LOCUS       sequence-300226-        8208 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 N (nucleocapsid).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8208)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 8208)
  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-m4134348
FEATURES             Location/Qualifiers
     source          1..8208
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        306..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1840..1860
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             2209..2226
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2319..2440
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2330..2349)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2384..2403
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2555..2601
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2631..2767
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2812..2883
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(3109..3697)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3189..3208)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        4197..5406
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     5243..5262
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             5410..6438
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    6465..6513
                     /note="synthetic polyadenylation signal"
     CDS             join(6779..6847,6848..7639)
                     /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(6997..7016)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      7652..7788
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7940..8011
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(8016..8062)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    8080..8128
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catgctgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatca gatctttgtc
      541 gatcctacca tccactcgac acacccgcca gcggccgcca ccatgtctga taatggaccc
      601 caaaatcagc gaaatgcacc ccgcattacg tttggtggac cctcagattc aactggcagt
      661 aaccagaatg gagaacgcag tggggcgcga tcaaaacaac gtcggcccca aggtttaccc
      721 aataatactg cgtcttggtt caccgctctc actcaacatg gcaaggaaga ccttaaattc
      781 cctcgaggac aaggcgttcc aattaacacc aatagcagtc cagatgacca aattggctac
      841 taccgaagag ctaccagacg aattcgtggt ggtgacggta aaatgaaaga tctcagtcca
      901 agatggtatt tctactacct aggaactggg ccagaagctg gacttcccta tggtgctaac
      961 aaagacggca tcatatgggt tgcaactgag ggagccttga atacaccaaa agatcacatt
     1021 ggcacccgca atcctgctaa caatgctgca atcgtgctac aacttcctca aggaacaaca
     1081 ttgccaaaag gcttctacgc agaagggagc agaggcggca gtcaagcctc ttctcgttcc
     1141 tcatcacgta gtcgcaacag ttcaagaaat tcaactccag gcagcagtag gggaacttct
     1201 cctgctagaa tggctggcaa tggcggtgat gctgctcttg ctttgctgct gcttgacaga
     1261 ttgaaccagc ttgagagcaa aatgtctggt aaaggccaac aacaacaagg ccaaactgtc
     1321 actaagaaat ctgctgctga ggcttctaag aagcctcggc aaaaacgtac tgccactaaa
     1381 gcatacaatg taacacaagc tttcggcaga cgtggtccag aacaaaccca aggaaatttt
     1441 ggggaccagg aactaatcag acaaggaact gattacaaac attggccgca aattgcacaa
     1501 tttgccccca gcgcttcagc gttcttcgga atgtcgcgca ttggcatgga agtcacacct
     1561 tcgggaacgt ggttgaccta cacaggtgcc atcaaattgg atgacaaaga tccaaatttc
     1621 aaagatcaag tcattttgct gaataagcat attgacgcat acaaaacatt cccaccaaca
     1681 gagcctaaaa aggacaaaaa gaagaaggct gatgaaactc aagccttacc gcagagacag
     1741 aagaaacagc aaactgtgac tcttcttcct gctgcagatt tggatgattt ctccaaacaa
     1801 ttgcaacaat ccatgagcag tgctgactca actcaggccg aaaaccttta ctttcagtcg
     1861 atgagtgaca aaatcattca tctaaccgat gactcctttg atacagacgt gttgaaggcc
     1921 gatggtgcga tactggtaga tttctgggct gaatggtgtg gcccgtgcaa gatgatcgca
