LOCUS       sequence-336409-        7160 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 7160)
  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 7160)
  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..7160
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     565..589
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          704..813
                     /label=CMV intron
                     /note="modified intron A from human cytomegalovirus (CMV)"
     primer_bind     759..778
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        759..777
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      814..823
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(2354..2371)
                     /codon_start=1
                     /product="tetracysteine peptide that binds biarsenical 
                     labeling reagents"
                     /label=tetracysteine tag
                     /translation="CCPGCC"
     CDS             3799..3825
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    3948..4069
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3985..4004)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4039..4058
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4076..4531)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4213..4234)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4425..4444
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4558..4662
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4664..5021
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4664..4855
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4687..4707)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4872..5007
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5056..5850
                     /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             5056..5850
                     /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(5110..5129)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5720..5739
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6038..6057)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6082..6129
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6458..7046
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6947..6966
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctcg tttagtgaac cgtcagatcg cctggagacg
      601 ccatccacgc tgttttgacc tccatagaag acaccgggac cgatccagcc tccgcggccg
      661 ggaacggtgc attggaacgc ggattccccg tgccaagagt gacgtaagta ccgcctatag
      721 actctatagg cacacccctt tggctcttat gcatgaatta atacgactca ctatagggag
      781 acagactgtt cctttcctgg gtcttttctg caggccacca tggaaaccga tacactgctg
      841 ctgtgggtgc tgttgctctg ggttccaggc tctaccggcg tgaccggcta ccggctgttc
      901 gaggagattc tcggaagttc cggtggagga gggagcggag gtggaggatc gtctggagga
      961 ggaggtggag ggtctagcgg tgtgaccggc taccggctgt tcgaggagat tctcggaagt
     1021 tccggtggag gagggagcgg aggtggagga tcgtctggag gaggaggtgg agggtctagc
     1081 cagtgtgtga acctgaccac caggacccaa cttcctcctg cctacaccaa ctccttcacc
     1141 aggggagtct actaccctga caaggtgttc aggtcctctg tgctgcacag cacccaggac
     1201 ctgttcctgc cattcttcag caatgtgacc tggttccatg ccatccatgt gtctggcacc
     1261 aatggcacca agaggtttga caaccctgtg ctgccattca atgatggagt ctactttgcc
     1321 agcacagaga agagcaacat catcaggggc tggatttttg gcaccaccct ggacagcaag
     1381 acccagtccc tgctgattgt gaacaatgcc accaatgtgg tgattaaggt gtgtgagttc
     1441 cagttctgta atgacccatt cctgggagtc tactaccaca agaacaacaa gtcctggatg
     1501 gagtctgagt tcagggtcta ctcctctgcc aacaactgta cctttgaata tgtgagccaa
     1561 ccattcctga tggacttgga gggcaagcag ggcaacttca agaacctgag ggagtttgtg
     1621 ttcaagaaca ttgatggcta cttcaagatt tacagcaaac acacaccaat caacctggtg
     1681 agggacctgc cacagggctt ctctgccttg gaaccactgg tggacctgcc aattggcatc
     1741 aacatcacca ggttccagac cctgctggct ctgcacaggt cctacctgac acctggagac
     1801 tcctcctctg gctggacagc aggagcagca gcctactatg tgggctacct ccaaccaagg
     1861 accttcctgc tgaaatacaa tgagaatggc accatcacag atgctgtgga ctgtgccctg
     1921 gacccactgt ctgagaccaa gtgtaccctg aaatccttca cagtggagaa gggcatctac
     1981 cagaccagca acttcagggt ccaaccaaca gagagcattg tgaggtttcc aaacatcacc
     2041 aacctgtgtc catttggaga ggtgttcaat gccaccaggt ttgcctctgt ctatgcctgg
     2101 aacaggaaga ggattagcaa ctgtgtggct gactactctg tgctctacaa ctctgcctcc
     2161 ttcagcacct tcaagtgtta tggagtgagc ccaaccaaac tgaatgacct gtgtttcacc
     2221 aatgtctatg ctgactcctt tgtgattagg ggagatgagg tgagacagat tgcccctgga
     2281 caaacaggca agattgctga ctacaactac aaactgcctg atgacttcac aggctgtgtg
     2341 attgcctgga acagcaacaa cctggacagc aaggtgggag gcaactacaa ctacctctac
     2401 agactgttca ggaagagcaa cctgaaacca tttgagaggg acatcagcac agagatttac
     2461 caggctggca gcacaccatg taatggagtg gagggcttca actgttactt tccactccaa
     2521 tcctatggct tccaaccaac caatggagtg ggctaccaac catacagggt ggtggtgctg
     2581 tcctttgaac tgctccatgc ccctgccaca gtgtgtggac caaagaagag caccaacctg
     2641 gtgaagaaca agtgtgtgaa cttcaacttc aatggactga caggcacagg agtgctgaca
