LOCUS       sequence-353883-        6086 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector for RaTG13 Nsp15, V5-tagged. Sequence 
            codon-optimized..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6086)
  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 6086)
  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 NSP15 C-V5-tag
FEATURES             Location/Qualifiers
     source          1..6086
                     /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             1714..1755
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     intron          1785..2357
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     complement(2489..2508)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2554..2609
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2608..2627)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      2963..3098
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3204..3221)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3375..3963)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3455..3474)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4134..4925,4926..4994))
                     /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     4757..4776
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4995..5099)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5167..5185
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5223..5245)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5345..5364
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(5431..5886)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5568..5589)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5780..5799
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6012..6034
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6026..6043
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6027..6043
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6050..6069
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6050..6068
                     /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 ccatgagcct cgaaaacgta gcttttaatg tagtcaataa aggccatttt
      721 gatggacagc agggagaagt ccccgtgtcc attattaata atactgtcta tacaaaagtc
      781 gacggcgtag acgtggagct cttcgagaac aagaccacgt tgccggtgaa cgttgcattc
      841 gaattgtggg ccaaacgtaa cataaaaccg gttccagaag tcaaaatcct taacaatctc
      901 ggagtggata tcgcagccaa caccgtgata tgggattata agagggacgc accggcacac
      961 atatccacga tcggggtctg ttccatgact gacatagcta agaagccaac agaaaatatc
     1021 tgcgccccct tgaccgtctt cttcgatggt cgggtgaatg ggcaggtcga tctctttcga
     1081 aacgctcgca atggtgtgtt gataactgaa ggtagtgtta aaggactgca gccgagtgtc
     1141 ggtcctaaac aagcttctct taacggagtc actcttatag gtgaagccct gaaaacgcaa
     1201 ttcaactatt acaagaaagt aaatggagtc gtgcagcaat tgcccgagac ttacttcacg
     1261 caaagtcgta atctgaaaga gttcaagcca aggagccaga tggaaattga tttcttggaa
     1321 cttgcaatgg acgaatttat tgaacgttac aaactggaag gatacgcatt tgaacacatt
     1381 gtgtatggcg acttttctca ccggcaactt ggcggccttc atctgttgat tggtcttgcc
     1441 aaaagatcca aggaaagtcc actcgaactc gaggacttca tccccatgga ttccacggtc
     1501 aagaactact tcattaccga cgcacagact ggtagctcta aatgtgtttg cagtgtgata
     1561 gatttgctcc tcgacgattt cgtggagata attaagagcc aagacctttc agttgttagc
     1621 aaggtggtga aggttactat tgattacaca gaaatatcat ttatgctttg gtgtaaagac
     1681 ggacatgtag agacctttta tccaaagctt caaggcaagc ccattcctaa ccctcttctc
     1741 ggtctcgact caacttagac gcgtcggatc ctgagaactt cagggtgagt ttggggaccc
     1801 ttgattgttc tttctttttc gctattgtaa aattcatgtt atatggaggg ggcaaagttt
     1861 tcagggtgtt gtttagaatg ggaagatgtc ccttgtatca ccatggaccc tcatgataat
     1921 tttgtttctt tcactttcta ctctgttgac aaccattgtc tcctcttatt ttcttttcat
     1981 tttctgtaac tttttcgtta aactttagct tgcatttgta acgaattttt aaattcactt
     2041 ttgtttattt gtcagattgt aagtactttc tctaatcact tttttttcaa ggcaatcagg
     2101 gtatattata ttgtacttca gcacagtttt agagaacaat tgttataatt aaatgataag
     2161 gtagaatatt tctgcatata aattctggct ggcgtggaaa tattcttatt ggtagaaaca
     2221 actacaccct ggtcatcatc ctgcctttct ctttatggtt acaatgatat acactgtttg
     2281 agatgaggat aaaatactct gagtccaaac cgggcccctc tgctaaccat gttcatgcct
     2341 tcttctcttt cctacagctc ctgggcaacg tgctggttgt tgtgctgtct catcattttg
     2401 gcaaagaatt cactcctcag gtgcaggctg cctatcagaa ggtggtggct ggtgtggcca
     2461 atgccctggc tcacaaatac cactgagatc tttttccctc tgccaaaaat tatggggaca
     2521 tcatgaagcc ccttgagcat ctgacttctg gctaataaag gaaatttatt ttcattgcaa
     2581 tagtgtgttg gaattttttg tgtctctcac tcggaaggac atatgggagg gcaaatcatt
     2641 taaaacatca gaatgagtat ttggtttaga gtttggcaac atatgcccat atgctggctg
     2701 ccatgaacaa aggttggcta taaagaggtc atcagtatat gaaacagccc cctgctgtcc
     2761 attccttatt ccatagaaaa gccttgactt gaggttagat tttttttata ttttgttttg
     2821 tgttattttt ttctttaaca tccctaaaat tttccttaca tgttttacta gccagatttt
     2881 tcctcctctc ctgactactc ccagtcatag ctgtccctct tctcttatgg agatccctcg
     2941 aggagctttt tgcaaaagcc taggcctcca aaaaagcctc ttcactactt ctggaatagc
     3001 tcagaggccg aggcggcctc ggcctctgca taaataaaaa aaattagtca gccatggggc
     3061 ggagaatggg cggaactggg cggagttagg ggcgggatgg gcggagttag gggcgggact
     3121 atggttgctg actaattgct gcattaatga atcggccaac gcgcggggag aggcggtttg
     3181 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     3241 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     3301 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     3361 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     3421 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     3481 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     3541 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     3601 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     3661 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     3721 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     3781 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     3841 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     3901 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     3961 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     4021 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     4081 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     4141 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     4201 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     4261 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     4321 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     4381 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     4441 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     4501 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     4561 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     4621 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     4681 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     4741 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     4801 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     4861 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     4921 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     4981 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     5041 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     5101 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
     5161 tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa
     5221 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg
     5281 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac
     5341 tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac
     5401 agatgcgtaa ggagaaaata ccgcatcagg aaattgtaag cgttaatatt ttgttaaaat
     5461 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     5521 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     5581 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     5641 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     5701 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     5761 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     5821 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     5881 gcgcgtccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
     5941 ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac
     6001 gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgaattgt aatacgactc
     6061 actatagggc gaattcgagc tcgccc
//
