LOCUS       sequence-300269-        9962 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 9962)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 9962)
  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-m4134154
FEATURES             Location/Qualifiers
     source          1..9962
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    138..209
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(214..260)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    278..326
                     /note="synthetic polyadenylation signal"
     enhancer        407..710
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        712..914
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     865..885
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             4808..4828
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             4829..4855
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    4908..5029
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4919..4938)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4973..4992
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      5144..5190
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      5220..5356
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5401..5472
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(5698..6286)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5778..5797)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        6786..7995
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     7832..7851
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             7999..8598
                     /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(7999..8018)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     8495..8515
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    8625..8673
                     /note="synthetic polyadenylation signal"
     CDS             join(8939..9007,9008..9799)
                     /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(9157..9176)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      9812..9948
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
ORIGIN
        1 taaactcgac tctggctcta tcgaatctcc gtcgtttcga gcttacgcga acagccgtgg
       61 cgctcatttg ctcgtcgggc atcgaatctc gtcagctatc gtcagcttac ctttttggca
      121 gcgatcgcgg ctcccgacat cttggaccat tagctccaca ggtatcttct tccctctagt
      181 ggtcataaca gcagcttcag ctacctctca attcaaaaaa cccctcaaga cccgtttaga
      241 ggccccaagg ggttatgcta tcaatcgttg cgttacacac acaaaaaacc aacacacatc
      301 catcttcgat ggatagcgat tttattatct aactgctgat cgagtgtagc cagatctagt
      361 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
      421 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
      481 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
      541 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
      601 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
      661 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg ctgatgcggt
      721 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
      781 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
      841 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
      901 atataagcag agctggttta gtgaaccgtc agatcagatc tttgtcgatc ctaccatcca
      961 ctcgacacac ccgccagcgg ccgccaccat gttcgtcttt cttgtgttac tccccctggt
     1021 ttcgtcccag tgcgtgaact tgactacgcg aacccaactg ccgccagcat atacaaatag
     1081 tttcacccgt ggagtatact accctgataa agtgtttaga tcaagcgtac tacactctac
     1141 tcaggacctg tttctcccct tcttctccaa tgtcacatgg tttcatgcga tccacgttag
     1201 cggcaccaac gggacaaagc gctttgacaa ccctgtgctg ccattcaatg atggtgtgta
     1261 ttttgcttct accgagaaga gcaacataat tcggggctgg atcttcggaa ctaccctcga
     1321 cagtaaaact cagtcattgc tgattgtgaa taacgccaca aatgtcgtta tcaaggtctg
     1381 tgaatttcaa ttctgcaacg atccattcct tggggtgtat taccataaaa ataataagtc
     1441 ctggatggag agcgaattta gggtatactc gtctgccaac aattgtactt tcgagtatgt
     1501 gtctcagccc ttcttaatgg atctggaagg taaacagggc aactttaaga acctccgcga
     1561 gtttgtattc aaaaatatag acggatactt taagatttat tccaagcaca caccaatcaa
     1621 tttggtgcgg gatctcccgc aagggttcag cgctctggaa cctctagtgg acctccccat
     1681 cggcattaac atcacaagat tccagacgct gttagccctg catagatcat accttactcc
     1741 cggtgacagt agttctgggt ggaccgcagg cgcggctgca tattacgtgg gatatttgca
     1801 gcctcggacg tttctgctga aatacaacga gaacggcacc attacagatg ccgtggattg
     1861 cgcccttgac cccctaagcg aaactaagtg tacgcttaaa agcttcaccg tcgagaaggg
     1921 aatttaccag acttccaact ttagggtcca gccgacagaa tcaatagtta ggttcccaaa
     1981 tatcaccaat ttgtgccctt ttggggaagt cttcaatgct acacgatttg cttcggttta
     2041 tgcctggaac aggaagcgta tctcaaattg cgtcgcggac tacagcgtgc tctataatag
     2101 tgcaagtttc tcgaccttta aatgttatgg cgtgtccccg accaaactaa atgacctctg
