LOCUS       sequence-363818-        8548 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses codon-optimized full length SARS-CoV-2 D614G Spike.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8548)
  AUTHORS   Benlarbi M, Laroche G, Fink C, Fu K, Mulloy RP, Phan A, Ariana A, 
            Stewart CM, Prevost J, Beaudoin-Bussieres G, Daniel R, Bo Y, El 
            Ferri O, Yockell-Lelievre J, Stanford WL, Giguere PM, Mubareka S, 
            Finzi A, Dekaban GA, Dikeakos JD, Cote M
  TITLE     Identification and differential usage of a host metalloproteinase 
            entry pathway by SARS-CoV-2 Delta and Omicron.
  JOURNAL   iScience. 2022 Nov 18;25(11):105316. doi: 
            10.1016/j.isci.2022.105316. Epub 2022 Oct 10.
  PUBMED    36254158
REFERENCE   2  (bases 1 to 8548)
  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: "iScience.";
            date: "2022-11-18"; volume: "25(11):105316. doi"; pages: " 
            10.1016/j.isci.2022.105316. Epub 2022 Oct 10"
COMMENT     Sequence Label: pCAGGS SARS-CoV-2 D614G Spike
FEATURES             Location/Qualifiers
     source          1..8548
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          661..1677
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1601..1623
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1685..1704
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(5569..5588)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5634..5689
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5688..5707)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6050..6066)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6050..6066)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6063..6085)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6074..6090
                     /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(6098..6104,6105..6122,6123..6128))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6143..6164
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6223..6418
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6269..6404
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6424..6558
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6461..6480)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6515..6534
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6626..6643)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6797..7385)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6877..6896)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7556..8347,8348..8416))
                     /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     8179..8198
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8417..8521)
                     /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 ggggcgcgcg ccaggsgggg sggggsgggg sgrggggsgg
      541 ggsggggsgr ggcggrgagg kgcggcggca gccaatcaga gcggcgcgct ccgaaagttt
      601 ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa agcgaagcgc gcggcgggcg
      661 ggagtcgctg cgcgctgcct tcgccccgtg ccccgctccg ccgccgcctc gcgccgcccg
      721 ccccggctct gactgaccgc gttactccca caggtgagcg ggcgggacgg cccttctcct
      781 ccgggctgta attagcgctt ggtttaatga cggcttgttt cttttctgtg gctgcgtgaa
      841 agccttgagg ggctccggga gggccctttg tgcgggggga gcggctcggg gggtgcgtgc
      901 gtgtgtgtgt gcgtggggag cgccgcgtgc ggctccgcgc tgcccggcgg ctgtgagcgc
      961 tgcgggcgcg gcgcggggct ttgtgcgctc cgcagtgtgc gcgaggggag cgcggccggg
     1021 ggcggtgccc cgcggtgcgg ggggggctgc gaggggaaca aaggctgcgt gcggggtgtg
     1081 tgcgtggggg ggtgagcagg gggtgtgggc gcgtcggtcg ggctgcaacc ccccctgcac
     1141 ccccctcccc gagttgctga gcacggcccg gcttcgggtg cggggctccg tacggggcgt
     1201 ggcgcggggc tcgccgtgcc gggcgggggg tggcggcagg tgggggtgcc gggcggggcg
     1261 gggccgcctc gggccggggr gggctcgggg gaggggcgcg gcggcccccg gagcgccggc
     1321 ggctgtcgag gcgcggcgag ccgcagccat tgccttttat ggtaatcgtg cgagagggcg
     1381 cagggacttc ctttgtccca aatctgtgcg gagccgaaat ctgggaggcg ccgccgcacc
     1441 ccctctagcg ggcgcggggc gaagcggtgc ggcgccggca ggaaggaaat gggcggggag
     1501 ggccttcgtg cgtcgccgcg ccgccgtccc cttctccctc tccagcctcg gggctgtccg
     1561 cggggggacg gctgccttcg ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg
