LOCUS       sequence-418138-        6809 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SARS-CoV-2 N protein fused with 2Strep in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6809)
  AUTHORS   Li J, Gui Q, Liang FX, Sall J, Zhang Q, Duan Y, Dhabaria A, Askenazi
            M, Ueberheide B, Stapleford KA, Pagano M
  TITLE     The REEP5/TRAM1 complex binds SARS-CoV-2 NSP3 and promotes virus 
            replication.
  JOURNAL   J Virol. 2023 Sep 28:e0050723. doi: 10.1128/jvi.00507-23.
  PUBMED    37768083
REFERENCE   2  (bases 1 to 6809)
  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: "J Virol. 
            2023 Sep 28:e0050723. doi: 10.1128/jvi.00507-23."
COMMENT     Sequence Label: SARS-CoV-2 N-2Strep
FEATURES             Location/Qualifiers
     source          1..6809
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(43..62)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        234..613
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        614..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     768..788
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     862..881
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        862..880
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2178..2187
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      2182..2191
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2199..2222
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             2259..2282
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             2307..2330
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     promoter        complement(2361..2379)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2362..2379)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2399..2416)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2405..2629
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2675..3103
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2762..2781)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2972..2993
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3112..3132)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3117..3446
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3297..3432
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3359..3378
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3513..4307
                     /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(3567..3586)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4177..4196
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4481..4602
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4518..4537)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4572..4591
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4651..4667)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4651..4667)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4664..4686)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4675..4691
                     /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(4699..4705,4706..4723,4724..4729))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4744..4765
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4882..4899)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5053..5641)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5133..5152)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5812..6603,6604..6672))
                     /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     6435..6454
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6673..6777)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct gctctgatgc
       61 cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc
      121 gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct
      181 tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc gttgacattg
      241 attattgact agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
      301 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
      361 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
      421 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
      481 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
      541 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
      601 cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga
      661 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
      721 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg
      781 taggcgtgta cggtgggagg tctatataag cagagctctc tggctaacta gagaacccac
      841 tgcttactgg cttatcgaaa ttaatacgac tcactatagg gagacccaag cttggtaccg
      901 agctcggatc catgtctgat aacgggccac aaaaccaaag gaatgcgccg agaattacgt
      961 tcggagggcc aagcgatagc accggtagta atcagaacgg tgagagaagc ggcgcacggt
     1021 caaagcaacg gagaccccag gggctgccta ataatactgc ctcttggttc accgcgctga
     1081 ctcaacacgg taaggaggac cttaagtttc cgcgaggaca gggcgtgcca atcaacacca
     1141 actccagccc tgatgatcaa atcggatatt atagaagggc aacgcgacga atccggggtg
     1201 gtgatgggaa gatgaaggac ctcagtcctc gctggtattt ttattacctt ggtacgggac
     1261 ctgaggctgg acttccgtat ggtgctaaca aggatgggat tatctgggtt gcaaccgaag
     1321 gagctctgaa caccccaaag gatcatatag gcacgagaaa cccagcgaac aacgctgcaa
     1381 ttgtcctgca actgccccaa ggcacaaccc tccccaaagg cttttacgcg gagggctcca
     1441 ggggcggctc acaagcctct tcccgatctt cctctaggtc taggaacagc tctcgaaatt
     1501 caacacccgg ttccagcaga ggtacatctc cggccagaat ggctggaaac gggggtgacg
     1561 cagctctggc gttgcttctt ctcgatcgac tgaaccagct ggaaagtaaa atgtccggta
     1621 aggggcaaca acaacaggga cagaccgtaa ctaaaaaaag tgctgctgag gccagcaaga
     1681 agccacgaca gaagcggacg gcgacaaagg cttacaacgt aacgcaggca tttggacggc
     1741 gcgggcctga gcagacacaa ggaaatttcg gggatcagga gctcattcgg cagggcaccg
     1801 actacaaaca ttggccacag atagcccagt ttgcgccatc tgcaagtgct tttttcggta
     1861 tgtcaagaat cggtatggaa gtgactccat caggcacatg gctcacctat actggtgcaa
     1921 tcaagctcga tgataaagac cctaacttta aagatcaggt cattctcctc aataaacata
     1981 ttgacgccta caagactttt cctcccacag agccgaaaaa agataagaag aagaaggccg
     2041 acgaaactca agcactgccg cagcgccaaa aaaagcaaca gacggttacg cttttgcccg
     2101 ccgcagacct ggatgacttt tctaagcagc tgcagcagtc aatgagctca gctgattcca
     2161 ctcaggcgga attcgccgcc accatggcgg ccgccagctg gagccaccct cagttcgaga
     2221 agggaggagg aagcggcgga ggcagcggag gaggaagctg gagccacccg cagttcgaga
     2281 aatgactcga gcatgcatct agaggctgga gccacccgca gttcgagaaa tgactcgagc
     2341 atgcatctag agggccctat tctatagtgt cacctaaatg ctagagctcg ctgatcagcc
     2401 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     2461 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     2521 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     2581 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg
     2641 gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc
     2701 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     2761 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     2821 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     2881 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     2941 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     3001 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     3061 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt
     3121 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     3181 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     3241 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     3301 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     3361 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct
     3421 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat
     3481 ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     3541 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     3601 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     3661 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc
     3721 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     3781 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     3841 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     3901 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     3961 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     4021 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg
     4081 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     4141 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     4201 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     4261 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     4321 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     4381 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     4441 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta
     4501 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     4561 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc
     4621 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     4681 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     4741 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     4801 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     4861 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     4921 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     4981 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     5041 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     5101 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     5161 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     5221 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     5281 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     5341 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     5401 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     5461 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     5521 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     5581 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     5641 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     5701 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     5761 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     5821 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     5881 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     5941 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     6001 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     6061 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     6121 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     6181 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     6241 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     6301 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     6361 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     6421 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     6481 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     6541 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     6601 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     6661 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     6721 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     6781 atttccccga aaagtgccac ctgacgtcg
//
