LOCUS       sequence-353722-        5273 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector for SARS-CoV-1 E, V5-tagged. Sequence 
            codon-optimized..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5273)
  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 5273)
  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 E C-V5-tag
FEATURES             Location/Qualifiers
     source          1..5273
                     /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             901..942
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     intron          972..1544
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     complement(1676..1695)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    1741..1796
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1795..1814)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      2150..2285
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2391..2408)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2562..3150)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2642..2661)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3321..4112,4113..4181))
                     /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     3944..3963
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4182..4286)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4354..4372
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4410..4432)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4532..4551
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(4618..5073)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4755..4776)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4967..4986
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5199..5221
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5213..5230
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5214..5230
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5237..5256
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5237..5255
                     /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 ctttgtgagc gaggagacag gaacactgat tgtgaattct
      721 gtgctgctgt ttctggcctt cgtggtgttt ctgctggtga cactggccat tctgacagcc
      781 ctgagactgt gtgcctattg ctgtaatatc gtgaatgtgt ctctggtgaa acctaccgtg
      841 tacgtgtaca gcagagtgaa gaatctgaat agctctgagg gcgtgcctga tctgctggtg
      901 ggcaagccta tccctaatcc tctgctgggc ctggattcta catgaacgcg tcggatcctg
      961 agaacttcag ggtgagtttg gggacccttg attgttcttt ctttttcgct attgtaaaat
     1021 tcatgttata tggagggggc aaagttttca gggtgttgtt tagaatggga agatgtccct
     1081 tgtatcacca tggaccctca tgataatttt gtttctttca ctttctactc tgttgacaac
     1141 cattgtctcc tcttattttc ttttcatttt ctgtaacttt ttcgttaaac tttagcttgc
     1201 atttgtaacg aatttttaaa ttcacttttg tttatttgtc agattgtaag tactttctct
     1261 aatcactttt ttttcaaggc aatcagggta tattatattg tacttcagca cagttttaga
     1321 gaacaattgt tataattaaa tgataaggta gaatatttct gcatataaat tctggctggc
     1381 gtggaaatat tcttattggt agaaacaact acaccctggt catcatcctg cctttctctt
     1441 tatggttaca atgatataca ctgtttgaga tgaggataaa atactctgag tccaaaccgg
     1501 gcccctctgc taaccatgtt catgccttct tctctttcct acagctcctg ggcaacgtgc
     1561 tggttgttgt gctgtctcat cattttggca aagaattcac tcctcaggtg caggctgcct
     1621 atcagaaggt ggtggctggt gtggccaatg ccctggctca caaataccac tgagatcttt
     1681 ttccctctgc caaaaattat ggggacatca tgaagcccct tgagcatctg acttctggct
     1741 aataaaggaa atttattttc attgcaatag tgtgttggaa ttttttgtgt ctctcactcg
     1801 gaaggacata tgggagggca aatcatttaa aacatcagaa tgagtatttg gtttagagtt
     1861 tggcaacata tgcccatatg ctggctgcca tgaacaaagg ttggctataa agaggtcatc
     1921 agtatatgaa acagccccct gctgtccatt ccttattcca tagaaaagcc ttgacttgag
     1981 gttagatttt ttttatattt tgttttgtgt tatttttttc tttaacatcc ctaaaatttt
     2041 ccttacatgt tttactagcc agatttttcc tcctctcctg actactccca gtcatagctg
     2101 tccctcttct cttatggaga tccctcgagg agctttttgc aaaagcctag gcctccaaaa
     2161 aagcctcttc actacttctg gaatagctca gaggccgagg cggcctcggc ctctgcataa
     2221 ataaaaaaaa ttagtcagcc atggggcgga gaatgggcgg aactgggcgg agttaggggc
     2281 gggatgggcg gagttagggg cgggactatg gttgctgact aattgctgca ttaatgaatc
     2341 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     2401 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     2461 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     2521 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     2581 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     2641 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     2701 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     2761 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     2821 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     2881 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     2941 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     3001 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     3061 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     3121 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     3181 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     3241 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     3301 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     3361 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     3421 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     3481 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     3541 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     3601 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     3661 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     3721 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     3781 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     3841 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     3901 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     3961 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     4021 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     4081 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     4141 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     4201 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     4261 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     4321 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
     4381 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
     4441 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
     4501 gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg
     4561 caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggaaa
     4621 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     4681 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
     4741 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     4801 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
     4861 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     4921 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     4981 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     5041 ccgccgcgct taatgcgccg ctacagggcg cgtccattcg ccattcaggc tgcgcaactg
     5101 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg
     5161 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac
     5221 gacggccagt gaattgtaat acgactcact atagggcgaa ttcgagctcg ccc
//
