LOCUS       sequence-297428-        6876 bp    DNA     circular SYN 15-DEC-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 6876)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6876)
  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-m4252906
FEATURES             Location/Qualifiers
     source          1..6876
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             251..271
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             275..340
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    409..530
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(420..439)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     474..493
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      645..691
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      721..857
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    902..973
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1199..1787)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1279..1298)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2287..3496
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3333..3352
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3500..4528
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    4555..4603
                     /note="synthetic polyadenylation signal"
     CDS             join(4869..4937,4938..5729)
                     /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(5087..5106)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5742..5878
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6030..6101
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6106..6152)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6170..6218
                     /note="synthetic polyadenylation signal"
     enhancer        6299..6602
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6604..6806
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6757..6777
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgaaa atgtcagatg taaagtgcac atcagtagtc ttactctcag ttttgcaaca
       61 actcagagta gaatcatcat ctaaattgtg ggctcaatgt gtccagttac acaatgacat
      121 tctcttagct aaagatacta ctgaagcctt tgaaaaaatg gtttcactac tttctgtttt
      181 gctttccatg cagggtgctg tagacataaa caagctttgt gaagaaatgc tggacaacag
      241 ggcaacctta gaaaaccttt actttcagtc gatggattat aaggaccacg atggagacta
      301 caaagatcat gacattgatt acaaggacga cgatgacaaa taaggatccg caggcctctg
      361 ctagcttgac tgactgagat acagcgtacc ttcagctcac agacatgata agatacattg
      421 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
      481 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
      541 attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt
      601 aaaacctcta caaatgtggt attggcccat ctctatcggt atcgtagcat aaccccttgg
      661 ggcctctaaa cgggtcttga ggggtttttt gtgcccctcg ggccggattg ctatctaccg
      721 gcattggcgc agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc ccacatatgc
      781 cagattcagc aacggatacg gcttccccaa cttgcccact tccatacgtg tcctccttac
      841 cagaaattta tccttaaggt cgtcagctat cctgcaggcg atctctcgat ttcgatcaag
      901 acattccttt aatggtcttt tctggacacc actaggggtc agaagtagtt catcaaactt
      961 tcttccctcc ctaatctcat tggttacctt gggctatcga aacttaatta accagtcaag
     1021 tcagctactt ggcgagatcg acttgtctgg gtttcgacta cgctcagaat tgcgtcagtc
     1081 aagttcgatc tggtccttgc tattgcaccc gttctccgat tacgagtttc atttaaatca
     1141 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     1201 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     1261 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     1321 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     1381 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     1441 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     1501 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     1561 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     1621 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     1681 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     1741 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     1801 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     1861 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     1921 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     1981 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcatttaaa tttccgaact
     2041 ctccaaggcc ctcgtcggaa aatcttcaaa cctttcgtcc gatccatctt gcaggctacc
     2101 tctcgaacga actatcgcaa gtctcttggc cggccttgcg ccttggctat tgcttggcag
     2161 cgcctatcgc caggtattac tccaatcccg aatatccgag atcgggatca cccgagagaa
     2221 gttcaaccta catcctcaat cccgatctat ccgagatccg aggaatatcg aaatcggggc
     2281 gcgcctggcc tccgcgccgg gttttggcgc ctcccgcggg cgcccccctc gtcacggcga
     2341 gcgctgccac gtcagacgaa gggcgcagga gcgtcctgat ccttccgccc ggacgctcag
     2401 gacagcggcc cgctgctcat aagactcggc cttagaaccc cagtatcagc agaaggacat
     2461 tttaggacgg gacttgggtg actctagggc actggttttc tttccagaga gcggaacagg
     2521 cgaggaaaag tagtcccttc tcggcgattc tgcggaggga tctccgtggg gcggtgaacg
     2581 ccgatgatta tataaggacg cgccgggtgt ggcacagcta gttccgtcgc agccgggatt
