LOCUS       sequence-353887-        5708 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector for RaTG13 M, V5-tagged. Sequence 
            codon-optimized..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5708)
  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 5708)
  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 M C-V5-tag
FEATURES             Location/Qualifiers
     source          1..5708
                     /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             1336..1377
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     intron          1407..1979
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     complement(2111..2130)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2176..2231
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2230..2249)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      2585..2720
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2826..2843)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2997..3585)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3077..3096)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3756..4547,4548..4616))
                     /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     4379..4398
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4617..4721)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4789..4807
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4845..4867)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4967..4986
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(5053..5508)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5190..5211)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5402..5421
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5634..5656
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5648..5665
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5649..5665
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5672..5691
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5672..5690
                     /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 ccatggcaga taacggcacc ataactgttg aagagctcaa gaagctgttg
      721 gaacaatgga acctcgtaat aggatttctg tttctcacat ggatctgttt gctgcaattt
      781 gcatacgcga atagaaatcg attcctgtac atcatcaagc tcatctttct ctggctcctc
      841 tggccagtca cgttggcatg cttcgtgctc gcagctgttt acagaataaa ctggattaca
      901 ggcgggatag ccatagctat ggcttgcctc gtcggtctca tgtggttgag ctactttata
      961 gctagctttc gactgttcgc cagaacccgg tcaatgtgga gttttaaccc tgaaaccaat
     1021 attctcctga acgtcccact ccacgggacc attctgacac ggccattgct cgagtcagaa
     1081 ttggtcattg gagccgtcat tcttcgcggc cacctgagga tcgctgggca tcacctgggg
     1141 cggtgtgaca taaaagattt gcccaaggaa atcacagttg ctacttccag gaccctgagc
     1201 tattataaac tgggagcaag tcaacgagtg gctggggaca gtggctttgc agcctatagc
     1261 cgctatcgca tcggtaatta caagttgaat acggaccact caagctccag cgataatatt
     1321 gcactcttgg tgcaaggaaa gcccatcccc aaccccctgc tgggccttga ttctacgtaa
     1381 acgcgtcgga tcctgagaac ttcagggtga gtttggggac ccttgattgt tctttctttt
     1441 tcgctattgt aaaattcatg ttatatggag ggggcaaagt tttcagggtg ttgtttagaa
     1501 tgggaagatg tcccttgtat caccatggac cctcatgata attttgtttc tttcactttc
     1561 tactctgttg acaaccattg tctcctctta ttttcttttc attttctgta actttttcgt
     1621 taaactttag cttgcatttg taacgaattt ttaaattcac ttttgtttat ttgtcagatt
     1681 gtaagtactt tctctaatca cttttttttc aaggcaatca gggtatatta tattgtactt
     1741 cagcacagtt ttagagaaca attgttataa ttaaatgata aggtagaata tttctgcata
     1801 taaattctgg ctggcgtgga aatattctta ttggtagaaa caactacacc ctggtcatca
     1861 tcctgccttt ctctttatgg ttacaatgat atacactgtt tgagatgagg ataaaatact
     1921 ctgagtccaa accgggcccc tctgctaacc atgttcatgc cttcttctct ttcctacagc
     1981 tcctgggcaa cgtgctggtt gttgtgctgt ctcatcattt tggcaaagaa ttcactcctc
     2041 aggtgcaggc tgcctatcag aaggtggtgg ctggtgtggc caatgccctg gctcacaaat
     2101 accactgaga tctttttccc tctgccaaaa attatgggga catcatgaag ccccttgagc
     2161 atctgacttc tggctaataa aggaaattta ttttcattgc aatagtgtgt tggaattttt
     2221 tgtgtctctc actcggaagg acatatggga gggcaaatca tttaaaacat cagaatgagt
     2281 atttggttta gagtttggca acatatgccc atatgctggc tgccatgaac aaaggttggc
     2341 tataaagagg tcatcagtat atgaaacagc cccctgctgt ccattcctta ttccatagaa
     2401 aagccttgac ttgaggttag atttttttta tattttgttt tgtgttattt ttttctttaa
     2461 catccctaaa attttcctta catgttttac tagccagatt tttcctcctc tcctgactac
     2521 tcccagtcat agctgtccct cttctcttat ggagatccct cgaggagctt tttgcaaaag
     2581 cctaggcctc caaaaaagcc tcttcactac ttctggaata gctcagaggc cgaggcggcc
     2641 tcggcctctg cataaataaa aaaaattagt cagccatggg gcggagaatg ggcggaactg
     2701 ggcggagtta ggggcgggat gggcggagtt aggggcggga ctatggttgc tgactaattg
     2761 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     2821 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     2881 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     2941 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     3001 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     3061 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     3121 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     3181 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     3241 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     3301 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     3361 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     3421 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     3481 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     3541 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     3601 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     3661 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     3721 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     3781 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     3841 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     3901 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     3961 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     4021 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     4081 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     4141 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     4201 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     4261 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     4321 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     4381 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     4441 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     4501 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     4561 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     4621 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     4681 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     4741 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     4801 acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
     4861 ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag
     4921 ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag
     4981 attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa
     5041 taccgcatca ggaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
     5101 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     5161 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     5221 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     5281 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     5341 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     5401 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
     5461 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc attcgccatt
     5521 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     5581 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     5641 acgacgttgt aaaacgacgg ccagtgaatt gtaatacgac tcactatagg gcgaattcga
     5701 gctcgccc
//
