LOCUS       sequence-300719-        6834 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 6834)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6834)
  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-m4252924
FEATURES             Location/Qualifiers
     source          1..6834
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        494..1703
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     1540..1559
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             1707..2735
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    2762..2810
                     /note="synthetic polyadenylation signal"
     CDS             join(3076..3144,3145..3936)
                     /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(3294..3313)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3949..4085
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4237..4308
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4313..4359)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4377..4425
                     /note="synthetic polyadenylation signal"
     enhancer        4506..4809
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4811..5013
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4964..4984
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             5334..5354
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             5355..5381
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    5450..5571
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5461..5480)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5515..5534
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      5686..5732
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      5762..5898
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5943..6014
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(6240..6828)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6320..6339)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
       61 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
      121 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
      181 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc atttaaattt
      241 ccgaactctc caaggccctc gtcggaaaat cttcaaacct ttcgtccgat ccatcttgca
      301 ggctacctct cgaacgaact atcgcaagtc tcttggccgg ccttgcgcct tggctattgc
      361 ttggcagcgc ctatcgccag gtattactcc aatcccgaat atccgagatc gggatcaccc
      421 gagagaagtt caacctacat cctcaatccc gatctatccg agatccgagg aatatcgaaa
      481 tcggggcgcg cctggcctcc gcgccgggtt ttggcgcctc ccgcgggcgc ccccctcgtc
      541 acggcgagcg ctgccacgtc agacgaaggg cgcaggagcg tcctgatcct tccgcccgga
      601 cgctcaggac agcggcccgc tgctcataag actcggcctt agaaccccag tatcagcaga
      661 aggacatttt aggacgggac ttgggtgact ctagggcact ggttttcttt ccagagagcg
      721 gaacaggcga ggaaaagtag tcccttctcg gcgattctgc ggagggatct ccgtggggcg
      781 gtgaacgccg atgattatat aaggacgcgc cgggtgtggc acagctagtt ccgtcgcagc
      841 cgggatttgg gtcgcggttc ttgtttgtgg atcgctgtga tcgtcacttg gtgagtagcg
      901 ggctgctggg ctggccgggg ctttcgtggc cgccgggccg ctcggtggga cggaagcgtg
      961 tggagagacc gccaagggct gtagtctggg tccgcgagca aggttgccct gaactggggg
     1021 ttggggggag cgcagcaaaa tggcggctgt tcccgagtct tgaatggaag acgcttgtga
     1081 ggcgggctgt gaggtcgttg aaacaaggtg gggggcatgg tgggcggcaa gaacccaagg
     1141 tcttgagccc ttcgctaatg cgggaaagct cttattcggg tgagatgggc tgggcaccat
     1201 ctggggaccc tgacgtgaag tttgtcactg actggagaac tcggtttgtc gtctgttgcg
     1261 ggggcggcag ttatggcggt gccgttgggc agtgcacccg tacctttggg agcgcgcgcc
     1321 ctcgtcgtgt cgtgacgtca cccgttctgt tggcttataa tgcagggtgg ggccaccagc
     1381 cggtaggtgt gcggtaggct tttctccgtc gcaggacgca gggttcgggc ctagggtagg
     1441 ctctcctgaa tcgacaggcg ccggacctct ggtgagggga gggataagtg aggcgtcagt
     1501 ttctttggtc ggttttatgt acctatcttc ttaagtagct gaagctccgg ttttgaacta
     1561 tgcgctcggg gttggcgagt gtgttttgtg aagtttttta ggcacctttt gaaatgtaat
     1621 catttgggtc aatatgtaat tttcagtgtt agacttgtaa attgtccgct aaattctggc
     1681 cgtttttggc ttttttgtta gacaacatgg gtaaaaagcc tgaactcacc gcgacgtctg
     1741 tcgagaagtt tctgatcgaa aagttcgaca gcgtctccga cctgatgcag ctctcggagg
     1801 gcgaagaatc tcgtgctttc agcttcgatg taggagggcg tggatatgtc ctgcgggtaa
     1861 atagctgcgc cgatggtttc tacaaagatc gttatgttta tcggcacttt gcatccgccg
     1921 cgctcccgat tccggaagtg cttgacattg gggagttcag cgagagcctg acctattgca
     1981 tctcccgccg tgcacagggt gtcacgttgc aagacctgcc tgaaaccgaa ctgcccgctg
     2041 ttctgcagcc ggtcgcggag gcaatggatg ccatcgctgc cgccgatctt agccagacga
     2101 gcgggttcgg cccattcgga ccgcaaggaa tcggtcaata cactacatgg cgtgatttca
     2161 tatgcgcgat tgctgatccc catgtgtatc actggcaaac tgtgatggac gacaccgtca
     2221 gtgcgtccgt cgcgcaggct ctcgatgagc tgatgctttg ggccgaggac tgccccgaag
     2281 tccggcacct cgtgcacgcg gatttcggct ccaacaatgt cctgacggac aatggccgca
     2341 taacagcggt cattgactgg agcgaggcga tgttcgggga ttcccaatac gaggtcgcca
     2401 acatcttctt ctggaggccg tggttggctt gtatggagca gcagacgcgc tacttcgagc
     2461 ggaggcatcc ggagcttgca ggatcgcccc ggctccgggc gtatatgctc cgcattggtc
