LOCUS       sequence-353720-        5953 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector for SARS-CoV-1 Nsp16, V5-tagged. 
            Sequence codon-optimized..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5953)
  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 5953)
  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 NSP16 C-V5-tag
FEATURES             Location/Qualifiers
     source          1..5953
                     /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             1570..1611
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     intron          1641..2213
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     complement(2345..2364)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2410..2465
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2464..2483)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      2819..2954
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3060..3077)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3242..3830)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3322..3341)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4001..4792,4793..4861))
                     /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     4624..4643
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4862..4966)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5034..5052
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5090..5112)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5212..5231
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(5298..5753)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5435..5456)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5647..5666
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5879..5901
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5893..5910
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5894..5910
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5917..5936
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5917..5935
                     /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 ccatggccag tcaagcatgg caaccaggag tagcaatgcc caatctgtat
      721 aaaatgcagc ggatgttgct ggaaaagtgc gacttgcaga attatggcga aaatgcggtc
      781 atcccgaagg gtattatgat gaatgtagca aaatacacac aactttgtca gtatttgaat
      841 accctgaccc tcgcagtacc ttataatatg agagtgattc attttggcgc agggagcgat
      901 aagggcgtgg cgcctgggac cgcggttctg cgccaatggc tgcccaccgg cacccttctg
      961 gtggactcag acttgaatga tttcgtcagt gatgccgact ccacccttat tggcgactgc
     1021 gccaccgtgc acactgctaa taagtgggat ttgatcatta gcgacatgta tgaccctcga
     1081 acaaaacatg tgaccaagga gaacgacagt aaggaaggtt tcttcacata tctgtgcggt
     1141 tttatcaagc aaaagctcgc ccttggcggg agtatagctg tgaaaataac cgaacactcc
     1201 tggaatgcag acctgtacaa gctcatgggg cacttctctt ggtggacagc ctttgtcact
     1261 aacgtgaatg ccagctcctc tgaagctttc ctcattggcg ccaactacct tggaaagccg
     1321 aaagaacaga tagacggtta cactatgcac gcgaactata tattttggag aaacactaat
     1381 cccattcagt tgagttcata cagcctcttt gacatgtcaa aattcccact gaagcttagg
     1441 ggaaccgcag tgatgagctt gaaggaaaac cagattaatg acatgatcta cagccttctg
     1501 gagaaaggta gactcatcat cagggaaaac aacagagtgg ttgtgtctag tgatattctg
     1561 gtgaataacg gcaagccaat accaaatccg ctcctgggcc ttgacagcac gtagacgcgt
     1621 cggatcctga gaacttcagg gtgagtttgg ggacccttga ttgttctttc tttttcgcta
     1681 ttgtaaaatt catgttatat ggagggggca aagttttcag ggtgttgttt agaatgggaa
     1741 gatgtccctt gtatcaccat ggaccctcat gataattttg tttctttcac tttctactct
     1801 gttgacaacc attgtctcct cttattttct tttcattttc tgtaactttt tcgttaaact
     1861 ttagcttgca tttgtaacga atttttaaat tcacttttgt ttatttgtca gattgtaagt
     1921 actttctcta atcacttttt tttcaaggca atcagggtat attatattgt acttcagcac
     1981 agttttagag aacaattgtt ataattaaat gataaggtag aatatttctg catataaatt
     2041 ctggctggcg tggaaatatt cttattggta gaaacaacta caccctggtc atcatcctgc
     2101 ctttctcttt atggttacaa tgatatacac tgtttgagat gaggataaaa tactctgagt
     2161 ccaaaccggg cccctctgct aaccatgttc atgccttctt ctctttccta cagctcctgg
     2221 gcaacgtgct ggttgttgtg ctgtctcatc attttggcaa agaattcact cctcaggtgc
     2281 aggctgccta tcagaaggtg gtggctggtg tggccaatgc cctggctcac aaataccact
     2341 gagatctttt tccctctgcc aaaaattatg gggacatcat gaagcccctt gagcatctga
     2401 cttctggcta ataaaggaaa tttattttca ttgcaatagt gtgttggaat tttttgtgtc
     2461 tctcactcgg aaggacatat gggagggcaa atcatttaaa acatcagaat gagtatttgg
     2521 tttagagttt ggcaacatat gcccatatgc tggctgccat gaacaaaggt tggctataaa
     2581 gaggtcatca gtatatgaaa cagccccctg ctgtccattc cttattccat agaaaagcct
     2641 tgacttgagg ttagattttt tttatatttt gttttgtgtt atttttttct ttaacatccc
     2701 taaaattttc cttacatgtt ttactagcca gatttttcct cctctcctga ctactcccag
     2761 tcatagctgt ccctcttctc ttatggagat ccctcgagga gctttttgca aaagcctagg
     2821 cctccaaaaa agcctcttca ctacttctgg aatagctcag aggccgaggc ggcctcggcc
     2881 tctgcataaa taaaaaaaat tagtcagcca tggggcggag aatgggcgga actgggcgga
     2941 gttaggggcg ggatgggcgg agttaggggc gggactatgg ttgctgacta attgctgcat
     3001 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     3061 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     3121 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     3181 aaaggccagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     3241 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     3301 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     3361 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     3421 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     3481 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     3541 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     3601 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     3661 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     3721 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     3781 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     3841 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     3901 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     3961 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     4021 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     4081 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     4141 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     4201 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     4261 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     4321 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     4381 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     4441 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     4501 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     4561 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     4621 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     4681 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     4741 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     4801 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     4861 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     4921 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     4981 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     5041 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     5101 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     5161 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag
     5221 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga
     5281 gaaaataccg catcaggaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt
     5341 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
     5401 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
     5461 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
     5521 acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
     5581 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
     5641 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac
     5701 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtccattcg
     5761 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
     5821 cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc
     5881 cagtcacgac gttgtaaaac gacggccagt gaattgtaat acgactcact atagggcgaa
     5941 ttcgagctcg ccc
//
