LOCUS       sequence-333040-        9122 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express SARS-CoV-1 WT spike protein with a 28aa deletion on the 
            C-terminal tail. Used for SARS1 pseudotyped virus production..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9122)
  AUTHORS   Rogers TF, Zhao F, Huang D, Beutler N, Burns A, He WT, Limbo O, 
            Smith C, Song G, Woehl J, Yang L, Abbott RK, Callaghan S, Garcia E, 
            Hurtado J, Parren M, Peng L, Ramirez S, Ricketts J, Ricciardi MJ, 
            Rawlings SA, Wu NC, Yuan M, Smith DM, Nemazee D, Teijaro JR, Voss 
            JE, Wilson IA, Andrabi R, Briney B, Landais E, Sok D, Jardine JG, 
            Burton DR
  TITLE     Isolation of potent SARS-CoV-2 neutralizing antibodies and 
            protection from disease in a small animal model.
  JOURNAL   Science. 2020 Aug 21;369(6506):956-963. doi: 
            10.1126/science.abc7520. Epub 2020 Jun 15.
  PUBMED    32540903
REFERENCE   2  (bases 1 to 9122)
  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"; doi: 
            "10.1126/science.abc7520"; journalName: "Science"; date: 
            "2020-08-21- 21"; volume: "369"; issue: "6506"; pages: "956-963"
COMMENT     Sequence Label: pcDNA3.3_CoV1_D28
FEATURES             Location/Qualifiers
     source          1..9122
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        64..443
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        444..647
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     598..618
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     644..668
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     complement(4522..4541)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4566..4614
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4816..5244
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4903..4922)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5113..5134
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5253..5273)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5258..5587
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5438..5573
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5500..5519
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5654..6448
                     /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(5708..5727)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6318..6337
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6624..6745
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6661..6680)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6715..6734
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6794..6810)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6794..6810)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6807..6829)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6818..6834
                     /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(6842..6848,6849..6866,6867..6872))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6887..6908
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7025..7042)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7196..7784)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7276..7295)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7955..8746,8747..8815))
                     /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     8578..8597
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8816..8920)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8995..9014)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc
       61 gttgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
      121 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      181 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      241 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      301 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      361 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      421 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat
      481 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
      541 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
      601 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctcgt ttagtgaacc
      661 gtcagatcgc ctggagacgc catccacgct gttttgacct ccatagaaga caccgggacc
      721 gatccagcct ccggactgag ctccttaagg ttaactctag agaattcgga tccatgttca
      781 tcttcctgct gttcctgacc ctgacctctg gctctgacct ggacaggtgt accacctttg
      841 atgatgtcca ggctccaaac tacacccaac acacctccag tatgagggga gtctactacc
      901 ctgatgagat tttcaggtct gacaccctct acctgaccca ggacctgttc ctgccattct
      961 acagcaatgt gacaggcttc cacaccatca accacacctt tgacaaccct gtgattccat
     1021 tcaaggatgg catctacttt gctgccacag agaagagcaa tgtggtgagg ggctgggtgt
     1081 ttggcagcac aatgaacaac aagagccagt ctgtgattat catcaacaac agcaccaatg
     1141 tggtgattag ggcttgtaac tttgaactgt gtgacaaccc attctttgct gtgagcaagc
     1201 ctatgggcac ccagacccac acaatgattt ttgacaatgc cttcaactgt acctttgaat
     1261 acatctctga tgccttctcc ctggatgtgt ctgagaagtc tggcaacttc aaacacctga
     1321 gggagtttgt gttcaagaac aaggatggct tcctctatgt ctacaagggc taccaaccaa
     1381 ttgatgtggt gagggacctg ccatctggct tcaacaccct gaaaccaatc ttcaaactgc
     1441 cactgggcat caacatcacc aacttcaggg ctatcctgac agccttcagc cctgcccagg
     1501 acacctgggg cacctctgct gctgcctact ttgtgggcta cctgaaacca accaccttta
     1561 tgctgaaata tgatgagaat ggcaccatca cagatgctgt ggactgtagc cagaacccac
     1621 tggctgaact gaaatgttct gtgaagtcct ttgagattga caagggcatc taccagacca
     1681 gcaacttcag ggtggtgcca tctggagatg tggtgaggtt tccaaacatc accaacctgt
     1741 gtccatttgg agaggtgttc aatgccacca agtttccatc tgtctatgcc tgggagagga
     1801 agaagattag caactgtgtg gctgactact ctgtgctcta caacagcacc ttcttcagca
     1861 ccttcaagtg ttatggagtg tctgccacca aactgaatga cctgtgtttc agcaatgtct
     1921 atgctgactc ctttgtggtg aagggagatg atgtgagaca gattgcccct ggacaaacag
     1981 gagtgattgc tgactacaac tacaaactgc ctgatgactt tatgggctgt gtgctggctt
     2041 ggaacaccag gaacattgat gccaccagca caggcaacta caactacaaa tacagatacc
     2101 tgagacatgg caaactgaga ccatttgaga gggacatcag caatgtgcca ttcagccctg
     2161 atggcaagcc atgtactcct cctgccctga actgttactg gccactgaat gactatggct
     2221 tctacaccac cacaggcatt ggctaccaac catacagggt ggtggtgctg tcctttgaac
     2281 tgctgaatgc ccctgccaca gtgtgtggac caaaactgag cacagacctg attaagaacc
     2341 agtgtgtgaa cttcaacttc aatggactga caggcacagg agtgctgaca ccatccagca
     2401 agaggttcca accattccaa cagtttggca gggatgtgtc tgacttcaca gactctgtga
     2461 gggacccaaa gacctctgag attctggaca tcagcccatg ttcctttgga ggagtgtctg
     2521 tgattacacc tggcaccaat gcctcctctg aggtggctgt gctctaccag gatgtgaact
     2581 gtactgatgt gagcacagcc atccatgctg accaacttac acctgcctgg aggatttaca
     2641 gcacaggcaa caatgtgttc cagacccagg ctggctgtct gattggagca gagcatgtgg
     2701 acacctccta tgagtgtgac atcccaattg gagcaggcat ctgtgcctcc taccacacag
     2761 tgtccctgct gaggagcacc agccagaaga gcattgtggc ttacacaatg agtctgggag
     2821 cagactccag cattgcctac agcaacaaca ccattgccat cccaaccaac ttcagcatca
