LOCUS       sequence-426315-        8257 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses hACE2 receptor with LgBiT tag in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8257)
  AUTHORS   Qing E, Kicmal T, Kumar B, Hawkins GM, Timm E, Perlman S, Gallagher 
            T
  TITLE     Dynamics of SARS-CoV-2 Spike Proteins in Cell Entry: Control 
            Elements in the Amino-Terminal Domains.
  JOURNAL   mBio. 2021 Aug 31;12(4):e0159021. doi: 10.1128/mBio.01590-21. Epub 
            2021 Aug 3.
  PUBMED    34340537
REFERENCE   2  (bases 1 to 8257)
  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: "mBio."; 
            date: "2021-08-31"; volume: "12(4):e0159021. doi"; pages: " 
            10.1128/mBio.01590-21. Epub 2021 Aug 3"
COMMENT     Sequence Label: pcDNA3.1-hACE2-LgBiT
FEATURES             Location/Qualifiers
     source          1..8257
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3616..3633)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3622..3846
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3892..4320
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3979..3998)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4189..4210
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4329..4349)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4334..4663
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4514..4649
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4576..4595
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     complement(4784..4803)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5392..5411
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5696..5817
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5733..5752)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5787..5806
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5866..5882)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5866..5882)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5879..5901)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5890..5906
                     /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(5914..5920,5921..5938,5939..5944))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5959..5980
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6097..6114)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6268..6856)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6348..6367)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7027..7818,7819..7887))
                     /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     7650..7669
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7888..7992)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8067..8086)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 gctagcgttt aaacgggccc tctagaccat gtcaagctct tcctggctcc ttctcagcct
      721 tgttgctgta actgctgctc agtccaccat tgaggaacag gccaagacat ttttggacaa
      781 gtttaaccac gaagccgaag acctgttcta tcaaagttca cttgcttctt ggaattataa
      841 caccaatatt actgaagaga atgtccaaaa catgaataat gctggggaca aatggtctgc
      901 ctttttaaag gaacagtcca cacttgccca aatgtatcca ctacaagaaa ttcagaatct
      961 cacagtcaag cttcagctgc aggctcttca gcaaaatggg tcttcagtgc tctcagaaga
     1021 caagagcaaa cggttgaaca caattctaaa tacaatgagc accatctaca gtactggaaa
     1081 agtttgtaac ccagataatc cacaagaatg cttattactt gaaccaggtt tgaatgaaat
     1141 aatggcaaac agtttagact acaatgagag gctctgggct tgggaaagct ggagatctga
     1201 ggtcggcaag cagctgaggc cattatatga agagtatgtg gtcttgaaaa atgagatggc
     1261 aagagcaaat cattatgagg actatgggga ttattggaga ggagactatg aagtaaatgg
     1321 ggtagatggc tatgactaca gccgcggcca gttgattgaa gatgtggaac atacctttga
     1381 agagattaaa ccattatatg aacatcttca tgcctatgtg agggcaaagt tgatgaatgc
     1441 ctatccttcc tatatcagtc caattggatg cctccctgct catttgcttg gtgatatgtg
     1501 gggtagattt tggacaaatc tgtactcttt gacagttccc tttggacaga aaccaaacat
     1561 agatgttact gatgcaatgg tggaccaggc ctgggatgca cagagaatat tcaaggaggc
     1621 cgagaagttc tttgtatctg ttggtcttcc taatatgact caaggattct gggaaaattc
     1681 catgctaacg gacccaggaa atgttcagaa agcagtctgc catcccacag cttgggacct
     1741 ggggaagggc gacttcagga tccttatgtg cacaaaggtg acaatggacg acttcctgac
     1801 agctcatcat gagatggggc atatccagta tgatatggca tatgctgcac aaccttttct
     1861 gctaagaaat ggagctaatg aaggattcca tgaagctgtt ggggaaatca tgtcactttc
     1921 tgcagccaca cctaagcatt taaaatccat tggtcttctg tcacccgatt ttcaagaaga
     1981 caatgaaaca gaaataaact tcctgctcaa acaagcactc acgattgttg ggactctgcc
     2041 atttacttac atgttagaga agtggaggtg gatggtcttt aaaggggaaa ttcccaaaga
     2101 ccagtggatg aaaaagtggt gggagatgaa gcgagagata gttggggtgg tggaacctgt
     2161 gccccatgat gaaacatact gtgaccccgc atctctgttc catgtttcta atgattactc
     2221 attcattcga tattacacaa ggacccttta ccaattccag tttcaagaag cactttgtca
     2281 agcagctaaa catgaaggcc ctctgcacaa atgtgacatc tcaaactcta cagaagctgg
     2341 acagaaactg ttcaatatgc tgaggcttgg aaaatcagaa ccctggaccc tagcattgga
     2401 aaatgttgta ggagcaaaga acatgaatgt aaggccactg ctcaactact ttgagccctt
     2461 atttacctgg ctgaaagacc agaacaagaa ttcttttgtg ggatggagta ccgactggag
     2521 tccatatgca gaccaaagca tcaaagtgag gataagccta aaatcagctc ttggagataa
     2581 agcatatgaa tggaacgaca atgaaatgta cctgttccga tcatctgttg catatgctat
     2641 gaggcagtac tttttaaaag taaaaaatca gatgattctt tttggggagg aggatgtgcg
     2701 agtggctaat ttgaaaccaa gaatctcctt taatttcttt gtcactgcac ctaaaaatgt
     2761 gtctgatatc attcctagaa ctgaagttga aaaggccatc aggatgtccc ggagccgtat
     2821 caatgatgct ttccgtctga atgacaacag cctagagttt ctggggatac agccaacact
     2881 tggacctcct aaccagcccc ctgtttccat atggctgatt gtttttggag ttgtgatggg
     2941 agtgatagtg gttggcattg tcatcctgat cttcactggg atcagagatc ggaagaagaa
