LOCUS       sequence-306280-        8316 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes SARS-CoV-2 Spike with 21 aa C-terminal deletions for 
            lentiviral pseudotyping.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8316)
  AUTHORS   Crawford KHD, Eguia R, Dingens AS, Loes AN, Malone KD, Wolf CR, Chu 
            HY, Tortorici MA, Veesler D, Murphy M, Pettie D, King NP, Balazs AB,
            Bloom JD
  TITLE     Protocol and Reagents for Pseudotyping Lentiviral Particles with 
            SARS-CoV-2 Spike Protein for Neutralization Assays.
  JOURNAL   Viruses. 2020 May 6;12(5). pii: v12050513. doi: 10.3390/v12050513.
  PUBMED    32384820
REFERENCE   2  (bases 1 to 8316)
  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: "Viruses."; 
            date: "2020-05-6"; volume: "12(5). pii: v12050513. doi"; pages: " 
            10.3390/v12050513"
COMMENT     Sequence Label: HDM-SARS2-Spike-delta21
FEATURES             Location/Qualifiers
     source          1..8316
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        265..644
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        645..848
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     799..819
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     845..869
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          962..1437
                     /label=beta-globin intron
                     /note="internally truncated intron from human beta-globin"
     primer_bind     complement(5269..5285)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5270..5286)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    5379..5773
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     primer_bind     complement(6107..6125)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6193..6297
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6298..6366,6367..7158)
                     /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(6516..6535)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7329..7917
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7818..7837
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcgggcatcc cgatgccgcc ggaagcgaga agaatcataa tggggaaggc catccagcct
       61 cgcgtcgggg agctttttgc aaaagcctag gcctccaaaa aagcctcctc actacttctg
      121 gaatagctca gaggccgagg cggcctcggc ctctgcataa ataaaaaaaa ttagtcagcc
      181 atgagcttgg cccattgcat acgttgtatc catatcataa tatgtacatt tatattggct
      241 catgtccaac attaccgcca tgttgacatt gattattgac tagttattaa tagtaatcaa
      301 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
      361 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
      421 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
      481 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
      541 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
      601 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
      661 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
      721 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
      781 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
      841 gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc
      901 tccatagaag acaccgggac cgatccagcc tcccctcgaa gctgatcctg agaacttcag
      961 ggtgagtcta tgggaccctt gatgttttct ttccccttct tttctatggt taagttcatg
     1021 tcataggaag gggagaagta acagggtaca catattgacc aaatcagggt aattttgcat
     1081 ttgtaatttt aaaaaatgct ttcttctttt aatatacttt tttgtttatc ttatttctaa
     1141 tactttccct aatctctttc tttcagggca ataatgatac aatgtatcat gcctctttgc
     1201 accattctaa agaataacag tgataatttc tgggttaagg caatagcaat atttctgcat
     1261 ataaatattt ctgcatataa attgtaactg atgtaagagg tttcatattg ctaatagcag
     1321 ctacaatcca gctaccattc tgcttttatt ttatggttgg gataaggctg gattattctg
     1381 agtccaagct aggccctttt gctaatcatg ttcatacctc ttatcttcct cccacagctc
     1441 ctgggcaacg tgctggtctg tgtgctggcc catcactttg gcaaagaatt ccgccaccat
     1501 gttcgtgttc ctggtactcc ttcctttggt gtcttcccag tgtgtaaatc ttaccactcg
     1561 gacccagctt ccacccgcct acaccaacag tttcacacgc ggcgtctact atcctgacaa