     1621 accggcggct ctagagcctc tgctaaccat gttcatgcct tcttcttttt cctacagctc
     1681 ctgggcaacg tgctggttat tgtgctgtct catcattttg gcaaagaatt cgccaccatg
     1741 ttcgtcttcc tcgtcctcct ccccctcgtc tcctcccagt gcgtcaacct caccacccgc
     1801 acccagctcc cccccgccta caccaactcc ttcacccgcg gcgtctacta ccccgacaag
     1861 gtcttccgct cctccgtcct ccactccacc caggacctct tcctcccctt cttctccaac
     1921 gtcacctggt tccacgccat ccacgtctcc ggcaccaacg gcaccaagcg cttcgacaac
     1981 cccgtcctcc ccttcaacga cggcgtctac ttcgcctcca ccgagaagtc caacatcatc
     2041 cgcggctgga tcttcggcac caccctcgac tccaagaccc agtccctcct catcgtcaac
     2101 aacgccacca acgtcgtcat caaggtctgc gagttccagt tctgcaacga ccccttcctc
     2161 ggcgtctact accacaagaa caacaagtcc tggatggagt ccgagttccg cgtctactcc
     2221 tccgccaaca actgcacctt cgagtacgtc tcccagccct tcctcatgga cctcgagggc
     2281 aagcagggca acttcaagaa cctccgcgag ttcgtcttca agaacatcga cggctacttc
     2341 aagatctact ccaagcacac ccccatcaac ctcgtccgcg acctccccca gggcttctcc
     2401 gccctcgagc ccctcgtcga cctccccatc ggcatcaaca tcacccgctt ccagaccctc
     2461 ctcgccctcc accgctccta cctcaccccc ggcgactcct cctccggctg gaccgccggc
     2521 gccgccgcct actacgtcgg ctacctccag ccccgcacct tcctcctcaa gtacaacgag
     2581 aacggcacca tcaccgacgc cgtcgactgc gccctcgacc ccctctccga gaccaagtgc
     2641 accctcaagt ccttcaccgt cgagaagggc atctaccaga cctccaactt ccgcgtccag
     2701 cccaccgagt ccatcgtccg cttccccaac atcaccaacc tctgcccctt cggcgaggtc
     2761 ttcaacgcca cccgcttcgc ctccgtctac gcctggaacc gcaagcgcat ctccaactgc
     2821 gtcgccgact actccgtcct ctacaactcc gcctccttct ccaccttcaa gtgctacggc
     2881 gtctccccca ccaagctcaa cgacctctgc ttcaccaacg tctacgccga ctccttcgtc
     2941 atccgcggcg acgaggtccg ccagatcgcc cccggccaga ccggcaagat cgccgactac
     3001 aactacaagc tccccgacga cttcaccggc tgcgtcatcg cctggaactc caacaacctc
     3061 gactccaagg tcggcggcaa ctacaactac ctctaccgcc tcttccgcaa gtccaacctc
     3121 aagcccttcg agcgcgacat ctccaccgag atctaccagg ccggctccac cccctgcaac
     3181 ggcgtcgagg gcttcaactg ctacttcccc ctccagtcct acggcttcca gcccaccaac
     3241 ggcgtcggct accagcccta ccgcgtcgtc gtcctctcct tcgagctcct ccacgccccc
     3301 gccaccgtct gcggccccaa gaagtccacc aacctcgtca agaacaagtg cgtcaacttc
     3361 aacttcaacg gcctcaccgg caccggcgtc ctcaccgagt ccaacaagaa gttcctcccc
     3421 ttccagcagt tcggccgcga catcgccgac accaccgacg ccgtccgcga cccccagacc
     3481 ctcgagatcc tcgacatcac cccctgctcc ttcggcggcg tctccgtcat cacccccggc
     3541 accaacacct ccaaccaggt cgccgtcctc taccagggtg tcaactgcac cgaggtcccc
     3601 gtcgccatcc acgccgacca gctcaccccc acctggcgcg tctactccac cggctccaac
     3661 gtcttccaga cccgcgccgg ctgcctcatc ggcgccgagc acgtcaacaa ctcctacgag
     3721 tgcgacatcc ccatcggcgc cggcatctgc gcctcctacc agacccagac caactccccc
     3781 cgccgcgccc gctccgtcgc ctcccagtcc atcatcgcct acaccatgtc cctcggcgcc
     3841 gagaactccg tcgcctactc caacaactcc atcgccatcc ccaccaactt caccatctcc
     3901 gtcaccaccg agatcctccc cgtctccatg accaagacct ccgtcgactg caccatgtac
     3961 atctgcggcg actccaccga gtgctccaac ctcctcctcc agtacggctc cttctgcacc
     4021 cagctcaacc gcgccctcac cggcatcgcc gtcgagcagg acaagaacac ccaggaggtc
     4081 ttcgcccagg tcaagcagat ctacaagacc ccccccatca aggacttcgg cggcttcaac
     4141 ttctcccaga tcctccccga cccctccaag ccctccaagc gctccttcat cgaggacctc
     4201 ctcttcaaca aggtcaccct cgccgacgcc ggcttcatca agcagtacgg cgactgcctc
     4261 ggcgacatcg ccgcccgcga cctcatctgc gcccagaagt tcaacggcct caccgtcctc
     4321 ccccccctcc tcaccgacga gatgatcgcc cagtacacct ccgccctcct cgccggcacc
     4381 atcacctccg gctggacctt cggcgccggc gccgccctcc agatcccctt cgccatgcag
     4441 atggcctacc gcttcaacgg catcggcgtc acccagaacg tcctctacga gaaccagaag
     4501 ctcatcgcca accagttcaa ctccgccatc ggcaagatcc aggactccct ctcctccacc
     4561 gcctccgccc tcggcaagct ccaggacgtc gtcaaccaga acgcccaggc cctcaacacc
     4621 ctcgtcaagc agctctcctc caacttcggc gccatctcct ccgtcctcaa cgacatcctc
     4681 tcccgcctcg acaaggtcga ggccgaggtc cagatcgacc gcctcatcac cggccgcctc
     4741 cagtccctcc agacctacgt cacccagcag ctcatccgcg ccgccgagat ccgcgcctcc
     4801 gccaacctcg ccgccaccaa gatgtccgag tgcgtcctcg gccagtccaa gcgcgtcgac
