LOCUS       sequence-299993-        6582 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 nsp11.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6582)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6582)
  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"
COMMENT     Sequence Label: pGBW-m4252994
FEATURES             Location/Qualifiers
     source          1..6582
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    115..236
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(126..145)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     180..199
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      351..397
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      427..563
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    608..679
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(905..1493)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(985..1004)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        1993..3202
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3039..3058
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3206..4234
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    4261..4309
                     /note="synthetic polyadenylation signal"
     CDS             join(4575..4643,4644..5435)
                     /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     complement(4793..4812)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5448..5584
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5736..5807
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5812..5858)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5876..5924
                     /note="synthetic polyadenylation signal"
     enhancer        6005..6308
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6310..6512
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6463..6483
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgagt gccgatgctc agtcctttct caacggtttc gcagtgtaag gatccgcagg
       61 cctctgctag cttgactgac tgagatacag cgtaccttca gctcacagac atgataagat
      121 acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg
      181 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca
      241 acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa
      301 gcaagtaaaa cctctacaaa tgtggtattg gcccatctct atcggtatcg tagcataacc
      361 ccttggggcc tctaaacggg tcttgagggg ttttttgtgc ccctcgggcc ggattgctat
      421 ctaccggcat tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt atactcccac
      481 atatgccaga ttcagcaacg gatacggctt ccccaacttg cccacttcca tacgtgtcct
      541 ccttaccaga aatttatcct taaggtcgtc agctatcctg caggcgatct ctcgatttcg
      601 atcaagacat tcctttaatg gtcttttctg gacaccacta ggggtcagaa gtagttcatc
      661 aaactttctt ccctccctaa tctcattggt taccttgggc tatcgaaact taattaacca
      721 gtcaagtcag ctacttggcg agatcgactt gtctgggttt cgactacgct cagaattgcg
      781 tcagtcaagt tcgatctggt ccttgctatt gcacccgttc tccgattacg agtttcattt
      841 aaatcatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
      901 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
      961 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     1021 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     1081 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     1141 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     1201 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     1261 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     1321 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     1381 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     1441 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     1501 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     1561 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     1621 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     1681 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgca tttaaatttc
     1741 cgaactctcc aaggccctcg tcggaaaatc ttcaaacctt tcgtccgatc catcttgcag
     1801 gctacctctc gaacgaacta tcgcaagtct cttggccggc cttgcgcctt ggctattgct
     1861 tggcagcgcc tatcgccagg tattactcca atcccgaata tccgagatcg ggatcacccg
     1921 agagaagttc aacctacatc ctcaatcccg atctatccga gatccgagga atatcgaaat
     1981 cggggcgcgc ctggcctccg cgccgggttt tggcgcctcc cgcgggcgcc cccctcgtca
     2041 cggcgagcgc tgccacgtca gacgaagggc gcaggagcgt cctgatcctt ccgcccggac
     2101 gctcaggaca gcggcccgct gctcataaga ctcggcctta gaaccccagt atcagcagaa
     2161 ggacatttta ggacgggact tgggtgactc tagggcactg gttttctttc cagagagcgg
     2221 aacaggcgag gaaaagtagt cccttctcgg cgattctgcg gagggatctc cgtggggcgg
     2281 tgaacgccga tgattatata aggacgcgcc gggtgtggca cagctagttc cgtcgcagcc
     2341 gggatttggg tcgcggttct tgtttgtgga tcgctgtgat cgtcacttgg tgagtagcgg
     2401 gctgctgggc tggccggggc tttcgtggcc gccgggccgc tcggtgggac ggaagcgtgt
     2461 ggagagaccg ccaagggctg tagtctgggt ccgcgagcaa ggttgccctg aactgggggt
     2521 tggggggagc gcagcaaaat ggcggctgtt cccgagtctt gaatggaaga cgcttgtgag