     1621 ggtgtttagg agttcagtct tgcactcaac tcaagacttg ttcctccctt tctttagcaa
     1681 tgtgacgtgg tttcatgcca ttcatgtctc cggcacaaac ggaacgaagc gctttgataa
     1741 tcctgtgctc ccgttcaacg atggagtgta cttcgcgtcc acagagaaga gcaatatcat
     1801 tcgaggttgg atcttcggaa cgacactcga ctcaaagacg cagtcccttc tcatcgtcaa
     1861 taatgccacg aacgtggtca tcaaagtgtg cgagtttcaa ttctgtaatg atcccttcct
     1921 gggcgtctat tatcacaaga acaacaaatc ctggatggag tccgaattta gagtctactc
     1981 cagcgccaac aactgcactt tcgaatacgt atcacagcca ttcttgatgg accttgaagg
     2041 aaagcagggt aatttcaaga acttgaggga gttcgtattc aagaatatcg acgggtactt
     2101 taagatttat agcaaacaca cacccattaa tttggtgcgg gatcttcctc agggatttag
     2161 tgctcttgag cctctcgttg acctccctat tggcattaac atcacccgct ttcaaaccct
     2221 gttggccctg catcggtcct acctgacacc gggcgactca agttccggat ggaccgcagg
     2281 tgccgccgca tactatgtgg gctaccttca gccaagaaca tttctgctga aatataatga
     2341 gaacgggacc attacagatg cggtggattg tgcactcgac cctctgtctg agacgaaatg
     2401 cacccttaag agcttcacgg tggagaaagg catttatcag acttctaact tcagagttca
     2461 acccaccgag tccattgtgc gattcccaaa tattacgaat ttgtgcccat ttggtgaggt
     2521 cttcaatgct actcgattcg cctcagttta tgcatggaac cgaaagagaa tttccaattg
     2581 tgtggcggac tactcagtat tgtataatag tgcaagcttt agcacattca aatgttacgg
     2641 cgtgtctcca acgaagctga acgatctctg tttcacaaac gtttatgcgg attccttcgt
     2701 gattcgcggc gatgaggtcc gacagattgc gcctgggcaa acgggtaaga tcgctgatta
     2761 caactataag ttgccggacg atttcacagg atgtgtcata gcttggaata gcaataattt
     2821 ggacagtaag gttggcggaa actacaatta tttgtacagg ttgtttcgca agtcaaattt
     2881 gaaaccattt gagagagata tatctacgga gatatatcaa gccggctcta caccatgtaa
     2941 tggtgtggag ggctttaact gctactttcc actccagtca tatggtttcc aacctacaaa
     3001 tggagtaggg tatcaaccgt acagagttgt ggtcttgagt ttcgaattgc tccacgctcc
     3061 agcaacggta tgcggtccta agaaatccac aaatcttgtg aagaacaagt gcgtaaattt
     3121 caacttcaat gggctgactg gaacaggcgt gctgactgag agtaacaaga agttcttgcc
     3181 tttccaacaa ttcgggcggg acatagctga taccactgac gccgtccgcg accctcagac
     3241 cctggagatt ctggacataa ctccttgttc tttcggtggc gtcagtgtta tcactcccgg
     3301 gaccaacacc tccaaccaag tcgcggtcct ctatcaagac gtcaactgta cggaagtacc
     3361 ggtagccatc catgcggacc aacttacacc gacttggagg gtttactcta caggaagcaa
     3421 tgtctttcaa acacgagccg ggtgtctgat cggagcagaa cacgttaaca acagctacga
     3481 atgtgacata ccaataggcg cagggatttg tgcttcatat cagacacaga ccaatagccc
     3541 gcggagagcg agaagcgtag caagccaaag catcatcgcg tacacgatga gcctcggagc
     3601 agagaacagc gtcgcgtata gcaataattc aatagctatc ccaacaaatt tcactatttc
     3661 ggtcaccact gagattctgc cggtctccat gaccaagaca tccgtcgatt gtactatgta
     3721 catatgcggc gacagcacgg agtgcagtaa cttgctcctt cagtacggtt ccttctgtac
     3781 gcagcttaac cgggcactga cgggtatcgc ggtagaacag gacaagaaca cacaggaggt
     3841 cttcgcgcag gtcaaacaaa tctacaagac accacccata aaggacttcg gcgggttcaa
     3901 tttcagccaa atcctgccgg acccttccaa acctagtaag aggtcattca ttgaggatct
     3961 tctgtttaac aaagttacgc ttgcggacgc gggattcatt aagcagtatg gtgactgcct
     4021 tggagatatt gccgccaggg atttgatatg tgcacagaaa tttaacggcc tcaccgttct
     4081 gccgcctctg ctcaccgatg agatgatagc gcagtacacg agcgcactcc tggcaggtac
     4141 aattacaagc ggatggacat tcggtgcagg agcagcgttg cagataccct ttgctatgca
     4201 gatggcttat cgatttaacg ggattggcgt cacgcagaac gtcctttatg agaatcagaa
     4261 attgattgca aatcagttca atagtgctat cggtaagatt caggacagct tgagcagtac
     4321 cgcgtctgca ctgggaaagt tgcaggacgt ggtgaatcag aatgcacaag cactgaatac
     4381 cttggttaag caattgagta gcaatttcgg cgccatatca agtgtactga atgatatcct
     4441 gtcacggttg gacaaggtag aagccgaagt tcagattgac cgcttgatca ccgggcgcct
     4501 ccaaagtctg cagacctacg tcacacaaca attgatcaga gcagcagaga taagagcatc
     4561 tgctaacctg gctgccacta agatgtctga atgtgtgctt gggcagtcaa agagggtaga
     4621 tttctgcgga aagggctacc accttatgtc tttccctcag agcgctccgc atggtgtggt
     4681 ctttctccat gtgacttatg tgcctgctca agagaagaac tttacgacgg cgcccgctat
     4741 atgccatgat ggtaaggcgc actttccaag ggagggagtg ttcgtgtcca acggcactca
