LOCUS       sequence-309495-        8316 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector for expressing SARS-2 Spike with 21 aa 
            del at C-terminus and pt mutation D to G at site 614.
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_del21_D614G
FEATURES             Location/Qualifiers
     source          1..8316
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        82..461
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        462..665
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     616..636
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     662..686
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          779..1254
                     /label=beta-globin intron
                     /note="internally truncated intron from human beta-globin"
     primer_bind     complement(5086..5102)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5087..5103)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    5196..5590
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     primer_bind     complement(5924..5942)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6010..6114
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6115..6183,6184..6975)
                     /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(6333..6352)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7146..7734
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7635..7654
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agcttggccc attgcatacg ttgtatccat atcataatat gtacatttat attggctcat
       61 gtccaacatt accgccatgt tgacattgat tattgactag ttattaatag taatcaatta
      121 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      181 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      241 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      301 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      361 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      421 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      481 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      541 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      601 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      661 gagctcgttt agtgaaccgt cagatcgcct ggagacgcca tccacgctgt tttgacctcc
      721 atagaagaca ccgggaccga tccagcctcc cctcgaagct gatcctgaga acttcagggt
      781 gagtctatgg gacccttgat gttttctttc cccttctttt ctatggttaa gttcatgtca
      841 taggaagggg agaagtaaca gggtacacat attgaccaaa tcagggtaat tttgcatttg
      901 taattttaaa aaatgctttc ttcttttaat atactttttt gtttatctta tttctaatac
      961 tttccctaat ctctttcttt cagggcaata atgatacaat gtatcatgcc tctttgcacc
     1021 attctaaaga ataacagtga taatttctgg gttaaggcaa tagcaatatt tctgcatata
     1081 aatatttctg catataaatt gtaactgatg taagaggttt catattgcta atagcagcta
     1141 caatccagct accattctgc ttttatttta tggttgggat aaggctggat tattctgagt
     1201 ccaagctagg cccttttgct aatcatgttc atacctctta tcttcctccc acagctcctg
     1261 ggcaacgtgc tggtctgtgt gctggcccat cactttggca aagaattccg ccaccatgtt
     1321 cgtgttcctg gtactccttc ctttggtgtc ttcccagtgt gtaaatctta ccactcggac
     1381 ccagcttcca cccgcctaca ccaacagttt cacacgcggc gtctactatc ctgacaaggt
     1441 gtttaggagt tcagtcttgc actcaactca agacttgttc ctccctttct ttagcaatgt
     1501 gacgtggttt catgccattc atgtctccgg cacaaacgga acgaagcgct ttgataatcc
     1561 tgtgctcccg ttcaacgatg gagtgtactt cgcgtccaca gagaagagca atatcattcg
     1621 aggttggatc ttcggaacga cactcgactc aaagacgcag tcccttctca tcgtcaataa
     1681 tgccacgaac gtggtcatca aagtgtgcga gtttcaattc tgtaatgatc ccttcctggg
     1741 cgtctattat cacaagaaca acaaatcctg gatggagtcc gaatttagag tctactccag
     1801 cgccaacaac tgcactttcg aatacgtatc acagccattc ttgatggacc ttgaaggaaa
     1861 gcagggtaat ttcaagaact tgagggagtt cgtattcaag aatatcgacg ggtactttaa
     1921 gatttatagc aaacacacac ccattaattt ggtgcgggat cttcctcagg gatttagtgc
