501 agacggatac tttaagattt attccaagca cacaccaatc aatttggtgc gggatctccc
     1561 gcaagggttc agcgctctgg aacctctagt ggacctcccc atcggcatta acatcacaag
     1621 attccagacg ctgttagccc tgcatagatc ataccttact cccggtgaca gtagttctgg
     1681 gtggaccgca ggcgcggctg catattacgt gggatatttg cagcctcgga cgtttctgct
     1741 gaaatacaac gagaacggca ccattacaga tgccgtggat tgcgcccttg accccctaag
     1801 cgaaactaag tgtacgctta aaagcttcac cgtcgagaag ggaatttacc agacttccaa
     1861 ctttagggtc cagccgacag aatcaatagt taggttccca aatatcacca atttgtgccc
     1921 ttttggggaa gtcttcaatg ctacacgatt tgcttcggtt tatgcctgga acaggaagcg
     1981 tatctcaaat tgcgtcgcgg actacagcgt gctctataat agtgcaagtt tctcgacctt
     2041 taaatgttat ggcgtgtccc cgaccaaact aaatgacctc tgttttacaa acgtttacgc
     2101 cgattctttt gtgattcgcg gtgatgaggt acgccaaatc gcaccgggcc agactggcaa
     2161 gattgccgat tacaactaca agttgcctga cgacttcacc ggatgtgtta tagcctggaa
     2221 tagcaataat ctggattcta aggtcggagg taactataac tacctgtata gactgttccg
     2281 taagtcaaac ctgaaaccct ttgagcggga catctccaca gagatttacc aagcagggtc
     2341 taccccctgc aatggggttg aaggattcaa ttgctacttt ccattacaga gctacgggtt
     2401 ccagcctacg aacggggttg gttaccagcc ttatagggta gtggtgctgt cgttcgaact
     2461 gttgcacgct ccagctacag tgtgcgggcc taaaaaatca actaacctgg ttaaaaataa
     2521 gtgtgttaat tttaacttca atggcttaac cggaactgga gtgctcacgg agtcaaacaa
     2581 gaaatttcta ccgtttcagc aattcgggag ggatatcgca gacactacgg acgcggtcag
     2641 agatccacag acccttgaga tccttgacat aacaccctgc tctttcggag gtgtctccgt
     2701 aattacacca ggcactaata cctctaatca ggtggccgtg ctttatcagg atgttaactg
     2761 tacagaggtc ccggtggcta tccatgccga ccaattgaca cctacgtggc gcgtctattc
     2821 taccggctcc aacgtcttcc agacccgggc tggatgcttg ataggcgcag aacacgtaaa
     2881 caactcttac gaatgtgata ttcccattgg agccgggatc tgcgcaagct accagaccca
     2941 aacaaacagt cccagacggg ctcgtagcgt ggcctctcag agcataatag cctataccat
     3001 gtcccttggc gcggagaata gtgtggcata ctcaaataat agcatcgcta tccccactaa
     3061 ctttactatt tccgtgacga ctgagattct gccagtgtct atgactaaga catccgtgga
     3121 ttgcacaatg tatatttgtg gggattccac agagtgtagc aacttgttgc tgcaatatgg
     3181 atcattctgt acgcagctga atcgcgccct caccggcatc gctgtggaac aggacaagaa
     3241 cactcaagag gtctttgctc aggtgaagca aatctacaag accccaccta taaaagactt
     3301 cgggggtttt aacttctcgc agattctccc cgaccccagc aagccttcaa aaaggagctt
     3361 tattgaggac ctgttgttca acaaggttac tttagccgat gccggtttta tcaaacagta
     3421 cggcgactgt cttggcgata tagcagcccg ggatctgatt tgcgcacaaa aattcaatgg
     3481 ccttaccgta ctgccacccc ttctcaccga cgaaatgatc gcgcagtaca cctccgcact
     3541 gctggctggg accatcactt cgggatggac ctttggggct ggtgctgctt tgcaaattcc
     3601 cttcgccatg cagatggcct atcgattcaa tggcataggg gtaacacaga acgtattgta
     3661 tgaaaatcaa aagctaatcg ctaaccagtt taattcagcc attggtaaga tccaggattc
     3721 actctccagt acagcctctg ccctgggaaa gctccaggac gtggtcaacc agaacgcgca
     3781 ggcattgaat acactcgtca aacagctgag tagcaatttc ggcgcaatca gtagtgtctt
     3841 aaacgatata ctgagtaggc ttgataaagt agaggccgag gtccagattg acagactaat
     3901 cacggggcgt cttcagagtt tacagaccta tgtgacccaa cagctgatcc gagcagcgga
     3961 aattcgagca agcgcgaacc tggccgctac taaaatgagt gagtgcgtgc tgggccaaag
     4021 taaacgagtc gacttttgcg gcaaagggta tcatctcatg tctttccctc agtccgcccc
     4081 tcacggtgtt gttttcctcc acgtgacata cgttcccgcg caggaaaaga attttactac
     4141 ggcacccgct atttgccatg acggaaaggc tcattttcca cgggagggag tgttcgtttc
     4201 aaacgggaca cactggtttg taactcagcg gaacttttat gagcctcaaa tcatcacgac