     2161 ttttacaaac gtttacgccg attcttttgt gattcgcggt gatgaggtac gccaaatcgc
     2221 accgggccag actggcaaga ttgccgatta caactacaag ttgcctgacg acttcaccgg
     2281 atgtgttata gcctggaata gcaataatct ggattctaag gtcggaggta actataacta
     2341 cctgtataga ctgttccgta agtcaaacct gaaacccttt gagcgggaca tctccacaga
     2401 gatttaccaa gcagggtcta ccccctgcaa tggggttgaa ggattcaatt gctactttcc
     2461 attacagagc tacgggttcc agcctacgaa cggggttggt taccagcctt atagggtagt
     2521 ggtgctgtcg ttcgaactgt tgcacgctcc agctacagtg tgcgggccta aaaaatcaac
     2581 taacctggtt aaaaataagt gtgttaattt taacttcaat ggcttaaccg gaactggagt
     2641 gctcacggag tcaaacaaga aatttctacc gtttcagcaa ttcgggaggg atatcgcaga
     2701 cactacggac gcggtcagag atccacagac ccttgagatc cttgacataa caccctgctc
     2761 tttcggaggt gtctccgtaa ttacaccagg cactaatacc tctaatcagg tggccgtgct
     2821 ttatcaggat gttaactgta cagaggtccc ggtggctatc catgccgacc aattgacacc
     2881 tacgtggcgc gtctattcta ccggctccaa cgtcttccag acccgggctg gatgcttgat
     2941 aggcgcagaa cacgtaaaca actcttacga atgtgatatt cccattggag ccgggatctg
     3001 cgcaagctac cagacccaaa caaacagtcc cagacgggct cgtagcgtgg cctctcagag
     3061 cataatagcc tataccatgt cccttggcgc ggagaatagt gtggcatact caaataatag
     3121 catcgctatc cccactaact ttactatttc cgtgacgact gagattctgc cagtgtctat
     3181 gactaagaca tccgtggatt gcacaatgta tatttgtggg gattccacag agtgtagcaa
     3241 cttgttgctg caatatggat cattctgtac gcagctgaat cgcgccctca ccggcatcgc
     3301 tgtggaacag gacaagaaca ctcaagaggt ctttgctcag gtgaagcaaa tctacaagac
     3361 cccacctata aaagacttcg ggggttttaa cttctcgcag attctccccg accccagcaa
     3421 gccttcaaaa aggagcttta ttgaggacct gttgttcaac aaggttactt tagccgatgc
     3481 cggttttatc aaacagtacg gcgactgtct tggcgatata gcagcccggg atctgatttg
     3541 cgcacaaaaa ttcaatggcc ttaccgtact gccacccctt ctcaccgacg aaatgatcgc
     3601 gcagtacacc tccgcactgc tggctgggac catcacttcg ggatggacct ttggggctgg
     3661 tgctgctttg caaattccct tcgccatgca gatggcctat cgattcaatg gcataggggt
     3721 aacacagaac gtattgtatg aaaatcaaaa gctaatcgct aaccagttta attcagccat
     3781 tggtaagatc caggattcac tctccagtac agcctctgcc ctgggaaagc tccaggacgt
     3841 ggtcaaccag aacgcgcagg cattgaatac actcgtcaaa cagctgagta gcaatttcgg
     3901 cgcaatcagt agtgtcttaa acgatatact gagtaggctt gataaagtag aggccgaggt
     3961 ccagattgac agactaatca cggggcgtct tcagagttta cagacctatg tgacccaaca
     4021 gctgatccga gcagcggaaa ttcgagcaag cgcgaacctg gccgctacta aaatgagtga
     4081 gtgcgtgctg ggccaaagta aacgagtcga cttttgcggc aaagggtatc atctcatgtc
     4141 tttccctcag tccgcccctc acggtgttgt tttcctccac gtgacatacg ttcccgcgca
     4201 ggaaaagaat tttactacgg cacccgctat ttgccatgac ggaaaggctc attttccacg
     4261 ggagggagtg ttcgtttcaa acgggacaca ctggtttgta actcagcgga acttttatga
     4321 gcctcaaatc atcacgacag acaatacctt cgtgagtggc aactgcgatg tagtcatagg
     4381 cattgtgaat aacactgttt atgatccact tcagccggaa ctcgacagct tcaaagagga
     4441 attggataag tactttaaaa atcataccag tccagacgta gatctcggcg atatatcggg
     4501 cattaacgcc agtgtggtca atatacaaaa ggagattgac agactgaatg aagtagctaa
     4561 gaacctcaac gaatctctga ttgacctgca agagctaggg aagtacgagc agtatatcaa
     4621 atggccatgg tacatatggc tgggttttat cgcgggtctt attgccatag tcatggtgac
     4681 gattatgttg tgctgtatga cctcctgctg ttcttgtttg aaaggctgct gctcctgtgg
     4741 tagctgttgc aaattcgatg aagatgactc tgaacccgtg ttgaaaggcg ttaagctaca
     4801 ctacacagaa aacctttact ttcagtcgta cccctatgac gtaccagatt atgcgtaagc
     4861 tagcttgact gactgagata cagcgtacct tcagctcaca gacatgataa gatacattga
     4921 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
     4981 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
     5041 ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta
     5101 aaacctctac aaatgtggta ttggcccatc tctatcggta tcgtagcata accccttggg
     5161 gcctctaaac gggtcttgag gggttttttg tgcccctcgg gccggattgc tatctaccgg
     5221 cattggcgca gaaaaaaatg cctgatgcga cgctgcgcgt cttatactcc cacatatgcc
     5281 agattcagca acggatacgg cttccccaac ttgcccactt ccatacgtgt cctccttacc
     5341 agaaatttat ccttaaggtc gtcagctatc ctgcaggcga tctctcgatt tcgatcaaga
     5401 cattccttta atggtctttt ctggacacca ctaggggtca gaagtagttc atcaaacttt
     5461 cttccctccc taatctcatt ggttaccttg ggctatcgaa acttaattaa ccagtcaagt
     5521 cagctacttg gcgagatcga cttgtctggg tttcgactac gctcagaatt gcgtcagtca
     5581 agttcgatct ggtccttgct attgcacccg ttctccgatt acgagtttca tttaaatcat