     1981 tcttgagcct ctcgttgacc tccctattgg cattaacatc acccgctttc aaaccctgtt
     2041 ggccctgcat cggtcctacc tgacaccggg cgactcaagt tccggatgga ccgcaggtgc
     2101 cgccgcatac tatgtgggct accttcagcc aagaacattt ctgctgaaat ataatgagaa
     2161 cgggaccatt acagatgcgg tggattgtgc actcgaccct ctgtctgaga cgaaatgcac
     2221 ccttaagagc ttcacggtgg agaaaggcat ttatcagact tctaacttca gagttcaacc
     2281 caccgagtcc attgtgcgat tcccaaatat tacgaatttg tgcccatttg gtgaggtctt
     2341 caatgctact cgattcgcct cagtttatgc atggaaccga aagagaattt ccaattgtgt
     2401 ggcggactac tcagtattgt ataatagtgc aagctttagc acattcaaat gttacggcgt
     2461 gtctccaacg aagctgaacg atctctgttt cacaaacgtt tatgcggatt ccttcgtgat
     2521 tcgcggcgat gaggtccgac agattgcgcc tgggcaaacg ggtaagatcg ctgattacaa
     2581 ctataagttg ccggacgatt tcacaggatg tgtcatagct tggaatagca ataatttgga
     2641 cagtaaggtt ggcggaaact acaattattt gtacaggttg tttcgcaagt caaatttgaa
     2701 accatttgag agagatatat ctacggagat atatcaagcc ggctctacac catgtaatgg
     2761 tgtggagggc tttaactgct actttccact ccagtcatat ggtttccaac ctacaaatgg
     2821 agtagggtat caaccgtaca gagttgtggt cttgagtttc gaattgctcc acgctccagc
     2881 aacggtatgc ggtcctaaga aatccacaaa tcttgtgaag aacaagtgcg taaatttcaa
     2941 cttcaatggg ctgactggaa caggcgtgct gactgagagt aacaagaagt tcttgccttt
     3001 ccaacaattc gggcgggaca tagctgatac cactgacgcc gtccgcgacc ctcagaccct
     3061 ggagattctg gacataactc cttgttcttt cggtggcgtc agtgttatca ctcccgggac
     3121 caacacctcc aaccaagtcg cggtcctcta tcaaggcgtc aactgtacgg aagtaccggt
     3181 agccatccat gcggaccaac ttacaccgac ttggagggtt tactctacag gaagcaatgt
     3241 ctttcaaaca cgagccgggt gtctgatcgg agcagaacac gttaacaaca gctacgaatg
     3301 tgacatacca ataggcgcag ggatttgtgc ttcatatcag acacagacca atagcccgcg
     3361 gagagcgaga agcgtagcaa gccaaagcat catcgcgtac acgatgagcc tcggagcaga
     3421 gaacagcgtc gcgtatagca ataattcaat agctatccca acaaatttca ctatttcggt
     3481 caccactgag attctgccgg tctccatgac caagacatcc gtcgattgta ctatgtacat
     3541 atgcggcgac agcacggagt gcagtaactt gctccttcag tacggttcct tctgtacgca
     3601 gcttaaccgg gcactgacgg gtatcgcggt agaacaggac aagaacacac aggaggtctt
     3661 cgcgcaggtc aaacaaatct acaagacacc acccataaag gacttcggcg ggttcaattt
     3721 cagccaaatc ctgccggacc cttccaaacc tagtaagagg tcattcattg aggatcttct
     3781 gtttaacaaa gttacgcttg cggacgcggg attcattaag cagtatggtg actgccttgg
     3841 agatattgcc gccagggatt tgatatgtgc acagaaattt aacggcctca ccgttctgcc
     3901 gcctctgctc accgatgaga tgatagcgca gtacacgagc gcactcctgg caggtacaat
     3961 tacaagcgga tggacattcg gtgcaggagc agcgttgcag ataccctttg ctatgcagat
     4021 ggcttatcga tttaacggga ttggcgtcac gcagaacgtc ctttatgaga atcagaaatt
     4081 gattgcaaat cagttcaata gtgctatcgg taagattcag gacagcttga gcagtaccgc
     4141 gtctgcactg ggaaagttgc aggacgtggt gaatcagaat gcacaagcac tgaatacctt
     4201 ggttaagcaa ttgagtagca atttcggcgc catatcaagt gtactgaatg atatcctgtc
     4261 acggttggac aaggtagaag ccgaagttca gattgaccgc ttgatcaccg ggcgcctcca
     4321 aagtctgcag acctacgtca cacaacaatt gatcagagca gcagagataa gagcatctgc
     4381 taacctggct gccactaaga tgtctgaatg tgtgcttggg cagtcaaaga gggtagattt
     4441 ctgcggaaag ggctaccacc ttatgtcttt ccctcagagc gctccgcatg gtgtggtctt
     4501 tctccatgtg acttatgtgc ctgctcaaga gaagaacttt acgacggcgc ccgctatatg
     4561 ccatgatggt aaggcgcact ttccaaggga gggagtgttc gtgtccaacg gcactcactg
     4621 gtttgtcacc caacgaaatt tctacgagcc tcaaattatt accaccgaca atacctttgt
     4681 tagcggtaac tgtgacgtcg taattgggat tgttaataat acagtctacg atcctctgca
     4741 gccggaactg gactccttta aagaggagct ggacaaatat ttcaagaacc acacatctcc
     4801 tgacgtagat cttggagaca taagcggtat aaatgcaagt gttgttaaca ttcagaaaga
     4861 aatagatagg ttgaacgaag ttgcgaagaa ccttaacgag tcactgatag acctccaaga
     4921 gcttgggaag tacgagcaat atatcaagtg gccttggtat atttggctcg ggttcatagc
