LOCUS       sequence-358584-        6635 bp    DNA     circular SYN 11-DEC-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 6635)
  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 6635)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 11, 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..6635
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        304..507
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     458..478
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     504..528
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     5'UTR           597..636
                     /label=HSV TK 5'-UTR
                     /note="5' untranslated region from the herpes simplex virus
                     thymidine kinase gene"
     CDS             924..965
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     intron          995..1567
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     complement(1699..1718)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    1764..1819
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1818..1837)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      2173..2308
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2414..2431)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2585..3173)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2665..2684)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4682..5473,5474..5542))
                     /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     5305..5324
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5543..5647)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5715..5733
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5771..5793)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5893..5912
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(5979..6434)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6116..6137)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6328..6347
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6560..6582
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6574..6591
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6575..6591
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6598..6617
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6598..6616
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     enhancer        join(6635,1..303)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
       61 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
      121 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
      181 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
      241 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
      301 atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga
      361 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
      421 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta
      481 cggtgggagg tctatataag cagagctcgt ttagtgaacc gtcagatcgc ctggagacgc
      541 catccacgct gttttgacct ccatagaaga caccgggacc gatccagcct ccgggggatc
      601 ttggtggcgt gaaactcccg cacctcttcg gccagcgcct tgtagaagcg cgtaggcctt
      661 ctagagccac catgtccaaa atgtctgacg tgaaatgcac ttctgtagtg ctcctttcag
      721 ttctccaaca gctcagagtg gaaagctcca gtaagttgtg ggcgcagtgt gttcaactgc
      781 acaatgatat attgcttgcc aaagacacca ccgaggcttt cgagaagatg gtctcacttc
      841 tcagcgtctt gctgtcaatg caaggggcag tggatataaa tcgcctctgt gaggaaatgc
      901 tcgacaacag ggccaccttg caagggaagc ccatcccaaa cccgcttctg gggctcgatt
      961 caacatgaac gcgtcggatc ctgagaactt cagggtgagt ttggggaccc ttgattgttc
     1021 tttctttttc gctattgtaa aattcatgtt atatggaggg ggcaaagttt tcagggtgtt
     1081 gtttagaatg ggaagatgtc ccttgtatca ccatggaccc tcatgataat tttgtttctt
     1141 tcactttcta ctctgttgac aaccattgtc tcctcttatt ttcttttcat tttctgtaac
     1201 tttttcgtta aactttagct tgcatttgta acgaattttt aaattcactt ttgtttattt
     1261 gtcagattgt aagtactttc tctaatcact tttttttcaa ggcaatcagg gtatattata
     1321 ttgtacttca gcacagtttt agagaacaat tgttataatt aaatgataag gtagaatatt
     1381 tctgcatata aattctggct ggcgtggaaa tattcttatt ggtagaaaca actacaccct
     1441 ggtcatcatc ctgcctttct ctttatggtt acaatgatat acactgtttg agatgaggat
     1501 aaaatactct gagtccaaac cgggcccctc tgctaaccat gttcatgcct tcttctcttt
     1561 cctacagctc ctgggcaacg tgctggttgt tgtgctgtct catcattttg gcaaagaatt
     1621 cactcctcag gtgcaggctg cctatcagaa ggtggtggct ggtgtggcca atgccctggc
     1681 tcacaaatac cactgagatc tttttccctc tgccaaaaat tatggggaca tcatgaagcc
     1741 ccttgagcat ctgacttctg gctaataaag gaaatttatt ttcattgcaa tagtgtgttg
     1801 gaattttttg tgtctctcac tcggaaggac atatgggagg gcaaatcatt taaaacatca
     1861 gaatgagtat ttggtttaga gtttggcaac atatgcccat atgctggctg ccatgaacaa
     1921 aggttggcta taaagaggtc atcagtatat gaaacagccc cctgctgtcc attccttatt
     1981 ccatagaaaa gccttgactt gaggttagat tttttttata ttttgttttg tgttattttt
     2041 ttctttaaca tccctaaaat tttccttaca tgttttacta gccagatttt tcctcctctc
     2101 ctgactactc ccagtcatag ctgtccctct tctcttatgg agatccctcg aggagctttt
     2161 tgcaaaagcc taggcctcca aaaaagcctc ttcactactt ctggaatagc tcagaggccg
