LOCUS       sequence-300643-        6621 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 6621)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6621)
  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-m4252963
FEATURES             Location/Qualifiers
     source          1..6621
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    20..141
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(57..76)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     111..130
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(210..227)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(228..248)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     promoter        complement(365..567)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(394..414)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        569..872
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    953..1001
                     /note="synthetic polyadenylation signal"
     terminator      1019..1065
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    1070..1141
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(1293..1429)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(join(1442..2233,2234..2302))
                     /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     2065..2084
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    2568..2616
                     /note="synthetic polyadenylation signal"
     CDS             complement(2643..3671)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     promoter        complement(3675..4884)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(3819..3838)
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     rep_origin      5384..5972
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5873..5892
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    6198..6269
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6314..6450)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     terminator      complement(6480..6526)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa
       61 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
      121 caaactcatc aatgtatctt atcatgtctg tgagctgaag gtacgctgta tctcagtcag
      181 tcaagctagc agaggcctgc ggatccttaa tggtgatggt gatggtgcga ctgaaagtaa
      241 aggttttcca ctgcgaaacc gttgagaaag gactgagcat cggcactcat ggtggcggcc
      301 gctggcgggt gtgtcgagtg gatggtagga tcgacaaaga tctgatctga cggttcacta
      361 aaccagctct gcttatatag acctcccacc gtacacgcct accgcccatt tgcgtcaatg
      421 gggcggagtt gttacgacat tttggaaagt cccgttgatt ttggtgccaa aacaaactcc
      481 cattgacgtc aatggggtgg agacttggaa atccccgtga gtcaaaccgc tatccacgcc
      541 cattgatgta ctgccaaaac cgcatcagca tggtaatagc gatgactaat acgtagatgt
      601 actgccaagt aggaaagtcc cataaggtca tgtactgggc ataatgccag gcgggccatt
      661 taccgtcatt gacgtcaata gggggcgtac ttggcatatg atacacttga tgtactgcca
      721 agtgggcagt ttaccgtaaa tactccaccc attgacgtca atggaaagtc cctattggcg
      781 ttactatggg aacatacgtc attattgacg tcaatgggcg ggggtcgttg ggcggtcagc
      841 caggcgggcc atttaccgta agttatgtaa cgcggaactc catatatggg ctatgaacta
      901 atgaccccgt aattgattac tagatctggc tacactcgat cagcagttag ataataaaat
      961 cgctatccat cgaagatgga tgtgtgttgg ttttttgtgt gtgtaacgca acgattgata
     1021 gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgaattg agaggtagct
     1081 gaagctgctg ttatgaccac tagagggaag aagatacctg tggagctaat ggtccaagat
     1141 gtcgggagcc gcgatcgctg ccaaaaaggt aagctgacga tagctgacga gattcgatgc
     1201 ccgacgagca aatgagcgcc acggctgttc gcgtaagctc gaaacgacgg agattcgata
     1261 gagccagagt cgagtttaaa cgattcggga tcttaaggat aaatttctgg taaggaggac
     1321 acgtatggaa gtgggcaagt tggggaagcc gtatccgttg ctgaatctgg catatgtggg
     1381 agtataagac gcgcagcgtc gcatcaggca tttttttctg cgccaatgca cctagaatcg
     1441 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     1501 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     1561 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     1621 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     1681 agtctatcaa ctgttgccgg gaagctagag taagtagttc gccagttaat agtttacgca
     1741 aggttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     1801 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     1861 cggttagctc cttcggtcct ccaatcgttg tcagaagtaa gttggccgca gtgttatcac
     1921 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     1981 ctgtgactgg tgaatactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     2041 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     2101 tcatcattgg aaagcgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     2161 ccagttcgat gtaacccact cgcgcaccca actgatcttc agcatctttt actttcacca
     2221 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     2281 cacggaaatg ttgaatactc atgcgaaagc ctcctgctga tcctgtctct tgatagatgt
     2341 ttgcaaaagc taggactcga ttatacgttg accgatcaga ctatcaatat ctaggtttaa
     2401 acagatcggt acgctgccgt gaccaggctt gagtacaata tctgacgatt ggacttcctg
     2461 ggtcccaagc tggattccga ctggctgact gccaagcaac gatatggatc gtcgagactc
