LOCUS       sequence-336400-        7259 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  protein expression plasmid of LgBiT-hACE2-IgG1 Fc.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7259)
  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 7259)
  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: LgBiT-hACE2
FEATURES             Location/Qualifiers
     source          1..7259
                     /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             3898..3924
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    4047..4168
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4084..4103)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4138..4157
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4175..4630)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4312..4333)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4524..4543
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4657..4761
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4763..5120
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4763..4954
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4786..4806)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4971..5106
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5155..5949
                     /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             5155..5949
                     /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(5209..5228)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5819..5838
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6137..6156)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6181..6228
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6557..7145
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7046..7065
                     /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 tgttcaccct ggaggacttc
      901 gtgggcgact gggagcagac cgccgcctac aacctggacc aggtgctgga gcagggcggc
      961 gtgagcagcc tgctgcagaa cctggccgtg agcgtgaccc ccatccagcg catcgtgcgc
     1021 agcggcgaga acgccctgaa gatcgacatc cacgtgatca tcccctacga gggcctgagc
     1081 gccgaccaga tggcccagat cgaggaggtg ttcaaggtgg tgtaccccgt ggacgaccac
     1141 cacttcaagg tgatcctgcc ctacggcacc ctggtgatcg acggcgtgac ccccaacatg
     1201 ctgaactact tcggccgccc ctacgagggc atcgccgtgt tcgacggcaa gaagatcacc
     1261 gtgaccggca ccctgtggaa cggcaacaag atcatcgacg agcgcctgat cacccccgac
     1321 ggcagcatgc tgttccgcgt gaccatcaac agcggcagca gcggtggcgg tggctctggc
     1381 ggtggcggca gctctggtgg ccagtccacc attgaggaac aggccaagac atttttggac
     1441 aagtttaacc acgaagccga agacctgttc tatcaaagtt cacttgcttc ttggaattat
     1501 aacaccaata ttactgaaga gaatgtccaa aacatgaata atgctgggga caaatggtct
     1561 gcctttttaa aggaacagtc cacacttgcc caaatgtatc cactacaaga aattcagaat
     1621 ctcacagtca agcttcagct gcaggctctt cagcaaaatg ggtcttcagt gctctcagaa
     1681 gacaagagca aacggttgaa cacaattcta aatacaatga gcaccatcta cagtactgga
     1741 aaagtttgta acccagataa tccacaagaa tgcttattac ttgaaccagg tttgaatgaa
     1801 ataatggcaa acagtttaga ctacaatgag aggctctggg cttgggaaag ctggagatct
     1861 gaggtcggca agcagctgag gccattatat gaagagtatg tggtcttgaa aaatgagatg
     1921 gcaagagcaa atcattatga ggactatggg gattattgga gaggagacta tgaagtaaat
     1981 ggggtagatg gctatgacta cagccgcggc cagttgattg aagatgtgga acataccttt
     2041 gaagagatta aaccattata tgaacatctt catgcctatg tgagggcaaa gttgatgaat
     2101 gcctatcctt cctatatcag tccaattgga tgcctccctg ctcatttgct tggtgatatg
     2161 tggggtagat tttggacaaa tctgtactct ttgacagttc cctttggaca gaaaccaaac
     2221 atagatgtta ctgatgcaat ggtggaccag gcctgggatg cacagagaat attcaaggag
     2281 gccgagaagt tctttgtatc tgttggtctt cctaatatga ctcaaggatt ctgggaaaat
     2341 tccatgctaa cggacccagg aaatgttcag aaagcagtct gccatcccac agcttgggac
     2401 ctggggaagg gcgacttcag gatccttatg tgcacaaagg tgacaatgga cgacttcctg
     2461 acagctcatc atgagatggg gcatatccag tatgatatgg catatgctgc acaacctttt
     2521 ctgctaagaa atggagctaa tgaaggattc catgaagctg ttggggaaat catgtcactt
     2581 tctgcagcca cacctaagca tttaaaatcc attggtcttc tgtcacccga ttttcaagaa
     2641 gacaatgaaa cagaaataaa cttcctgctc aaacaagcac tcacgattgt tgggactctg
     2701 ccatttactt acatgttaga gaagtggagg tggatggtct ttaaagggga aattcccaaa
     2761 gaccagtgga tgaaaaagtg gtgggagatg aagcgagaga tagttggggt ggtggaacct
     2821 gtgccccatg atgaaacata ctgtgacccc gcatctctgt tccatgtttc taatgattac
     2881 tcattcattc gatattacac aaggaccctt taccaattcc agtttcaaga agcactttgt
     2941 caagcagcta aacatgaagg ccctctgcac aaatgtgaca tctcaaactc tacagaagct
     3001 ggacagaaac tgttcaatat gctgaggctt ggaaaatcag aaccctggac cctagcattg
     3061 gaaaatgttg taggagcaaa gaacatgaat gtaaggccac tgctcaacta ctttgagccc
     3121 ttatttacct ggctgaaaga ccagaacaag aattcttttg tgggatggag taccgactgg
     3181 agtccatatg cagaccccaa gagctgcgac aagacccaca cctgtcctcc atgtcctgct
     3241 ccagaactgc tcggcggacc ttccgtgttc ctgtttcctc caaagcctaa ggacaccctg
     3301 atgatcagca gaacccctga agtgacctgc gtggtggtgg atgtgtccca cgaggatccc
     3361 gaagtgaagt tcaattggta cgtggacggc gtggaagtgc acaacgccaa gaccaagcct
     3421 agagaggaac agtacaacag cacctacaga gtggtgtccg tgctgaccgt gctgcaccag
     3481 gattggctga acggcaaaga gtacaagtgc aaggtgtcca acaaggccct gcctgctcct
     3541 atcgagaaaa ccatcagcaa ggccaagggc cagcctaggg aaccccaggt ttacacactg
     3601 cctccaagca gggacgagct gaccaagaat caggtgtccc tgacctgcct ggtcaagggc
     3661 ttctaccctt ccgatatcgc cgtggaatgg gagagcaatg gccagcctga gaacaactac
     3721 aagacaaccc ctcctgtgct ggacagcgac ggctcattct tcctgtacag caagctgaca
     3781 gtggacaaga gcagatggca gcagggcaac gtgttcagct gcagcgtgat gcacgaggcc
     3841 ctgcacaacc actacaccca gaagtccctg agcctgtctc ctggcaaatg aggatcctac
     3901 ccatacgatg ttccggatta cgcttaaggc cgcgactcta gatcataatc agccatacca
     3961 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     4021 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     4081 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     4141 gtttgtccaa actcatcaat gtatcttaag gcgtaaattg taagcgttaa tattttgtta
     4201 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     4261 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     4321 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     4381 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     4441 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     4501 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     4561 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     4621 cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     4681 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     4741 taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt
     4801 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4861 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     4921 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     4981 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     5041 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     5101 gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt
     5161 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     5221 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     5281 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac
     5341 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     5401 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     5461 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     5521 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     5581 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     5641 caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag
     5701 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     5761 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     5821 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     5881 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     5941 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     6001 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     6061 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     6121 ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga
     6181 cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg
     6241 ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg
     6301 cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg
     6361 cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga
     6421 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     6481 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     6541 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     6601 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     6661 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     6721 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     6781 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     6841 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     6901 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     6961 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     7021 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     7081 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     7141 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     7201 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcat
//
