LOCUS       sequence-301940-        8592 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of SARS-CoV-2 S protein (D614) with the flag 
            tag at C-terminus. 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 8592)
  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 8592)
  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 (C-flag)
FEATURES             Location/Qualifiers
     source          1..8592
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..661
                     /label=chicken beta-actin promoter
     intron          662..1674
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1594..1616
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1682..1701
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             5563..5586
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(5614..5633)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5679..5734
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5733..5752)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6095..6111)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6095..6111)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6108..6130)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6119..6135
                     /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(6143..6149,6150..6167,6168..6173))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6188..6209
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6268..6463
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6314..6449
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6469..6603
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6506..6525)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6560..6579
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6671..6688)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6842..7430)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6922..6941)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7601..8392,8393..8461))
                     /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     8224..8243
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8462..8566)
                     /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 tgggtggact atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atgggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg
      541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgttgcctt cgccccgtgc cccgctccgc gccgcctcgc gccgcccgcc
      721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
      781 gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag
      841 ccttaaaggg ctccgggagg gccctttgtg cgggggggag cggctcgggg ggtgcgtgcg
      901 tgtgtgtgtg cgtggggagc gccgcgtgcg gcccgcgctg cccggcggct gtgagcgctg
      961 cgggcgcggc gcggggcttt gtgcgctccg cgtgtgcgcg aggggagcgc ggccgggggc
     1021 ggtgccccgc ggtgcggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg
     1081 tgggggggtg agcagggggt gtgggcgcgg cggtcgggct gtaacccccc cctgcacccc
     1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtgc ggggcgtggc
     1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
     1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gccccggagc gccggcggct
     1321 gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg
     1381 gacttccttt gtcccaaatc tggcggagcc gaaatctggg aggcgccgcc gcaccccctc
     1441 tagcgggcgc gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt
     1501 cgtgcgtcgc cgcgccgccg tccccttctc catctccagc ctcggggctg ccgcaggggg
     1561 acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg
     1621 gctctagcta gagcctctgc taaccatgtt catgccttct tctttttcct acagctcctg
     1681 ggcaacgtgc tggttattgt gctgtctcat cattttggca aagaattcgc cgccaccatg
     1741 tttgtgttcc tggtcctgct cccactggtt tccagccaat gtgttaacct cacaacccgc
     1801 acccagctgc ctcccgccta taccaacagt tttactcggg gagtgtatta ccctgacaaa
     1861 gttttcagga gctcagttct gcactcaacc caggatctct ttctgccttt tttttccaac
     1921 gtgacttggt tccatgcaat tcacgtttcc gggaccaacg gcaccaaaag gttcgataac
     1981 cccgtgctgc cctttaacga cggcgtctat ttcgcaagca ctgagaaaag caatatcatc
     2041 aggggctgga tttttgggac aaccttggat agcaagaccc agtcactgct gatcgttaat
     2101 aatgcaacca atgtggtgat caaagtgtgc gaatttcagt tttgcaatga cccctttctg
     2161 ggcgtctact atcataaaaa caataaatct tggatggagt ctgagtttag ggtgtattct
     2221 agcgcgaata attgtacctt tgaatatgta tcacaaccat ttctcatgga tctcgaaggc
     2281 aaacaaggca atttcaaaaa cttgagagag ttcgtcttca aaaacatcga tggatatttt
     2341 aaaatataca gcaaacatac cccaataaac ctggtccgcg acctccctca gggtttcagc
     2401 gccctcgagc ctctcgtcga tcttcctatc gggatcaaca tcacaagatt tcaaaccttg
     2461 ctggctcttc atcgatccta cctgacccct ggcgattcct cttccgggtg gaccgccggc
     2521 gccgctgcct attatgtggg gtatctgcaa cctcgcacgt ttctcctcaa gtataatgag
     2581 aatggaacta tcaccgatgc ggtagactgt gcgttggacc cactctctga gaccaagtgc
     2641 actttgaagt cctttaccgt tgaaaagggg atctatcaga catccaattt tagagtccaa
     2701 cccacggaat ccatcgttcg gttccccaat atcacaaatc tctgcccttt cggagaggta
     2761 ttcaacgcaa ctcgcttcgc ttctgtgtat gcgtggaaca gaaaacgcat ctccaactgc
     2821 gtcgctgatt actctgttct gtataactcc gcctccttta gcaccttcaa gtgttacggc
     2881 gtcagcccca caaagctcaa cgacttgtgt ttcactaacg tctacgccga ctccttcgtg
     2941 atccgggggg acgaagttcg gcagattgcg cctggacaga ctggtaaaat cgctgactat
     3001 aactataagc tcccagatga ctttaccggc tgtgtgattg cttggaactc taataatctc
     3061 gacagcaaag ttggaggcaa ctataattat ctctaccggt tgtttcggaa gagcaatctc
     3121 aagccattcg agagagacat aagcacggag atctaccaag ccggctccac tccatgcaat
     3181 ggggttgaag gctttaattg ttattttccc ctccaatcct atggttttca accgactaat
     3241 ggagtcgggt atcaacccta cagggttgtc gtcttgagct tcgagctctt gcacgctcct
     3301 gctacggtgt gtggtccgaa aaaatcaaca aacctcgtca aaaacaaatg tgtgaatttt
     3361 aactttaacg gactgactgg caccggggtg ctgactgaga gcaataagaa gtttcttcca
     3421 tttcaacaat tcggtcggga tatcgcggac accactgatg ctgtgcgcga tccgcaaacc
     3481 ttggagatct tggacataac tccgtgcagc ttcggaggcg tctctgttat cacccctgga
