LOCUS       sequence-313164-        9202 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express SARS-CoV-2 spike protein with C9 tag at C-terminal in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9202)
  AUTHORS   Shang J, Ye G, Shi K, Wan Y, Luo C, Aihara H, Geng Q, Auerbach A, Li
            F
  TITLE     Structural basis of receptor recognition by SARS-CoV-2.
  JOURNAL   Nature. 2020 Mar 30. pii: 10.1038/s41586-020-2179-y. doi: 
            10.1038/s41586-020-2179-y.
  PUBMED    32225175
REFERENCE   2  (bases 1 to 9202)
  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: "Nature. 
            2020 Mar 30. pii: 10.1038/s41586-020-2179-y. doi: 
            10.1038/s41586-020-2179-y."
COMMENT     Sequence Label: pcDNA3.1-SARS2-Spike
FEATURES             Location/Qualifiers
     source          1..9202
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        167..546
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        547..750
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     701..721
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     795..814
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        795..813
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4725..4742)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4731..4955
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5001..5429
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5088..5107)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5298..5319
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5438..5458)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5443..5772
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5623..5758
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5685..5704
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5839..6633
                     /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(5893..5912)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6503..6522
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6807..6928
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6844..6863)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6898..6917
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6977..6993)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6977..6993)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6990..7012)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7001..7017
                     /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(join(7025..7031,7032..7049,7050..7055))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7070..7091
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7208..7225)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7379..7967)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7459..7478)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8138..8929,8930..8998))
                     /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     8761..8780
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8999..9103)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(9178..9197)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa
       61 atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagggtt
      121 aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca ttgattattg
      181 actagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
      241 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
      301 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
      361 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
      421 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
      481 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
      541 accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg
      601 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
      661 cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt
      721 gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc cactgcttac
      781 tggcttatcg aaattaatac gactcactat agggagaccc aagctggcta gcatgtttgt
      841 cttcctggtc ctgctgcctc tggtctcgtc tcagtgcgtg aacctgacta ctagaaccca
      901 gctgcctcct gcctatacta actccttcac ccgcggcgtg tactacccag acaaggtgtt
      961 ccgcagctcc gtgctgcact ccacccagga tctgttcctg cccttcttca gcaacgtgac
     1021 ctggttccac gccatccacg tgagcggcac caatggcacc aagcggttcg acaatcccgt
     1081 gctgccattc aacgatggcg tgtacttcgc ctccaccgag aagagcaaca tcatccgcgg
     1141 ctggatcttc ggcaccaccc tggactccaa gacccagagc ctgctgatcg tgaacaatgc
     1201 caccaacgtg gtcatcaagg tgtgcgagtt ccagttctgc aatgatccat tcctgggcgt
     1261 gtactaccac aagaacaata agtcctggat ggagagcgag ttccgcgtgt acagctccgc
     1321 caacaattgc accttcgagt acgtgtccca gcccttcctg atggacctgg agggcaagca
     1381 gggcaatttc aagaacctgc gcgagttcgt gttcaagaat atcgatggct acttcaagat
     1441 ctactccaag cacaccccca tcaacctggt gcgcgacctg ccacagggct tcagcgccct
     1501 ggagccactg gtggatctgc caatcggcat caacatcacc aggttccaga ccctgctggc
     1561 cctgcaccgc agctacctga ccccaggcga cagctccagc ggatggaccg ctggagctgc
     1621 tgcctactac gtgggctacc tgcagccccg caccttcctg ctgaagtaca acgagaatgg
     1681 caccatcacc gacgccgtgg attgcgccct ggatccactg tccgagacaa agtgcaccct
     1741 gaagagcttc accgtggaga agggcatcta ccagacctcc aatttccgcg tgcagccaac
     1801 cgagagcatc gtgcgcttcc ccaatatcac caacctgtgc ccattcggcg aggtgttcaa
     1861 cgctaccagg ttcgccagcg tgtacgcttg gaatcgcaag cgcatctcca actgcgtggc
     1921 cgactacagc gtgctgtaca actccgccag cttctccacc ttcaagtgct acggcgtgtc
     1981 ccccaccaag ctgaatgatc tgtgcttcac caacgtgtac gccgatagct tcgtgatcag
     2041 gggcgacgag gtgcgccaga tcgctccagg acagaccggc aagatcgctg actacaatta
     2101 caagctgccc gacgatttca ccggctgcgt gatcgcctgg aactccaaca atctggatag
     2161 caaagtgggc ggcaactaca attacctgta ccgcctgttc cgcaagtcca atctgaagcc
     2221 attcgagcgc gacatctcca ccgagatcta ccaggctgga agcaccccat gcaatggagt
     2281 ggagggcttc aactgctact tccccctgca gagctacggc ttccagccaa ccaacggagt
     2341 gggataccag ccatacaggg tggtggtgct gtccttcgag ctgctgcacg ctccagctac
     2401 cgtgtgcgga ccaaagaaga gcaccaatct ggtgaagaac aagtgcgtga acttcaattt
     2461 caacggcctg accggaaccg gcgtgctgac cgagtccaac aagaagttcc tgccattcca
     2521 gcagttcgga agggacatcg ctgataccac cgacgccgtg cgcgacccac agaccctgga
     2581 gatcctggat atcaccccat gctccttcgg cggcgtgagc gtgatcaccc caggaaccaa
     2641 taccagcaac caggtggccg tgctgtacca ggacgtgaat tgcaccgagg tgccagtggc
     2701 tatccacgct gatcagctga ccccaacctg gcgcgtgtac agcaccggat ccaacgtgtt
     2761 ccagacccgc gccggatgcc tgatcggagc tgagcacgtg aacaattcct acgagtgcga
     2821 catcccaatc ggagctggaa tctgcgccag ctaccagacc cagaccaact ccccaaggag
     2881 ggctcgcagc gtggccagcc agtccatcat cgcctacacc atgtccctgg gcgccgagaa
     2941 tagcgtggcc tacagcaaca attccatcgc catcccaacc aacttcacca tctccgtgac
     3001 caccgagatc ctgcccgtgt ccatgaccaa gaccagcgtg gactgcacca tgtacatctg
