LOCUS       sequence-350069-        6848 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Furin-cleavage site deficient S.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6848)
  AUTHORS   Yu S, Zheng X, Zhou B, Li J, Chen M, Deng R, Wong G, Lavillette D, 
            Meng G
  TITLE     SARS-CoV-2 spike engagement of ACE2 primes S2' site cleavage and 
            fusion initiation.
  JOURNAL   Proc Natl Acad Sci U S A. 2022 Jan 4;119(1). pii: 2111199119. doi: 
            10.1073/pnas.2111199119.
  PUBMED    34930824
REFERENCE   2  (bases 1 to 6848)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2022-01-4"; volume: "119(1). pii: 
            2111199119. doi"; pages: " 10.1073/pnas.2111199119"
COMMENT     Sequence Label: pVAX1-SARS-CoV-2 R685A S HA-tag
FEATURES             Location/Qualifiers
     source          1..6848
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        36..415
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        416..619
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     570..590
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     664..683
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        664..682
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4549..4575
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(4672..4689)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4678..4902
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     CDS             5075..5869
                     /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             5075..5869
                     /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(5129..5148)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5739..5758
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      6195..6783
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6684..6703
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gactcttcgc gatgtacggg ccagatatac gcgttgacat tgattattga ctagttatta
       61 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
      121 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
      181 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
      241 ctatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
      301 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
      361 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
      421 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag
      481 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
      541 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga
      601 ggtctatata agcagagctc tctggctaac tagagaaccc actgcttact ggcttatcga
      661 aattaatacg actcactata gggagaccca agctggctag cgtttaaact taagcttggt
      721 accgccacca tgttcgtgtt tctggtgctg ctgcctctgg tgagctccca gtgcgtgaat
      781 ctgaccacaa ggacccagct gccccctgcc tataccaact ccttcacacg gggcgtgtac
      841 tatcccgaca aggtgttccg gagcagcgtg ctgcactcca cacaggatct gtttctgcct
      901 ttcttttcta acgtgacctg gttccacgcc atccacgtga gcggcaccaa tggcacaaag
      961 aggttcgaca acccagtgct gcccttcaat gatggcgtgt acttcgcctc caccgagaag
     1021 tctaatatca tccgcggctg gatctttggc accacactgg acagcaagac acagtccctg
     1081 ctgatcgtga acaatgccac caacgtggtc atcaaggtgt gcgagttcca gttttgtaac
     1141 gatccattcc tgggcgtgta ctatcacaag aacaataagt cttggatgga gagcgagttt
     1201 cgcgtgtatt cctctgccaa caattgcaca tttgagtacg tgtcccagcc cttcctgatg
     1261 gacctggagg gcaagcaggg caatttcaag aacctgcggg agttcgtgtt taagaacatc
     1321 gatggctact tcaaaatcta ctccaagcac accccaatca atctggtgag agacctgcca
     1381 cagggcttct ctgccctgga gccactggtg gatctgccca tcggcatcaa catcacccgg
     1441 tttcagacac tgctggccct gcacagaagc tacctgacac caggcgacag ctcctctgga
     1501 tggaccgcag gagcagcagc ctactatgtg ggctatctgc agcccaggac cttcctgctg
     1561 aagtacaacg agaatggcac catcacagac gcagtggatt gcgcactgga ccccctgtct
     1621 gagaccaagt gtacactgaa gagctttacc gtggagaagg gcatctatca gacaagcaac
     1681 ttcagggtgc agcctaccga gtccatcgtg cgctttccca atatcacaaa cctgtgccct
     1741 tttggcgagg tgttcaatgc aaccaggttc gcaagcgtgt acgcatggaa taggaagcgc
     1801 atctccaact gcgtggccga ctattctgtg ctgtacaata gcgcctcctt ctctaccttt
     1861 aagtgctacg gcgtgagccc cacaaagctg aatgacctgt gctttaccaa cgtgtacgcc
     1921 gattccttcg tgatcagggg cgacgaggtg cgccagatcg caccaggaca gacaggcaag
     1981 atcgcagact acaactataa gctgcctgac gatttcaccg gctgcgtgat cgcctggaac
     2041 agcaacaatc tggatagcaa agtgggcggc aactacaatt atctgtaccg gctgtttaga
     2101 aagtctaacc tgaagccatt cgagagggac atctccacag aaatctacca ggccggctct
     2161 accccctgca atggcgtgga gggctttaac tgttatttcc ctctgcagag ctacggcttc
     2221 cagccaacaa atggcgtggg ctatcagccc taccgcgtgg tggtgctgtc ttttgagctg
     2281 ctgcacgccc ctgcaacagt gtgcggacca aagaagagca ccaatctggt gaagaacaag
     2341 tgcgtgaact tcaacttcaa cggactgacc ggcacaggcg tgctgaccga gtccaacaag
     2401 aagttcctgc cttttcagca gttcggcagg gacatcgcag ataccacaga cgccgtgcgc
     2461 gaccctcaga ccctggagat cctggatatc acaccatgct ccttcggcgg cgtgtctgtg
     2521 atcacaccag gcaccaatac aagcaaccag gtggccgtgc tgtatcagga cgtgaactgt
