LOCUS       sequence-426313-        6805 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses N protein with HiBiT tag in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6805)
  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 6805)
  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-SARS-CoV-2-HiBiT-N
FEATURES             Location/Qualifiers
     source          1..6805
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      949..958
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2309..2338
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2354..2371
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2394..2411)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2400..2624
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2670..3098
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2757..2776)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2967..2988
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3107..3127)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3112..3441
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3292..3427
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3354..3373
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3508..4302
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3562..3581)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4172..4191
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4478..4599
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4515..4534)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4569..4588
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4648..4664)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4648..4664)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4661..4683)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4672..4688
                     /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(4696..4702,4703..4720,4721..4726))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4741..4762
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4879..4896)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5050..5638)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5130..5149)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5809..6600,6601..6669))
                     /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     6432..6451
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6670..6774)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagt
      901 taagcttggt accgagctcg gatccactag tccagtgtgg tggaattcgc caccatggtg
      961 agcggctggc ggctgttcaa gaagattagc ggctcgaccg gtggctcgag cggtgccacc
     1021 atgtccgaca acggccccca gaatcagaga aatgccccta gaatcacctt cggcggccct
     1081 agcgatagca ccggcagcaa ccagaacggc gagagaagcg gcgccagaag caagcagcgg
     1141 cggcctcagg gcctgcctaa caacaccgcc agctggttca ccgccctgac acagcacggc
     1201 aaggaagatc tgaagttccc ccgcggccag ggcgtgccta ttaacaccaa cagctctccc
     1261 gacgaccaga tcggctacta cagaagagcc accagacgga tcagaggcgg cgacggcaag
     1321 atgaaggacc tgagccctag atggtacttc tactacctgg gcaccggccc ggaagccgga
     1381 ctgccttacg gcgccaacaa ggacggcatc atctgggtcg ccaccgaggg cgctctgaac
     1441 acacctaaag accacatcgg cacccggaac cccgccaaca acgccgccat cgtgctgcag
     1501 ctgccacagg gtacgaccct gcccaagggc ttctacgccg aaggcagcag gggaggaagc
     1561 caggctagca gccggtcttc ctccagatct agaaacagct ctcggaacag tacccctggc
     1621 tcttctcggg gaaccagccc cgctagaatg gcgggaaatg gcggcgacgc cgccctggcc
     1681 ctgctcctgc ttgatagact gaatcaactg gaaagcaaga tgtccggcaa aggccagcag
     1741 caacagggac agaccgtgac caagaaaagc gccgctgagg ccagcaagaa acctagacag
     1801 aagcggaccg ccacaaaggc ctacaacgtg acacaggctt ttggaagaag aggccctgag
     1861 cagacccaag gcaacttcgg agatcaggag ctgatccggc agggcacaga ctacaagcac
     1921 tggcctcaga tcgcccagtt cgcccctagc gcctctgcct ttttcggcat gtctagaatc
     1981 ggcatggaag tgacacccag cggaacatgg ctgacctaca ccggcgctat caagctggac
     2041 gataaggacc ccaatttcaa ggatcaagtg atcctgctga acaagcatat cgacgcttat
     2101 aagacctttc cacctacaga gcctaagaag gataagaaga agaaagccga cgagacacag
     2161 gccctgccac agcggcaaaa gaagcagcag acagtgaccc tgctgcctgc tgccgatctg
     2221 gacgacttca gcaaacagct gcagcagagc atgagctccg ccgacagcac acaggcctga
     2281 gcggccgctc gagtctagag ggcccttcga acaaaaactc atctcagaag aggatctgaa
     2341 tatgcatacc ggtcatcatc accatcacca ttgagtttaa acccgctgat cagcctcgac
     2401 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     2461 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     2521 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     2581 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag
     2641 aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc
     2701 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     2761 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     2821 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     2881 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     2941 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     3001 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     3061 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
     3121 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3181 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3241 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     3301 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     3361 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt
     3421 ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat
     3481 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
     3541 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     3601 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     3661 gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc
     3721 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     3781 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     3841 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     3901 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     3961 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     4021 gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     4081 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     4141 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     4201 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     4261 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
     4321 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     4381 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     4441 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     4501 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     4561 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     4621 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     4681 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     4741 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     4801 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     4861 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     4921 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     4981 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     5041 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     5101 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     5161 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     5221 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     5281 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     5341 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     5401 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     5461 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     5521 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     5581 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     5641 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     5701 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     5761 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     5821 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     5881 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     5941 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     6001 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     6061 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     6121 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     6181 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     6241 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     6301 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     6361 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     6421 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     6481 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     6541 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     6601 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     6661 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     6721 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     6781 tccccgaaaa gtgccacctg acgtc
//
