LOCUS       Exported                9206 bp ds-DNA     circular SYN 19-JUL-2021
DEFINITION  Express SARS-CoV-2 WT spike protein with an 18aa deletion on the 
            C-terminal tail. Used for SARS2 pseudotyped virus production..
ACCESSION   .
VERSION     .
KEYWORDS    pcDNA3.3_CoV2_D18
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9206)
  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 9206)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 19, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9206
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        102..481
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        482..685
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     636..656
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     682..706
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             complement(2124..2141)
                     /codon_start=1
                     /product="tetracysteine peptide that binds biarsenical 
                     labeling reagents"
                     /label=tetracysteine tag
                     /translation="CCPGCC"
     primer_bind     complement(4644..4663)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4688..4736
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4938..5366
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5025..5044)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5235..5256
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5375..5395)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5380..5709
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5560..5695
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5622..5641
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5776..6570
                     /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(5830..5849)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6440..6459
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6746..6867
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6783..6802)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6837..6856
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6916..6932)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6916..6932)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6929..6951)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6940..6956
                     /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(6964..6994)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7009..7030
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7147..7164)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7318..7906)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7398..7417)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(8077..8937)
                     /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     8700..8719
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8938..9042)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(9117..9136)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gggttaggcg
       61 ttttgcgctg cttcgcgatg tacgggccag atatacgcgt tgacattgat tattgactag
      121 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
      181 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
      241 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
      301 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
      361 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
      421 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
      481 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
      541 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
      601 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
      661 gtgggaggtc tatataagca gagctcgttt agtgaaccgt cagatcgcct ggagacgcca
      721 tccacgctgt tttgacctcc atagaagaca ccgggaccga tccagcctcc ggactgagct
      781 ccttaaggtt aactctagag aattcggatc catgtttgtg ttcctggtgc tgctgccact
      841 ggtgtccagc cagtgtgtga acctgaccac caggacccaa cttcctcctg cctacaccaa
      901 ctccttcacc aggggagtct actaccctga caaggtgttc aggtcctctg tgctgcacag
      961 cacccaggac ctgttcctgc cattcttcag caatgtgacc tggttccatg ccatccatgt
     1021 gtctggcacc aatggcacca agaggtttga caaccctgtg ctgccattca atgatggagt
     1081 ctactttgcc agcacagaga agagcaacat catcaggggc tggatttttg gcaccaccct
     1141 ggacagcaag acccagtccc tgctgattgt gaacaatgcc accaatgtgg tgattaaggt
     1201 gtgtgagttc cagttctgta atgacccatt cctgggagtc tactaccaca agaacaacaa
     1261 gtcctggatg gagtctgagt tcagggtcta ctcctctgcc aacaactgta cctttgaata
     1321 tgtgagccaa ccattcctga tggacttgga gggcaagcag ggcaacttca agaacctgag
     1381 ggagtttgtg ttcaagaaca ttgatggcta cttcaagatt tacagcaaac acacaccaat
     1441 caacctggtg agggacctgc cacagggctt ctctgccttg gaaccactgg tggacctgcc
     1501 aattggcatc aacatcacca ggttccagac cctgctggct ctgcacaggt cctacctgac
     1561 acctggagac tcctcctctg gctggacagc aggagcagca gcctactatg tgggctacct
     1621 ccaaccaagg accttcctgc tgaaatacaa tgagaatggc accatcacag atgctgtgga
     1681 ctgtgccctg gacccactgt ctgagaccaa gtgtaccctg aaatccttca cagtggagaa
     1741 gggcatctac cagaccagca acttcagggt ccaaccaaca gagagcattg tgaggtttcc
     1801 aaacatcacc aacctgtgtc catttggaga ggtgttcaat gccaccaggt ttgcctctgt
     1861 ctatgcctgg aacaggaaga ggattagcaa ctgtgtggct gactactctg tgctctacaa
     1921 ctctgcctcc ttcagcacct tcaagtgtta tggagtgagc ccaaccaaac tgaatgacct
     1981 gtgtttcacc aatgtctatg ctgactcctt tgtgattagg ggagatgagg tgagacagat
     2041 tgcccctgga caaacaggca agattgctga ctacaactac aaactgcctg atgacttcac
     2101 aggctgtgtg attgcctgga acagcaacaa cctggacagc aaggtgggag gcaactacaa
     2161 ctacctctac agactgttca ggaagagcaa cctgaaacca tttgagaggg acatcagcac
     2221 agagatttac caggctggca gcacaccatg taatggagtg gagggcttca actgttactt
     2281 tccactccaa tcctatggct tccaaccaac caatggagtg ggctaccaac catacagggt
     2341 ggtggtgctg tcctttgaac tgctccatgc ccctgccaca gtgtgtggac caaagaagag
     2401 caccaacctg gtgaagaaca agtgtgtgaa cttcaacttc aatggactga caggcacagg
     2461 agtgctgaca gagagcaaca agaagttcct gccattccaa cagtttggca gggacattgc
     2521 tgacaccaca gatgctgtga gggacccaca gaccttggag attctggaca tcacaccatg
     2581 ttcctttgga ggagtgtctg tgattacacc tggcaccaac accagcaacc aggtggctgt
     2641 gctctaccag gatgtgaact gtactgaggt gcctgtggct atccatgctg accaacttac
     2701 accaacctgg agggtctaca gcacaggcag caatgtgttc cagaccaggg ctggctgtct
     2761 gattggagca gagcatgtga acaactccta tgagtgtgac atcccaattg gagcaggcat
     2821 ctgtgcctcc taccagaccc agaccaacag cccaaggagg gcaaggtctg tggcaagcca
     2881 gagcatcatt gcctacacaa tgagtctggg agcagagaac tctgtggctt acagcaacaa
     2941 cagcattgcc atcccaacca acttcaccat ctctgtgacc acagagattc tgcctgtgag
     3001 tatgaccaag acctctgtgg actgtacaat gtatatctgt ggagacagca cagagtgtag
     3061 caacctgctg ctccaatatg gctccttctg tacccaactt aacagggctc tgacaggcat
