LOCUS       sequence-322362-        8980 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A mammalian expression vector to express codon-optimized SARS-CoV-2 
            Spike protein, with a C terminal truncation of 18 amino acids to 
            improve pseudoparticle generation..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8980)
  TITLE     SARS-CoV-2 Spike plasmid to generate pseudovirus
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8980)
  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: 
            "Unpublished"
COMMENT     Sequence Label: SARS-CoV-2 Spike-d18
FEATURES             Location/Qualifiers
     source          1..8980
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    42..62
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     promoter        63..1241
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          294..1232
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1189..1209
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    1241..1265
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(2584..2601)
                     /codon_start=1
                     /product="tetracysteine peptide that binds biarsenical 
                     labeling reagents"
                     /label=tetracysteine tag
                     /translation="CCPGCC"
     protein_bind    complement(5040..5064)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    5093..5214
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5104..5123)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5158..5177
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     enhancer        5312..5691
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5692..5895
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5846..5866
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             5928..6722
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             5928..6722
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5982..6001)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6592..6611
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6760..6777)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6766..6990
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(7015..7032)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7186..7774)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7266..7285)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7945..8736,8737..8805))
                     /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     8568..8587
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8806..8910)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     join(8978..8980,1..16)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 aggcgtatca cgaggccctt tcgtcggcgc gccgcggccg ccaactttgt atagaaaagt
       61 tgggctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg
      121 ggggaggggt cggcaattga accggtgcct agagaaggtg gcgcggggta aactgggaaa
      181 gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg
      241 cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca caggtaagtg
      301 ccgtgtgtgg ttcccgcggg cctggcctct ttacgggtta tggcccttgc gtgccttgaa
      361 ttacttccac ctggctgcag tacgtgattc ttgatcccga gcttcgggtt ggaagtgggt
      421 gggagagttc gaggccttgc gcttaaggag ccccttcgcc tcgtgcttga gttgaggcct
      481 ggcctgggcg ctggggccgc cgcgtgcgaa tctggtggca ccttcgcgcc tgtctcgctg
      541 ctttcgataa gtctctagcc atttaaaatt tttgatgacc tgctgcgacg ctttttttct
      601 ggcaagatag tcttgtaaat gcgggccaag atctgcacac tggtatttcg gtttttgggg
      661 ccgcgggcgg cgacggggcc cgtgcgtccc agcgcacatg ttcggcgagg cggggcctgc
      721 gagcgcggcc accgagaatc ggacgggggt agtctcaagc tggccggcct gctctggtgc
      781 ctggtctcgc gccgccgtgt atcgccccgc cctgggcggc aaggctggcc cggtcggcac
      841 cagttgcgtg agcggaaaga tggccgcttc ccggccctgc tgcagggagc tcaaaatgga
      901 ggacgcggcg ctcgggagag cgggcgggtg agtcacccac acaaaggaaa agggcctttc
      961 cgtcctcagc cgtcgcttca tgtgactcca cggagtaccg ggcgccgtcc aggcacctcg
     1021 attagttctc gagcttttgg agtacgtcgt ctttaggttg gggggagggg ttttatgcga
     1081 tggagtttcc ccacactgag tgggtggaga ctgaagttag gccagcttgg cacttgatgt
     1141 aattctcctt ggaatttgcc ctttttgagt ttggatcttg gttcattctc aagcctcaga
     1201 cagtggttca aagttttttt cttccatttc aggtgtcgtg acaagtttgt acaaaaaagc
     1261 aggctgccac catgtttgtg ttcctggtgc tgctgccact ggtgtccagc cagtgtgtga
     1321 acctgaccac caggacccaa cttcctcctg cctacaccaa ctccttcacc aggggagtct