     3001 aaataaagca agaagtggag aaaatcctta tgcctccatc gatattagca aaggagaaaa
     3061 taatccagga ttccaaaaca ctgatgatgt tcagacctcc tttgtcttca cactcgaaga
     3121 tttcgttggg gactgggaac agacagccgc ctacaacctg gaccaagtcc ttgaacaggg
     3181 aggtgtgtcc agtttgctgc agaatctcgc cgtgtccgta actccgatcc aaaggattgt
     3241 ccggagcggt gaaaatgccc tgaagatcga catccatgtc atcatcccgt atgaaggtct
     3301 gagcgccgac caaatggccc agatcgaaga ggtgtttaag gtggtgtacc ctgtggatga
     3361 tcatcacttt aaggtgatcc tgccctatgg cacactggta atcgacgggg ttacgccgaa
     3421 catgctgaac tatttcggac ggccgtatga aggcatcgcc gtgttcgacg gcaaaaagat
     3481 cactgtaaca gggaccctgt ggaacggcaa caaaattatc gacgagcgcc tgatcacccc
     3541 cgacggctcc atgctgttcc gagtaaccat caacagttaa gcttggatcc gcgttaagtt
     3601 taaaccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
     3661 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
     3721 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
     3781 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct
     3841 ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct
     3901 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     3961 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     4021 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     4081 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     4141 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     4201 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     4261 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     4321 aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     4381 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     4441 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     4501 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     4561 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca
     4621 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg
     4681 tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca
     4741 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     4801 ggcaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     4861 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag
     4921 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     4981 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     5041 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     5101 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     5161 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     5221 tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg
     5281 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     5341 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     5401 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     5461 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     5521 gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     5581 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     5641 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt
     5701 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     5761 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     5821 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
     5881 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     5941 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     6001 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     6061 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     6121 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     6181 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     6241 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     6301 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     6361 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     6421 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     6481 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     6541 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     6601 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     6661 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     6721 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     6781 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     6841 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     6901 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     6961 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     7021 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     7081 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     7141 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     7201 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     7261 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     7321 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     7381 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     7441 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     7501 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     7561 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     7621 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     7681 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     7741 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     7801 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     7861 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     7921 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     7981 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct
     8041 cccgatcccc tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     8101 tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct
     8161 acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagggt taggcgtttt
     8221 gcgctgcttc gcgatgtacg ggccagatat acgcgtt
//
