LOCUS       sequence-353892-        5408 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector for RaTG13 ORF7a, V5-tagged. Sequence 
            codon-optimized..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5408)
  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 5408)
  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_batCoV RaTG13 ORF7a C-V5-tag
FEATURES             Location/Qualifiers
     source          1..5408
                     /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             1036..1077
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     intron          1107..1679
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     complement(1811..1830)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    1876..1931
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1930..1949)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      2285..2420
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2526..2543)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2697..3285)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2777..2796)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3456..4247,4248..4316))
                     /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     4079..4098
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4317..4421)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4489..4507
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4545..4567)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4667..4686
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(4753..5208)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4890..4911)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5102..5121
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5334..5356
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5348..5365
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5349..5365
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5372..5391
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5372..5390
                     /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 ccatgaaaat tatccttttc ctcgtgctgg tcacgttggc tacctgcgaa
      721 ctctatcact atcaagaatg cgtaagagga actacggtgc tcctcaagga accctgttcc
      781 agtggcacct atgaaggtaa ttctcctttc catccgctcg ccgataataa gtttgcactg
      841 acttgcttta gtactcagtt cgccttcgca tgcccggatg gtgtaaagca tgtttaccaa
      901 ctgagagccc ggagcgtgag tccaaaattg tttatccgac aggaagaggt tcaagaactt
      961 tatagcccta ttttccttat tattgccgcc atagtattca tcactttgtg tttcaccctg
     1021 aaaaggaaaa ccgaaggtaa acccatacct aatcctttgc ttggcctcga ttctacttaa
     1081 acgcgtcgga tcctgagaac ttcagggtga gtttggggac ccttgattgt tctttctttt
     1141 tcgctattgt aaaattcatg ttatatggag ggggcaaagt tttcagggtg ttgtttagaa
     1201 tgggaagatg tcccttgtat caccatggac cctcatgata attttgtttc tttcactttc
     1261 tactctgttg acaaccattg tctcctctta ttttcttttc attttctgta actttttcgt
     1321 taaactttag cttgcatttg taacgaattt ttaaattcac ttttgtttat ttgtcagatt
     1381 gtaagtactt tctctaatca cttttttttc aaggcaatca gggtatatta tattgtactt
     1441 cagcacagtt ttagagaaca attgttataa ttaaatgata aggtagaata tttctgcata
     1501 taaattctgg ctggcgtgga aatattctta ttggtagaaa caactacacc ctggtcatca
     1561 tcctgccttt ctctttatgg ttacaatgat atacactgtt tgagatgagg ataaaatact
     1621 ctgagtccaa accgggcccc tctgctaacc atgttcatgc cttcttctct ttcctacagc
     1681 tcctgggcaa cgtgctggtt gttgtgctgt ctcatcattt tggcaaagaa ttcactcctc
     1741 aggtgcaggc tgcctatcag aaggtggtgg ctggtgtggc caatgccctg gctcacaaat
     1801 accactgaga tctttttccc tctgccaaaa attatgggga catcatgaag ccccttgagc
     1861 atctgacttc tggctaataa aggaaattta ttttcattgc aatagtgtgt tggaattttt
     1921 tgtgtctctc actcggaagg acatatggga gggcaaatca tttaaaacat cagaatgagt
     1981 atttggttta gagtttggca acatatgccc atatgctggc tgccatgaac aaaggttggc
     2041 tataaagagg tcatcagtat atgaaacagc cccctgctgt ccattcctta ttccatagaa
     2101 aagccttgac ttgaggttag atttttttta tattttgttt tgtgttattt ttttctttaa
     2161 catccctaaa attttcctta catgttttac tagccagatt tttcctcctc tcctgactac
     2221 tcccagtcat agctgtccct cttctcttat ggagatccct cgaggagctt tttgcaaaag
     2281 cctaggcctc caaaaaagcc tcttcactac ttctggaata gctcagaggc cgaggcggcc
     2341 tcggcctctg cataaataaa aaaaattagt cagccatggg gcggagaatg ggcggaactg
     2401 ggcggagtta ggggcgggat gggcggagtt aggggcggga ctatggttgc tgactaattg
     2461 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     2521 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     2581 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     2641 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     2701 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     2761 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     2821 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     2881 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     2941 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     3001 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     3061 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     3121 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     3181 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     3241 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     3301 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     3361 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     3421 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     3481 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     3541 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     3601 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     3661 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     3721 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     3781 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     3841 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     3901 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     3961 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     4021 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     4081 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     4141 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     4201 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     4261 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     4321 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     4381 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     4441 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     4501 acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
     4561 ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag
     4621 ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag
     4681 attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa
     4741 taccgcatca ggaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
     4801 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     4861 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     4921 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     4981 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     5041 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     5101 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
     5161 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc attcgccatt
     5221 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     5281 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     5341 acgacgttgt aaaacgacgg ccagtgaatt gtaatacgac tcactatagg gcgaattcga
     5401 gctcgccc
//
