LOCUS       sequence-353803-        6311 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector for SARS-CoV-1 N, V5-tagged. Sequence 
            codon-optimized..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6311)
  AUTHORS   Hayn M, Hirschenberger M, Koepke L, Nchioua R, Straub JH, Klute S, 
            Hunszinger V, Zech F, Prelli Bozzo C, Aftab W, Christensen MH, 
            Conzelmann C, Muller JA, Srinivasachar Badarinarayan S, Sturzel CM, 
            Forne I, Stenger S, Conzelmann KK, Munch J, Schmidt FI, Sauter D, 
            Imhof A, Kirchhoff F, Sparrer KMJ
  TITLE     Systematic functional analysis of SARS-CoV-2 proteins uncovers viral
            innate immune antagonists and remaining vulnerabilities.
  JOURNAL   Cell Rep. 2021 May 18;35(7):109126.
  PUBMED    33974846
REFERENCE   2  (bases 1 to 6311)
  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: "Cell Rep.";
            date: "2021-05-18"; volume: "35(7)"; pages: "109126"
COMMENT     Sequence Label: pCG_SARS-CoV1 N C-V5-tag
FEATURES             Location/Qualifiers
     source          1..6311
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..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"
     5'UTR           598..637
                     /label=HSV TK 5'-UTR
                     /note="5' untranslated region from the herpes simplex virus
                     thymidine kinase gene"
     CDS             1939..1980
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     intron          2010..2582
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     complement(2714..2733)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2779..2834
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2833..2852)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      3188..3323
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3429..3446)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3600..4188)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3680..3699)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4359..5150,5151..5219))
                     /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     4982..5001
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5220..5324)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5392..5410
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5448..5470)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5570..5589
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(5656..6111)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5793..5814)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6005..6024
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6237..6259
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6251..6268
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6252..6268
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6275..6294
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6275..6293
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac 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 tccgggggat
      601 cttggtggcg tgaaactccc gcacctcttc ggccagcgcc ttgtagaagc gcgtaggcct
      661 tctagagcca ccatgagcga taatggacca caatctaatc aaagatccgc tccccgcatt
      721 accttcggtg ggcccacaga tagtacggac aataatcaaa acggcgggcg aaacggggcg
      781 cgacctaagc agcgacgccc tcaagggctg cccaacaaca ccgcgagctg gtttaccgct
      841 ctgactcaac atgggaaaga agagctccgg tttccaaggg gacaaggagt tcctataaat
      901 actaacagcg gtccagacga tcagataggc tactaccgga gagccacacg tcgtgtcagg
      961 ggtggtgacg gcaaaatgaa agaattgtcc ccacggtggt acttttatta cctcggaacc
     1021 gggcctgaag caagcctgcc ctatggcgct aataaagagg gaatcgtctg ggtagccacc
     1081 gagggcgctc ttaatacacc taaggatcac attgggacac gaaatcccaa taacaacgct
     1141 gcaactgtct tgcaactgcc ccaagggacc actctgccga aggggtttta tgctgaaggt
     1201 agcagaggtg gatctcaagc gtcttctcgc tcctcatcca gatctcgggg taattccaga
     1261 aactccactc ccggaagctc acggggaaac agtccagccc ggatggcatc aggcggcgga
     1321 gaaaccgccc tggctctgct gctgctcgac aggcttaatc aacttgaaag caaggtgtct
     1381 gggaaagggc aacagcagca gggtcaaaca gtgacaaaga aatctgctgc tgaggcgagt
     1441 aagaagcccc ggcagaaacg gacagcaaca aaacaataca acgtaactca agccttcgga
     1501 cgccgcggcc cagagcagac acaggggaac tttggcgatc aagatttgat ccgccaggga
     1561 actgattaca aacattggcc tcaaattgcg cagttcgctc ctagcgcatc agcgttcttc
     1621 ggcatgagta gaataggcat ggaagtcact ccaagcggta cttggctgac ctaccatggc
     1681 gctatcaaac tcgatgacaa agatccgcaa tttaaagata atgtcatcct tcttaacaaa
     1741 cacatagatg cgtacaaaac ctttccacct acagaaccga agaaggataa gaagaagaag
     1801 actgatgaag cacaacctct gccacaacgg caaaagaagc agcccactgt gaccctgctt
     1861 cccgctgcgg acatggacga cttttctcgt cagttgcaaa acagtatgag cggtgcgtca
     1921 gctgattcta cacaggcagg taaaccgatc cctaaccccc tgctcggcct tgattccact
     1981 tgaacgcgtc ggatcctgag aacttcaggg tgagtttggg gacccttgat tgttctttct