     2521 ttgaccaact ctatcagagc ttggttgacg gcaatttcga tgatgcagct tgggcgcagg
     2581 gtcgatgcga cgcaatcgtc cgatccggag ccgggactgt cgggcgtaca caaatcgccc
     2641 gcagaagcgc cgccgtctgg accgatggct gtgtagaagt actcgccgat agtggaaacc
     2701 gacgccccag cactcgtccg agggcaaagg aataagctag tatgtaagcc tagtcttaga
     2761 taataaaatc gctatccatc gaagatggat gtgtgttggt tttttgtgtg tgtaacgcta
     2821 ggcgcgcctg gtgtaccgag aacgatcctc tcagtgcgag tctcgacgat ccatatcgtt
     2881 gcttggcagt cagccagtcg gaatccagct tgggacccag gaagtccaat cgtcagatat
     2941 tgtactcaag cctggtcacg gcagcgtacc gatctgttta aacctagata ttgatagtct
     3001 gatcggtcaa cgtataatcg agtcctagct tttgcaaaca tctatcaaga gacaggatca
     3061 gcaggaggct ttcgcatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     3121 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     3181 gatcagttgg gtgcgcgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     3241 gagagttttc gccccgaaga acgctttcca atgatgagca cttttaaagt tctgctatgt
     3301 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     3361 tctcagaatg acttggttga gtattcacca gtcacagaaa agcatcttac ggatggcatg
     3421 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     3481 cttctgacaa cgattggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     3541 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     3601 cgtgacacca cgatgcctgt agcaatggca acaaccttgc gtaaactatt aactggcgaa
     3661 ctacttactc tagcttcccg gcaacagttg atagactgga tggaggcgga taaagttgca
     3721 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     3781 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     3841 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     3901 gctgagatag gtgcctcact gattaagcat tggtaaccga ttctaggtgc attggcgcag
     3961 aaaaaaatgc ctgatgcgac gctgcgcgtc ttatactccc acatatgcca gattcagcaa
     4021 cggatacggc ttccccaact tgcccacttc catacgtgtc ctccttacca gaaatttatc
     4081 cttaagatcc cgaatcgttt aaactcgact ctggctctat cgaatctccg tcgtttcgag
     4141 cttacgcgaa cagccgtggc gctcatttgc tcgtcgggca tcgaatctcg tcagctatcg
     4201 tcagcttacc tttttggcag cgatcgcggc tcccgacatc ttggaccatt agctccacag
     4261 gtatcttctt ccctctagtg gtcataacag cagcttcagc tacctctcaa ttcaaaaaac
     4321 ccctcaagac ccgtttagag gccccaaggg gttatgctat caatcgttgc gttacacaca
     4381 caaaaaacca acacacatcc atcttcgatg gatagcgatt ttattatcta actgctgatc
     4441 gagtgtagcc agatctagta atcaattacg gggtcattag ttcatagccc atatatggag
     4501 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
     4561 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
     4621 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
     4681 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
     4741 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
     4801 attaccatgc tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
     4861 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
     4921 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
     4981 cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca gatcagatct
     5041 ttgtcgatcc taccatccac tcgacacacc cgccagcggc cgccaccatg aaaatgtcag
     5101 atgtaaagtg cacatcagta gtcttactct cagttttgca acaactcaga gtagaatcat
     5161 catctaaatt gtgggctcaa tgtgtccagt tacacaatga cattctctta gctaaagata
     5221 ctactgaagc ctttgaaaaa atggtttcac tactttctgt tttgctttcc atgcagggtg
     5281 ctgtagacat aaacaagctt tgtgaagaaa tgctggacaa cagggcaacc ttagaaaacc
     5341 tttactttca gtcgtacccc tatgacgtac cagattatgc gtaaggatcc gcaggcctct
     5401 gctagcttga ctgactgaga tacagcgtac cttcagctca cagacatgat aagatacatt
     5461 gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt
     5521 tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac
     5581 aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag
     5641 taaaacctct acaaatgtgg tattggccca tctctatcgg tatcgtagca taaccccttg
     5701 gggcctctaa acgggtcttg aggggttttt tgtgcccctc gggccggatt gctatctacc
     5761 ggcattggcg cagaaaaaaa tgcctgatgc gacgctgcgc gtcttatact cccacatatg
     5821 ccagattcag caacggatac ggcttcccca acttgcccac ttccatacgt gtcctcctta
     5881 ccagaaattt atccttaagg tcgtcagcta tcctgcaggc gatctctcga tttcgatcaa
     5941 gacattcctt taatggtctt ttctggacac cactaggggt cagaagtagt tcatcaaact
     6001 ttcttccctc cctaatctca ttggttacct tgggctatcg aaacttaatt aaccagtcaa
     6061 gtcagctact tggcgagatc gacttgtctg ggtttcgact acgctcagaa ttgcgtcagt
     6121 caagttcgat ctggtccttg ctattgcacc cgttctccga ttacgagttt catttaaatc
     6181 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     6241 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     6301 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     6361 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     6421 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     6481 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     6541 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     6601 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     6661 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     6721 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     6781 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agat
//