     4861 ttctgcggca agggctacca cctcatgtcc ttcccccagt ccgcccccca cggcgtcgtc
     4921 ttcctccacg tcacctacgt ccccgcccag gagaagaact tcaccaccgc ccccgccatc
     4981 tgccacgacg gcaaggccca cttcccccgc gagggcgtct tcgtctccaa cggcacccac
     5041 tggttcgtca cccagcgcaa cttctacgag ccccagatca tcaccaccga caacaccttc
     5101 gtctccggca actgcgacgt cgtcatcggc atcgtcaaca acaccgtcta cgaccccctc
     5161 cagcccgagc tcgactcctt caaggaggag ctcgacaagt acttcaagaa ccacacctcc
     5221 cccgacgtcg acctcggcga catctccggc atcaacgcct ccgtcgtcaa catccagaag
     5281 gagatcgacc gcctcaacga ggtcgccaag aacctcaacg agtccctcat cgacctccag
     5341 gagctcggca agtacgagca gtacatcaag tggccctggt acatctggct cggcttcatc
     5401 gccggcctca tcgccatcgt catggtcacc atcatgctct gctgcatgac ctcctgctgc
     5461 tcctgcctca agggctgctg ctcctgcggc tcctgctgca agttcgacga ggacgactcc
     5521 gagcccgtcc tcaagggcgt caagctccac tacacctaag ctagcagatc tttttccctc
     5581 tgccaaaaat tatggggaca tcatgaagcc ccttgagcat ctgacttctg gctaataaag
     5641 gaaatttatt ttcattgcaa tagtgtgttg gaattttttg tgtctctcac tcggaaggac
     5701 atatgggagg gcaaatcatt taaaacatca gaatgagtat ttggtttaga gtttggcaac
     5761 atatgcccat atgctggctg ccatgaacaa aggttggcta taaagaggtc atcagtatat
     5821 gaaacagccc cctgctgtcc attccttatt ccatagaaaa gccttgactt gaggttagat
     5881 tttttttata ttttgttttg tgttattttt ttctttaaca tccctaaaat tttccttaca
     5941 tgttttacta gccagatttt tcctcctctc ctgactactc ccagtcatag ctgtccctct
     6001 tctcttatgg agatccctcg acctgcagcc caagcttggc gtaatcatgg tcatagctgt
     6061 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     6121 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     6181 tgcccgcttt ccagtcggga aacctgtcgt gccagcggat ccgcatctca attagtcagc
     6241 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     6301 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     6361 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
     6421 gctaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     6481 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     6541 tcttatcatg tctggatccg ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     6601 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     6661 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     6721 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     6781 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     6841 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     6901 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     6961 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     7021 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     7081 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     7141 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     7201 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     7261 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     7321 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     7381 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     7441 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     7501 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     7561 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     7621 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     7681 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     7741 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     7801 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     7861 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     7921 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     7981 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     8041 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     8101 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     8161 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     8221 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     8281 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     8341 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     8401 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     8461 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     8521 gcacatttcc ccgaaaagtg ccacctgg
//