     4801 ctggtttgtc acccaacgaa atttctacga gcctcaaatt attaccaccg acaatacctt
     4861 tgttagcggt aactgtgacg tcgtaattgg gattgttaat aatacagtct acgatcctct
     4921 gcagccggaa ctggactcct ttaaagagga gctggacaaa tatttcaaga accacacatc
     4981 tcctgacgta gatcttggag acataagcgg tataaatgca agtgttgtta acattcagaa
     5041 agaaatagat aggttgaacg aagttgcgaa gaaccttaac gagtcactga tagacctcca
     5101 agagcttggg aagtacgagc aatatatcaa gtggccttgg tatatttggc tcgggttcat
     5161 agcaggactt atcgctatag tcatggtgac tataatgctg tgctgcatga caagctgctg
     5221 cagctgtctc aaaggctgtt gctcttgcgg ctcttaatga aagcttatcg ataccgtcga
     5281 cctcgagggc ccagatctaa ttcaccccac cagtgcaggc tgcctatcag aaagtggtgg
     5341 ctggtgtggc taatgccctg gcccacaagt atcactaagc tcgctttctt gctgtccaat
     5401 ttctattaaa ggttcctttg ttccctaagt ccaactacta aactggggga tattatgaag
     5461 ggccttgagc atctggattc tgcctaataa aaaacattta ttttcattgc aatgatgtat
     5521 ttaaattatt tctgaatatt ttactaaaaa gggaatgtgg gaggtcagtg catttaaaac
     5581 ataaagaaat gaagagctag ttcaaacctt gggaaaatac actatatctt aaactccatg
     5641 aaagaaggtg aggctgcaaa cagctaatgc acattggcaa cagcccctga tgcctatgcc
     5701 ttattcatcc ctcagaaaag gattcaagta gaggcttgat ttggaggtta aagttttgct
     5761 atgctgtatt ttacattact tattgtttta gctgtcctca tgaatgtctt ttcactaccc
     5821 atttgcttat cctgcatctc tcagccttga ctccactcag ttctcttgct tagagatacc
     5881 acctttcccc tgaagtgttc cttccatgtt ttacggcgag atggtttctc ctcgcctggc
     5941 cactcagcct tagttgtctc tgttgtctta tagaggtcta cttgaagaag gaaaaacagg
     6001 gggcatggtt tgactgtcct gtgagccctt cttccctgcc tcccccactc acagtgaccc
     6061 ggaatccctc gacatggcag tctagatcat tcttgaagac gaaagggcct cgtgatacgc
     6121 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     6181 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     6241 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     6301 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     6361 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     6421 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     6481 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     6541 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     6601 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     6661 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     6721 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     6781 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     6841 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     6901 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     6961 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     7021 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     7081 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     7141 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     7201 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     7261 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     7321 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     7381 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     7441 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     7501 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     7561 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     7621 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     7681 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     7741 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     7801 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     7861 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     7921 gccagcaacg gagatgcgcc gcgtgcggct gctggagatg gcggacgcga tggatatgtt
     7981 ctgccaaggg ttggtttgcg cattcacagt tctccgcaag aattgattgg ctccaattct
     8041 tggagtggtg aatccgttag cgaggtgccg ccggcttcca ttcaggtcga ggtggcccgg
     8101 ctccatgcac cgcgacgcaa cgcggggagg cagacaaggt atagggcggc gcctacaatc
     8161 catgccaacc cgttccatgt gctcgccgag gcggcataaa tcgccgtgac gatcagcggt
     8221 ccaatgatcg aagttaggct ggtaagagcc gcgagcgatc cttgaagctg tccctgatgg
     8281 tcgtcatcta cctgcctgga cagcatggcc tgcaac
//
