LOCUS       sequence-353818-        5234 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Mammalian expression vector for SARS-CoV-1 ORF6, V5-tagged. Sequence
            codon-optimized..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5234)
  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 5234)
  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 ORF6 C-V5-tag
FEATURES             Location/Qualifiers
     source          1..5234
                     /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             862..903
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     intron          933..1505
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     complement(1637..1656)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    1702..1757
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1756..1775)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      2111..2246
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2352..2369)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2523..3111)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2603..2622)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3282..4073,4074..4142))
                     /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     3905..3924
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4143..4247)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4315..4333
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4371..4393)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4493..4512
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(4579..5034)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4716..4737)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4928..4947
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5160..5182
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5174..5191
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5175..5191
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5198..5217
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5198..5216
                     /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 ccatgttcca cttggtggat tttcaagtga ccatcgctga aatccttatc
      721 attatcatgc gcacatttag aatagcaatc tggaacttgg atgtgatcat tagctctatt
      781 gtaaggcagt tgttcaaacc attgacaaag aagaactact ccgaactgga cgacgaggag
      841 cccatggaac tggactaccc cggcaaaccc attccaaatc cccttcttgg actggacagt
      901 acttaaacgc gtcggatcct gagaacttca gggtgagttt ggggaccctt gattgttctt
      961 tctttttcgc tattgtaaaa ttcatgttat atggaggggg caaagttttc agggtgttgt
     1021 ttagaatggg aagatgtccc ttgtatcacc atggaccctc atgataattt tgtttctttc
     1081 actttctact ctgttgacaa ccattgtctc ctcttatttt cttttcattt tctgtaactt
     1141 tttcgttaaa ctttagcttg catttgtaac gaatttttaa attcactttt gtttatttgt
     1201 cagattgtaa gtactttctc taatcacttt tttttcaagg caatcagggt atattatatt
     1261 gtacttcagc acagttttag agaacaattg ttataattaa atgataaggt agaatatttc
     1321 tgcatataaa ttctggctgg cgtggaaata ttcttattgg tagaaacaac tacaccctgg
     1381 tcatcatcct gcctttctct ttatggttac aatgatatac actgtttgag atgaggataa
     1441 aatactctga gtccaaaccg ggcccctctg ctaaccatgt tcatgccttc ttctctttcc
     1501 tacagctcct gggcaacgtg ctggttgttg tgctgtctca tcattttggc aaagaattca
     1561 ctcctcaggt gcaggctgcc tatcagaagg tggtggctgg tgtggccaat gccctggctc
     1621 acaaatacca ctgagatctt tttccctctg ccaaaaatta tggggacatc atgaagcccc
     1681 ttgagcatct gacttctggc taataaagga aatttatttt cattgcaata gtgtgttgga
     1741 attttttgtg tctctcactc ggaaggacat atgggagggc aaatcattta aaacatcaga
     1801 atgagtattt ggtttagagt ttggcaacat atgcccatat gctggctgcc atgaacaaag
     1861 gttggctata aagaggtcat cagtatatga aacagccccc tgctgtccat tccttattcc
     1921 atagaaaagc cttgacttga ggttagattt tttttatatt ttgttttgtg ttattttttt
     1981 ctttaacatc cctaaaattt tccttacatg ttttactagc cagatttttc ctcctctcct
     2041 gactactccc agtcatagct gtccctcttc tcttatggag atccctcgag gagctttttg
     2101 caaaagccta ggcctccaaa aaagcctctt cactacttct ggaatagctc agaggccgag
     2161 gcggcctcgg cctctgcata aataaaaaaa attagtcagc catggggcgg agaatgggcg
     2221 gaactgggcg gagttagggg cgggatgggc ggagttaggg gcgggactat ggttgctgac
     2281 taattgctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     2341 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     2401 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     2461 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     2521 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     2581 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     2641 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     2701 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     2761 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     2821 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     2881 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     2941 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     3001 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     3061 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     3121 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     3181 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     3241 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     3301 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     3361 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     3421 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     3481 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     3541 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     3601 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     3661 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     3721 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     3781 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     3841 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     3901 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     3961 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     4021 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     4081 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     4141 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     4201 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     4261 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     4321 cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
     4381 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
     4441 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca
     4501 gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg
     4561 agaaaatacc gcatcaggaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     4621 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     4681 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     4741 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     4801 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     4861 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     4921 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     4981 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtccattc
     5041 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     5101 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
     5161 ccagtcacga cgttgtaaaa cgacggccag tgaattgtaa tacgactcac tatagggcga
     5221 attcgagctc gccc
//