     2641 tgggtcgcgg ttcttgtttg tggatcgctg tgatcgtcac ttggtgagta gcgggctgct
     2701 gggctggccg gggctttcgt ggccgccggg ccgctcggtg ggacggaagc gtgtggagag
     2761 accgccaagg gctgtagtct gggtccgcga gcaaggttgc cctgaactgg gggttggggg
     2821 gagcgcagca aaatggcggc tgttcccgag tcttgaatgg aagacgcttg tgaggcgggc
     2881 tgtgaggtcg ttgaaacaag gtggggggca tggtgggcgg caagaaccca aggtcttgag
     2941 cccttcgcta atgcgggaaa gctcttattc gggtgagatg ggctgggcac catctgggga
     3001 ccctgacgtg aagtttgtca ctgactggag aactcggttt gtcgtctgtt gcgggggcgg
     3061 cagttatggc ggtgccgttg ggcagtgcac ccgtaccttt gggagcgcgc gccctcgtcg
     3121 tgtcgtgacg tcacccgttc tgttggctta taatgcaggg tggggccacc agccggtagg
     3181 tgtgcggtag gcttttctcc gtcgcaggac gcagggttcg ggcctagggt aggctctcct
     3241 gaatcgacag gcgccggacc tctggtgagg ggagggataa gtgaggcgtc agtttctttg
     3301 gtcggtttta tgtacctatc ttcttaagta gctgaagctc cggttttgaa ctatgcgctc
     3361 ggggttggcg agtgtgtttt gtgaagtttt ttaggcacct tttgaaatgt aatcatttgg
     3421 gtcaatatgt aattttcagt gttagacttg taaattgtcc gctaaattct ggccgttttt
     3481 ggcttttttg ttagacaaca tgggtaaaaa gcctgaactc accgcgacgt ctgtcgagaa
     3541 gtttctgatc gaaaagttcg acagcgtctc cgacctgatg cagctctcgg agggcgaaga
     3601 atctcgtgct ttcagcttcg atgtaggagg gcgtggatat gtcctgcggg taaatagctg
     3661 cgccgatggt ttctacaaag atcgttatgt ttatcggcac tttgcatccg ccgcgctccc
     3721 gattccggaa gtgcttgaca ttggggagtt cagcgagagc ctgacctatt gcatctcccg
     3781 ccgtgcacag ggtgtcacgt tgcaagacct gcctgaaacc gaactgcccg ctgttctgca
     3841 gccggtcgcg gaggcaatgg atgccatcgc tgccgccgat cttagccaga cgagcgggtt
     3901 cggcccattc ggaccgcaag gaatcggtca atacactaca tggcgtgatt tcatatgcgc
     3961 gattgctgat ccccatgtgt atcactggca aactgtgatg gacgacaccg tcagtgcgtc
     4021 cgtcgcgcag gctctcgatg agctgatgct ttgggccgag gactgccccg aagtccggca
     4081 cctcgtgcac gcggatttcg gctccaacaa tgtcctgacg gacaatggcc gcataacagc
     4141 ggtcattgac tggagcgagg cgatgttcgg ggattcccaa tacgaggtcg ccaacatctt
     4201 cttctggagg ccgtggttgg cttgtatgga gcagcagacg cgctacttcg agcggaggca
     4261 tccggagctt gcaggatcgc cccggctccg ggcgtatatg ctccgcattg gtcttgacca
     4321 actctatcag agcttggttg acggcaattt cgatgatgca gcttgggcgc agggtcgatg
     4381 cgacgcaatc gtccgatccg gagccgggac tgtcgggcgt acacaaatcg cccgcagaag
     4441 cgccgccgtc tggaccgatg gctgtgtaga agtactcgcc gatagtggaa accgacgccc
     4501 cagcactcgt ccgagggcaa aggaataagc tagtatgtaa gcctagtctt agataataaa
     4561 atcgctatcc atcgaagatg gatgtgtgtt ggttttttgt gtgtgtaacg ctaggcgcgc
     4621 ctggtgtacc gagaacgatc ctctcagtgc gagtctcgac gatccatatc gttgcttggc
     4681 agtcagccag tcggaatcca gcttgggacc caggaagtcc aatcgtcaga tattgtactc
     4741 aagcctggtc acggcagcgt accgatctgt ttaaacctag atattgatag tctgatcggt
     4801 caacgtataa tcgagtccta gcttttgcaa acatctatca agagacagga tcagcaggag
     4861 gctttcgcat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     4921 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     4981 tgggtgcgcg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     5041 ttcgccccga agaacgcttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     5101 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     5161 atgacttggt tgagtattca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     5221 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     5281 caacgattgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     5341 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     5401 ccacgatgcc tgtagcaatg gcaacaacct tgcgtaaact attaactggc gaactactta
     5461 ctctagcttc ccggcaacag ttgatagact ggatggaggc ggataaagtt gcaggaccac
     5521 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     5581 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     5641 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     5701 taggtgcctc actgattaag cattggtaac cgattctagg tgcattggcg cagaaaaaaa
     5761 tgcctgatgc gacgctgcgc gtcttatact cccacatatg ccagattcag caacggatac
     5821 ggcttcccca acttgcccac ttccatacgt gtcctcctta ccagaaattt atccttaaga
     5881 tcccgaatcg tttaaactcg actctggctc tatcgaatct ccgtcgtttc gagcttacgc
     5941 gaacagccgt ggcgctcatt tgctcgtcgg gcatcgaatc tcgtcagcta tcgtcagctt
     6001 acctttttgg cagcgatcgc ggctcccgac atcttggacc attagctcca caggtatctt
     6061 cttccctcta gtggtcataa cagcagcttc agctacctct caattcaaaa aacccctcaa
     6121 gacccgttta gaggccccaa ggggttatgc tatcaatcgt tgcgttacac acacaaaaaa
     6181 ccaacacaca tccatcttcg atggatagcg attttattat ctaactgctg atcgagtgta
     6241 gccagatcta gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
     6301 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
     6361 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
     6421 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
     6481 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
     6541 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
     6601 tgctgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
     6661 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
     6721 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
     6781 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatcaga tctttgtcga
     6841 tcctaccatc cactcgacac acccgccagc ggccgc
//
