LOCUS       sequence-307819-        8625 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of SARS-CoV-2 S protein (D614) with the flag 
            tag at N- and C-termini. Pseudotypes onto the MLV vector but not 
            efficiently to VSV or lentiviral vectors..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8625)
  AUTHORS   Zhang L, Jackson CB, Mou H, Ojha A, Peng H, Quinlan BD, Rangarajan 
            ES, Pan A, Vanderheiden A, Suthar MS, Li W, Izard T, Rader C, Farzan
            M, Choe H
  TITLE     SARS-CoV-2 spike-protein D614G mutation increases virion spike 
            density and infectivity.
  JOURNAL   Nat Commun. 2020 Nov 26;11(1):6013. doi: 10.1038/s41467-020-19808-4.
  PUBMED    33243994
REFERENCE   2  (bases 1 to 8625)
  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"; journalName: "Nat 
            Commun."; date: "2020-11-26"; volume: "11(1):6013. doi"; pages: " 
            10.1038/s41467-020-19808-4"
COMMENT     Sequence Label: pCAGGS-SARS2-S-D614 (NC-flag)
FEATURES             Location/Qualifiers
     source          1..8625
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        385..661
                     /label=chicken beta-actin promoter
     intron          662..1682
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1602..1624
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1690..1709
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1791..1814
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             5595..5618
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(5646..5665)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5711..5766
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5765..5784)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6127..6143)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6127..6143)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6140..6162)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6151..6167
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(6175..6181,6182..6199,6200..6205))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6220..6241
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6300..6495
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6346..6481
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6501..6635
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6538..6557)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6592..6611
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6703..6720)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6874..7462)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6954..6973)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7633..8424,8425..8493))
                     /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     8256..8275
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8494..8598)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca
      421 tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag
      481 cgatgggggc gggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg
      541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc
      721 gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc
      781 tccgggctgt aattagcgct tggtttaatg acggcttgtt tcttttctgt ggctgcgtga
      841 aagccttgag gggctccggg agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg
      901 cgtgtgtgtg tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg
      961 ctgcgggcgc ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg
     1021 gggcggtgcc ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg tgcggggtgt
     1081 gtgcgtgggg gggtgagcag ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca
     1141 cccccctccc cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtacggggcg
     1201 tggcgcgggg ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc cgggcggggc
     1261 ggggccgcct cgggccgggg agggctcggg ggaggggcgc ggcggccccc ggagcgccgg
     1321 cggctgtcga ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt gcgagagggc
     1381 gcagggactt cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac
     1441 cccctctagc gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga
     1501 gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc
