LOCUS       sequence-341136-        7167 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  protein expression plasmid of 2XSmBiT-SARS-CoV-2 S1-IgG1 Fc.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7167)
  AUTHORS   Huang D, Tran JT, Peng L, Yang L, Suhandynata RT, Hoffman MA, Zhao 
            F, Song G, He WT, Limbo O, Callaghan S, Landais E, Andrabi R, Sok D,
            Jardine JG, Burton DR, Voss JE, Fitzgerald RL, Nemazee D
  TITLE     A Rapid Assay for SARS-CoV-2 Neutralizing Antibodies That Is 
            Insensitive to Antiretroviral Drugs.
  JOURNAL   J Immunol. 2021 Jul 1;207(1):344-351. doi: 10.4049/jimmunol.2100155.
            Epub 2021 Jun 28.
  PUBMED    34183368
REFERENCE   2  (bases 1 to 7167)
  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"; doi: 
            "10.4049/jimmunol.2100155"; journalName: "J Immunol"; date: 
            "2021-07-1- 1"; volume: "207"; issue: "1"; pages: "344-351"
COMMENT     Sequence Label: 2XSmBiT-SARS-CoV-2 S1
FEATURES             Location/Qualifiers
     source          1..7167
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        2..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     505..529
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          644..753
                     /label=CMV intron
                     /note="modified intron A from human cytomegalovirus (CMV)"
     primer_bind     699..718
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        699..717
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      754..763
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(2303..2320)
                     /codon_start=1
                     /product="tetracysteine peptide that binds biarsenical 
                     labeling reagents"
                     /label=tetracysteine tag
                     /translation="CCPGCC"
     CDS             3747..3773
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    3896..4017
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3933..3952)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3987..4006
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4024..4479)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4161..4182)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4373..4392
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4506..4610
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4612..4969
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4612..4803
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4635..4655)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4820..4955
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5004..5798
                     /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             5004..5798
                     /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(5058..5077)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5668..5687
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5986..6005)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6030..6077
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6406..6994
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6895..6914
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
       61 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
      121 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
      181 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
      241 acatgacctt atgggacttt cctacttggc agtacatcta gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctcg tttagtgaac cgtcagatcg cctggagacg
      541 ccatccacgc tgttttgacc tccatagaag acaccgggac cgatccagcc tccgcggccg
      601 ggaacggtgc attggaacgc ggattccccg tgccaagagt gacgtaagta ccgcctatag
      661 actctatagg cacacccctt tggctcttat gcatgaatta atacgactca ctatagggag
      721 acagactgtt cctttcctgg gtcttttctg caggccacca tggaaaccga tacactgctg
      781 ctgtgggtgc tgttgctctg ggttccaggc tctaccggcg tgaccggcta ccggctgttc
      841 gaggagattc tcggaagttc cggtggagga gggagcggag gtggaggatc gtctggagga
      901 ggaggtggag ggtctagcgg tggcggaggt gtgaccggct accggctgtt cgaggagatt
      961 ctcggaagtt ccggtggagg agggagcgga ggtggaggat cgtctggagg aggaggtgga
     1021 gggtctagcc agtgtgtgaa cctgaccacc aggacccaac ttcctcctgc ctacaccaac
     1081 tccttcacca ggggagtcta ctaccctgac aaggtgttca ggtcctctgt gctgcacagc
     1141 acccaggacc tgttcctgcc attcttcagc aatgtgacct ggttccatgc catccatgtg
     1201 tctggcacca atggcaccaa gaggtttgac aaccctgtgc tgccattcaa tgatggagtc
     1261 tactttgcca gcacagagaa gagcaacatc atcaggggct ggatttttgg caccaccctg
     1321 gacagcaaga cccagtccct gctgattgtg aacaatgcca ccaatgtggt gattaaggtg
     1381 tgtgagttcc agttctgtaa tgacccattc ctgggagtct actaccacaa gaacaacaag
     1441 tcctggatgg agtctgagtt cagggtctac tcctctgcca acaactgtac ctttgaatat
     1501 gtgagccaac cattcctgat ggacttggag ggcaagcagg gcaacttcaa gaacctgagg
     1561 gagtttgtgt tcaagaacat tgatggctac ttcaagattt acagcaaaca cacaccaatc
     1621 aacctggtga gggacctgcc acagggcttc tctgccttgg aaccactggt ggacctgcca
     1681 attggcatca acatcaccag gttccagacc ctgctggctc tgcacaggtc ctacctgaca
     1741 cctggagact cctcctctgg ctggacagca ggagcagcag cctactatgt gggctacctc
     1801 caaccaagga ccttcctgct gaaatacaat gagaatggca ccatcacaga tgctgtggac
     1861 tgtgccctgg acccactgtc tgagaccaag tgtaccctga aatccttcac agtggagaag
     1921 ggcatctacc agaccagcaa cttcagggtc caaccaacag agagcattgt gaggtttcca
     1981 aacatcacca acctgtgtcc atttggagag gtgttcaatg ccaccaggtt tgcctctgtc
     2041 tatgcctgga acaggaagag gattagcaac tgtgtggctg actactctgt gctctacaac
     2101 tctgcctcct tcagcacctt caagtgttat ggagtgagcc caaccaaact gaatgacctg
     2161 tgtttcacca atgtctatgc tgactccttt gtgattaggg gagatgaggt gagacagatt