     4261 agacaatacc ttcgtgagtg gcaactgcga tgtagtcata ggcattgtga ataacactgt
     4321 ttatgatcca cttcagccgg aactcgacag cttcaaagag gaattggata agtactttaa
     4381 aaatcatacc agtccagacg tagatctcgg cgatatatcg ggcattaacg ccagtgtggt
     4441 caatatacaa aaggagattg acagactgaa tgaagtagct aagaacctca acgaatctct
     4501 gattgacctg caagagctag ggaagtacga gcagtatatc aaatggccat ggtacatatg
     4561 gctgggtttt atcgcgggtc ttattgccat agtcatggtg acgattatgt tgtgctgtat
     4621 gacctcctgc tgttcttgtt tgaaaggctg ctgctcctgt ggtagctgtt gcaaattcga
     4681 tgaagatgac tctgaacccg tgttgaaagg cgttaagcta cactacacag aaaaccttta
     4741 ctttcagtcg atggattata aggaccacga tggagactac aaagatcatg acattgatta
     4801 caaggacgac gatgacaaat aagctagctt gactgactga gatacagcgt accttcagct
     4861 cacagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     4921 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     4981 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt
     5041 gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtattggcc catctctatc
     5101 ggtatcgtag cataacccct tggggcctct aaacgggtct tgaggggttt tttgtgcccc
     5161 tcgggccgga ttgctatcta ccggcattgg cgcagaaaaa aatgcctgat gcgacgctgc
     5221 gcgtcttata ctcccacata tgccagattc agcaacggat acggcttccc caacttgccc
     5281 acttccatac gtgtcctcct taccagaaat ttatccttaa ggtcgtcagc tatcctgcag
     5341 gcgatctctc gatttcgatc aagacattcc tttaatggtc ttttctggac accactaggg
     5401 gtcagaagta gttcatcaaa ctttcttccc tccctaatct cattggttac cttgggctat
     5461 cgaaacttaa ttaaccagtc aagtcagcta cttggcgaga tcgacttgtc tgggtttcga
     5521 ctacgctcag aattgcgtca gtcaagttcg atctggtcct tgctattgca cccgttctcc
     5581 gattacgagt ttcatttaaa tcatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     5641 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     5701 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     5761 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     5821 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     5881 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     5941 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     6001 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     6061 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     6121 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     6181 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     6241 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     6301 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     6361 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     6421 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     6481 agttgcattt aaatttccga actctccaag gccctcgtcg gaaaatcttc aaacctttcg
     6541 tccgatccat cttgcaggct acctctcgaa cgaactatcg caagtctctt ggccggcctt
     6601 gcgccttggc tattgcttgg cagcgcctat cgccaggtat tactccaatc ccgaatatcc
     6661 gagatcggga tcacccgaga gaagttcaac ctacatcctc aatcccgatc tatccgagat
     6721 ccgaggaata tcgaaatcgg ggcgcgcctg gcctccgcgc cgggttttgg cgcctcccgc
     6781 gggcgccccc ctcgtcacgg cgagcgctgc cacgtcagac gaagggcgca ggagcgtcct
     6841 gatccttccg cccggacgct caggacagcg gcccgctgct cataagactc ggccttagaa
     6901 ccccagtatc agcagaagga cattttagga cgggacttgg gtgactctag ggcactggtt
     6961 ttctttccag agagcggaac aggcgaggaa aagtagtccc ttctcggcga ttctgcggag
     7021 ggatctccgt ggggcggtga acgccgatga ttatataagg acgcgccggg tgtggcacag
     7081 ctagttccgt cgcagccggg atttgggtcg cggttcttgt ttgtggatcg ctgtgatcgt
