LOCUS       sequence-295112-        4539 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 nsp7.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4539)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4539)
  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-m4134403
FEATURES             Location/Qualifiers
     source          1..4539
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             257..277
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             278..355
                     /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"
     polyA_signal    408..529
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(419..438)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     473..492
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      644..690
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      720..856
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    901..972
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1198..1786)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1278..1297)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(2532..2600,2601..3392)
                     /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(2750..2769)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3405..3541
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3693..3764
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(3769..3815)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    3833..3881
                     /note="synthetic polyadenylation signal"
     enhancer        3962..4265
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4267..4469
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4420..4440
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgagc aaaatgtctg acgtcaagtg tacatccgta gttctgttat cagtgcttca
       61 acagctgcgg gtggagagtt cgtcaaagct ctgggcccag tgcgtgcagc tgcacaatga
      121 tatattgctc gcgaaagata ctacggaagc tttcgagaag atggtcagct tgctatctgt
      181 tctactgagc atgcaaggag cagtagacat caacaaactt tgcgaagaga tgttagacaa
      241 ccgagccacc ctgcaagaaa acctttactt tcagtcgaag aggcgatgga aaaagaattt
      301 tatcgcggtg agtgcagcca acagattcaa aaagatatcc tcgtctggtg ccctctaagc
      361 tagcttgact gactgagata cagcgtacct tcagctcaca gacatgataa gatacattga
      421 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
      481 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
      541 ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta
      601 aaacctctac aaatgtggta ttggcccatc tctatcggta tcgtagcata accccttggg
      661 gcctctaaac gggtcttgag gggttttttg tgcccctcgg gccggattgc tatctaccgg
      721 cattggcgca gaaaaaaatg cctgatgcga cgctgcgcgt cttatactcc cacatatgcc
      781 agattcagca acggatacgg cttccccaac ttgcccactt ccatacgtgt cctccttacc
      841 agaaatttat ccttaaggtc gtcagctatc ctgcaggcga tctctcgatt tcgatcaaga
      901 cattccttta atggtctttt ctggacacca ctaggggtca gaagtagttc atcaaacttt
      961 cttccctccc taatctcatt ggttaccttg ggctatcgaa acttaattaa ccagtcaagt
     1021 cagctacttg gcgagatcga cttgtctggg tttcgactac gctcagaatt gcgtcagtca
     1081 agttcgatct ggtccttgct attgcacccg ttctccgatt acgagtttca tttaaatcat
     1141 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     1201 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     1261 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     1321 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     1381 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     1441 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     1501 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     1561 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     1621 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     1681 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     1741 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     1801 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     1861 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     1921 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     1981 acctatctca gcgatctgtc tatttcgttc atccatagtt gcatttaaat ttccgaactc
     2041 tccaaggccc tcgtcggaaa atcttcaaac ctttcgtccg atccatcttg caggctacct
     2101 ctcgaacgaa ctatcgcaag tctcttggcc ggccttgcgc cttggctatt gcttggcagc
     2161 gcctatcgcc aggtattact ccaatcccga atatccgaga tcgggatcac ccgagagaag
     2221 ttcaacctac atcctcaatc ccgatctatc cgagatccga ggaatatcga aatcggggcg
     2281 cgcctggtgt accgagaacg atcctctcag tgcgagtctc gacgatccat atcgttgctt
     2341 ggcagtcagc cagtcggaat ccagcttggg acccaggaag tccaatcgtc agatattgta
     2401 ctcaagcctg gtcacggcag cgtaccgatc tgtttaaacc tagatattga tagtctgatc
     2461 ggtcaacgta taatcgagtc ctagcttttg caaacatcta tcaagagaca ggatcagcag
     2521 gaggctttcg catgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     2581 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     2641 agttgggtgc gcgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     2701 gttttcgccc cgaagaacgc tttccaatga tgagcacttt taaagttctg ctatgtggcg
     2761 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     2821 agaatgactt ggttgagtat tcaccagtca cagaaaagca tcttacggat ggcatgacag
     2881 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     2941 tgacaacgat tggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     3001 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     3061 acaccacgat gcctgtagca atggcaacaa ccttgcgtaa actattaact ggcgaactac
     3121 ttactctagc ttcccggcaa cagttgatag actggatgga ggcggataaa gttgcaggac
     3181 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     3241 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     3301 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     3361 agataggtgc ctcactgatt aagcattggt aaccgattct aggtgcattg gcgcagaaaa
     3421 aaatgcctga tgcgacgctg cgcgtcttat actcccacat atgccagatt cagcaacgga
     3481 tacggcttcc ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa tttatcctta
     3541 agatcccgaa tcgtttaaac tcgactctgg ctctatcgaa tctccgtcgt ttcgagctta
     3601 cgcgaacagc cgtggcgctc atttgctcgt cgggcatcga atctcgtcag ctatcgtcag
     3661 cttacctttt tggcagcgat cgcggctccc gacatcttgg accattagct ccacaggtat
     3721 cttcttccct ctagtggtca taacagcagc ttcagctacc tctcaattca aaaaacccct
     3781 caagacccgt ttagaggccc caaggggtta tgctatcaat cgttgcgtta cacacacaaa
     3841 aaaccaacac acatccatct tcgatggata gcgattttat tatctaactg ctgatcgagt
     3901 gtagccagat ctagtaatca attacggggt cattagttca tagcccatat atggagttcc
     3961 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
     4021 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
     4081 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
     4141 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
     4201 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
     4261 ccatgctgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
     4321 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
     4381 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
     4441 tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc agatctttgt
     4501 cgatcctacc atccactcga cacacccgcc agcggccgc
//