     4981 aggacttatc gctatagtca tggtgactat aatgctgtgc tgcatgacaa gctgctgcag
     5041 ctgtctcaaa ggctgttgct cttgcggctc ttaatgaaag cttatcgata ccgtcgacct
     5101 cgagggccca gatctaattc accccaccag tgcaggctgc ctatcagaaa gtggtggctg
     5161 gtgtggctaa tgccctggcc cacaagtatc actaagctcg ctttcttgct gtccaatttc
     5221 tattaaaggt tcctttgttc cctaagtcca actactaaac tgggggatat tatgaagggc
     5281 cttgagcatc tggattctgc ctaataaaaa acatttattt tcattgcaat gatgtattta
     5341 aattatttct gaatatttta ctaaaaaggg aatgtgggag gtcagtgcat ttaaaacata
     5401 aagaaatgaa gagctagttc aaaccttggg aaaatacact atatcttaaa ctccatgaaa
     5461 gaaggtgagg ctgcaaacag ctaatgcaca ttggcaacag cccctgatgc ctatgcctta
     5521 ttcatccctc agaaaaggat tcaagtagag gcttgatttg gaggttaaag ttttgctatg
     5581 ctgtatttta cattacttat tgttttagct gtcctcatga atgtcttttc actacccatt
     5641 tgcttatcct gcatctctca gccttgactc cactcagttc tcttgcttag agataccacc
     5701 tttcccctga agtgttcctt ccatgtttta cggcgagatg gtttctcctc gcctggccac
     5761 tcagccttag ttgtctctgt tgtcttatag aggtctactt gaagaaggaa aaacaggggg
     5821 catggtttga ctgtcctgtg agcccttctt ccctgcctcc cccactcaca gtgacccgga
     5881 atccctcgac atggcagtct agatcattct tgaagacgaa agggcctcgt gatacgccta
     5941 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     6001 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     6061 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     6121 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     6181 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     6241 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     6301 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     6361 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     6421 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     6481 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     6541 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     6601 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     6661 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     6721 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     6781 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     6841 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     6901 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     6961 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     7021 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     7081 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     7141 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     7201 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     7261 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     7321 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     7381 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     7441 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     7501 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     7561 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     7621 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     7681 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     7741 agcaacggag atgcgccgcg tgcggctgct ggagatggcg gacgcgatgg atatgttctg
     7801 ccaagggttg gtttgcgcat tcacagttct ccgcaagaat tgattggctc caattcttgg
     7861 agtggtgaat ccgttagcga ggtgccgccg gcttccattc aggtcgaggt ggcccggctc
     7921 catgcaccgc gacgcaacgc ggggaggcag acaaggtata gggcggcgcc tacaatccat
     7981 gccaacccgt tccatgtgct cgccgaggcg gcataaatcg ccgtgacgat cagcggtcca
     8041 atgatcgaag ttaggctggt aagagccgcg agcgatcctt gaagctgtcc ctgatggtcg
     8101 tcatctacct gcctggacag catggcctgc aacgcgggca tcccgatgcc gccggaagcg
     8161 agaagaatca taatggggaa ggccatccag cctcgcgtcg gggagctttt tgcaaaagcc
     8221 taggcctcca aaaaagcctc ctcactactt ctggaatagc tcagaggccg aggcggcctc
     8281 ggcctctgca taaataaaaa aaattagtca gccatg
//
