LOCUS       sequence-353889-        5870 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector for RaTG13 ORF3a, V5-tagged. Sequence 
            codon-optimized..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5870)
  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 5870)
  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 ORF3a C-V5-tag
FEATURES             Location/Qualifiers
     source          1..5870
                     /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             1498..1539
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     intron          1569..2141
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     complement(2273..2292)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2338..2393
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2392..2411)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      2747..2882
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2988..3005)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3159..3747)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3239..3258)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3918..4709,4710..4778))
                     /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     4541..4560
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4779..4883)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4951..4969
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5007..5029)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5129..5148
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(5215..5670)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5352..5373)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5564..5583
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5796..5818
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5810..5827
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5811..5827
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5834..5853
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5834..5852
                     /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 ctttatgcgg attttcacgc tgggcactgt aactctgaag
      721 caaggcgaaa taaaagatgc tactccctct gactctgtca gagcaactgc aactataccg
      781 atccaagcgt cactgccttt tgggtggctg atcgtcggcg tcgcattctt ggccgttttc
      841 cagtcagcca gtaaaatcat aaccctcaag aaaagatggc aacttgcact gagcaaagga
      901 atccatttca tctgcaatct cctgctcctt ttcgttaccg tgtactccca tctgttgctc
      961 gtcgctgcgg gtctggaagc cccttttctg tacctttatg ccctggttta cttcttgcaa
     1021 agcatcaact tcgtccggat catcatgagg ctctggctgt gctggaaatg cagaagtaag
     1081 aatccgcttc tttatgacgc aaattatttc ctttgttggc atacaaattg ctatgactac
     1141 tgtattcctt acaacagcgt cacatccagc attgttatca cgtccggcga tggaactacc
     1201 agtcctattt cagaacacga ttatcaaata ggcgggtata ccgagaagtg ggagtctggt
     1261 gtgaaggact gcgtcgtcct gcattcatac tttacatcag actattacca gctctattcc
     1321 acccaactca gcacggacac cggcgtggag catgtgacat tctttattta taataaaatc
     1381 gtagacgaac ctgaagagca tgttcaaatt cacaccatcg acgggagctc aggtgtcgtg
     1441 aatcctgcta tggagcctat ctacgacgag ccgacaacaa ccacgagcgt cccattggga
     1501 aaaccaattc ccaacccact cctgggcctg gacagcacct aaacgcgtcg gatcctgaga
     1561 acttcagggt gagtttgggg acccttgatt gttctttctt tttcgctatt gtaaaattca
     1621 tgttatatgg agggggcaaa gttttcaggg tgttgtttag aatgggaaga tgtcccttgt
     1681 atcaccatgg accctcatga taattttgtt tctttcactt tctactctgt tgacaaccat
     1741 tgtctcctct tattttcttt tcattttctg taactttttc gttaaacttt agcttgcatt
     1801 tgtaacgaat ttttaaattc acttttgttt atttgtcaga ttgtaagtac tttctctaat
     1861 cacttttttt tcaaggcaat cagggtatat tatattgtac ttcagcacag ttttagagaa
     1921 caattgttat aattaaatga taaggtagaa tatttctgca tataaattct ggctggcgtg
     1981 gaaatattct tattggtaga aacaactaca ccctggtcat catcctgcct ttctctttat
     2041 ggttacaatg atatacactg tttgagatga ggataaaata ctctgagtcc aaaccgggcc
     2101 cctctgctaa ccatgttcat gccttcttct ctttcctaca gctcctgggc aacgtgctgg
     2161 ttgttgtgct gtctcatcat tttggcaaag aattcactcc tcaggtgcag gctgcctatc
     2221 agaaggtggt ggctggtgtg gccaatgccc tggctcacaa ataccactga gatctttttc
     2281 cctctgccaa aaattatggg gacatcatga agccccttga gcatctgact tctggctaat
     2341 aaaggaaatt tattttcatt gcaatagtgt gttggaattt tttgtgtctc tcactcggaa
     2401 ggacatatgg gagggcaaat catttaaaac atcagaatga gtatttggtt tagagtttgg
     2461 caacatatgc ccatatgctg gctgccatga acaaaggttg gctataaaga ggtcatcagt
     2521 atatgaaaca gccccctgct gtccattcct tattccatag aaaagccttg acttgaggtt
     2581 agattttttt tatattttgt tttgtgttat ttttttcttt aacatcccta aaattttcct
     2641 tacatgtttt actagccaga tttttcctcc tctcctgact actcccagtc atagctgtcc
     2701 ctcttctctt atggagatcc ctcgaggagc tttttgcaaa agcctaggcc tccaaaaaag
     2761 cctcttcact acttctggaa tagctcagag gccgaggcgg cctcggcctc tgcataaata
     2821 aaaaaaatta gtcagccatg gggcggagaa tgggcggaac tgggcggagt taggggcggg
     2881 atgggcggag ttaggggcgg gactatggtt gctgactaat tgctgcatta atgaatcggc
     2941 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     3001 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     3061 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     3121 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     3181 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     3241 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     3301 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     3361 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     3421 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     3481 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     3541 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     3601 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     3661 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     3721 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     3781 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     3841 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     3901 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     3961 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     4021 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     4081 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     4141 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     4201 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     4261 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     4321 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     4381 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     4441 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     4501 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     4561 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     4621 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     4681 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     4741 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     4801 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     4861 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     4921 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc
     4981 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
     5041 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
     5101 ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac
     5161 catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggaaattg
     5221 taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta
     5281 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt
     5341 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
     5401 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa
     5461 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
     5521 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
     5581 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
     5641 ccgcgcttaa tgcgccgcta cagggcgcgt ccattcgcca ttcaggctgc gcaactgttg
     5701 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc
     5761 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac
     5821 ggccagtgaa ttgtaatacg actcactata gggcgaattc gagctcgccc
//