     3061 cggcgattcc accgagtgca gcaacctgct gctgcagtac ggcagcttct gcacccagct
     3121 gaatcgcgcc ctgaccggaa tcgctgtgga gcaggataag aacacccagg aggtgttcgc
     3181 ccaggtgaag cagatctaca agaccccccc aatcaaggac ttcggcggct tcaatttcag
     3241 ccagatcctg cccgatccaa gcaagccctc caagcgcagc ttcatcgagg acctgctgtt
     3301 caacaaggtg accctggccg atgccggctt catcaagcag tacggcgatt gcctgggcga
     3361 catcgctgcc cgcgacctga tctgcgccca gaagttcaat ggcctgaccg tgctgccacc
     3421 actgctgacc gatgagatga tcgctcagta cacctccgcc ctgctggccg gaaccatcac
     3481 cagcggatgg accttcggcg ctggagccgc cctgcagatc cccttcgcca tgcagatggc
     3541 ctaccgcttc aacggcatcg gcgtgaccca gaatgtgctg tacgagaacc agaagctgat
     3601 cgccaatcag ttcaactccg ccatcggcaa gatccaggac tccctgtcca gcaccgccag
     3661 cgccctgggc aagctgcagg atgtggtgaa tcagaacgcc caggccctga ataccctggt
     3721 gaagcagctg tccagcaact tcggcgccat ctccagcgtg ctgaatgata tcctgagccg
     3781 cctggacaag gtggaggctg aggtgcagat cgataggctg atcaccggcc gcctgcagtc
     3841 cctgcagacc tacgtgaccc agcagctgat cagggctgct gagatcaggg ccagcgccaa
     3901 tctggctgct accaagatgt ccgagtgcgt gctgggacag agcaagaggg tggacttctg
     3961 cggcaagggc taccacctga tgtccttccc acagagcgcc ccacacggag tggtgttcct
     4021 gcacgtgacc tacgtgccag cccaggagaa gaacttcacc accgctccag ctatctgcca
     4081 cgatggcaag gctcacttcc cacgcgaggg cgtgttcgtg tccaacggca cccactggtt
     4141 cgtgacccag cgcaatttct acgagcccca gatcatcacc accgacaata ccttcgtgag
     4201 cggcaactgc gacgtggtca tcggaatcgt gaacaatacc gtgtacgatc ccctgcagcc
     4261 agagctggac tccttcaagg aggagctgga taagtacttc aagaatcaca ccagccccga
     4321 cgtggatctg ggcgacatct ccggcatcaa tgccagcgtg gtgaacatcc agaaggagat
     4381 cgaccgcctg aacgaggtgg ccaagaatct gaacgagtcc ctgatcgatc tgcaggagct
     4441 gggcaagtac gagcagtaca tcaagtggcc atggtacatc tggctgggct tcatcgccgg
     4501 cctgatcgcc atcgtgatgg tgaccatcat gctgtgctgc atgacctcct gctgcagctg
     4561 cctgaagggc tgctgctcct gcggcagctg ctgcaagttc gatgaggacg atagcgagcc
     4621 cgtgctgaag ggcgtcaaac tgcactatac aggctccacc gagacatccc aggtcgctcc
     4681 cgcttagctc gagtctagag ggcccgttta aacccgctga tcagcctcga ctgtgccttc
     4741 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     4801 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     4861 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     4921 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
     4981 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     5041 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     5101 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     5161 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     5221 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     5281 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     5341 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     5401 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
     5461 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     5521 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     5581 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     5641 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     5701 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     5761 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca
     5821 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     5881 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     5941 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     6001 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc
     6061 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     6121 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     6181 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     6241 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     6301 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     6361 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     6421 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     6481 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     6541 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     6601 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     6661 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     6721 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     6781 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
     6841 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     6901 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
     6961 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     7021 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     7081 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     7141 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     7201 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     7261 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     7321 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     7381 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     7441 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     7501 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     7561 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     7621 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     7681 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     7741 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     7801 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     7861 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     7921 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     7981 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     8041 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     8101 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     8161 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     8221 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     8281 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     8341 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     8401 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     8461 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     8521 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     8581 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     8641 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     8701 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     8761 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     8821 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     8881 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     8941 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     9001 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     9061 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     9121 tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtgca ctctcagtac
     9181 aatctgctct gatgccgcat ag
//
