LOCUS       sequence-373991-        5297 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Mammalian expression vector for SARS-CoV-1 Nsp7, V5-tagged. Sequence
            codon-optimized..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5297)
  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 5297)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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 NSP7 C-V5-tag
FEATURES             Location/Qualifiers
     source          1..5297
                     /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             925..966
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     intron          996..1568
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     complement(1700..1719)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    1765..1820
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1819..1838)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      2174..2309
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2415..2432)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2586..3174)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2666..2685)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3345..4136,4137..4205))
                     /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     3968..3987
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4206..4310)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4378..4396
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4434..4456)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4556..4575
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(4642..5097)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4779..4800)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4991..5010
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5223..5245
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5237..5254
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5238..5254
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5261..5280
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5261..5279
                     /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 ccatgtccaa aatgtctgac gtgaaatgca cttctgtagt gctcctttca
      721 gttctccaac agctcagagt ggaaagctcc agtaagttgt gggcgcagtg tgttcaactg
      781 cacaatgata tattgcttgc caaagacacc accgaggctt tcgagaagat ggtctcactt
      841 ctcagcgtct tgctgtcaat gcaaggggca gtggatataa atcgcctctg tgaggaaatg
      901 ctcgacaaca gggccacctt gcaagggaag cccatcccaa acccgcttct ggggctcgat
      961 tcaacatgaa cgcgtcggat cctgagaact tcagggtgag tttggggacc cttgattgtt
     1021 ctttcttttt cgctattgta aaattcatgt tatatggagg gggcaaagtt ttcagggtgt
     1081 tgtttagaat gggaagatgt cccttgtatc accatggacc ctcatgataa ttttgtttct
     1141 ttcactttct actctgttga caaccattgt ctcctcttat tttcttttca ttttctgtaa
     1201 ctttttcgtt aaactttagc ttgcatttgt aacgaatttt taaattcact tttgtttatt
     1261 tgtcagattg taagtacttt ctctaatcac ttttttttca aggcaatcag ggtatattat
     1321 attgtacttc agcacagttt tagagaacaa ttgttataat taaatgataa ggtagaatat
     1381 ttctgcatat aaattctggc tggcgtggaa atattcttat tggtagaaac aactacaccc
     1441 tggtcatcat cctgcctttc tctttatggt tacaatgata tacactgttt gagatgagga
     1501 taaaatactc tgagtccaaa ccgggcccct ctgctaacca tgttcatgcc ttcttctctt
     1561 tcctacagct cctgggcaac gtgctggttg ttgtgctgtc tcatcatttt ggcaaagaat
     1621 tcactcctca ggtgcaggct gcctatcaga aggtggtggc tggtgtggcc aatgccctgg
     1681 ctcacaaata ccactgagat ctttttccct ctgccaaaaa ttatggggac atcatgaagc
     1741 cccttgagca tctgacttct ggctaataaa ggaaatttat tttcattgca atagtgtgtt
     1801 ggaatttttt gtgtctctca ctcggaagga catatgggag ggcaaatcat ttaaaacatc
     1861 agaatgagta tttggtttag agtttggcaa catatgccca tatgctggct gccatgaaca
     1921 aaggttggct ataaagaggt catcagtata tgaaacagcc ccctgctgtc cattccttat
     1981 tccatagaaa agccttgact tgaggttaga ttttttttat attttgtttt gtgttatttt
     2041 tttctttaac atccctaaaa ttttccttac atgttttact agccagattt ttcctcctct
     2101 cctgactact cccagtcata gctgtccctc ttctcttatg gagatccctc gaggagcttt
     2161 ttgcaaaagc ctaggcctcc aaaaaagcct cttcactact tctggaatag ctcagaggcc
     2221 gaggcggcct cggcctctgc ataaataaaa aaaattagtc agccatgggg cggagaatgg
     2281 gcggaactgg gcggagttag gggcgggatg ggcggagtta ggggcgggac tatggttgct
     2341 gactaattgc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     2401 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     2461 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     2521 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     2581 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     2641 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     2701 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     2761 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     2821 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     2881 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     2941 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     3001 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     3061 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     3121 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     3181 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     3241 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     3301 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     3361 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     3421 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     3481 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     3541 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     3601 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     3661 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     3721 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     3781 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     3841 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     3901 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     3961 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     4021 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     4081 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     4141 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     4201 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     4261 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     4321 cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat
     4381 aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga
     4441 cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa
     4501 gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca
     4561 tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta
     4621 aggagaaaat accgcatcag gaaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     4681 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     4741 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     4801 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     4861 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     4921 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     4981 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     5041 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcca
     5101 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
     5161 acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt
     5221 ttcccagtca cgacgttgta aaacgacggc cagtgaattg taatacgact cactataggg
     5281 cgaattcgag ctcgccc
//