     2221 aggcggcctc ggcctctgca taaataaaaa aaattagtca gccatggggc ggagaatggg
     2281 cggaactggg cggagttagg ggcgggatgg gcggagttag gggcgggact atggttgctg
     2341 actaattgct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     2401 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     2461 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     2521 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     2581 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     2641 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     2701 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     2761 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     2821 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     2881 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     2941 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     3001 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     3061 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     3121 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     3181 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     3241 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     3301 ttaaatcaat ctaaagtact gagagatccc ctcataattt ccccaaagcg taaccatgtg
     3361 tgaataaatt ttgagctagt agggttgcag ccacgagtaa gtcttccctt gttattgtgt
     3421 agccagaatg ccgcaaaact tccatgccta agcgaactgt tgagagtacg tttcgatttc
     3481 tgactgtgtt agcctggaag tgcttgtccc aaccttgttt ctgagcatga acgcccgcaa
     3541 gccaacatgt tagttgaagc atcagggcga ttagcagcat gatatcaaaa cgctctgagc
     3601 tgctcgttcg gctatggcgt aggcctagtc cgtaggcagg acttttcaag tctcggaagg
     3661 tttcttcaat ctgcattcgc ttcgaataga tattaacaag ttgtttgggt gttcgaattt
     3721 caacaggtaa gttagttgct agaatccatg gctcctttgc cgacgctgag tagattttag
     3781 gtgacgggtg gtgacaatga gtccgtgtcg agcgctgatt ttttcggcct ttagagcgag
     3841 atttatacaa tagaatttgg catgagattg gattgctttt agtcagcctc ttatagccta
     3901 aagtctttga gtgactagat gacatatcat gtaagttgct gataggtttc cagttttccg
     3961 ctcctaggtc tgcatattgt acttttcctc ttactcgact taaccagtac caacccagct
     4021 tctcaacgga tttataccat ggcactttaa agccagcatc actgacaatg agcggtgtgg
     4081 tgttactcgg tagaatgctc gcaaggtcgg ctagaaattg gtcatgagct ttctttgaac
     4141 attgctctga aagcgggaac gctttctcat aaagagtaac agaacgaccg tgtagtgcga
     4201 ctgaagctcg caataccata agccgttttt gctcacggat atcagaccag tcaacaagta
     4261 caatgggcat cgtattgccc gaacagataa agctagcatg ccaacggtat acagcgagtc
     4321 gctctttgtg gaggtgacga ttacctaaca atcggtcgat tcgtttgatg ttatgttttg
     4381 ttctcgcttt ggttggcagg ttacggccaa gttcggtaag agtgagagtt ttacagtcaa
     4441 gtaaggcgtg gcaagccaac gttaagctgt tgagtcgttt taagtgtaat tcggggcaga
     4501 attggtaaag agagtcgtgt aaaatatcga gttcgcacat tttgttgtct gattattgat
     4561 ttttggcgaa accatttgat catatgacaa gatgtgtatc taccttaact taatgatttt
     4621 gataaaaatc attaggggat tcatcagtct aaagtatata tgagtaaact tggtctgaca
     4681 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     4741 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     4801 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     4861 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     4921 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     4981 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     5041 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     5101 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     5161 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     5221 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     5281 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     5341 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     5401 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     5461 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     5521 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     5581 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     5641 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     5701 cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg
     5761 acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg
     5821 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct
     5881 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa
     5941 taccgcacag atgcgtaagg agaaaatacc gcatcaggaa attgtaagcg ttaatatttt
     6001 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     6061 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     6121 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     6181 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     6241 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     6301 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     6361 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     6421 gctacagggc gcgtccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
     6481 cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt
     6541 tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgaattgtaa
     6601 tacgactcac tatagggcga attcgagctc gcccc
//