     3541 acgaatacat ctaaccaagt tgcggtcctt tatcaggatg ttaactgcac agaagtaccc
     3601 gtcgcgatac atgcggatca gctgacccct acttggcggg tctactcaac cggctctaat
     3661 gtgtttcaaa ctcgggcggg atgtttgatc ggagctgagc acgtgaataa ctcttatgaa
     3721 tgcgatatac caatcggcgc agggatttgc gcatcttacc aaacccaaac caactctcct
     3781 cggcgggctc gctccgtcgc atcccaatcc attatcgcct ataccatgag tctcggagct
     3841 gagaactctg tcgcttacag caacaattct atagccatcc caaccaactt cacgattagc
     3901 gttacaacag agatactccc cgtcagcatg acgaaaactt ccgttgattg tacgatgtat
     3961 atctgtgggg attctacaga gtgtagcaat ctgctgttgc agtatggctc cttctgtacc
     4021 caattgaatc gggcactcac cggcatcgct gtggaacaag acaaaaatac gcaagaagtt
     4081 ttcgcacagg tgaagcaaat ctacaagacg cctccaatca aagactttgg cggattcaat
     4141 ttttctcaga ttctgcctga tccctcaaag cccagcaaaa gaagcttcat cgaggacctg
     4201 ctcttcaaca aggtcactct cgcagatgcc gggttcatca aacaatatgg agattgcttg
     4261 ggcgacattg ctgcgcggga tcttatttgt gctcaaaagt tcaatgggct gaccgtcctg
     4321 ccgcccctct tgaccgatga aatgattgct caatacacct ccgcgttgct cgcgggaact
     4381 atcacttccg gctggacctt cggcgccggc gctgccctcc agataccgtt tgccatgcaa
     4441 atggcctaca gattcaacgg catcggagtc acacagaatg tcctgtacga aaatcaaaaa
     4501 ttgatagcca atcagtttaa ttctgcaatc ggcaaaatac aagactccct gtcctctact
     4561 gcctccgcac ttgggaaatt gcaagacgtc gtgaaccaaa acgcacaggc ccttaacact
     4621 ctggtgaaac agttgagctc caacttcggc gctataagct ccgtcttgaa cgatatactc
     4681 tctcgcctgg acaaagtcga ggctgaagtc cagatcgaca gattgataac aggcaggctt
     4741 caatccctgc aaacctacgt cacgcagcag ctgatcagag ccgcagagat cagggcttcc
     4801 gcaaatcttg ctgcaacaaa aatgtccgaa tgcgtcttgg gccaatcaaa gcgcgtcgat
     4861 ttctgtggaa agggctacca tctcatgagc tttccccaaa gcgccccgca cggcgtggtt
     4921 tttctccacg tcacctacgt ccccgctcaa gagaagaact ttacaaccgc tcctgcaatt
     4981 tgccatgatg ggaaagcgca ctttcctcgc gaaggcgtct ttgtctccaa tggcacacat
     5041 tggtttgtta ctcaacggaa cttttatgaa ccacaaatca ttaccacgga caacacgttc
     5101 gtgtccggca actgtgacgt tgtcatcgga attgttaaca acacggttta cgatcctctg
     5161 caacccgagc ttgactcctt taaggaagaa ctggacaagt acttcaaaaa tcacacctca
     5221 ccggatgtgg acttgggcga tatctctggc ataaatgcgt cagtcgtgaa cattcaaaag
     5281 gaaattgatc gactcaacga ggtcgcaaag aatcttaatg aatccctgat agatctgcag
     5341 gagttgggga agtacgagca gtatataaag tggccttggt atatatggct tggctttatc
     5401 gccggcctca ttgcaatcgt gatggtcacc atcatgctgt gctgcatgac ctcatgttgc
     5461 tcctgtctta aggggtgctg ctcctgcggt tcctgttgca aatttgacga ggacgacagc
     5521 gaaccggtgc ttaagggtgt gaagctgcat tacaccggag gcgattacaa ggatgacgat
     5581 gacaagtgac ggccgcctgc aggttaatta agatcttttt ccctctgcca aaaattatgg
     5641 ggacatcatg aagccccttg agcatctgac ttctggctaa taaaggaaat ttattttcat
     5701 tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga aggacatatg ggagggcaaa
     5761 tcatttaaaa catcagaatg agtatttggt ttagagtttg gcaacatatg cccatatgct
     5821 ggctgccatg aacaaaggtt ggctataaag aggtcatcag tatatgaaac agccccctgc
     5881 tgtccattcc ttattccata gaaaagcctt gacttgaggt tagatttttt ttatattttg
     5941 ttttgtgtta tttttttctt taacatccct aaaattttcc ttacatgttt tactagccag
     6001 atttttcctc ctctcctgac tactcccagt catagctgtc cctcttctct tatggagatc
     6061 cctcgacctg cagcccaagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     6121 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     6181 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     6241 cgggaaacct gtcgtgccag cggatccgca tctcaattag tcagcaacca tagtcccgcc
     6301 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     6361 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     6421 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctaa cttgtttatt
     6481 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     6541 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg
     6601 atccgctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     6661 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     6721 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     6781 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     6841 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     6901 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     6961 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     7021 gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     7081 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     7141 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     7201 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     7261 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
     7321 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     7381 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     7441 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     7501 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     7561 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     7621 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     7681 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     7741 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     7801 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     7861 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     7921 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     7981 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     8041 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     8101 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     8161 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     8221 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     8281 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     8341 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     8401 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     8461 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     8521 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     8581 aagtgccacc tg
//