     1981 cccattctgg acgagatcgc cgacgagtat caaggaaaat taactgtcgc taagcttaat
     2041 atagatcaga acccagggac ggcacctaaa tacggcatca ggggaatacc taccctcctg
     2101 ctctttaaga atggggaagt tgccgcgaca aaagtgggag ccctgtctaa gggccagcta
     2161 aaagagttct tggacgctaa cctggcaggt tcaggctcgg ggcacatgca ccatcaccat
     2221 caccatagca gcgggctcgt gccacgcggc taaggatccg caggcctctg ctagcttgac
     2281 tgactgagat acagcgtacc ttcagctcac agacatgata agatacattg atgagtttgg
     2341 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     2401 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     2461 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta
     2521 caaatgtggt attggcccat ctctatcggt atcgtagcat aaccccttgg ggcctctaaa
     2581 cgggtcttga ggggtttttt gtgcccctcg ggccggattg ctatctaccg gcattggcgc
     2641 agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc ccacatatgc cagattcagc
     2701 aacggatacg gcttccccaa cttgcccact tccatacgtg tcctccttac cagaaattta
     2761 tccttaaggt cgtcagctat cctgcaggcg atctctcgat ttcgatcaag acattccttt
     2821 aatggtcttt tctggacacc actaggggtc agaagtagtt catcaaactt tcttccctcc
     2881 ctaatctcat tggttacctt gggctatcga aacttaatta accagtcaag tcagctactt
     2941 ggcgagatcg acttgtctgg gtttcgacta cgctcagaat tgcgtcagtc aagttcgatc
     3001 tggtccttgc tattgcaccc gttctccgat tacgagtttc atttaaatca tgtgagcaaa
     3061 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     3121 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     3181 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     3241 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     3301 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     3361 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     3421 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     3481 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     3541 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     3601 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     3661 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     3721 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     3781 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     3841 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     3901 agcgatctgt ctatttcgtt catccatagt tgcatttaaa tttccgaact ctccaaggcc
     3961 ctcgtcggaa aatcttcaaa cctttcgtcc gatccatctt gcaggctacc tctcgaacga
     4021 actatcgcaa gtctcttggc cggccttgcg ccttggctat tgcttggcag cgcctatcgc
     4081 caggtattac tccaatcccg aatatccgag atcgggatca cccgagagaa gttcaaccta
     4141 catcctcaat cccgatctat ccgagatccg aggaatatcg aaatcggggc gcgcctggcc
     4201 tccgcgccgg gttttggcgc ctcccgcggg cgcccccctc gtcacggcga gcgctgccac
     4261 gtcagacgaa gggcgcagga gcgtcctgat ccttccgccc ggacgctcag gacagcggcc
     4321 cgctgctcat aagactcggc cttagaaccc cagtatcagc agaaggacat tttaggacgg
     4381 gacttgggtg actctagggc actggttttc tttccagaga gcggaacagg cgaggaaaag
     4441 tagtcccttc tcggcgattc tgcggaggga tctccgtggg gcggtgaacg ccgatgatta
     4501 tataaggacg cgccgggtgt ggcacagcta gttccgtcgc agccgggatt tgggtcgcgg
     4561 ttcttgtttg tggatcgctg tgatcgtcac ttggtgagta gcgggctgct gggctggccg
     4621 gggctttcgt ggccgccggg ccgctcggtg ggacggaagc gtgtggagag accgccaagg
     4681 gctgtagtct gggtccgcga gcaaggttgc cctgaactgg gggttggggg gagcgcagca
     4741 aaatggcggc tgttcccgag tcttgaatgg aagacgcttg tgaggcgggc tgtgaggtcg
     4801 ttgaaacaag gtggggggca tggtgggcgg caagaaccca aggtcttgag cccttcgcta
     4861 atgcgggaaa gctcttattc gggtgagatg ggctgggcac catctgggga ccctgacgtg
     4921 aagtttgtca ctgactggag aactcggttt gtcgtctgtt gcgggggcgg cagttatggc
     4981 ggtgccgttg ggcagtgcac ccgtaccttt gggagcgcgc gccctcgtcg tgtcgtgacg
     5041 tcacccgttc tgttggctta taatgcaggg tggggccacc tgccggtagg tgtgcggtag