     2581 accgaggtgc ccgtggcaat ccacgcagat cagctgaccc ctacatggcg ggtgtactct
     2641 accggcagca acgtgttcca gacaagagcc ggatgcctga tcggagcaga gcacgtgaac
     2701 aatagctatg agtgcgacat ccctatcggc gccggcatct gtgcctccta ccagacccag
     2761 acaaactccc caaggagagc agcgtctgtg gcaagccagt ccatcatcgc ctataccatg
     2821 agcctgggcg ccgagaactc cgtggcctac tccaacaatt ctatcgccat ccctaccaat
     2881 ttcacaatct ccgtgaccac agagatcctg ccagtgagca tgaccaagac atccgtggac
     2941 tgcacaatgt atatctgtgg cgattccacc gagtgctcta atctgctgct gcagtacggc
     3001 tctttttgta cccagctgaa cagagccctg acaggcatcg ccgtggagca ggacaagaat
     3061 acacaggagg tgttcgccca ggtgaagcaa atctacaaga ccccacccat caaggacttt
     3121 ggcggcttca actttagcca gatcctgccc gatcctagca agccatccaa gcggtctttt
     3181 atcgaggacc tgctgttcaa taaggtgacc ctggccgatg ccggcttcat caagcagtat
     3241 ggcgattgcc tgggcgacat cgccgccaga gacctgatct gtgcccagaa gtttaacggc
     3301 ctgaccgtgc tgcctccact gctgacagat gagatgatcg cccagtacac atctgccctg
     3361 ctggcaggca ccatcacaag cggatggacc ttcggcgcag gagccgccct gcagatcccc
     3421 tttgccatgc agatggccta tcggttcaat ggcatcggcg tgacccagaa tgtgctgtac
     3481 gagaaccaga agctgatcgc caatcagttt aactccgcca tcggcaagat ccaggactct
     3541 ctgagctcca cagcaagcgc cctgggcaag ctgcaggatg tggtgaatca gaacgcccag
     3601 gccctgaaca ccctggtgaa gcagctgtct agcaatttcg gcgccatctc ctctgtgctg
     3661 aacgatatcc tgagccggct ggacaaggtg gaggcagagg tgcagatcga ccggctgatc
     3721 acaggcagac tgcagtccct gcagacctac gtgacacagc agctgatcag ggcagcagag
     3781 atcagggcat ctgccaacct ggcagcaacc aagatgagcg agtgcgtgct gggccagtcc
     3841 aagagagtgg acttttgtgg caagggctat cacctgatga gcttcccaca gtccgcccct
     3901 cacggagtgg tgtttctgca cgtgacctac gtgccagccc aggagaagaa cttcaccaca
     3961 gcaccagcaa tctgccacga tggcaaggca cactttccta gggagggcgt gttcgtgagc
     4021 aatggcaccc actggtttgt gacacagcgc aacttctacg agccacagat catcaccaca
     4081 gacaatacat tcgtgtccgg caactgtgac gtggtcatcg gcatcgtgaa caataccgtg
     4141 tatgatcctc tgcagccaga gctggactct tttaaggagg agctggataa gtacttcaag
     4201 aaccacacca gccccgacgt ggatctgggc gacatctctg gcatcaatgc cagcgtggtg
     4261 aacatccaga aggagatcga caggctgaat gaggtggcca agaatctgaa cgagtccctg
     4321 atcgatctgc aggagctggg caagtatgag cagtacatca agtggccctg gtatatctgg
     4381 ctgggcttca tcgccggcct gatcgccatc gtgatggtga ccatcatgct gtgctgtatg
     4441 acaagctgct gttcctgcct gaagggctgc tgttcttgtg gcagctgctg taagtttgat
     4501 gaggacgata gcgagcctgt gctgaagggc gtgaagctgc actacaccta cccctacgac
     4561 gtgccagatt atgcctgagg atccactagt ccagtgtggt ggaattctgc agatatccag
     4621 cacagtggcg gccgctcgag tctagagggc ccgtttaaac ccgctgatca gcctcgactg
     4681 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     4741 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     4801 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     4861 aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctact gggcggtttt
     4921 atggacagca agcgaaccgg aattgccagc tggggcgccc tctggtaagg ttgggaagcc
     4981 ctgcaaagta aactggatgg ctttctcgcc gccaaggatc tgatggcgca ggggatcaag
     5041 ctctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc
     5101 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     5161 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     5221 caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg
     5281 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     5341 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     5401 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     5461 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     5521 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     5581 actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg
     5641 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     5701 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     5761 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     5821 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgaa ttattaacgc
     5881 ttacaatttc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     5941 acaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     6001 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatagcac
     6061 gtgctaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     6121 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     6181 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     6241 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     6301 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
     6361 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     6421 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     6481 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     6541 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     6601 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     6661 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     6721 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     6781 aaaaacgcca gcaacgcggc ctttttacgg ttcctgggct tttgctggcc ttttgctcac
     6841 atgttctt
//