     1561 gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt
     1621 gaccggcggc tctagctaga gcctctgcta accatgttca tgccttcttc tttttcctac
     1681 agctcctggg caacgtgctg gttattgtgc tgtctcatca ttttggcaaa gaattcgccg
     1741 ccaccatgtt tgtgttcctg gtcctgctcc cactggtttc cagccaatgt gattacaagg
     1801 atgacgatga caaggttaac ctcacaaccc gcacccagct gcctcccgcc tataccaaca
     1861 gttttactcg gggagtgtat taccctgaca aagttttcag gagctcagtt ctgcactcaa
     1921 cccaggatct ctttctgcct tttttttcca acgtgacttg gttccatgca attcacgttt
     1981 ccgggaccaa cggcaccaaa aggttcgata accccgtgct gccctttaac gacggcgtct
     2041 atttcgcaag cactgagaaa agcaatatca tcaggggctg gatttttggg acaaccttgg
     2101 atagcaagac ccagtcactg ctgatcgtta ataatgcaac caatgtggtg atcaaagtgt
     2161 gcgaatttca gttttgcaat gacccctttc tgggcgtcta ctatcataaa aacaataaat
     2221 cttggatgga gtctgagttt agggtgtatt ctagcgcgaa taattgtacc tttgaatatg
     2281 tatcacaacc atttctcatg gatctcgaag gcaaacaagg caatttcaaa aacttgagag
     2341 agttcgtctt caaaaacatc gatggatatt ttaaaatata cagcaaacat accccaataa
     2401 acctggtccg cgacctccct cagggtttca gcgccctcga gcctctcgtc gatcttccta
     2461 tcgggatcaa catcacaaga tttcaaacct tgctggctct tcatcgatcc tacctgaccc
     2521 ctggcgattc ctcttccggg tggaccgccg gcgccgctgc ctattatgtg gggtatctgc
     2581 aacctcgcac gtttctcctc aagtataatg agaatggaac tatcaccgat gcggtagact
     2641 gtgcgttgga cccactctct gagaccaagt gcactttgaa gtcctttacc gttgaaaagg
     2701 ggatctatca gacatccaat tttagagtcc aacccacgga atccatcgtt cggttcccca
     2761 atatcacaaa tctctgccct ttcggagagg tattcaacgc aactcgcttc gcttctgtgt
     2821 atgcgtggaa cagaaaacgc atctccaact gcgtcgctga ttactctgtt ctgtataact
     2881 ccgcctcctt tagcaccttc aagtgttacg gcgtcagccc cacaaagctc aacgacttgt
     2941 gtttcactaa cgtctacgcc gactccttcg tgatccgggg ggacgaagtt cggcagattg
     3001 cgcctggaca gactggtaaa atcgctgact ataactataa gctcccagat gactttaccg
     3061 gctgtgtgat tgcttggaac tctaataatc tcgacagcaa agttggaggc aactataatt
     3121 atctctaccg gttgtttcgg aagagcaatc tcaagccatt cgagagagac ataagcacgg
     3181 agatctacca agccggctcc actccatgca atggggttga aggctttaat tgttattttc
     3241 ccctccaatc ctatggtttt caaccgacta atggagtcgg gtatcaaccc tacagggttg
     3301 tcgtcttgag cttcgagctc ttgcacgctc ctgctacggt gtgtggtccg aaaaaatcaa
     3361 caaacctcgt caaaaacaaa tgtgtgaatt ttaactttaa cggactgact ggcaccgggg
     3421 tgctgactga gagcaataag aagtttcttc catttcaaca attcggtcgg gatatcgcgg
     3481 acaccactga tgctgtgcgc gatccgcaaa ccttggagat cttggacata actccgtgca
     3541 gcttcggagg cgtctctgtt atcacccctg gaacgaatac atctaaccaa gttgcggtcc
     3601 tttatcagga tgttaactgc acagaagtac ccgtcgcgat acatgcggat cagctgaccc
     3661 ctacttggcg ggtctactca accggctcta atgtgtttca aactcgggcg ggatgtttga
     3721 tcggagctga gcacgtgaat aactcttatg aatgcgatat accaatcggc gcagggattt
     3781 gcgcatctta ccaaacccaa accaactctc ctcggcgggc tcgctccgtc gcatcccaat
     3841 ccattatcgc ctataccatg agtctcggag ctgagaactc tgtcgcttac agcaacaatt
     3901 ctatagccat cccaaccaac ttcacgatta gcgttacaac agagatactc cccgtcagca
     3961 tgacgaaaac ttccgttgat tgtacgatgt atatctgtgg ggattctaca gagtgtagca
     4021 atctgctgtt gcagtatggc tccttctgta cccaattgaa tcgggcactc accggcatcg
     4081 ctgtggaaca agacaaaaat acgcaagaag ttttcgcaca ggtgaagcaa atctacaaga
     4141 cgcctccaat caaagacttt ggcggattca atttttctca gattctgcct gatccctcaa
     4201 agcccagcaa aagaagcttc atcgaggacc tgctcttcaa caaggtcact ctcgcagatg
     4261 ccgggttcat caaacaatat ggagattgct tgggcgacat tgctgcgcgg gatcttattt
     4321 gtgctcaaaa gttcaatggg ctgaccgtcc tgccgcccct cttgaccgat gaaatgattg
     4381 ctcaatacac ctccgcgttg ctcgcgggaa ctatcacttc cggctggacc ttcggcgccg
     4441 gcgctgccct ccagataccg tttgccatgc aaatggccta cagattcaac ggcatcggag
     4501 tcacacagaa tgtcctgtac gaaaatcaaa aattgatagc caatcagttt aattctgcaa
     4561 tcggcaaaat acaagactcc ctgtcctcta ctgcctccgc acttgggaaa ttgcaagacg
     4621 tcgtgaacca aaacgcacag gcccttaaca ctctggtgaa acagttgagc tccaacttcg
     4681 gcgctataag ctccgtcttg aacgatatac tctctcgcct ggacaaagtc gaggctgaag
     4741 tccagatcga cagattgata acaggcaggc ttcaatccct gcaaacctac gtcacgcagc
     4801 agctgatcag agccgcagag atcagggctt ccgcaaatct tgctgcaaca aaaatgtccg
     4861 aatgcgtctt gggccaatca aagcgcgtcg atttctgtgg aaagggctac catctcatga