     1381 actaccctga caaggtgttc aggtcctctg tgctgcacag cacccaggac ctgttcctgc
     1441 cattcttcag caatgtgacc tggttccatg ccatccatgt gtctggcacc aatggcacca
     1501 agaggtttga caaccctgtg ctgccattca atgatggagt ctactttgcc agcacagaga
     1561 agagcaacat catcaggggc tggatttttg gcaccaccct ggacagcaag acccagtccc
     1621 tgctgattgt gaacaatgcc accaatgtgg tgattaaggt gtgtgagttc cagttctgta
     1681 atgacccatt cctgggagtc tactaccaca agaacaacaa gtcctggatg gagtctgagt
     1741 tcagggtcta ctcctctgcc aacaactgta cctttgaata tgtgagccaa ccattcctga
     1801 tggacttgga gggcaagcag ggcaacttca agaacctgag ggagtttgtg ttcaagaaca
     1861 ttgatggcta cttcaagatt tacagcaaac acacaccaat caacctggtg agggacctgc
     1921 cacagggctt ctctgccttg gaaccactgg tggacctgcc aattggcatc aacatcacca
     1981 ggttccagac cctgctggct ctgcacaggt cctacctgac acctggagac tcctcctctg
     2041 gctggacagc aggagcagca gcctactatg tgggctacct ccaaccaagg accttcctgc
     2101 tgaaatacaa tgagaatggc accatcacag atgctgtgga ctgtgccctg gacccactgt
     2161 ctgagaccaa gtgtaccctg aaatccttca cagtggagaa gggcatctac cagaccagca
     2221 acttcagggt ccaaccaaca gagagcattg tgaggtttcc aaacatcacc aacctgtgtc
     2281 catttggaga ggtgttcaat gccaccaggt ttgcctctgt ctatgcctgg aacaggaaga
     2341 ggattagcaa ctgtgtggct gactactctg tgctctacaa ctctgcctcc ttcagcacct
     2401 tcaagtgtta tggagtgagc ccaaccaaac tgaatgacct gtgtttcacc aatgtctatg
     2461 ctgactcctt tgtgattagg ggagatgagg tgagacagat tgcccctgga caaacaggca
     2521 agattgctga ctacaactac aaactgcctg atgacttcac aggctgtgtg attgcctgga
     2581 acagcaacaa cctggacagc aaggtgggag gcaactacaa ctacctctac agactgttca
     2641 ggaagagcaa cctgaaacca tttgagaggg acatcagcac agagatttac caggctggca
     2701 gcacaccatg taatggagtg gagggcttca actgttactt tccactccaa tcctatggct
     2761 tccaaccaac caatggagtg ggctaccaac catacagggt ggtggtgctg tcctttgaac
     2821 tgctccatgc ccctgccaca gtgtgtggac caaagaagag caccaacctg gtgaagaaca
     2881 agtgtgtgaa cttcaacttc aatggactga caggcacagg agtgctgaca gagagcaaca
     2941 agaagttcct gccattccaa cagtttggca gggacattgc tgacaccaca gatgctgtga
     3001 gggacccaca gaccttggag attctggaca tcacaccatg ttcctttgga ggagtgtctg
     3061 tgattacacc tggcaccaac accagcaacc aggtggctgt gctctaccag gatgtgaact
     3121 gtactgaggt gcctgtggct atccatgctg accaacttac accaacctgg agggtctaca
     3181 gcacaggcag caatgtgttc cagaccaggg ctggctgtct gattggagca gagcatgtga
     3241 acaactccta tgagtgtgac atcccaattg gagcaggcat ctgtgcctcc taccagaccc
     3301 agaccaacag cccaaggagg gcaaggtctg tggcaagcca gagcatcatt gcctacacaa
     3361 tgagtctggg agcagagaac tctgtggctt acagcaacaa cagcattgcc atcccaacca
     3421 acttcaccat ctctgtgacc acagagattc tgcctgtgag tatgaccaag acctctgtgg
     3481 actgtacaat gtatatctgt ggagacagca cagagtgtag caacctgctg ctccaatatg
     3541 gctccttctg tacccaactt aacagggctc tgacaggcat tgctgtggaa caggacaaga
     3601 acacccagga ggtgtttgcc caggtgaagc agatttacaa gacacctcca atcaaggact
     3661 ttggaggctt caacttcagc cagattctgc ctgacccaag caagccaagc aagaggtcct
     3721 tcattgagga cctgctgttc aacaaggtga ccctggctga tgctggcttc atcaagcaat
     3781 atggagactg tctgggagac attgctgcca gggacctgat ttgtgcccag aagttcaatg
     3841 gactgacagt gctgcctcca ctgctgacag atgagatgat tgcccaatac acctctgccc
     3901 tgctggctgg caccatcacc tctggctgga cctttggagc aggagcagcc ctccaaatcc
     3961 catttgctat gcagatggct tacaggttca atggcattgg agtgacccag aatgtgctct
     4021 atgagaacca gaaactgatt gccaaccagt tcaactctgc cattggcaag attcaggact
     4081 ccctgtccag cacagcctct gccctgggca aactccaaga tgtggtgaac cagaatgccc
     4141 aggctctgaa caccctggtg aagcaacttt ccagcaactt tggagccatc tcctctgtgc
     4201 tgaatgacat cctgagcaga ctggacaagg tggaggctga ggtccagatt gacagactga
     4261 ttacaggcag actccaatcc ctccaaacct atgtgaccca acaacttatc agggctgctg
     4321 agattagggc atctgccaac ctggctgcca ccaagatgag tgagtgtgtg ctgggacaaa
     4381 gcaagagggt ggacttctgt ggcaagggct accacctgat gagttttcca cagtctgccc
     4441 ctcatggagt ggtgttcctg catgtgacct atgtgcctgc ccaggagaag aacttcacca
     4501 cagcccctgc catctgccat gatggcaagg ctcactttcc aagggaggga gtgtttgtga
     4561 gcaatggcac ccactggttt gtgacccaga ggaacttcta tgaaccacag attatcacca
     4621 cagacaacac ctttgtgtct ggcaactgtg atgtggtgat tggcattgtg aacaacacag
     4681 tctatgaccc actccaacct gaactggact ccttcaagga ggaactggac aaatacttca
     4741 agaaccacac cagccctgat gtggacctgg gagacatctc tggcatcaat gcctctgtgg
     4801 tgaacatcca gaaggagatt gacagactga atgaggtggc taagaacctg aatgagtccc
