LOCUS       sequence-334274-        9202 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For expression of SARS-CoV-2 Spike D614G in mammalian cells (C9 
            tagged at C-term).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9202)
  AUTHORS   Daniloski Z, Jordan TX, Ilmain JK, Guo X, Bhabha G, tenOever BR, 
            Sanjana NE
  TITLE     The Spike D614G mutation increases SARS-CoV-2 infection of multiple 
            human cell types.
  JOURNAL   Elife. 2021 Feb 11;10. pii: 65365. doi: 10.7554/eLife.65365.
  PUBMED    33570490
REFERENCE   2  (bases 1 to 9202)
  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: "Elife."; 
            date: "2021-02-11"; volume: "10. pii: 65365. doi"; pages: " 
            10.7554/eLife.65365"
COMMENT     Sequence Label: pcDNA3.1-SARS2-Spike-D614G
FEATURES             Location/Qualifiers
     source          1..9202
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     241..261
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(270..698)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(380..401)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     592..611
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    744..968
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     957..974
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     primer_bind     complement(4885..4904)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4886..4904)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(4949..5152)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4978..4998)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        5153..5532
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     5704..5723
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        5798..5902
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5903..5971,5972..6763)
                     /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(6121..6140)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6934..7522
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7423..7442
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7676..7693
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7810..7831
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7846..7851,7852..7869,7870..7876)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7884..7900
                     /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)."
     primer_bind     7889..7911
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7908..7924
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7908..7924
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    7973..8094
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7984..8003)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     8038..8057
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(8268..9062)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(8379..8398)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     8989..9008
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(9129..9202,1..256))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(9143..9202,1..76)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(join(9197..9202,1..14))
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
ORIGIN
        1 cctctgcata aataaaaaaa attagtcagc catggggcgg agaatgggcg gaactgggcg
       61 gagttagggg cgggatgggc ggagttaggg gcgggactat ggttgctgac taattgagat
      121 gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac ctggttgctg
      181 actaattgag atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac
      241 accctaactg acacacattc cacagaatta attcgcgtta aatttttgtt aaatcagctc
      301 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      361 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      421 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      481 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      541 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      601 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      661 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtgg ggataccccc tagagcccca
      721 gctggttctt tccgcctcag aagccataga gcccaccgca tccccagcat gcctgctatt
      781 gtcttcccaa tcctccccct tgctgtcctg ccccacccca ccccccagaa tagaatgaca
      841 cctactcaga caatgcgatg caatttcctc attttattag gaaaggacag tgggagtggc
      901 accttccagg gtcaaggaag gcacggggga ggggcaaaca acagatggct ggcaactaga
      961 aggcacagtc gaggctgatc agcgggttta aacgggccct ctagactcga gctaagcggg
     1021 agcgacctgg gatgtctcgg tggagcctgt atagtgcagt ttgacgccct tcagcacggg
     1081 ctcgctatcg tcctcatcga acttgcagca gctgccgcag gagcagcagc ccttcaggca
     1141 gctgcagcag gaggtcatgc agcacagcat gatggtcacc atcacgatgg cgatcaggcc
     1201 ggcgatgaag cccagccaga tgtaccatgg ccacttgatg tactgctcgt acttgcccag