     5641 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     5701 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     5761 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     5821 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     5881 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     5941 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     6001 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     6061 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     6121 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     6181 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     6241 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     6301 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     6361 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     6421 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     6481 acctatctca gcgatctgtc tatttcgttc atccatagtt gcatttaaat ttccgaactc
     6541 tccaaggccc tcgtcggaaa atcttcaaac ctttcgtccg atccatcttg caggctacct
     6601 ctcgaacgaa ctatcgcaag tctcttggcc ggccttgcgc cttggctatt gcttggcagc
     6661 gcctatcgcc aggtattact ccaatcccga atatccgaga tcgggatcac ccgagagaag
     6721 ttcaacctac atcctcaatc ccgatctatc cgagatccga ggaatatcga aatcggggcg
     6781 cgcctggcct ccgcgccggg ttttggcgcc tcccgcgggc gcccccctcg tcacggcgag
     6841 cgctgccacg tcagacgaag ggcgcaggag cgtcctgatc cttccgcccg gacgctcagg
     6901 acagcggccc gctgctcata agactcggcc ttagaacccc agtatcagca gaaggacatt
     6961 ttaggacggg acttgggtga ctctagggca ctggttttct ttccagagag cggaacaggc
     7021 gaggaaaagt agtcccttct cggcgattct gcggagggat ctccgtgggg cggtgaacgc
     7081 cgatgattat ataaggacgc gccgggtgtg gcacagctag ttccgtcgca gccgggattt
     7141 gggtcgcggt tcttgtttgt ggatcgctgt gatcgtcact tggtgagtag cgggctgctg
     7201 ggctggccgg ggctttcgtg gccgccgggc cgctcggtgg gacggaagcg tgtggagaga
     7261 ccgccaaggg ctgtagtctg ggtccgcgag caaggttgcc ctgaactggg ggttgggggg
     7321 agcgcagcaa aatggcggct gttcccgagt cttgaatgga agacgcttgt gaggcgggct
     7381 gtgaggtcgt tgaaacaagg tggggggcat ggtgggcggc aagaacccaa ggtcttgagc
     7441 ccttcgctaa tgcgggaaag ctcttattcg ggtgagatgg gctgggcacc atctggggac
     7501 cctgacgtga agtttgtcac tgactggaga actcggtttg tcgtctgttg cgggggcggc
     7561 agttatggcg gtgccgttgg gcagtgcacc cgtacctttg ggagcgcgcg ccctcgtcgt
     7621 gtcgtgacgt cacccgttct gttggcttat aatgcagggt ggggccacct gccggtaggt
     7681 gtgcggtagg cttttctccg tcgcaggacg cagggttcgg gcctagggta ggctctcctg
     7741 aatcgacagg cgccggacct ctggtgaggg gagggataag tgaggcgtca gtttctttgg
     7801 tcggttttat gtacctatct tcttaagtag ctgaagctcc ggttttgaac tatgcgctcg
     7861 gggttggcga gtgtgttttg tgaagttttt taggcacctt ttgaaatgta atcatttggg
     7921 tcaatatgta attttcagtg ttagacttgt aaattgtccg ctaaattctg gccgtttttg
     7981 gcttttttgt tagacaacat gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac
     8041 gacgtcccca gggccgtacg caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc
     8101 cacaccgtcg atccggaccg ccacatcgag cgggtcaccg agctgcaaga actcttcctc
     8161 acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg
     8221 gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc
     8281 atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaggg cctcctggcg
     8341 ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac
     8401 cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggccgc cgagcgcgcc
     8461 ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta cgagcggctc
     8521 ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc
     8581 cgcaagcccg gtgcctaggc tagtatgtaa gcctagtctt agataataaa atcgctatcc
     8641 atcgaagatg gatgtgtgtt ggttttttgt gtgtgtaacg ctaggcgcgc ctggtgtacc
     8701 gagaacgatc ctctcagtgc gagtctcgac gatccatatc gttgcttggc agtcagccag
     8761 tcggaatcca gcttgggacc caggaagtcc aatcgtcaga tattgtactc aagcctggtc
     8821 acggcagcgt accgatctgt ttaaacctag atattgatag tctgatcggt caacgtataa
     8881 tcgagtccta gcttttgcaa acatctatca agagacagga tcagcaggag gctttcgcat
     8941 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     9001 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcgcg
     9061 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     9121 agaacgcttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     9181 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     9241 tgagtattca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     9301 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgattgg
     9361 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     9421 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     9481 tgtagcaatg gcaacaacct tgcgtaaact attaactggc gaactactta ctctagcttc
     9541 ccggcaacag ttgatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     9601 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     9661 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     9721 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     9781 actgattaag cattggtaac cgattctagg tgcattggcg cagaaaaaaa tgcctgatgc
     9841 gacgctgcgc gtcttatact cccacatatg ccagattcag caacggatac ggcttcccca
     9901 acttgcccac ttccatacgt gtcctcctta ccagaaattt atccttaaga tcccgaatcg
     9961 tt
//