     5101 gcttttctcc gtcgcaggac gcagggttcg ggcctagggt aggctctcct gaatcgacag
     5161 gcgccggacc tctggtgagg ggagggataa gtgaggcgtc agtttctttg gtcggtttta
     5221 tgtacctatc ttcttaagta gctgaagctc cggttttgaa ctatgcgctc ggggttggcg
     5281 agtgtgtttt gtgaagtttt ttaggcacct tttgaaatgt aatcatttgg gtcaatatgt
     5341 aattttcagt gttagacttg taaattgtcc gctaaattct ggccgttttt ggcttttttg
     5401 ttagacaaca tgggtaaaaa gcctgaactc accgcgacgt ctgtcgagaa gtttctgatc
     5461 gaaaagttcg acagcgtctc cgacctgatg cagctctcgg agggcgaaga atctcgtgct
     5521 ttcagcttcg atgtaggagg gcgtggatat gtcctgcggg taaatagctg cgccgatggt
     5581 ttctacaaag atcgttatgt ttatcggcac tttgcatccg ccgcgctccc gattccggaa
     5641 gtgcttgaca ttggggagtt cagcgagagc ctgacctatt gcatctcccg ccgtgcacag
     5701 ggtgtcacgt tgcaagacct gcctgaaacc gaactgcccg ctgttctgca gccggtcgcg
     5761 gaggcaatgg atgccatcgc tgccgccgat cttagccaga cgagcgggtt cggcccattc
     5821 ggaccgcaag gaatcggtca atacactaca tggcgtgatt tcatatgcgc gattgctgat
     5881 ccccatgtgt atcactggca aactgtgatg gacgacaccg tcagtgcgtc cgtcgcgcag
     5941 gctctcgatg agctgatgct ttgggccgag gactgccccg aagtccggca cctcgtgcac
     6001 gcggatttcg gctccaacaa tgtcctgacg gacaatggcc gcataacagc ggtcattgac
     6061 tggagcgagg cgatgttcgg ggattcccaa tacgaggtcg ccaacatctt cttctggagg
     6121 ccgtggttgg cttgtatgga gcagcagacg cgctacttcg agcggaggca tccggagctt
     6181 gcaggatcgc cccggctccg ggcgtatatg ctccgcattg gtcttgacca actctatcag
     6241 agcttggttg acggcaattt cgatgatgca gcttgggcgc agggtcgatg cgacgcaatc
     6301 gtccgatccg gagccgggac tgtcgggcgt acacaaatcg cccgcagaag cgccgccgtc
     6361 tggaccgatg gctgtgtaga agtactcgcc gatagtggaa accgacgccc cagcactcgt
     6421 ccgagggcaa aggaataagc tagtatgtaa gcctagtctt agataataaa atcgctatcc
     6481 atcgaagatg gatgtgtgtt ggttttttgt gtgtgtaacg ctaggcgcgc ctggtgtacc
     6541 gagaacgatc ctctcagtgc gagtctcgac gatccatatc gttgcttggc agtcagccag
     6601 tcggaatcca gcttgggacc caggaagtcc aatcgtcaga tattgtactc aagcctggtc
     6661 acggcagcgt accgatctgt ttaaacctag atattgatag tctgatcggt caacgtataa
     6721 tcgagtccta gcttttgcaa acatctatca agagacagga tcagcaggag gctttcgcat
     6781 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     6841 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcgcg
     6901 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     6961 agaacgcttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     7021 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     7081 tgagtattca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     7141 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgattgg
     7201 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     7261 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     7321 tgtagcaatg gcaacaacct tgcgtaaact attaactggc gaactactta ctctagcttc
     7381 ccggcaacag ttgatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     7441 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     7501 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     7561 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     7621 actgattaag cattggtaac cgattctagg tgcattggcg cagaaaaaaa tgcctgatgc
     7681 gacgctgcgc gtcttatact cccacatatg ccagattcag caacggatac ggcttcccca
     7741 acttgcccac ttccatacgt gtcctcctta ccagaaattt atccttaaga tcccgaatcg
     7801 tttaaactcg actctggctc tatcgaatct ccgtcgtttc gagcttacgc gaacagccgt
     7861 ggcgctcatt tgctcgtcgg gcatcgaatc tcgtcagcta tcgtcagctt acctttttgg
     7921 cagcgatcgc ggctcccgac atcttggacc attagctcca caggtatctt cttccctcta
     7981 gtggtcataa cagcagcttc agctacctct caattcaaaa aacccctcaa gacccgttta
     8041 gaggccccaa ggggttatgc tatcaatcgt tgcgttacac acacaaaaaa ccaacacaca
     8101 tccatcttcg atggatagcg attttattat ctaactgctg atcgagtgta gccagatcta
     8161 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccg
//