     7141 cacttggtga gtagcgggct gctgggctgg ccggggcttt cgtggccgcc gggccgctcg
     7201 gtgggacgga agcgtgtgga gagaccgcca agggctgtag tctgggtccg cgagcaaggt
     7261 tgccctgaac tgggggttgg ggggagcgca gcaaaatggc ggctgttccc gagtcttgaa
     7321 tggaagacgc ttgtgaggcg ggctgtgagg tcgttgaaac aaggtggggg gcatggtggg
     7381 cggcaagaac ccaaggtctt gagcccttcg ctaatgcggg aaagctctta ttcgggtgag
     7441 atgggctggg caccatctgg ggaccctgac gtgaagtttg tcactgactg gagaactcgg
     7501 tttgtcgtct gttgcggggg cggcagttat ggcggtgccg ttgggcagtg cacccgtacc
     7561 tttgggagcg cgcgccctcg tcgtgtcgtg acgtcacccg ttctgttggc ttataatgca
     7621 gggtggggcc acctgccggt aggtgtgcgg taggcttttc tccgtcgcag gacgcagggt
     7681 tcgggcctag ggtaggctct cctgaatcga caggcgccgg acctctggtg aggggaggga
     7741 taagtgaggc gtcagtttct ttggtcggtt ttatgtacct atcttcttaa gtagctgaag
     7801 ctccggtttt gaactatgcg ctcggggttg gcgagtgtgt tttgtgaagt tttttaggca
     7861 ccttttgaaa tgtaatcatt tgggtcaata tgtaattttc agtgttagac ttgtaaattg
     7921 tccgctaaat tctggccgtt tttggctttt ttgttagaca acatgaccga gtacaagccc
     7981 acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg
     8041 ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc
     8101 accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc
     8161 gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg
     8221 gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag
     8281 caacagatgg agggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc
     8341 accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc
     8401 ggagtggagg ccgccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc
     8461 aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa
     8521 ggaccgcgca cctggtgcat gacccgcaag cccggtgcct aggctagtat gtaagcctag
     8581 tcttagataa taaaatcgct atccatcgaa gatggatgtg tgttggtttt ttgtgtgtgt
     8641 aacgctaggc gcgcctggtg taccgagaac gatcctctca gtgcgagtct cgacgatcca
     8701 tatcgttgct tggcagtcag ccagtcggaa tccagcttgg gacccaggaa gtccaatcgt
     8761 cagatattgt actcaagcct ggtcacggca gcgtaccgat ctgtttaaac ctagatattg
     8821 atagtctgat cggtcaacgt ataatcgagt cctagctttt gcaaacatct atcaagagac
     8881 aggatcagca ggaggctttc gcatgagtat tcaacatttc cgtgtcgccc ttattccctt
     8941 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     9001 tgctgaagat cagttgggtg cgcgagtggg ttacatcgaa ctggatctca acagcggtaa
     9061 gatccttgag agttttcgcc ccgaagaacg ctttccaatg atgagcactt ttaaagttct
     9121 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     9181 acactattct cagaatgact tggttgagta ttcaccagtc acagaaaagc atcttacgga
     9241 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     9301 caacttactt ctgacaacga ttggaggacc gaaggagcta accgcttttt tgcacaacat
     9361 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     9421 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca accttgcgta aactattaac
     9481 tggcgaacta cttactctag cttcccggca acagttgata gactggatgg aggcggataa
     9541 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     9601 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     9661 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     9721 acagatcgct gagataggtg cctcactgat taagcattgg taaccgattc taggtgcatt
     9781 ggcgcagaaa aaaatgcctg atgcgacgct gcgcgtctta tactcccaca tatgccagat
     9841 tcagcaacgg atacggcttc cccaacttgc ccacttccat acgtgtcctc cttaccagaa
     9901 atttatcctt aagatcccga atcgtttaaa ctcgactctg gctctatcga atctccgtcg
     9961 tttcgagctt acgcgaacag ccgtggcgct catttgctcg tcgg
//