     2221 gcccctggac aaacaggcaa gattgctgac tacaactaca aactgcctga tgacttcaca
     2281 ggctgtgtga ttgcctggaa cagcaacaac ctggacagca aggtgggagg caactacaac
     2341 tacctctaca gactgttcag gaagagcaac ctgaaaccat ttgagaggga catcagcaca
     2401 gagatttacc aggctggcag cacaccatgt aatggagtgg agggcttcaa ctgttacttt
     2461 ccactccaat cctatggctt ccaaccaacc aatggagtgg gctaccaacc atacagggtg
     2521 gtggtgctgt cctttgaact gctccatgcc cctgccacag tgtgtggacc aaagaagagc
     2581 accaacctgg tgaagaacaa gtgtgtgaac ttcaacttca atggactgac aggcacagga
     2641 gtgctgacag agagcaacaa gaagttcctg ccattccaac agtttggcag ggacattgct
     2701 gacaccacag atgctgtgag ggacccacag accttggaga ttctggacat cacaccatgt
     2761 tcctttggag gagtgtctgt gattacacct ggcaccaaca ccagcaacca ggtggctgtg
     2821 ctctaccagg atgtgaactg tactgaggtg cctgtggcta tccatgctga ccaacttaca
     2881 ccaacctgga gggtctacag cacaggcagc aatgtgttcc agaccagggc tggctgtctg
     2941 attggagcag agcatgtgaa caactcctat gagtgtgaca tcccaattgg agcaggcatc
     3001 tgtgcctcct accagaccca gaccaacagc ccaggcagcg ccagccccaa gagctgcgac
     3061 aagacccaca cctgtcctcc atgtcctgct ccagaactgc tcggcggacc ttccgtgttc
     3121 ctgtttcctc caaagcctaa ggacaccctg atgatcagca gaacccctga agtgacctgc
     3181 gtggtggtgg atgtgtccca cgaggatccc gaagtgaagt tcaattggta cgtggacggc
     3241 gtggaagtgc acaacgccaa gaccaagcct agagaggaac agtacaacag cacctacaga
     3301 gtggtgtccg tgctgaccgt gctgcaccag gattggctga acggcaaaga gtacaagtgc
     3361 aaggtgtcca acaaggccct gcctgctcct atcgagaaaa ccatcagcaa ggccaagggc
     3421 cagcctaggg aaccccaggt ttacacactg cctccaagca gggacgagct gaccaagaat
     3481 caggtgtccc tgacctgcct ggtcaagggc ttctaccctt ccgatatcgc cgtggaatgg
     3541 gagagcaatg gccagcctga gaacaactac aagacaaccc ctcctgtgct ggacagcgac
     3601 ggctcattct tcctgtacag caagctgaca gtggacaaga gcagatggca gcagggcaac
     3661 gtgttcagct gcagcgtgat gcacgaggcc ctgcacaacc actacaccca gaagtccctg
     3721 agcctgtctc ctggcaaatg aggatctacc catacgatgt tccggattac gcttaaggcc
     3781 gcgactctag atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa
     3841 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
     3901 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     3961 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg
     4021 cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     4081 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     4141 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     4201 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     4261 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     4321 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     4381 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     4441 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa
     4501 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     4561 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg
     4621 aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     4681 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     4741 aggctcccca gcaggcaaaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     4801 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     4861 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     4921 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag
     4981 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     5041 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     5101 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     5161 tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga
     5221 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     5281 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     5341 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     5401 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     5461 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     5521 ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     5581 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     5641 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     5701 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     5761 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     5821 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     5881 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     5941 ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga
     6001 gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg
     6061 tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac
     6121 cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc
     6181 cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat
     6241 agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     6301 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     6361 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     6421 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     6481 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     6541 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     6601 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     6661 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     6721 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     6781 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     6841 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     6901 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     6961 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     7021 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     7081 tgtggataac cgtattaccg ccatgcatta gttattaata gtaatcaatt acggggtcat
     7141 tagttcatag cccatatatg gagttcc
//
