LOCUS       sequence-353866-        5270 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector for SARS-CoV-2 E or bat RaTG13 E, 
            V5-tagged (SARS-CoV-2 E and RaTG13 E have the same amino acid 
            sequence and codon optimized sequence)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5270)
  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 5270)
  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/bat-CoV RaTG13 E C-V5-tag
FEATURES             Location/Qualifiers
     source          1..5270
                     /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             898..939
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     intron          969..1541
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     complement(1673..1692)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    1738..1793
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1792..1811)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      2147..2282
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2388..2405)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2559..3147)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2639..2658)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3318..4109,4110..4178))
                     /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     3941..3960
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4179..4283)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4351..4369
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4407..4429)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4529..4548
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(4615..5070)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4752..4773)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4964..4983
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5196..5218
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5210..5227
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5211..5227
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5234..5253
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5234..5252
                     /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 ccatgtatag ttttgtctct gaagaaacag gtacattgat tgtgaatagc
      721 gtgcttttgt ttctggcctt tgttgtattt ctgcttgtga ccctggcaat cttgacagcg
      781 ctccggctgt gtgcctattg ttgcaatatc gtgaacgtct cattggttaa gccctcattc
      841 tacgtttatt ccagagtgaa gaatctcaat agctctcggg tgccagatct cctggtcggc
      901 aagcctatac caaaccctct ccttggcctt gatagcacct agacgcgtcg gatcctgaga
      961 acttcagggt gagtttgggg acccttgatt gttctttctt tttcgctatt gtaaaattca
     1021 tgttatatgg agggggcaaa gttttcaggg tgttgtttag aatgggaaga tgtcccttgt
     1081 atcaccatgg accctcatga taattttgtt tctttcactt tctactctgt tgacaaccat
     1141 tgtctcctct tattttcttt tcattttctg taactttttc gttaaacttt agcttgcatt
     1201 tgtaacgaat ttttaaattc acttttgttt atttgtcaga ttgtaagtac tttctctaat
     1261 cacttttttt tcaaggcaat cagggtatat tatattgtac ttcagcacag ttttagagaa
     1321 caattgttat aattaaatga taaggtagaa tatttctgca tataaattct ggctggcgtg
     1381 gaaatattct tattggtaga aacaactaca ccctggtcat catcctgcct ttctctttat
     1441 ggttacaatg atatacactg tttgagatga ggataaaata ctctgagtcc aaaccgggcc
     1501 cctctgctaa ccatgttcat gccttcttct ctttcctaca gctcctgggc aacgtgctgg
     1561 ttgttgtgct gtctcatcat tttggcaaag aattcactcc tcaggtgcag gctgcctatc
     1621 agaaggtggt ggctggtgtg gccaatgccc tggctcacaa ataccactga gatctttttc
     1681 cctctgccaa aaattatggg gacatcatga agccccttga gcatctgact tctggctaat
     1741 aaaggaaatt tattttcatt gcaatagtgt gttggaattt tttgtgtctc tcactcggaa
     1801 ggacatatgg gagggcaaat catttaaaac atcagaatga gtatttggtt tagagtttgg
     1861 caacatatgc ccatatgctg gctgccatga acaaaggttg gctataaaga ggtcatcagt
     1921 atatgaaaca gccccctgct gtccattcct tattccatag aaaagccttg acttgaggtt
     1981 agattttttt tatattttgt tttgtgttat ttttttcttt aacatcccta aaattttcct
     2041 tacatgtttt actagccaga tttttcctcc tctcctgact actcccagtc atagctgtcc
     2101 ctcttctctt atggagatcc ctcgaggagc tttttgcaaa agcctaggcc tccaaaaaag
     2161 cctcttcact acttctggaa tagctcagag gccgaggcgg cctcggcctc tgcataaata
     2221 aaaaaaatta gtcagccatg gggcggagaa tgggcggaac tgggcggagt taggggcggg
     2281 atgggcggag ttaggggcgg gactatggtt gctgactaat tgctgcatta atgaatcggc
     2341 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     2401 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     2461 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     2521 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     2581 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     2641 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     2701 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     2761 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     2821 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     2881 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     2941 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     3001 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     3061 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     3121 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     3181 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     3241 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     3301 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     3361 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     3421 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     3481 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     3541 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     3601 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     3661 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     3721 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     3781 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     3841 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     3901 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     3961 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     4021 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     4081 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     4141 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     4201 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     4261 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     4321 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc
     4381 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
     4441 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
     4501 ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac
     4561 catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggaaattg
     4621 taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta
     4681 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt
     4741 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
     4801 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa
     4861 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
     4921 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
     4981 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
     5041 ccgcgcttaa tgcgccgcta cagggcgcgt ccattcgcca ttcaggctgc gcaactgttg
     5101 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc
     5161 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac
     5221 ggccagtgaa ttgtaatacg actcactata gggcgaattc gagctcgccc
//