     4921 gctttcccca aagcgccccg cacggcgtgg tttttctcca cgtcacctac gtccccgctc
     4981 aagagaagaa ctttacaacc gctcctgcaa tttgccatga tgggaaagcg cactttcctc
     5041 gcgaaggcgt ctttgtctcc aatggcacac attggtttgt tactcaacgg aacttttatg
     5101 aaccacaaat cattaccacg gacaacacgt tcgtgtccgg caactgtgac gttgtcatcg
     5161 gaattgttaa caacacggtt tacgatcctc tgcaacccga gcttgactcc tttaaggaag
     5221 aactggacaa gtacttcaaa aatcacacct caccggatgt ggacttgggc gatatctctg
     5281 gcataaatgc gtcagtcgtg aacattcaaa aggaaattga tcgactcaac gaggtcgcaa
     5341 agaatcttaa tgaatccctg atagatctgc aggagttggg gaagtacgag cagtatataa
     5401 agtggccttg gtatatatgg cttggcttta tcgccggcct cattgcaatc gtgatggtca
     5461 ccatcatgct gtgctgcatg acctcatgtt gctcctgtct taaggggtgc tgctcctgcg
     5521 gttcctgttg caaatttgac gaggacgaca gcgaaccggt gcttaagggt gtgaagctgc
     5581 attacaccgg aggcgattac aaggatgacg atgacaagtg acggccgcct gcaggttaat
     5641 taagatcttt ttccctctgc caaaaattat ggggacatca tgaagcccct tgagcatctg
     5701 acttctggct aataaaggaa atttattttc attgcaatag tgtgttggaa ttttttgtgt
     5761 ctctcactcg gaaggacata tgggagggca aatcatttaa aacatcagaa tgagtatttg
     5821 gtttagagtt tggcaacata tgcccatatg ctggctgcca tgaacaaagg ttggctataa
     5881 agaggtcatc agtatatgaa acagccccct gctgtccatt ccttattcca tagaaaagcc
     5941 ttgacttgag gttagatttt ttttatattt tgttttgtgt tatttttttc tttaacatcc
     6001 ctaaaatttt ccttacatgt tttactagcc agatttttcc tcctctcctg actactccca
     6061 gtcatagctg tccctcttct cttatggaga tccctcgacc tgcagcccaa gcttggcgta
     6121 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     6181 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     6241 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agcggatccg
     6301 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     6361 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     6421 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     6481 ctaggctttt gcaaaaagct aacttgttta ttgcagctta taatggttac aaataaagca
     6541 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     6601 ccaaactcat caatgtatct tatcatgtct ggatccgctg cattaatgaa tcggccaacg
     6661 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     6721 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     6781 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     6841 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     6901 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     6961 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     7021 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     7081 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     7141 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     7201 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     7261 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     7321 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     7381 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     7441 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     7501 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     7561 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     7621 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     7681 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     7741 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     7801 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     7861 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     7921 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     7981 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     8041 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     8101 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     8161 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     8221 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     8281 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     8341 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     8401 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     8461 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     8521 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     8581 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgg
//
