LOCUS       sequence-301677-        6230 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 6230)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6230)
  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-m4252783
FEATURES             Location/Qualifiers
     source          1..6230
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1427..2015
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1916..1935
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    2241..2312
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(2357..2493)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     terminator      complement(2523..2569)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    2684..2805
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2721..2740)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2775..2794
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(2858..2935)
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     CDS             complement(2936..2956)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     regulatory      2989..2998
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3067..3269)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3096..3116)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3271..3574
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    3655..3703
                     /note="synthetic polyadenylation signal"
     terminator      3721..3767
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    3772..3843
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(3995..4131)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(join(4144..4935,4936..5004))
                     /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     4767..4786
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    5270..5318
                     /note="synthetic polyadenylation signal"
     CDS             complement(5345..5944)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(5428..5448)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     5925..5944
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(join(5948..6230,1..927))
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(6092..6111)
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
ORIGIN
        1 tgcgtcctgc gacggagaaa agcctaccgc acacctaccg gctggtggcc ccaccctgca
       61 ttataagcca acagaacggg tgacgtcacg acacgacgag ggcgcgcgct cccaaaggta
      121 cgggtgcact gcccaacggc accgccataa ctgccgcccc cgcaacagac gacaaaccga
      181 gttctccagt cagtgacaaa cttcacgtca gggtccccag atggtgccca gcccatctca
      241 cccgaataag agctttcccg cattagcgaa gggctcaaga ccttgggttc ttgccgccca
      301 ccatgccccc caccttgttt caacgacctc acagcccgcc tcacaagcgt cttccattca
      361 agactcggga acagccgcca ttttgctgcg ctccccccaa cccccagttc agggcaacct
      421 tgctcgcgga cccagactac agcccttggc ggtctctcca cacgcttccg tcccaccgag
      481 cggcccggcg gccacgaaag ccccggccag cccagcagcc cgctactcac caagtgacga
      541 tcacagcgat ccacaaacaa gaaccgcgac ccaaatcccg gctgcgacgg aactagctgt
      601 gccacacccg gcgcgtcctt atataatcat cggcgttcac cgccccacgg agatccctcc
      661 gcagaatcgc cgagaaggga ctacttttcc tcgcctgttc cgctctctgg aaagaaaacc
      721 agtgccctag agtcacccaa gtcccgtcct aaaatgtcct tctgctgata ctggggttct
      781 aaggccgagt cttatgagca gcgggccgct gtcctgagcg tccgggcgga aggatcagga
      841 cgctcctgcg cccttcgtct gacgtggcag cgctcgccgt gacgaggggg gcgcccgcgg
      901 gaggcgccaa aacccggcgc ggaggccagg cgcgccccga tttcgatatt cctcggatct
      961 cggatagatc gggattgagg atgtaggttg aacttctctc gggtgatccc gatctcggat
     1021 attcgggatt ggagtaatac ctggcgatag gcgctgccaa gcaatagcca aggcgcaagg
     1081 ccggccaaga gacttgcgat agttcgttcg agaggtagcc tgcaagatgg atcggacgaa
     1141 aggtttgaag attttccgac gagggccttg gagagttcgg aaatttaaat gcaactatgg
     1201 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     1261 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     1321 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     1381 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     1441 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     1501 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     1561 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     1621 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     1681 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     1741 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     1801 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     1861 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     1921 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     1981 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     2041 ggttcctggc cttttgctgg ccttttgctc acatgattta aatgaaactc gtaatcggag
     2101 aacgggtgca atagcaagga ccagatcgaa cttgactgac gcaattctga gcgtagtcga
     2161 aacccagaca agtcgatctc gccaagtagc tgacttgact ggttaattaa gtttcgatag