     2581 gcgggctgtg aggtcgttga aacaaggtgg ggggcatggt gggcggcaag aacccaaggt
     2641 cttgagccct tcgctaatgc gggaaagctc ttattcgggt gagatgggct gggcaccatc
     2701 tggggaccct gacgtgaagt ttgtcactga ctggagaact cggtttgtcg tctgttgcgg
     2761 gggcggcagt tatggcggtg ccgttgggca gtgcacccgt acctttggga gcgcgcgccc
     2821 tcgtcgtgtc gtgacgtcac ccgttctgtt ggcttataat gcagggtggg gccaccagcc
     2881 ggtaggtgtg cggtaggctt ttctccgtcg caggacgcag ggttcgggcc tagggtaggc
     2941 tctcctgaat cgacaggcgc cggacctctg gtgaggggag ggataagtga ggcgtcagtt
     3001 tctttggtcg gttttatgta cctatcttct taagtagctg aagctccggt tttgaactat
     3061 gcgctcgggg ttggcgagtg tgttttgtga agttttttag gcaccttttg aaatgtaatc
     3121 atttgggtca atatgtaatt ttcagtgtta gacttgtaaa ttgtccgcta aattctggcc
     3181 gtttttggct tttttgttag acaacatggg taaaaagcct gaactcaccg cgacgtctgt
     3241 cgagaagttt ctgatcgaaa agttcgacag cgtctccgac ctgatgcagc tctcggaggg
     3301 cgaagaatct cgtgctttca gcttcgatgt aggagggcgt ggatatgtcc tgcgggtaaa
     3361 tagctgcgcc gatggtttct acaaagatcg ttatgtttat cggcactttg catccgccgc
     3421 gctcccgatt ccggaagtgc ttgacattgg ggagttcagc gagagcctga cctattgcat
     3481 ctcccgccgt gcacagggtg tcacgttgca agacctgcct gaaaccgaac tgcccgctgt
     3541 tctgcagccg gtcgcggagg caatggatgc catcgctgcc gccgatctta gccagacgag
     3601 cgggttcggc ccattcggac cgcaaggaat cggtcaatac actacatggc gtgatttcat
     3661 atgcgcgatt gctgatcccc atgtgtatca ctggcaaact gtgatggacg acaccgtcag
     3721 tgcgtccgtc gcgcaggctc tcgatgagct gatgctttgg gccgaggact gccccgaagt
     3781 ccggcacctc gtgcacgcgg atttcggctc caacaatgtc ctgacggaca atggccgcat
     3841 aacagcggtc attgactgga gcgaggcgat gttcggggat tcccaatacg aggtcgccaa
     3901 catcttcttc tggaggccgt ggttggcttg tatggagcag cagacgcgct acttcgagcg
     3961 gaggcatccg gagcttgcag gatcgccccg gctccgggcg tatatgctcc gcattggtct
     4021 tgaccaactc tatcagagct tggttgacgg caatttcgat gatgcagctt gggcgcaggg
     4081 tcgatgcgac gcaatcgtcc gatccggagc cgggactgtc gggcgtacac aaatcgcccg
     4141 cagaagcgcc gccgtctgga ccgatggctg tgtagaagta ctcgccgata gtggaaaccg
     4201 acgccccagc actcgtccga gggcaaagga ataagctagt atgtaagcct agtcttagat
     4261 aataaaatcg ctatccatcg aagatggatg tgtgttggtt ttttgtgtgt gtaacgctag
     4321 gcgcgcctgg tgtaccgaga acgatcctct cagtgcgagt ctcgacgatc catatcgttg
     4381 cttggcagtc agccagtcgg aatccagctt gggacccagg aagtccaatc gtcagatatt
     4441 gtactcaagc ctggtcacgg cagcgtaccg atctgtttaa acctagatat tgatagtctg
     4501 atcggtcaac gtataatcga gtcctagctt ttgcaaacat ctatcaagag acaggatcag
     4561 caggaggctt tcgcatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     4621 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     4681 atcagttggg tgcgcgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     4741 agagttttcg ccccgaagaa cgctttccaa tgatgagcac ttttaaagtt ctgctatgtg
     4801 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     4861 ctcagaatga cttggttgag tattcaccag tcacagaaaa gcatcttacg gatggcatga
     4921 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     4981 ttctgacaac gattggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     5041 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     5101 gtgacaccac gatgcctgta gcaatggcaa caaccttgcg taaactatta actggcgaac
     5161 tacttactct agcttcccgg caacagttga tagactggat ggaggcggat aaagttgcag
     5221 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     5281 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     5341 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     5401 ctgagatagg tgcctcactg attaagcatt ggtaaccgat tctaggtgca ttggcgcaga
     5461 aaaaaatgcc tgatgcgacg ctgcgcgtct tatactccca catatgccag attcagcaac
     5521 ggatacggct tccccaactt gcccacttcc atacgtgtcc tccttaccag aaatttatcc
     5581 ttaagatccc gaatcgttta aactcgactc tggctctatc gaatctccgt cgtttcgagc
     5641 ttacgcgaac agccgtggcg ctcatttgct cgtcgggcat cgaatctcgt cagctatcgt
     5701 cagcttacct ttttggcagc gatcgcggct cccgacatct tggaccatta gctccacagg
     5761 tatcttcttc cctctagtgg tcataacagc agcttcagct acctctcaat tcaaaaaacc
     5821 cctcaagacc cgtttagagg ccccaagggg ttatgctatc aatcgttgcg ttacacacac
     5881 aaaaaaccaa cacacatcca tcttcgatgg atagcgattt tattatctaa ctgctgatcg
     5941 agtgtagcca gatctagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
     6001 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
     6061 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
     6121 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
     6181 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
     6241 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
     6301 ttaccatgct gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
     6361 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
     6421 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
     6481 gtgtacggtg ggaggtctat ataagcagag ctggtttagt gaaccgtcag atcagatctt
     6541 tgtcgatcct accatccact cgacacaccc gccagcggcc gc
//