     1261 ctcctgcaga tcgatcaggg actcgttcag attcttggcc acctcgttca ggcggtcgat
     1321 ctccttctgg atgttcacca cgctggcatt gatgccggag atgtcgccca gatccacgtc
     1381 ggggctggtg tgattcttga agtacttatc cagctcctcc ttgaaggagt ccagctctgg
     1441 ctgcagggga tcgtacacgg tattgttcac gattccgatg accacgtcgc agttgccgct
     1501 cacgaaggta ttgtcggtgg tgatgatctg gggctcgtag aaattgcgct gggtcacgaa
     1561 ccagtgggtg ccgttggaca cgaacacgcc ctcgcgtggg aagtgagcct tgccatcgtg
     1621 gcagatagct ggagcggtgg tgaagttctt ctcctgggct ggcacgtagg tcacgtgcag
     1681 gaacaccact ccgtgtgggg cgctctgtgg gaaggacatc aggtggtagc ccttgccgca
     1741 gaagtccacc ctcttgctct gtcccagcac gcactcggac atcttggtag cagccagatt
     1801 ggcgctggcc ctgatctcag cagccctgat cagctgctgg gtcacgtagg tctgcaggga
     1861 ctgcaggcgg ccggtgatca gcctatcgat ctgcacctca gcctccacct tgtccaggcg
     1921 gctcaggata tcattcagca cgctggagat ggcgccgaag ttgctggaca gctgcttcac
     1981 cagggtattc agggcctggg cgttctgatt caccacatcc tgcagcttgc ccagggcact
     2041 ggcggtgctg gacagggagt cctggatctt gccgatggcg gagttgaact gattggcgat
     2101 cagcttctgg ttctcgtaca gcacattctg ggtcacgccg atgccgttga agcggtaggc
     2161 catctgcatg gcgaagggga tctgcagggc ggctccagcg ccgaaggtcc atccgctggt
     2221 gatggttccg gccagcaggg cggaggtgta ctgagcgatc atctcatcgg tcagcagtgg
     2281 tggcagcacg gtcaggccat tgaacttctg ggcgcagatc aggtcgcggg cagcgatgtc
     2341 gcccaggcaa tcgccgtact gcttgatgaa gccggcatcg gccagggtca ccttgttgaa
     2401 cagcaggtcc tcgatgaagc tgcgcttgga gggcttgctt ggatcgggca ggatctggct
     2461 gaaattgaag ccgccgaagt ccttgattgg gggggtcttg tagatctgct tcacctgggc
     2521 gaacacctcc tgggtgttct tatcctgctc cacagcgatt ccggtcaggg cgcgattcag
     2581 ctgggtgcag aagctgccgt actgcagcag caggttgctg cactcggtgg aatcgccgca
     2641 gatgtacatg gtgcagtcca cgctggtctt ggtcatggac acgggcagga tctcggtggt
     2701 cacggagatg gtgaagttgg ttgggatggc gatggaattg ttgctgtagg ccacgctatt
     2761 ctcggcgccc agggacatgg tgtaggcgat gatggactgg ctggccacgc tgcgagccct
     2821 ccttggggag ttggtctggg tctggtagct ggcgcagatt ccagctccga ttgggatgtc
     2881 gcactcgtag gaattgttca cgtgctcagc tccgatcagg catccggcgc gggtctggaa
     2941 cacgttggat ccggtgctgt acacgcgcca ggttggggtc agctgatcag cgtggatagc
     3001 cactggcacc tcggtgcaat tcacgccctg gtacagcacg gccacctggt tgctggtatt
     3061 ggttcctggg gtgatcacgc tcacgccgcc gaaggagcat ggggtgatat ccaggatctc
     3121 cagggtctgt gggtcgcgca cggcgtcggt ggtatcagcg atgtcccttc cgaactgctg
     3181 gaatggcagg aacttcttgt tggactcggt cagcacgccg gttccggtca ggccgttgaa
     3241 attgaagttc acgcacttgt tcttcaccag attggtgctc ttctttggtc cgcacacggt
     3301 agctggagcg tgcagcagct cgaaggacag caccaccacc ctgtatggct ggtatcccac
     3361 tccgttggtt ggctggaagc cgtagctctg cagggggaag tagcagttga agccctccac
     3421 tccattgcat ggggtgcttc cagcctggta gatctcggtg gagatgtcgc gctcgaatgg
     3481 cttcagattg gacttgcgga acaggcggta caggtaattg tagttgccgc ccactttgct
     3541 atccagattg ttggagttcc aggcgatcac gcagccggtg aaatcgtcgg gcagcttgta
     3601 attgtagtca gcgatcttgc cggtctgtcc tggagcgatc tggcgcacct cgtcgcccct
     3661 gatcacgaag ctatcggcgt acacgttggt gaagcacaga tcattcagct tggtggggga
     3721 cacgccgtag cacttgaagg tggagaagct ggcggagttg tacagcacgc tgtagtcggc
     3781 cacgcagttg gagatgcgct tgcgattcca agcgtacacg ctggcgaacc tggtagcgtt
     3841 gaacacctcg ccgaatgggc acaggttggt gatattgggg aagcgcacga tgctctcggt
     3901 tggctgcacg cggaaattgg aggtctggta gatgcccttc tccacggtga agctcttcag
     3961 ggtgcacttt gtctcggaca gtggatccag ggcgcaatcc acggcgtcgg tgatggtgcc
     4021 attctcgttg tacttcagca ggaaggtgcg gggctgcagg tagcccacgt agtaggcagc
     4081 agctccagcg gtccatccgc tggagctgtc gcctggggtc aggtagctgc ggtgcagggc
     4141 cagcagggtc tggaacctgg tgatgttgat gccgattggc agatccacca gtggctccag
     4201 ggcgctgaag ccctgtggca ggtcgcgcac caggttgatg ggggtgtgct tggagtagat
     4261 cttgaagtag ccatcgatat tcttgaacac gaactcgcgc aggttcttga aattgccctg
     4321 cttgccctcc aggtccatca ggaagggctg ggacacgtac tcgaaggtgc aattgttggc
     4381 ggagctgtac acgcggaact cgctctccat ccaggactta ttgttcttgt ggtagtacac
     4441 gcccaggaat ggatcattgc agaactggaa ctcgcacacc ttgatgacca cgttggtggc
     4501 attgttcacg atcagcaggc tctgggtctt ggagtccagg gtggtgccga agatccagcc
     4561 gcggatgatg ttgctcttct cggtggaggc gaagtacacg ccatcgttga atggcagcac
     4621 gggattgtcg aaccgcttgg tgccattggt gccgctcacg tggatggcgt ggaaccaggt
     4681 cacgttgctg aagaagggca ggaacagatc ctgggtggag tgcagcacgg agctgcggaa
     4741 caccttgtct gggtagtaca cgccgcgggt gaaggagtta gtataggcag gaggcagctg
     4801 ggttctagta gtcaggttca cgcactgaga cgagaccaga ggcagcagga ccaggaagac
     4861 aaacatgcta gccagcttgg gtctccctat agtgagtcgt attaatttcg ataagccagt
     4921 aagcagtggg ttctctagtt agccagagag ctctgcttat atagacctcc caccgtacac
     4981 gcctaccgcc catttgcgtc aatggggcgg agttgttacg acattttgga aagtcccgtt