     4861 tgattgacct ccaagaactg ggcaaatatg aacaatacat caagtggcca tggtacatct
     4921 ggctgggctt cattgctgga ctgattgcca ttgtgatggt gaccataatg ctgtgttgta
     4981 tgacctcctg ttgttcctgt ctgaaaggct gttgttcctg tggctcctgt tgtaagtgaa
     5041 cccagctttc ttgtacaaag tggtgatggc cggccgcttc gagcagacat gataagatac
     5101 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     5161 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     5221 aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc
     5281 aagtaaaacc tctacaaatg tggtacgcgt tgacattgat tattgactag ttattaatag
     5341 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
     5401 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
     5461 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
     5521 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct
     5581 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg
     5641 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg
     5701 ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc
     5761 caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa
     5821 tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc
     5881 tatataagca gagctctctg gctaactaga gaacccactg cgccaccatg attgaacaag
     5941 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     6001 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     6061 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag
     6121 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     6181 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     6241 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     6301 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     6361 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     6421 tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg
     6481 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     6541 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     6601 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     6661 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     6721 gactcgagtc tagagggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt
     6781 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     6841 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     6901 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     6961 ggcatgctgg ggatgcggtg ggctctatgg gcggccgcgg cgctcttccg cttcctcgct
     7021 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     7081 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     7141 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     7201 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     7261 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     7321 cctgccgctt accggatacc tgtccgcctt tctctcttcg ggaagcgtgg cgctttctca
     7381 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     7441 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     7501 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     7561 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     7621 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     7681 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     7741 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     7801 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     7861 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     7921 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     7981 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     8041 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     8101 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     8161 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     8221 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     8281 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     8341 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     8401 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     8461 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     8521 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     8581 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     8641 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     8701 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     8761 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     8821 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     8881 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     8941 ctaagaaacc attattatca tgacattaac ctataaaaat
//
