LOCUS       Exported                5942 bp ds-DNA     circular SYN 07-FEB-2022
DEFINITION  Mammalian expression vector for SARS-CoV-2 Nsp16, V5-tagged. 
            Sequence codon-optimized..
ACCESSION   .
VERSION     .
KEYWORDS    pCG_SARS-CoV2 NSP16 C-V5-tag
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5942)
  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 5942)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Feb 7, 2022 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5942
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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             1458..1469
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             1570..1611
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     intron          1641..2213
                     /label=-beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     complement(2345..2364)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2410..2465
                     /label=-beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2464..2483)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      2819..2954
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2873..2892)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     complement(3060..3077)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3231..3819)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3311..3330)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3990..4850)
                     /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     4613..4632
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4851..4955)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5023..5041
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5079..5101)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5201..5220
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(5287..5742)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5424..5445)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5636..5655
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5868..5890
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5882..5899
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5883..5899
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5906..5925
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5906..5924
                     /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 ccatgtcatc tcaagcatgg caacccggag tcgcaatgcc caatctctat
      721 aaaatgcaaa gaatgttgtt ggaaaagtgt gatctgcaaa actacggcga ttctgccacc
      781 ctgccaaagg gtatcatgat gaacgtggct aagtacactc aattgtgtca atatctgaat
      841 acgctgacac tcgctgttcc atacaatatg agagtgattc acttcggggc cggctctgat
      901 aaaggcgttg ctcccggcac cgctgttctc aggcaatggc ttccaacagg gactctgctc
      961 gtggatagtg acctgaacga ttttgtgagc gatgcagata gcacgctcat tggagactgt
     1021 gccaccgttc acaccgcgaa taagtgggat ctcattatct ctgacatgta cgacccaaag
     1081 actaagaacg taactaaaga gaacgatagc aaggaaggtt tcttcactta catatgcggt
     1141 ttcatccagc agaaactggc gctgggcggg tccgtagcta taaagatcac tgagcactca
     1201 tggaatgctg atctgtataa actgatggga cattttgcat ggtggacagc cttcgtgacc
     1261 aatgtgaacg caagttcatc agaagctttt cttattgggt gtaattacct gggtaaacca
     1321 agagaacaga tcgacggcta tgtaatgcac gcaaactata ttttctggag aaatactaat
     1381 ccgattcagc tgagctcata cagcttgttc gacatgtcta aattcccgct caaactgcgt
     1441 ggaacggctg ttatgtcatt gaaggaagga caaataaatg atatgatctt gagcctgctc
     1501 tctaaaggga gactgataat tagagagaac aacagagtgg tcattagctc agatgtgctc
     1561 gtcaacaacg gaaaaccgat acctaatcca ctgctgggac tcgattccac ctaaacgcgt
     1621 cggatcctga gaacttcagg gtgagtttgg ggacccttga ttgttctttc tttttcgcta
     1681 ttgtaaaatt catgttatat ggagggggca aagttttcag ggtgttgttt agaatgggaa
     1741 gatgtccctt gtatcaccat ggaccctcat gataattttg tttctttcac tttctactct
     1801 gttgacaacc attgtctcct cttattttct tttcattttc tgtaactttt tcgttaaact
     1861 ttagcttgca tttgtaacga atttttaaat tcacttttgt ttatttgtca gattgtaagt
     1921 actttctcta atcacttttt tttcaaggca atcagggtat attatattgt acttcagcac
     1981 agttttagag aacaattgtt ataattaaat gataaggtag aatatttctg catataaatt
     2041 ctggctggcg tggaaatatt cttattggta gaaacaacta caccctggtc atcatcctgc
     2101 ctttctcttt atggttacaa tgatatacac tgtttgagat gaggataaaa tactctgagt
     2161 ccaaaccggg cccctctgct aaccatgttc atgccttctt ctctttccta cagctcctgg
     2221 gcaacgtgct ggttgttgtg ctgtctcatc attttggcaa agaattcact cctcaggtgc
     2281 aggctgccta tcagaaggtg gtggctggtg tggccaatgc cctggctcac aaataccact
     2341 gagatctttt tccctctgcc aaaaattatg gggacatcat gaagcccctt gagcatctga
     2401 cttctggcta ataaaggaaa tttattttca ttgcaatagt gtgttggaat tttttgtgtc
     2461 tctcactcgg aaggacatat gggagggcaa atcatttaaa acatcagaat gagtatttgg
     2521 tttagagttt ggcaacatat gcccatatgc tggctgccat gaacaaaggt tggctataaa
     2581 gaggtcatca gtatatgaaa cagccccctg ctgtccattc cttattccat agaaaagcct
     2641 tgacttgagg ttagattttt tttatatttt gttttgtgtt atttttttct ttaacatccc
     2701 taaaattttc cttacatgtt ttactagcca gatttttcct cctctcctga ctactcccag
     2761 tcatagctgt ccctcttctc ttatggagat ccctcgagga gctttttgca aaagcctagg
     2821 cctccaaaaa agcctcttca ctacttctgg aatagctcag aggccgaggc ggcctcggcc
     2881 tctgcataaa taaaaaaaat tagtcagcca tggggcggag aatgggcgga actgggcgga
     2941 gttaggggcg ggatgggcgg agttaggggc gggactatgg ttgctgacta attgctgcat
     3001 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     3061 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     3121 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     3181 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     3241 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     3301 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     3361 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     3421 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     3481 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     3541 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     3601 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     3661 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     3721 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     3781 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     3841 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     3901 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     3961 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     4021 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     4081 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     4141 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     4201 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     4261 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     4321 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     4381 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     4441 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     4501 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     4561 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     4621 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     4681 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     4741 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     4801 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     4861 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     4921 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     4981 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     5041 ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     5101 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     5161 tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta
     5221 ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc
     5281 atcaggaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     5341 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     5401 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     5461 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     5521 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     5581 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     5641 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     5701 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtccattcgc cattcaggct
     5761 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
     5821 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
     5881 ttgtaaaacg acggccagtg aattgtaata cgactcacta tagggcgaat tcgagctcgc
     5941 cc
//
