LOCUS       sequence-297622-        6792 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 6792)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6792)
  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-m4252983
FEATURES             Location/Qualifiers
     source          1..6792
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    325..446
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(336..355)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     390..409
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      561..607
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      637..773
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    818..889
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1115..1703)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1195..1214)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2203..3412
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3249..3268
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3416..4444
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    4471..4519
                     /note="synthetic polyadenylation signal"
     CDS             join(4785..4853,4854..5645)
                     /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(5003..5022)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5658..5794
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5946..6017
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6022..6068)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6086..6134
                     /note="synthetic polyadenylation signal"
     enhancer        6215..6518
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6520..6722
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6673..6693
                     /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 ctgcaataag gatccgcagg cctctgctag cttgactgac tgagatacag
      301 cgtaccttca gctcacagac atgataagat acattgatga gtttggacaa accacaacta
      361 gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa
      421 ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg
      481 ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtattg
      541 gcccatctct atcggtatcg tagcataacc ccttggggcc tctaaacggg tcttgagggg
      601 ttttttgtgc ccctcgggcc ggattgctat ctaccggcat tggcgcagaa aaaaatgcct
      661 gatgcgacgc tgcgcgtctt atactcccac atatgccaga ttcagcaacg gatacggctt
      721 ccccaacttg cccacttcca tacgtgtcct ccttaccaga aatttatcct taaggtcgtc
      781 agctatcctg caggcgatct ctcgatttcg atcaagacat tcctttaatg gtcttttctg
      841 gacaccacta ggggtcagaa gtagttcatc aaactttctt ccctccctaa tctcattggt
      901 taccttgggc tatcgaaact taattaacca gtcaagtcag ctacttggcg agatcgactt
      961 gtctgggttt cgactacgct cagaattgcg tcagtcaagt tcgatctggt ccttgctatt
     1021 gcacccgttc tccgattacg agtttcattt aaatcatgtg agcaaaaggc cagcaaaagg
     1081 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     1141 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     1201 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     1261 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     1321 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     1381 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     1441 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     1501 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     1561 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     1621 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     1681 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     1741 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     1801 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     1861 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     1921 ttcgttcatc catagttgca tttaaatttc cgaactctcc aaggccctcg tcggaaaatc
     1981 ttcaaacctt tcgtccgatc catcttgcag gctacctctc gaacgaacta tcgcaagtct
     2041 cttggccggc cttgcgcctt ggctattgct tggcagcgcc tatcgccagg tattactcca
     2101 atcccgaata tccgagatcg ggatcacccg agagaagttc aacctacatc ctcaatcccg
     2161 atctatccga gatccgagga atatcgaaat cggggcgcgc ctggcctccg cgccgggttt
     2221 tggcgcctcc cgcgggcgcc cccctcgtca cggcgagcgc tgccacgtca gacgaagggc
     2281 gcaggagcgt cctgatcctt ccgcccggac gctcaggaca gcggcccgct gctcataaga
     2341 ctcggcctta gaaccccagt atcagcagaa ggacatttta ggacgggact tgggtgactc
     2401 tagggcactg gttttctttc cagagagcgg aacaggcgag gaaaagtagt cccttctcgg
     2461 cgattctgcg gagggatctc cgtggggcgg tgaacgccga tgattatata aggacgcgcc
     2521 gggtgtggca cagctagttc cgtcgcagcc gggatttggg tcgcggttct tgtttgtgga
     2581 tcgctgtgat cgtcacttgg tgagtagcgg gctgctgggc tggccggggc tttcgtggcc
     2641 gccgggccgc tcggtgggac ggaagcgtgt ggagagaccg ccaagggctg tagtctgggt
     2701 ccgcgagcaa ggttgccctg aactgggggt tggggggagc gcagcaaaat ggcggctgtt
     2761 cccgagtctt gaatggaaga cgcttgtgag gcgggctgtg aggtcgttga aacaaggtgg
