LOCUS       sequence-353804-        5867 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector for SARS-CoV-1 ORF3a, V5-tagged. 
            Sequence codon-optimized..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5867)
  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 5867)
  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 C-V5-tag ORF3a
FEATURES             Location/Qualifiers
     source          1..5867
                     /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             1495..1536
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     intron          1566..2138
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     complement(2270..2289)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2335..2390
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2389..2408)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      2744..2879
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2985..3002)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3156..3744)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3236..3255)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3915..4706,4707..4775))
                     /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     4538..4557
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4776..4880)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4948..4966
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5004..5026)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5126..5145
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(5212..5667)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5349..5370)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5561..5580
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5793..5815
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5807..5824
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5808..5824
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5831..5850
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5831..5849
                     /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 ccatggacct gttcatgaga ttcttcaccc tgagatccat caccgcccag
      721 cccgtgaaga ttgacaacgc cagccccgcc agcaccgtgc acgccaccgc caccatcccc
      781 ctgcaggcct ccctgccctt cggctggctc gtgatcggcg tggccttcct ggccgtgttc
      841 cagagcgcca ccaagatcat cgccctgaac aagcggtggc agctggccct gtacaaaggc
      901 ttccagttca tctgcaacct gctgctgctg ttcgtgacca tctacagcca cctgctgctg
      961 gtggccgctg gcatggaggc ccagttcctg tacctgtacg ccctgatcta cttcctgcag
     1021 tgcatcaacg cctgcaggat catcatgagg tgctggctgt gctggaagtg caagagcaag
     1081 aaccccctgc tgtacgacgc caactacttc gtgtgctggc acacacacaa ctacgactac
     1141 tgcatcccct acaacagcgt gaccgacacc atcgtggtga ccgagggcga cggcatcagc
     1201 acccccaagc tgaaggagga ctaccagatc ggcggctaca gcgaggatag acattccggc
     1261 gtgaaggatt atgtggtggt gcatggatat tttacagagg tctattacca gctggagagt
     1321 acacagatca caacagacac aggcatcgag aatgccacat ttttcatctt caataagctg
     1381 gtgaaggatc ctccaaatgt gcagatccac accatcgatg gatctagcgg agtggccaat
     1441 ccagccatgg accccatcta cgatgagccc accaccacca ccagcgtgcc cctgggcaag
     1501 cctatcccca accccctgct gggcctggac agcacctgaa cgcgtcggat cctgagaact
     1561 tcagggtgag tttggggacc cttgattgtt ctttcttttt cgctattgta aaattcatgt
     1621 tatatggagg gggcaaagtt ttcagggtgt tgtttagaat gggaagatgt cccttgtatc
     1681 accatggacc ctcatgataa ttttgtttct ttcactttct actctgttga caaccattgt
     1741 ctcctcttat tttcttttca ttttctgtaa ctttttcgtt aaactttagc ttgcatttgt
     1801 aacgaatttt taaattcact tttgtttatt tgtcagattg taagtacttt ctctaatcac
     1861 ttttttttca aggcaatcag ggtatattat attgtacttc agcacagttt tagagaacaa
     1921 ttgttataat taaatgataa ggtagaatat ttctgcatat aaattctggc tggcgtggaa
     1981 atattcttat tggtagaaac aactacaccc tggtcatcat cctgcctttc tctttatggt
     2041 tacaatgata tacactgttt gagatgagga taaaatactc tgagtccaaa ccgggcccct
     2101 ctgctaacca tgttcatgcc ttcttctctt tcctacagct cctgggcaac gtgctggttg
     2161 ttgtgctgtc tcatcatttt ggcaaagaat tcactcctca ggtgcaggct gcctatcaga
     2221 aggtggtggc tggtgtggcc aatgccctgg ctcacaaata ccactgagat ctttttccct
     2281 ctgccaaaaa ttatggggac atcatgaagc cccttgagca tctgacttct ggctaataaa
     2341 ggaaatttat tttcattgca atagtgtgtt ggaatttttt gtgtctctca ctcggaagga
     2401 catatgggag ggcaaatcat ttaaaacatc agaatgagta tttggtttag agtttggcaa
     2461 catatgccca tatgctggct gccatgaaca aaggttggct ataaagaggt catcagtata
     2521 tgaaacagcc ccctgctgtc cattccttat tccatagaaa agccttgact tgaggttaga
     2581 ttttttttat attttgtttt gtgttatttt tttctttaac atccctaaaa ttttccttac
     2641 atgttttact agccagattt ttcctcctct cctgactact cccagtcata gctgtccctc
     2701 ttctcttatg gagatccctc gaggagcttt ttgcaaaagc ctaggcctcc aaaaaagcct
     2761 cttcactact tctggaatag ctcagaggcc gaggcggcct cggcctctgc ataaataaaa
     2821 aaaattagtc agccatgggg cggagaatgg gcggaactgg gcggagttag gggcgggatg
     2881 ggcggagtta ggggcgggac tatggttgct gactaattgc tgcattaatg aatcggccaa
     2941 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     3001 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     3061 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     3121 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     3181 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     3241 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     3301 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     3361 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     3421 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     3481 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     3541 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     3601 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     3661 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     3721 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     3781 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     3841 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     3901 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     3961 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     4021 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     4081 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     4141 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     4201 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     4261 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     4321 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     4381 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     4441 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     4501 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     4561 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     4621 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     4681 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     4741 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     4801 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     4861 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc
     4921 attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg
     4981 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct
     5041 tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc
     5101 gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat
     5161 atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag gaaattgtaa
     5221 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     5281 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     5341 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     5401 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
     5461 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     5521 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     5581 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     5641 cgcttaatgc gccgctacag ggcgcgtcca ttcgccattc aggctgcgca actgttggga
     5701 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc
     5761 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc
     5821 cagtgaattg taatacgact cactataggg cgaattcgag ctcgccc
//