     3121 tgctgtggaa caggacaaga acacccagga ggtgtttgcc caggtgaagc agatttacaa
     3181 gacacctcca atcaaggact ttggaggctt caacttcagc cagattctgc ctgacccaag
     3241 caagccaagc aagaggtcct tcattgagga cctgctgttc aacaaggtga ccctggctga
     3301 tgctggcttc atcaagcaat atggagactg tctgggagac attgctgcca gggacctgat
     3361 ttgtgcccag aagttcaatg gactgacagt gctgcctcca ctgctgacag atgagatgat
     3421 tgcccaatac acctctgccc tgctggctgg caccatcacc tctggctgga cctttggagc
     3481 aggagcagcc ctccaaatcc catttgctat gcagatggct tacaggttca atggcattgg
     3541 agtgacccag aatgtgctct atgagaacca gaaactgatt gccaaccagt tcaactctgc
     3601 cattggcaag attcaggact ccctgtccag cacagcctct gccctgggca aactccaaga
     3661 tgtggtgaac cagaatgccc aggctctgaa caccctggtg aagcaacttt ccagcaactt
     3721 tggagccatc tcctctgtgc tgaatgacat cctgagcaga ctggacaagg tggaggctga
     3781 ggtccagatt gacagactga ttacaggcag actccaatcc ctccaaacct atgtgaccca
     3841 acaacttatc agggctgctg agattagggc atctgccaac ctggctgcca ccaagatgag
     3901 tgagtgtgtg ctgggacaaa gcaagagggt ggacttctgt ggcaagggct accacctgat
     3961 gagttttcca cagtctgccc ctcatggagt ggtgttcctg catgtgacct atgtgcctgc
     4021 ccaggagaag aacttcacca cagcccctgc catctgccat gatggcaagg ctcactttcc
     4081 aagggaggga gtgtttgtga gcaatggcac ccactggttt gtgacccaga ggaacttcta
     4141 tgaaccacag attatcacca cagacaacac ctttgtgtct ggcaactgtg atgtggtgat
     4201 tggcattgtg aacaacacag tctatgaccc actccaacct gaactggact ccttcaagga
     4261 ggaactggac aaatacttca agaaccacac cagccctgat gtggacctgg gagacatctc
     4321 tggcatcaat gcctctgtgg tgaacatcca gaaggagatt gacagactga atgaggtggc
     4381 taagaacctg aatgagtccc tgattgacct ccaagaactg ggcaaatatg aacaatacat
     4441 caagtggcca tggtacatct ggctgggctt cattgctgga ctgattgcca ttgtgatggt
     4501 gaccataatg ctgtgttgta tgacctcctg ttgttcctgt ctgaaaggct gttgttcctg
     4561 tggctcctgt tgtaagtgac tcgagccatg gaagcttgat atctaactga ctgaaccggt
     4621 ggtaccaaac gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc
     4681 gctatgacgg caataaaaag acagaataaa acgcacgggt gttgggtcgt ttgttcataa
     4741 acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg
     4801 ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc
     4861 agggctcgca gccaacgtcg gggcggcagg ccctgccata gcagatctgc gcagctgggg
     4921 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     4981 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     5041 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     5101 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     5161 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     5221 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     5281 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     5341 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga
     5401 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     5461 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     5521 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     5581 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     5641 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     5701 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca agagacagga
     5761 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     5821 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     5881 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     5941 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     6001 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     6061 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     6121 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     6181 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     6241 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     6301 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     6361 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     6421 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     6481 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     6541 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgcg aaatgaccga
     6601 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     6661 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     6721 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     6781 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     6841 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     6901 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     6961 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     7021 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     7081 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     7141 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     7201 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     7261 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     7321 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     7381 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     7441 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     7501 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     7561 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     7621 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     7681 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     7741 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     7801 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     7861 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     7921 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     7981 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     8041 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     8101 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     8161 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     8221 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     8281 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     8341 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     8401 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     8461 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     8521 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     8581 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     8641 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     8701 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     8761 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     8821 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     8881 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     8941 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     9001 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     9061 gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca
     9121 atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc
     9181 gctgagtagt gcgcgagcaa aattta
//
