LOCUS       sequence-353867-        5711 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector for SARS-CoV-2 M, V5-tagged. Sequence 
            codon-optimized..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5711)
  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 5711)
  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-CoV2 M C-V5-tag
FEATURES             Location/Qualifiers
     source          1..5711
                     /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"
     regulatory      667..676
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1339..1380
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     intron          1410..1982
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     complement(2114..2133)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2179..2234
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2233..2252)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      2588..2723
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2829..2846)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3000..3588)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3080..3099)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3759..4550,4551..4619))
                     /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     4382..4401
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4620..4724)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4792..4810
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4848..4870)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4970..4989
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(5056..5511)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5193..5214)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5405..5424
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5637..5659
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5651..5668
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5652..5668
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5675..5694
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5675..5693
                     /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 ccatggcaga ttcaaacggg acgatcacag tagaggagct gaagaaactg
      721 ctggagcaat ggaatctggt cataggcttc cttttcctta catggatttg tctgctgcaa
      781 ttcgcgtacg caaaccgcaa ccgctttctc tatattatca aacttatatt cctctggttg
      841 ctctggccag taactctggc ttgtttcgtt ctggcggccg tgtacaggat taattggatt
      901 accggcggaa tcgcaatcgc aatggcatgc ctggtgggtc tgatgtggct gagctacttt
      961 atcgcaagct tcagattgtt cgccagaact cgctctatgt ggtctttcaa tcctgagact
     1021 aacattctgc tgaacgttcc tctgcatgga actattctca cccgccccct gctcgaatcc
     1081 gagctcgtca tcggcgcagt aattctgcgg ggacatctcc gaatcgctgg acaccatctt
     1141 ggacgttgtg atatcaagga tctgccgaaa gagattacag ttgcaactag tcgaacactg
     1201 tcctattata aactgggcgc ctctcaacga gtggccggtg atagtgggtt cgcagcttac
     1261 tctcgttaca gaatcggaaa ctataaactt aacacggacc acagctcttc ctctgataat
     1321 attgccctgc tggtgcaagg aaaacctatt cccaatccac tgctcggatt ggactctact
     1381 tagacgcgtc ggatcctgag aacttcaggg tgagtttggg gacccttgat tgttctttct
     1441 ttttcgctat tgtaaaattc atgttatatg gagggggcaa agttttcagg gtgttgttta
     1501 gaatgggaag atgtcccttg tatcaccatg gaccctcatg ataattttgt ttctttcact
     1561 ttctactctg ttgacaacca ttgtctcctc ttattttctt ttcattttct gtaacttttt
     1621 cgttaaactt tagcttgcat ttgtaacgaa tttttaaatt cacttttgtt tatttgtcag
     1681 attgtaagta ctttctctaa tcactttttt ttcaaggcaa tcagggtata ttatattgta
     1741 cttcagcaca gttttagaga acaattgtta taattaaatg ataaggtaga atatttctgc
     1801 atataaattc tggctggcgt ggaaatattc ttattggtag aaacaactac accctggtca
     1861 tcatcctgcc tttctcttta tggttacaat gatatacact gtttgagatg aggataaaat
     1921 actctgagtc caaaccgggc ccctctgcta accatgttca tgccttcttc tctttcctac
     1981 agctcctggg caacgtgctg gttgttgtgc tgtctcatca ttttggcaaa gaattcactc
     2041 ctcaggtgca ggctgcctat cagaaggtgg tggctggtgt ggccaatgcc ctggctcaca
     2101 aataccactg agatcttttt ccctctgcca aaaattatgg ggacatcatg aagccccttg
     2161 agcatctgac ttctggctaa taaaggaaat ttattttcat tgcaatagtg tgttggaatt
     2221 ttttgtgtct ctcactcgga aggacatatg ggagggcaaa tcatttaaaa catcagaatg
     2281 agtatttggt ttagagtttg gcaacatatg cccatatgct ggctgccatg aacaaaggtt
     2341 ggctataaag aggtcatcag tatatgaaac agccccctgc tgtccattcc ttattccata
     2401 gaaaagcctt gacttgaggt tagatttttt ttatattttg ttttgtgtta tttttttctt
     2461 taacatccct aaaattttcc ttacatgttt tactagccag atttttcctc ctctcctgac
     2521 tactcccagt catagctgtc cctcttctct tatggagatc cctcgaggag ctttttgcaa
     2581 aagcctaggc ctccaaaaaa gcctcttcac tacttctgga atagctcaga ggccgaggcg
     2641 gcctcggcct ctgcataaat aaaaaaaatt agtcagccat ggggcggaga atgggcggaa
     2701 ctgggcggag ttaggggcgg gatgggcgga gttaggggcg ggactatggt tgctgactaa
     2761 ttgctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     2821 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     2881 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     2941 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     3001 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     3061 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     3121 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     3181 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     3241 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     3301 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     3361 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     3421 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     3481 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     3541 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     3601 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     3661 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     3721 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     3781 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     3841 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     3901 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     3961 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     4021 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     4081 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     4141 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     4201 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     4261 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     4321 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     4381 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     4441 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     4501 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     4561 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     4621 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     4681 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     4741 gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt
     4801 atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg
     4861 cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt
     4921 cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag
     4981 cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga
     5041 aaataccgca tcaggaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     5101 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     5161 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     5221 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     5281 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     5341 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     5401 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     5461 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tccattcgcc
     5521 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
     5581 gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca
     5641 gtcacgacgt tgtaaaacga cggccagtga attgtaatac gactcactat agggcgaatt
     5701 cgagctcgcc c
//
