LOCUS       sequence-353876-        5585 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector for RaTG13 Nsp1, V5-tagged. Sequence 
            codon-optimized..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5585)
  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 5585)
  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_batCoV RaTG13 NSP1 C-V5-tag
FEATURES             Location/Qualifiers
     source          1..5585
                     /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             1213..1254
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     intron          1284..1856
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     complement(1988..2007)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2053..2108
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2107..2126)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      2462..2597
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2703..2720)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2874..3462)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2954..2973)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3633..4424,4425..4493))
                     /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     4256..4275
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4494..4598)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4666..4684
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4722..4744)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4844..4863
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(4930..5385)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5067..5088)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5279..5298
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5511..5533
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5525..5542
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5526..5542
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5549..5568
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5549..5567
                     /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 ccatggagag ccttgtcccc ggttttaatg agaagactca tgtgcaactc
      721 agtttgcccg ttctccaagt gagagacgtg cttgtgaggg gatttggtga tagtgtcgag
      781 gaagccttgt ctgaagcacg ccaacacctc aaagacggca cttgcgggct ggtcgaggtc
      841 gagaagggtg tactccccca actggaacaa ccttatgtct ttatcaaaag gtccgatgcg
      901 cgcactgccc cacatggcca tgttatggtt gagctcgtgg cagaacttaa tggcattcag
      961 tacggccgct caggcgaaac actcggcgta ctcgttccct atgtgggcga aacccccgtt
     1021 gtctaccgga aagtcctgct gaggaagaac ggaaacaaag gagctggcgg ccatagttat
     1081 ggtgcagatt tgaaatcatt cgaccttggg gacgaactcg ggaccgaccc ttatgaggat
     1141 tttcaagaga attggaatac taaacattct tcaggcgtta ctagggatct gatgcgcgaa
     1201 ttgaatggtg gcgggaaacc aattcccaat ccattgttgg ggcttgatag cacctaaacg
     1261 cgtcggatcc tgagaacttc agggtgagtt tggggaccct tgattgttct ttctttttcg
     1321 ctattgtaaa attcatgtta tatggagggg gcaaagtttt cagggtgttg tttagaatgg
     1381 gaagatgtcc cttgtatcac catggaccct catgataatt ttgtttcttt cactttctac
     1441 tctgttgaca accattgtct cctcttattt tcttttcatt ttctgtaact ttttcgttaa
     1501 actttagctt gcatttgtaa cgaattttta aattcacttt tgtttatttg tcagattgta
     1561 agtactttct ctaatcactt ttttttcaag gcaatcaggg tatattatat tgtacttcag
     1621 cacagtttta gagaacaatt gttataatta aatgataagg tagaatattt ctgcatataa
     1681 attctggctg gcgtggaaat attcttattg gtagaaacaa ctacaccctg gtcatcatcc
     1741 tgcctttctc tttatggtta caatgatata cactgtttga gatgaggata aaatactctg
     1801 agtccaaacc gggcccctct gctaaccatg ttcatgcctt cttctctttc ctacagctcc
     1861 tgggcaacgt gctggttgtt gtgctgtctc atcattttgg caaagaattc actcctcagg
     1921 tgcaggctgc ctatcagaag gtggtggctg gtgtggccaa tgccctggct cacaaatacc
     1981 actgagatct ttttccctct gccaaaaatt atggggacat catgaagccc cttgagcatc
     2041 tgacttctgg ctaataaagg aaatttattt tcattgcaat agtgtgttgg aattttttgt
     2101 gtctctcact cggaaggaca tatgggaggg caaatcattt aaaacatcag aatgagtatt
     2161 tggtttagag tttggcaaca tatgcccata tgctggctgc catgaacaaa ggttggctat
     2221 aaagaggtca tcagtatatg aaacagcccc ctgctgtcca ttccttattc catagaaaag
     2281 ccttgacttg aggttagatt ttttttatat tttgttttgt gttatttttt tctttaacat
     2341 ccctaaaatt ttccttacat gttttactag ccagattttt cctcctctcc tgactactcc
     2401 cagtcatagc tgtccctctt ctcttatgga gatccctcga ggagcttttt gcaaaagcct
     2461 aggcctccaa aaaagcctct tcactacttc tggaatagct cagaggccga ggcggcctcg
     2521 gcctctgcat aaataaaaaa aattagtcag ccatggggcg gagaatgggc ggaactgggc
     2581 ggagttaggg gcgggatggg cggagttagg ggcgggacta tggttgctga ctaattgctg
     2641 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     2701 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     2761 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     2821 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     2881 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     2941 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     3001 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     3061 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     3121 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     3181 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     3241 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     3301 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     3361 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     3421 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     3481 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     3541 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     3601 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     3661 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     3721 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     3781 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     3841 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     3901 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     3961 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     4021 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     4081 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     4141 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     4201 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     4261 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     4321 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     4381 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     4441 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     4501 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     4561 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     4621 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     4681 aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
     4741 ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
     4801 gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt
     4861 gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
     4921 cgcatcagga aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
     4981 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
     5041 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
     5101 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
     5161 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
     5221 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     5281 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
     5341 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtccatt cgccattcag
     5401 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc
     5461 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
     5521 acgttgtaaa acgacggcca gtgaattgta atacgactca ctatagggcg aattcgagct
     5581 cgccc
//