     2821 ggggcatggt gggcggcaag aacccaaggt cttgagccct tcgctaatgc gggaaagctc
     2881 ttattcgggt gagatgggct gggcaccatc tggggaccct gacgtgaagt ttgtcactga
     2941 ctggagaact cggtttgtcg tctgttgcgg gggcggcagt tatggcggtg ccgttgggca
     3001 gtgcacccgt acctttggga gcgcgcgccc tcgtcgtgtc gtgacgtcac ccgttctgtt
     3061 ggcttataat gcagggtggg gccaccagcc ggtaggtgtg cggtaggctt ttctccgtcg
     3121 caggacgcag ggttcgggcc tagggtaggc tctcctgaat cgacaggcgc cggacctctg
     3181 gtgaggggag ggataagtga ggcgtcagtt tctttggtcg gttttatgta cctatcttct
     3241 taagtagctg aagctccggt tttgaactat gcgctcgggg ttggcgagtg tgttttgtga
     3301 agttttttag gcaccttttg aaatgtaatc atttgggtca atatgtaatt ttcagtgtta
     3361 gacttgtaaa ttgtccgcta aattctggcc gtttttggct tttttgttag acaacatggg
     3421 taaaaagcct gaactcaccg cgacgtctgt cgagaagttt ctgatcgaaa agttcgacag
     3481 cgtctccgac ctgatgcagc tctcggaggg cgaagaatct cgtgctttca gcttcgatgt
     3541 aggagggcgt ggatatgtcc tgcgggtaaa tagctgcgcc gatggtttct acaaagatcg
     3601 ttatgtttat cggcactttg catccgccgc gctcccgatt ccggaagtgc ttgacattgg
     3661 ggagttcagc gagagcctga cctattgcat ctcccgccgt gcacagggtg tcacgttgca
     3721 agacctgcct gaaaccgaac tgcccgctgt tctgcagccg gtcgcggagg caatggatgc
     3781 catcgctgcc gccgatctta gccagacgag cgggttcggc ccattcggac cgcaaggaat
     3841 cggtcaatac actacatggc gtgatttcat atgcgcgatt gctgatcccc atgtgtatca
     3901 ctggcaaact gtgatggacg acaccgtcag tgcgtccgtc gcgcaggctc tcgatgagct
     3961 gatgctttgg gccgaggact gccccgaagt ccggcacctc gtgcacgcgg atttcggctc
     4021 caacaatgtc ctgacggaca atggccgcat aacagcggtc attgactgga gcgaggcgat
     4081 gttcggggat tcccaatacg aggtcgccaa catcttcttc tggaggccgt ggttggcttg
     4141 tatggagcag cagacgcgct acttcgagcg gaggcatccg gagcttgcag gatcgccccg
     4201 gctccgggcg tatatgctcc gcattggtct tgaccaactc tatcagagct tggttgacgg
     4261 caatttcgat gatgcagctt gggcgcaggg tcgatgcgac gcaatcgtcc gatccggagc
     4321 cgggactgtc gggcgtacac aaatcgcccg cagaagcgcc gccgtctgga ccgatggctg
     4381 tgtagaagta ctcgccgata gtggaaaccg acgccccagc actcgtccga gggcaaagga
     4441 ataagctagt atgtaagcct agtcttagat aataaaatcg ctatccatcg aagatggatg
     4501 tgtgttggtt ttttgtgtgt gtaacgctag gcgcgcctgg tgtaccgaga acgatcctct
     4561 cagtgcgagt ctcgacgatc catatcgttg cttggcagtc agccagtcgg aatccagctt
     4621 gggacccagg aagtccaatc gtcagatatt gtactcaagc ctggtcacgg cagcgtaccg
     4681 atctgtttaa acctagatat tgatagtctg atcggtcaac gtataatcga gtcctagctt
     4741 ttgcaaacat ctatcaagag acaggatcag caggaggctt tcgcatgagt attcaacatt
     4801 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     4861 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcgcgagtg ggttacatcg
     4921 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgctttccaa
     4981 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     5041 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tattcaccag
     5101 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     5161 ccatgagtga taacactgcg gccaacttac ttctgacaac gattggagga ccgaaggagc
     5221 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     5281 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     5341 caaccttgcg taaactatta actggcgaac tacttactct agcttcccgg caacagttga
     5401 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     5461 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     5521 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     5581 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     5641 ggtaaccgat tctaggtgca ttggcgcaga aaaaaatgcc tgatgcgacg ctgcgcgtct
     5701 tatactccca catatgccag attcagcaac ggatacggct tccccaactt gcccacttcc
     5761 atacgtgtcc tccttaccag aaatttatcc ttaagatccc gaatcgttta aactcgactc
     5821 tggctctatc gaatctccgt cgtttcgagc ttacgcgaac agccgtggcg ctcatttgct
     5881 cgtcgggcat cgaatctcgt cagctatcgt cagcttacct ttttggcagc gatcgcggct
     5941 cccgacatct tggaccatta gctccacagg tatcttcttc cctctagtgg tcataacagc
     6001 agcttcagct acctctcaat tcaaaaaacc cctcaagacc cgtttagagg ccccaagggg
     6061 ttatgctatc aatcgttgcg ttacacacac aaaaaaccaa cacacatcca tcttcgatgg
     6121 atagcgattt tattatctaa ctgctgatcg agtgtagcca gatctagtaa tcaattacgg
     6181 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
     6241 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
     6301 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
     6361 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
     6421 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
     6481 ggcagtacat ctacgtatta gtcatcgcta ttaccatgct gatgcggttt tggcagtaca
     6541 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
     6601 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
     6661 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
     6721 ctggtttagt gaaccgtcag atcagatctt tgtcgatcct accatccact cgacacaccc
     6781 gccagcggcc gc
//