     2221 cccaaggtaa ccaatgagat tagggaggga agaaagtttg atgaactact tctgacccct
     2281 agtggtgtcc agaaaagacc attaaaggaa tgtcttgatc gaaatcgaga gatcgcctgc
     2341 aggatagctg acgaccttaa ggataaattt ctggtaagga ggacacgtat ggaagtgggc
     2401 aagttgggga agccgtatcc gttgctgaat ctggcatatg tgggagtata agacgcgcag
     2461 cgtcgcatca ggcatttttt tctgcgccaa tgccggtaga tagcaatccg gcccgagggg
     2521 cacaaaaaac ccctcaagac ccgtttagag gccccaaggg gttatgctac gataccgata
     2581 gagatgggcc aataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct
     2641 ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc
     2701 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     2761 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtgagct
     2821 gaaggtacgc tgtatctcag tcagtcaagc tagcttagag ggcaccagac gaggatatct
     2881 ttttgaatct gttggctgca ctcaccgcga taaaattctt tttccatcgc ctcttcgact
     2941 gaaagtaaag gttttccgca aacccgttta aaaacgattg tgcatcagcc atggtggcgg
     3001 ccgctggcgg gtgtgtcgag tggatggtag gatcgacaaa gatctgatct gacggttcac
     3061 taaaccagct ctgcttatat agacctccca ccgtacacgc ctaccgccca tttgcgtcaa
     3121 tggggcggag ttgttacgac attttggaaa gtcccgttga ttttggtgcc aaaacaaact
     3181 cccattgacg tcaatggggt ggagacttgg aaatccccgt gagtcaaacc gctatccacg
     3241 cccattgatg tactgccaaa accgcatcag catggtaata gcgatgacta atacgtagat
     3301 gtactgccaa gtaggaaagt cccataaggt catgtactgg gcataatgcc aggcgggcca
     3361 tttaccgtca ttgacgtcaa tagggggcgt acttggcata tgatacactt gatgtactgc
     3421 caagtgggca gtttaccgta aatactccac ccattgacgt caatggaaag tccctattgg
     3481 cgttactatg ggaacatacg tcattattga cgtcaatggg cgggggtcgt tgggcggtca
     3541 gccaggcggg ccatttaccg taagttatgt aacgcggaac tccatatatg ggctatgaac
     3601 taatgacccc gtaattgatt actagatctg gctacactcg atcagcagtt agataataaa
     3661 atcgctatcc atcgaagatg gatgtgtgtt ggttttttgt gtgtgtaacg caacgattga
     3721 tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgaat tgagaggtag
     3781 ctgaagctgc tgttatgacc actagaggga agaagatacc tgtggagcta atggtccaag
     3841 atgtcgggag ccgcgatcgc tgccaaaaag gtaagctgac gatagctgac gagattcgat
     3901 gcccgacgag caaatgagcg ccacggctgt tcgcgtaagc tcgaaacgac ggagattcga
     3961 tagagccaga gtcgagttta aacgattcgg gatcttaagg ataaatttct ggtaaggagg
     4021 acacgtatgg aagtgggcaa gttggggaag ccgtatccgt tgctgaatct ggcatatgtg
     4081 ggagtataag acgcgcagcg tcgcatcagg catttttttc tgcgccaatg cacctagaat
     4141 cggttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     4201 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     4261 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     4321 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     4381 ccagtctatc aactgttgcc gggaagctag agtaagtagt tcgccagtta atagtttacg
     4441 caaggttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     4501 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     4561 agcggttagc tccttcggtc ctccaatcgt tgtcagaagt aagttggccg cagtgttatc
     4621 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     4681 ttctgtgact ggtgaatact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     4741 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     4801 gctcatcatt ggaaagcgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     4861 atccagttcg atgtaaccca ctcgcgcacc caactgatct tcagcatctt ttactttcac
     4921 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     4981 gacacggaaa tgttgaatac tcatgcgaaa gcctcctgct gatcctgtct cttgatagat
     5041 gtttgcaaaa gctaggactc gattatacgt tgaccgatca gactatcaat atctaggttt
     5101 aaacagatcg gtacgctgcc gtgaccaggc ttgagtacaa tatctgacga ttggacttcc
     5161 tgggtcccaa gctggattcc gactggctga ctgccaagca acgatatgga tcgtcgagac
     5221 tcgcactgag aggatcgttc tcggtacacc aggcgcgcct agcgttacac acacaaaaaa
     5281 ccaacacaca tccatcttcg atggatagcg attttattat ctaagactag gcttacatac
     5341 tagcctaggc accgggcttg cgggtcatgc accaggtgcg cggtccttcg ggcacctcga
     5401 cgtcggcggt gacggtgaag ccgagccgct cgtagaaggg gaggttgcgg ggcgcggagg
     5461 tctccaggaa ggcgggcacc ccggcgcgct cggcggcctc cactccgggg agcacgacgg
     5521 cgctgcccag acccttgccc tggtggtcgg gcgagacgcc gacggtggcc aggaaccacg
     5581 cgggctcctt gggccggtgc ggcgccagga ggccctccat ctgttgctgc gcggccagcc
     5641 gggaaccgct caactcggcc atgcgcgggc cgatctcggc gaacaccgcc cccgcttcga
     5701 cgctctccgg cgtggtccag accgccaccg cggcgccgtc gtccgcgacc cacaccttgc
     5761 cgatgtcgag cccgacgcgc gtgaggaaga gttcttgcag ctcggtgacc cgctcgatgt
     5821 ggcggtccgg atcgacggtg tggcgcgtgg cggggtagtc ggcgaacgcg gcggcgaggg
     5881 tgcgtacggc cctggggacg tcgtcgcggg tggcgaggcg caccgtgggc ttgtactcgg
     5941 tcatgttgtc taacaaaaaa gccaaaaacg gccagaattt agcggacaat ttacaagtct
     6001 aacactgaaa attacatatt gacccaaatg attacatttc aaaaggtgcc taaaaaactt
     6061 cacaaaacac actcgccaac cccgagcgca tagttcaaaa ccggagcttc agctacttaa
     6121 gaagataggt acataaaacc gaccaaagaa actgacgcct cacttatccc tcccctcacc
     6181 agaggtccgg cgcctgtcga ttcaggagag cctaccctag gcccgaaccc
//