     2521 gcactgagag gatcgttctc ggtacaccag gcgcgcctag cgttacacac acaaaaaacc
     2581 aacacacatc catcttcgat ggatagcgat tttattatct aagactaggc ttacatacta
     2641 gcttattcct ttgccctcgg acgagtgctg gggcgtcggt ttccactatc ggcgagtact
     2701 tctacacagc catcggtcca gacggcggcg cttctgcggg cgatttgtgt acgcccgaca
     2761 gtcccggctc cggatcggac gattgcgtcg catcgaccct gcgcccaagc tgcatcatcg
     2821 aaattgccgt caaccaagct ctgatagagt tggtcaagac caatgcggag catatacgcc
     2881 cggagccggg gcgatcctgc aagctccgga tgcctccgct cgaagtagcg cgtctgctgc
     2941 tccatacaag ccaaccacgg cctccagaag aagatgttgg cgacctcgta ttgggaatcc
     3001 ccgaacatcg cctcgctcca gtcaatgacc gctgttatgc ggccattgtc cgtcaggaca
     3061 ttgttggagc cgaaatccgc gtgcacgagg tgccggactt cggggcagtc ctcggcccaa
     3121 agcatcagct catcgagagc ctgcgcgacg gacgcactga cggtgtcgtc catcacagtt
     3181 tgccagtgat acacatgggg atcagcaatc gcgcatatga aatcacgcca tgtagtgtat
     3241 tgaccgattc cttgcggtcc gaatgggccg aacccgctcg tctggctaag atcggcggca
     3301 gcgatggcat ccattgcctc cgcgaccggc tgcagaacag cgggcagttc ggtttcaggc
     3361 aggtcttgca acgtgacacc ctgtgcacgg cgggagatgc aataggtcag gctctcgctg
     3421 aactccccaa tgtcaagcac ttccggaatc gggagcgcgg cggatgcaaa gtgccgataa
     3481 acataacgat ctttgtagaa accatcggcg cagctattta cccgcaggac atatccacgc
     3541 cctcctacat cgaagctgaa agcacgagat tcttcgccct ccgagagctg catcaggtcg
     3601 gagacgctgt cgaacttttc gatcagaaac ttctcgacag acgtcgcggt gagttcaggc
     3661 tttttaccca tgttgtctaa caaaaaagcc aaaaacggcc agaatttagc ggacaattta
     3721 caagtctaac actgaaaatt acatattgac ccaaatgatt acatttcaaa aggtgcctaa
     3781 aaaacttcac aaaacacact cgccaacccc gagcgcatag ttcaaaaccg gagcttcagc
     3841 tacttaagaa gataggtaca taaaaccgac caaagaaact gacgcctcac ttatccctcc
     3901 cctcaccaga ggtccggcgc ctgtcgattc aggagagcct accctaggcc cgaaccctgc
     3961 gtcctgcgac ggagaaaagc ctaccgcaca cctaccggct ggtggcccca ccctgcatta
     4021 taagccaaca gaacgggtga cgtcacgaca cgacgagggc gcgcgctccc aaaggtacgg
     4081 gtgcactgcc caacggcacc gccataactg ccgcccccgc aacagacgac aaaccgagtt
     4141 ctccagtcag tgacaaactt cacgtcaggg tccccagatg gtgcccagcc catctcaccc
     4201 gaataagagc tttcccgcat tagcgaaggg ctcaagacct tgggttcttg ccgcccacca
     4261 tgccccccac cttgtttcaa cgacctcaca gcccgcctca caagcgtctt ccattcaaga
     4321 ctcgggaaca gccgccattt tgctgcgctc cccccaaccc ccagttcagg gcaaccttgc
     4381 tcgcggaccc agactacagc ccttggcggt ctctccacac gcttccgtcc caccgagcgg
     4441 cccggcggcc acgaaagccc cggccagccc agcagcccgc tactcaccaa gtgacgatca
     4501 cagcgatcca caaacaagaa ccgcgaccca aatcccggct gcgacggaac tagctgtgcc
     4561 acacccggcg cgtccttata taatcatcgg cgttcaccgc cccacggaga tccctccgca
     4621 gaatcgccga gaagggacta cttttcctcg cctgttccgc tctctggaaa gaaaaccagt
     4681 gccctagagt cacccaagtc ccgtcctaaa atgtccttct gctgatactg gggttctaag
     4741 gccgagtctt atgagcagcg ggccgctgtc ctgagcgtcc gggcggaagg atcaggacgc
     4801 tcctgcgccc ttcgtctgac gtggcagcgc tcgccgtgac gaggggggcg cccgcgggag
     4861 gcgccaaaac ccggcgcgga ggccaggcgc gccccgattt cgatattcct cggatctcgg
     4921 atagatcggg attgaggatg taggttgaac ttctctcggg tgatcccgat ctcggatatt
     4981 cgggattgga gtaatacctg gcgataggcg ctgccaagca atagccaagg cgcaaggccg
     5041 gccaagagac ttgcgatagt tcgttcgaga ggtagcctgc aagatggatc ggacgaaagg
     5101 tttgaagatt ttccgacgag ggccttggag agttcggaaa tttaaatgca actatggatg
     5161 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     5221 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     5281 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     5341 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     5401 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     5461 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     5521 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     5581 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     5641 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     5701 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     5761 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     5821 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     5881 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     5941 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     6001 tcctggcctt ttgctggcct tttgctcaca tgatttaaat gaaactcgta atcggagaac
     6061 gggtgcaata gcaaggacca gatcgaactt gactgacgca attctgagcg tagtcgaaac
     6121 ccagacaagt cgatctcgcc aagtagctga cttgactggt taattaagtt tcgatagccc
     6181 aaggtaacca atgagattag ggagggaaga aagtttgatg aactacttct gacccctagt
     6241 ggtgtccaga aaagaccatt aaaggaatgt cttgatcgaa atcgagagat cgcctgcagg
     6301 atagctgacg accttaagga taaatttctg gtaaggagga cacgtatgga agtgggcaag
     6361 ttggggaagc cgtatccgtt gctgaatctg gcatatgtgg gagtataaga cgcgcagcgt
     6421 cgcatcaggc atttttttct gcgccaatgc cggtagatag caatccggcc cgaggggcac
     6481 aaaaaacccc tcaagacccg tttagaggcc ccaaggggtt atgctacgat accgatagag
     6541 atgggccaat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
     6601 cctgaacctg aaacataaaa t
//