     2881 gcatcaccac agaggtgatg cctgtgagta tggctaagac ctctgtggac tgtaatatgt
     2941 atatctgtgg agacagcaca gagtgtgcca acctgctgct ccaatatggc tccttctgta
     3001 cccaacttaa cagggctctg tctggcattg ctgctgaaca ggacaggaac accagggagg
     3061 tgtttgccca ggtgaagcag atgtataaga caccaaccct gaaatacttt ggaggcttca
     3121 acttcagcca gattctgcct gacccactga aaccaaccaa gaggtccttc attgaggacc
     3181 tgctgttcaa caaggtgacc ctggctgatg ctggctttat gaagcaatat ggagagtgtc
     3241 tgggagacat caatgccagg gacctgattt gtgcccagaa gttcaatgga ctgacagtgc
     3301 tgcctccact gctgacagat gatatgattg ctgcctacac agcagccctg gtgtctggca
     3361 cagccacagc aggctggacc tttggagcag gagcagccct ccaaatccca tttgctatgc
     3421 agatggctta caggttcaat ggcattggag tgacccagaa tgtgctctat gagaaccaga
     3481 agcagattgc caaccagttc aacaaggcta tcagccagat tcaggagtcc ctgaccacca
     3541 ccagcacagc cctgggcaaa ctccaagatg tggtgaacca gaatgcccag gctctgaaca
     3601 ccctggtgaa gcaactttcc agcaactttg gagccatctc ctctgtgctg aatgacatcc
     3661 tgagcagact ggacaaggtg gaggctgagg tccagattga cagactgatt acaggcagac
     3721 tccaatccct ccaaacctat gtgacccaac aacttatcag ggctgctgag attagggcat
     3781 ctgccaacct ggctgccacc aagatgagtg agtgtgtgct gggacaaagc aagagggtgg
     3841 acttctgtgg caagggctac cacctgatga gttttccaca ggctgcccct catggagtgg
     3901 tgttcctgca tgtgacctat gtgccaagcc aggagaggaa cttcaccaca gcccctgcca
     3961 tctgccatga gggcaaggct tactttccaa gggagggagt gtttgtgttc aatggcacct
     4021 cctggttcat cacccagagg aacttcttca gcccacagat tatcaccaca gacaacacct
     4081 ttgtgtctgg caactgtgat gtggtgattg gcatcatcaa caacacagtc tatgacccac
     4141 tccaacctga actggactcc ttcaaggagg aactggacaa atacttcaag aaccacacca
     4201 gccctgatgt ggacctggga gacatctctg gcatcaatgc ctctgtggtg aacatccaga
     4261 aggagattga cagactgaat gaggtggcta agaacctgaa tgagtccctg attgacctcc
     4321 aagaactggg caaatatgaa caatacatca agtggccatg gtatgtgtgg ctgggcttca
     4381 ttgctggact gattgccatt gtgatggtga ccatcctgct gtgttgtatg acctcctgtt
     4441 gttcctgtct gaaataactc gagccatgga agcttgatat ctaactgact gaaccggtgg
     4501 taccaaacgg gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc
     4561 tatgacggca ataaaaagac agaataaaac gcacgggtgt tgggtcgttt gttcataaac
     4621 gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc
     4681 aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag
     4741 ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc agatctgcgc agctggggct
     4801 ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     4861 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     4921 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     4981 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     5041 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     5101 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     5161 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     5221 tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa
     5281 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     5341 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     5401 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     5461 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     5521 cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     5581 ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg
     5641 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     5701 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     5761 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     5821 cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     5881 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     5941 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     6001 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     6061 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     6121 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     6181 gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     6241 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     6301 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     6361 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     6421 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgcgaa atgaccgacc
     6481 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     6541 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     6601 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa
     6661 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     6721 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct
     6781 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     6841 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     6901 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     6961 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     7021 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     7081 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     7141 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     7201 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     7261 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     7321 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     7381 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     7441 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     7501 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     7561 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     7621 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     7681 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     7741 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     7801 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     7861 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     7921 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     7981 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     8041 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     8101 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     8161 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     8221 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     8281 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     8341 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     8401 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     8461 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     8521 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     8581 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     8641 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     8701 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     8761 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     8821 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     8881 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     8941 cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtgcactc tcagtacaat
     9001 ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc
     9061 tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat
     9121 ga
//
