LOCUS       sequence-353888-        6302 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector for RaTG13 N, V5-tagged. Sequence 
            codon-optimized..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6302)
  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 6302)
  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 N C-V5-tag
FEATURES             Location/Qualifiers
     source          1..6302
                     /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             1930..1971
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     intron          2001..2573
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     complement(2705..2724)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2770..2825
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2824..2843)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      3179..3314
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3420..3437)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3591..4179)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3671..3690)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4350..5141,5142..5210))
                     /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     4973..4992
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5211..5315)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5383..5401
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5439..5461)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5561..5580
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(5647..6102)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5784..5805)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5996..6015
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6228..6250
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6242..6259
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6243..6259
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6266..6285
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6266..6284
                     /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 ccatgtctga caatggtccc cagaatcagc gaaatgctcc ccggatcacc
      721 tttggtgggc cctctgacag caccggaagt aatcaaaatg gagagcggag tggtgccagg
      781 cctaagcaaa ggagacctca agggctgccg aacaatacag catcctggtt tacagctttg
      841 actcaacatg gcaaagaaga cttgaaattt cctcgaggcc agggagtgcc aataaacacc
      901 aattcctctc ctgacgacca aatcggatat tataggcgcg ccactcgcag aatccggggc
      961 ggagatggga agatgaaaga cctctctcca cgttggtatt tctattatct gggaactggc
     1021 cctgaagccg ggctccctta cggggctaat aaagacggga tcatctgggt agctacagaa
     1081 ggggcactta acacccctaa agatcacata ggcactagga atcccgcgaa caatgctgcg
     1141 atagtgcttc agcttcctca aggaacaact ctccccaaag gattttacgc cgagggtagc
     1201 agaggcggta gtcaagccag ttcaaggagc tctagccgct cccgcaactc aagtcgtaac
     1261 tccacgccag gtagctctcg gggcacaagc ccggcacgaa tggcgggaaa cggaagcgat
     1321 gctgcactcg ccttgctgct cctggaccgc ctgaaccaac tggaatctaa gatgtccggg
     1381 aaaggacaac aacaacaatc tcagacagtg accaagaaga gtgctgctga agcgagtaag
     1441 aagccgagac agaaacggac cgcaacaaag cagtataacg taacccaagc attcggccgc
     1501 cgcggccctg aacaaactca gggcaacttc ggcgaccaag aactcattcg gcagggaacc
     1561 gactataagc attggccgca gattgcacag tttgcgccga gcgcgtccgc cttctttggg
     1621 atgtctcgga tcggaatgga agtgaccccg tccggtacgt ggctgaccta tacaggagcg
     1681 ataaaactcg acgacaagga ccctaatttt aaggatcaag taattttgct gaataaacac
     1741 atagacgcat acaagacatt tccccctacg gaacctaaga aggacaagaa gaagaaagcc
     1801 gatgagactc aagctctgcc acaacgccaa aagaaacaac aaaccgttac gctcctgcct
     1861 gccgctgatc ttgatgactt tagtaagcaa ctccaacaat ccatgagtag cgcggacagc
     1921 actcaggccg gaaaaccaat tcctaatcct ctcctcggcc tcgatagcac gtaaacgcgt
     1981 cggatcctga gaacttcagg gtgagtttgg ggacccttga ttgttctttc tttttcgcta
     2041 ttgtaaaatt catgttatat ggagggggca aagttttcag ggtgttgttt agaatgggaa
     2101 gatgtccctt gtatcaccat ggaccctcat gataattttg tttctttcac tttctactct
     2161 gttgacaacc attgtctcct cttattttct tttcattttc tgtaactttt tcgttaaact
     2221 ttagcttgca tttgtaacga atttttaaat tcacttttgt ttatttgtca gattgtaagt
     2281 actttctcta atcacttttt tttcaaggca atcagggtat attatattgt acttcagcac
     2341 agttttagag aacaattgtt ataattaaat gataaggtag aatatttctg catataaatt
     2401 ctggctggcg tggaaatatt cttattggta gaaacaacta caccctggtc atcatcctgc
     2461 ctttctcttt atggttacaa tgatatacac tgtttgagat gaggataaaa tactctgagt
     2521 ccaaaccggg cccctctgct aaccatgttc atgccttctt ctctttccta cagctcctgg
     2581 gcaacgtgct ggttgttgtg ctgtctcatc attttggcaa agaattcact cctcaggtgc
     2641 aggctgccta tcagaaggtg gtggctggtg tggccaatgc cctggctcac aaataccact
     2701 gagatctttt tccctctgcc aaaaattatg gggacatcat gaagcccctt gagcatctga
     2761 cttctggcta ataaaggaaa tttattttca ttgcaatagt gtgttggaat tttttgtgtc
     2821 tctcactcgg aaggacatat gggagggcaa atcatttaaa acatcagaat gagtatttgg
     2881 tttagagttt ggcaacatat gcccatatgc tggctgccat gaacaaaggt tggctataaa
     2941 gaggtcatca gtatatgaaa cagccccctg ctgtccattc cttattccat agaaaagcct
     3001 tgacttgagg ttagattttt tttatatttt gttttgtgtt atttttttct ttaacatccc
     3061 taaaattttc cttacatgtt ttactagcca gatttttcct cctctcctga ctactcccag
     3121 tcatagctgt ccctcttctc ttatggagat ccctcgagga gctttttgca aaagcctagg
     3181 cctccaaaaa agcctcttca ctacttctgg aatagctcag aggccgaggc ggcctcggcc
     3241 tctgcataaa taaaaaaaat tagtcagcca tggggcggag aatgggcgga actgggcgga
     3301 gttaggggcg ggatgggcgg agttaggggc gggactatgg ttgctgacta attgctgcat
     3361 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     3421 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     3481 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     3541 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     3601 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     3661 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     3721 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     3781 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     3841 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     3901 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     3961 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     4021 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     4081 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     4141 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     4201 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     4261 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     4321 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     4381 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     4441 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     4501 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     4561 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     4621 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     4681 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     4741 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     4801 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     4861 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     4921 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     4981 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     5041 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     5101 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     5161 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     5221 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     5281 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     5341 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     5401 ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     5461 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     5521 tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta
     5581 ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc
     5641 atcaggaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     5701 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     5761 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     5821 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     5881 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     5941 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     6001 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     6061 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtccattcgc cattcaggct
     6121 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
     6181 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
     6241 ttgtaaaacg acggccagtg aattgtaata cgactcacta tagggcgaat tcgagctcgc
     6301 cc
//
