LOCUS       sequence-340335-        7257 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 7257)
  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 7257)
  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..7257
                     /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             3837..3863
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    3986..4107
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4023..4042)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4077..4096
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4114..4569)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4251..4272)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4463..4482
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4596..4700
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4702..5059
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4702..4893
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4725..4745)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4910..5045
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5094..5888
                     /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             5094..5888
                     /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(5148..5167)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5758..5777
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6076..6095)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6120..6167
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6496..7084
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6985..7004
                     /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 tgttcaccct ggaggacttc
      841 gtgggcgact gggagcagac cgccgcctac aacctggacc aggtgctgga gcagggcggc
      901 gtgagcagcc tgctgcagaa cctggccgtg agcgtgaccc ccatccagcg catcgtgcgc
      961 agcggcgaga acgccctgaa gatcgacatc cacgtgatca tcccctacga gggcctgagc
     1021 gccgaccaga tggcccagat cgaggaggtg ttcaaggtgg tgtaccccgt ggacgaccac
     1081 cacttcaagg tgatcctgcc ctacggcacc ctggtgatcg acggcgtgac ccccaacatg
     1141 ctgaactact tcggccgccc ctacgagggc atcgccgtgt tcgacggcaa gaagatcacc
     1201 gtgaccggca ccctgtggaa cggcaacaag atcatcgacg agcgcctgat cacccccgac
     1261 ggcagcatgc tgttccgcgt gaccatcaac agcggcagca gcggtggcgg tggctctggc
     1321 ggtggcggca gctctggtgg ccagtccacc attgaggaac aggccaagac atttttggac
     1381 aagtttaacc acgaagccga agacctgttc tatcaaagtt cacttgcttc ttggaattat
     1441 aacaccaata ttactgaaga gaatgtccaa aacatgaata atgctgggga caaatggtct
     1501 gcctttttaa aggaacagtc cacacttgcc caaatgtatc cactacaaga aattcagaat
     1561 ctcacagtca agcttcagct gcaggctctt cagcaaaatg ggtcttcagt gctctcagaa
     1621 gacaagagca aacggttgaa cacaattcta aatacaatga gcaccatcta cagtactgga
     1681 aaagtttgta acccagataa tccacaagaa tgcttattac ttgaaccagg tttgaatgaa
     1741 ataatggcaa acagtttaga ctacaatgag aggctctggg cttgggaaag ctggagatct
     1801 gaggtcggca agcagctgag gccattatat gaagagtatg tggtcttgaa aaatgagatg
     1861 gcaagagcaa atcattatga ggactatggg gattattgga gaggagacta tgaagtaaat
     1921 ggggtagatg gctatgacta cagccgcggc cagttgattg aagatgtgga acataccttt
     1981 gaagagatta aaccattata tgaacatctt catgcctatg tgagggcaaa gttgatgaat
     2041 gcctatcctt cctatatcag tccaattgga tgcctccctg ctcatttgct tggtgatatg
     2101 tggggtagat tttggacaaa tctgtactct ttgacagttc cctttggaca gaaaccaaac
     2161 atagatgtta ctgatgcaat ggtggaccag gcctgggatg cacagagaat attcaaggag
     2221 gccgagaagt tctttgtatc tgttggtctt cctaatatga ctcaaggatt ctgggaaaat
     2281 tccatgctaa cggacccagg aaatgttcag aaagcagtct gccatcccac agcttgggac
     2341 ctggggaagg gcgacttcag gatccttatg tgcacaaagg tgacaatgga cgacttcctg
     2401 acagctcatc atgagatggg gcatatccag tatgatatgg catatgctgc acaacctttt
     2461 ctgctaagaa atggagctaa tgaaggattc catgaagctg ttggggaaat catgtcactt
     2521 tctgcagcca cacctaagca tttaaaatcc attggtcttc tgtcacccga ttttcaagaa
     2581 gacaatgaaa cagaaataaa cttcctgctc aaacaagcac tcacgattgt tgggactctg
     2641 ccatttactt acatgttaga gaagtggagg tggatggtct ttaaagggga aattcccaaa
     2701 gaccagtgga tgaaaaagtg gtgggagatg aagcgagaga tagttggggt ggtggaacct
     2761 gtgccccatg atgaaacata ctgtgacccc gcatctctgt tccatgtttc taatgattac
     2821 tcattcattc gatattacac aaggaccctt taccaattcc agtttcaaga agcactttgt
     2881 caagcagcta aacatgaagg ccctctgcac aaatgtgaca tctcaaactc tacagaagct
     2941 ggacagaaac tgttcaatat gctgaggctt ggaaaatcag aaccctggac cctagcattg
     3001 gaaaatgttg taggagcaaa gaacatgaat gtaaggccac tgctcaacta ctttgagccc
     3061 ttatttacct ggctgaaaga ccagaacaag aattcttttg tgggatggag taccgactgg
     3121 agtccatatg cagaccccaa gagctgcgac aagacccaca cctgtcctcc atgtcctgct
     3181 ccagaactgc tcggcggacc ttccgtgttc ctgtttcctc caaagcctaa ggacaccctg
     3241 atgatcagca gaacccctga agtgacctgc gtggtggtgg atgtgtccca cgaggatccc
     3301 gaagtgaagt tcaattggta cgtggacggc gtggaagtgc acaacgccaa gaccaagcct
     3361 agagaggaac agtacaacag cacctacaga gtggtgtccg tgctgaccgt gctgcaccag
     3421 gattggctga acggcaaaga gtacaagtgc aaggtgtcca acaaggccct gcctgctcct
     3481 atcgagaaaa ccatcagcaa ggccaagggc cagcctaggg aaccccaggt ttacacactg
     3541 cctccaagca gggacgagct gaccaagaat caggtgtccc tgacctgcct ggtcaagggc
     3601 ttctaccctt ccgatatcgc cgtggaatgg gagagcaatg gccagcctga gaacaactac
     3661 aagacaaccc ctcctgtgct ggacagcgac ggctcattct tcctgtacag caagctgaca
     3721 gtggacaaga gcagatggca gcagggcaac gtgttcagct gcagcgtgat gcacgaggcc
     3781 ctgcacaacc actacaccca gaagtccctg agcctgtctc ctggcaaatg aggatctacc
     3841 catacgatgt tccggattac gcttaaggcc gcgactctag atcataatca gccataccac
     3901 atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca
     3961 taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata
     4021 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     4081 tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt aagcgttaat attttgttaa
     4141 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
     4201 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
     4261 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     4321 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
     4381 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     4441 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
     4501 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     4561 agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     4621 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     4681 aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt
     4741 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     4801 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcaaaa gtatgcaaag
     4861 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     4921 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     4981 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     5041 aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt cgcatgattg
     5101 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     5161 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     5221 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg
     5281 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     5341 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     5401 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     5461 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     5521 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     5581 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg
     5641 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     5701 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     5761 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     5821 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     5881 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc
     5941 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     6001 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     6061 tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac
     6121 ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata aacgcggggt
     6181 tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc
     6241 ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc
     6301 agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata tactttagat
     6361 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     6421 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     6481 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     6541 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     6601 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     6661 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     6721 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     6781 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     6841 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     6901 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     6961 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     7021 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     7081 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     7141 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcatta
     7201 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttcc
//
