LOCUS       sequence-300254-        4230 bp    DNA     circular SYN 15-APR-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 4230)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4230)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pGBW-m4136059
FEATURES             Location/Qualifiers
     source          1..4230
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    99..220
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(110..129)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     164..183
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      335..381
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      411..547
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    592..663
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(889..1477)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(969..988)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(2223..2291,2292..3083)
                     /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(2441..2460)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3096..3232
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3384..3455
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(3460..3506)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    3524..3572
                     /note="synthetic polyadenylation signal"
     enhancer        3653..3956
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3958..4160
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4111..4131
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgagt gccgatgctc agtcctttct caacggtttc gcagtgtaag ctagcttgac
       61 tgactgagat acagcgtacc ttcagctcac agacatgata agatacattg atgagtttgg
      121 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
      181 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
      241 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta
      301 caaatgtggt attggcccat ctctatcggt atcgtagcat aaccccttgg ggcctctaaa
      361 cgggtcttga ggggtttttt gtgcccctcg ggccggattg ctatctaccg gcattggcgc
      421 agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc ccacatatgc cagattcagc
      481 aacggatacg gcttccccaa cttgcccact tccatacgtg tcctccttac cagaaattta
      541 tccttaaggt cgtcagctat cctgcaggcg atctctcgat ttcgatcaag acattccttt
      601 aatggtcttt tctggacacc actaggggtc agaagtagtt catcaaactt tcttccctcc
      661 ctaatctcat tggttacctt gggctatcga aacttaatta accagtcaag tcagctactt
      721 ggcgagatcg acttgtctgg gtttcgacta cgctcagaat tgcgtcagtc aagttcgatc
      781 tggtccttgc tattgcaccc gttctccgat tacgagtttc atttaaatca tgtgagcaaa
      841 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
      901 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
      961 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     1021 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     1081 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     1141 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     1201 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     1261 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     1321 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     1381 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     1441 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     1501 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     1561 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     1621 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     1681 agcgatctgt ctatttcgtt catccatagt tgcatttaaa tttccgaact ctccaaggcc
     1741 ctcgtcggaa aatcttcaaa cctttcgtcc gatccatctt gcaggctacc tctcgaacga
     1801 actatcgcaa gtctcttggc cggccttgcg ccttggctat tgcttggcag cgcctatcgc
     1861 caggtattac tccaatcccg aatatccgag atcgggatca cccgagagaa gttcaaccta
     1921 catcctcaat cccgatctat ccgagatccg aggaatatcg aaatcggggc gcgcctggtg
     1981 taccgagaac gatcctctca gtgcgagtct cgacgatcca tatcgttgct tggcagtcag
     2041 ccagtcggaa tccagcttgg gacccaggaa gtccaatcgt cagatattgt actcaagcct
     2101 ggtcacggca gcgtaccgat ctgtttaaac ctagatattg atagtctgat cggtcaacgt
     2161 ataatcgagt cctagctttt gcaaacatct atcaagagac aggatcagca ggaggctttc
     2221 gcatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     2281 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     2341 cgcgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     2401 ccgaagaacg ctttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     2461 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     2521 tggttgagta ttcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     2581 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     2641 ttggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     2701 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     2761 tgcctgtagc aatggcaaca accttgcgta aactattaac tggcgaacta cttactctag
     2821 cttcccggca acagttgata gactggatgg aggcggataa agttgcagga ccacttctgc
     2881 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     2941 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     3001 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     3061 cctcactgat taagcattgg taaccgattc taggtgcatt ggcgcagaaa aaaatgcctg
     3121 atgcgacgct gcgcgtctta tactcccaca tatgccagat tcagcaacgg atacggcttc
     3181 cccaacttgc ccacttccat acgtgtcctc cttaccagaa atttatcctt aagatcccga
     3241 atcgtttaaa ctcgactctg gctctatcga atctccgtcg tttcgagctt acgcgaacag
     3301 ccgtggcgct catttgctcg tcgggcatcg aatctcgtca gctatcgtca gcttaccttt
     3361 ttggcagcga tcgcggctcc cgacatcttg gaccattagc tccacaggta tcttcttccc
     3421 tctagtggtc ataacagcag cttcagctac ctctcaattc aaaaaacccc tcaagacccg
     3481 tttagaggcc ccaaggggtt atgctatcaa tcgttgcgtt acacacacaa aaaaccaaca
     3541 cacatccatc ttcgatggat agcgatttta ttatctaact gctgatcgag tgtagccaga
     3601 tctagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
     3661 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
     3721 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
     3781 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
     3841 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
     3901 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatgctga
     3961 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
     4021 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
     4081 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
     4141 aggtctatat aagcagagct ggtttagtga accgtcagat cagatctttg tcgatcctac
     4201 catccactcg acacacccgc cagcggccgc
//