     5041 gattttggtg ccaaaacaaa ctcccattga cgtcaatggg gtggagactt ggaaatcccc
     5101 gtgagtcaaa ccgctatcca cgcccattga tgtactgcca aaaccgcatc accatggtaa
     5161 tagcgatgac taatacgtag atgtactgcc aagtaggaaa gtcccataag gtcatgtact
     5221 gggcataatg ccaggcgggc catttaccgt cattgacgtc aatagggggc gtacttggca
     5281 tatgatacac ttgatgtact gccaagtggg cagtttaccg taaatactcc acccattgac
     5341 gtcaatggaa agtccctatt ggcgttacta tgggaacata cgtcattatt gacgtcaatg
     5401 ggcgggggtc gttgggcggt cagccaggcg ggccatttac cgtaagttat gtaacgcgga
     5461 actccatata tgggctatga actaatgacc ccgtaattga ttactattaa taactagtca
     5521 ataatcaatg tcaacgcgta tatctggccc gtacatcgcg aagcagcgca aaacgcctaa
     5581 ccctaagcag attcttcatg caattgtcgg tcaagccttg ccttgttgta gcttaaattt
     5641 tgctcgcgca ctactcagcg acctccaaca cacaagcagg gagcagatac tggcttaact
     5701 atgcggcatc agagcagatt gtactgagag tgcaccatag gggatcggga gatctcccga
     5761 tccgtcgacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     5821 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     5881 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     5941 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     6001 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     6061 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     6121 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     6181 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     6241 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     6301 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     6361 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     6421 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     6481 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     6541 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     6601 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     6661 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     6721 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     6781 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     6841 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     6901 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     6961 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     7021 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     7081 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     7141 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     7201 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     7261 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     7321 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     7381 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     7441 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     7501 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     7561 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     7621 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     7681 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     7741 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
     7801 caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct
     7861 tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta
     7921 tgaccatgat tacgccaagc tctagctaga ggtcgacggt atacagacat gataagatac
     7981 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     8041 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttggggtg
     8101 ggcgaagaac tccagcatga gatccccgcg ctggaggatc atccagccgg cgtcccggaa
     8161 aacgattccg aagcccaacc tttcatagaa ggcggcggtg gaatcgaaat ctcgtgatgg
     8221 caggttgggc gtcgcttggt cggtcatttc gaaccccaga gtcccgctca gaagaactcg
     8281 tcaagaaggc gatagaaggc gatgcgctgc gaatcgggag cggcgatacc gtaaagcacg
     8341 aggaagcggt cagcccattc gccgccaagc tcttcagcaa tatcacgggt agccaacgct
     8401 atgtcctgat agcggtccgc cacacccagc cggccacagt cgatgaatcc agaaaagcgg
     8461 ccattttcca ccatgatatt cggcaagcag gcatcgccat gggtcacgac gagatcctcg
     8521 ccgtcgggca tgcgcgcctt gagcctggcg aacagttcgg ctggcgcgag cccctgatgc
     8581 tcttcgtcca gatcatcctg atcgacaaga ccggcttcca tccgagtacg tgctcgctcg
     8641 atgcgatgtt tcgcttggtg gtcgaatggg caggtagccg gatcaagcgt atgcagccgc
     8701 cgcattgcat cagccatgat ggatactttc tcggcaggag caaggtgaga tgacaggaga
     8761 tcctgccccg gcacttcgcc caatagcagc cagtcccttc ccgcttcagt gacaacgtcg
     8821 agcacagctg cgcaaggaac gcccgtcgtg gccagccacg atagccgcgc tgcctcgtcc
     8881 tgcagttcat tcagggcacc ggacaggtcg gtcttgacaa aaagaaccgg gcgcccctgc
     8941 gctgacagcc ggaacacggc ggcatcagag cagccgattg tctgttgtgc ccagtcatag
     9001 ccgaatagcc tctccaccca agcggccgga gaacctgcgt gcaatccatc ttgttcaatc
     9061 atgcgaaacg atcctcatcc tgtctcttga tcagatccga aaatggatat acaagctccc
     9121 gggagctttt tgcaaaagcc taggcctcca aaaaagcctc ctcactactt ctggaatagc
     9181 tcagaggcag aggcggcctc gg
//