     2701 gagagcaaca agaagttcct gccattccaa cagtttggca gggacattgc tgacaccaca
     2761 gatgctgtga gggacccaca gaccttggag attctggaca tcacaccatg ttcctttgga
     2821 ggagtgtctg tgattacacc tggcaccaac accagcaacc aggtggctgt gctctaccag
     2881 gatgtgaact gtactgaggt gcctgtggct atccatgctg accaacttac accaacctgg
     2941 agggtctaca gcacaggcag caatgtgttc cagaccaggg ctggctgtct gattggagca
     3001 gagcatgtga acaactccta tgagtgtgac atcccaattg gagcaggcat ctgtgcctcc
     3061 taccagaccc agaccaacag cccaggcagc gccagcccca agagctgcga caagacccac
     3121 acctgtcctc catgtcctgc tccagaactg ctcggcggac cttccgtgtt cctgtttcct
     3181 ccaaagccta aggacaccct gatgatcagc agaacccctg aagtgacctg cgtggtggtg
     3241 gatgtgtccc acgaggatcc cgaagtgaag ttcaattggt acgtggacgg cgtggaagtg
     3301 cacaacgcca agaccaagcc tagagaggaa cagtacaaca gcacctacag agtggtgtcc
     3361 gtgctgaccg tgctgcacca ggattggctg aacggcaaag agtacaagtg caaggtgtcc
     3421 aacaaggccc tgcctgctcc tatcgagaaa accatcagca aggccaaggg ccagcctagg
     3481 gaaccccagg tttacacact gcctccaagc agggacgagc tgaccaagaa tcaggtgtcc
     3541 ctgacctgcc tggtcaaggg cttctaccct tccgatatcg ccgtggaatg ggagagcaat
     3601 ggccagcctg agaacaacta caagacaacc cctcctgtgc tggacagcga cggctcattc
     3661 ttcctgtaca gcaagctgac agtggacaag agcagatggc agcagggcaa cgtgttcagc
     3721 tgcagcgtga tgcacgaggc cctgcacaac cactacaccc agaagtccct gagcctgtct
     3781 cctggcaaat gaggatccta cccatacgat gttccggatt acgcttaagg ccgcgactct
     3841 agatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
     3901 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
     3961 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     4021 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt
     4081 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     4141 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     4201 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     4261 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     4321 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     4381 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     4441 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     4501 gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc
     4561 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     4621 taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc
     4681 agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     4741 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     4801 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     4861 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     4921 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     4981 agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga
     5041 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     5101 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     5161 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     5221 gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     5281 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     5341 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     5401 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     5461 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     5521 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     5581 gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     5641 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     5701 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     5761 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     5821 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     5881 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     5941 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     6001 tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac
     6061 cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt
     6121 cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga
     6181 gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt
     6241 cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag
     6301 gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     6361 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     6421 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     6481 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     6541 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     6601 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     6661 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     6721 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     6781 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     6841 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     6901 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     6961 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     7021 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     7081 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     7141 accgtattac cgccatgcat
//