     2041 ttttcgctat tgtaaaattc atgttatatg gagggggcaa agttttcagg gtgttgttta
     2101 gaatgggaag atgtcccttg tatcaccatg gaccctcatg ataattttgt ttctttcact
     2161 ttctactctg ttgacaacca ttgtctcctc ttattttctt ttcattttct gtaacttttt
     2221 cgttaaactt tagcttgcat ttgtaacgaa tttttaaatt cacttttgtt tatttgtcag
     2281 attgtaagta ctttctctaa tcactttttt ttcaaggcaa tcagggtata ttatattgta
     2341 cttcagcaca gttttagaga acaattgtta taattaaatg ataaggtaga atatttctgc
     2401 atataaattc tggctggcgt ggaaatattc ttattggtag aaacaactac accctggtca
     2461 tcatcctgcc tttctcttta tggttacaat gatatacact gtttgagatg aggataaaat
     2521 actctgagtc caaaccgggc ccctctgcta accatgttca tgccttcttc tctttcctac
     2581 agctcctggg caacgtgctg gttgttgtgc tgtctcatca ttttggcaaa gaattcactc
     2641 ctcaggtgca ggctgcctat cagaaggtgg tggctggtgt ggccaatgcc ctggctcaca
     2701 aataccactg agatcttttt ccctctgcca aaaattatgg ggacatcatg aagccccttg
     2761 agcatctgac ttctggctaa taaaggaaat ttattttcat tgcaatagtg tgttggaatt
     2821 ttttgtgtct ctcactcgga aggacatatg ggagggcaaa tcatttaaaa catcagaatg
     2881 agtatttggt ttagagtttg gcaacatatg cccatatgct ggctgccatg aacaaaggtt
     2941 ggctataaag aggtcatcag tatatgaaac agccccctgc tgtccattcc ttattccata
     3001 gaaaagcctt gacttgaggt tagatttttt ttatattttg ttttgtgtta tttttttctt
     3061 taacatccct aaaattttcc ttacatgttt tactagccag atttttcctc ctctcctgac
     3121 tactcccagt catagctgtc cctcttctct tatggagatc cctcgaggag ctttttgcaa
     3181 aagcctaggc ctccaaaaaa gcctcttcac tacttctgga atagctcaga ggccgaggcg
     3241 gcctcggcct ctgcataaat aaaaaaaatt agtcagccat ggggcggaga atgggcggaa
     3301 ctgggcggag ttaggggcgg gatgggcgga gttaggggcg ggactatggt tgctgactaa
     3361 ttgctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     3421 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     3481 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     3541 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     3601 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     3661 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     3721 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     3781 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     3841 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     3901 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     3961 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     4021 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     4081 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     4141 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     4201 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     4261 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     4321 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     4381 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     4441 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     4501 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     4561 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     4621 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     4681 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     4741 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     4801 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     4861 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     4921 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     4981 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     5041 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     5101 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     5161 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     5221 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     5281 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     5341 gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt
     5401 atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg
     5461 cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt
     5521 cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag
     5581 cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga
     5641 aaataccgca tcaggaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     5701 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     5761 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     5821 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     5881 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     5941 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     6001 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     6061 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tccattcgcc
     6121 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
     6181 gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca
     6241 gtcacgacgt tgtaaaacga cggccagtga attgtaatac gactcactat agggcgaatt
     6301 cgagctcgcc c
//
