LOCUS       sequence-300497-        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"
     misc_feature    222..293
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(298..344)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    362..410
                     /note="synthetic polyadenylation signal"
     enhancer        491..794
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        796..998
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     949..969
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     polyA_signal    1393..1514
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1404..1423)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1458..1477
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1629..1675
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1705..1841
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1886..1957
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2183..2771)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2263..2282)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3271..4480
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4317..4336
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4484..5512
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5539..5587
                     /note="synthetic polyadenylation signal"
     CDS             join(5853..5921,5922..6713)
                     /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(6071..6090)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      join(6726..6792,1..70)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
ORIGIN
        1 agcaacggat acggcttccc caacttgccc acttccatac gtgtcctcct taccagaaat
       61 ttatccttaa gatcccgaat cgtttaaact cgactctggc tctatcgaat ctccgtcgtt
      121 tcgagcttac gcgaacagcc gtggcgctca tttgctcgtc gggcatcgaa tctcgtcagc
      181 tatcgtcagc ttaccttttt ggcagcgatc gcggctcccg acatcttgga ccattagctc
      241 cacaggtatc ttcttccctc tagtggtcat aacagcagct tcagctacct ctcaattcaa
      301 aaaacccctc aagacccgtt tagaggcccc aaggggttat gctatcaatc gttgcgttac
      361 acacacaaaa aaccaacaca catccatctt cgatggatag cgattttatt atctaactgc
      421 tgatcgagtg tagccagatc tagtaatcaa ttacggggtc attagttcat agcccatata
      481 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      541 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      601 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      661 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      721 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      781 tcgctattac catgctgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      841 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      901 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      961 gtaggcgtgt acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatca
     1021 gatctttgtc gatcctacca tccactcgac acacccgcca gcggccgcca ccatgagcaa
     1081 aatgtctgac gtcaagtgta catccgtagt tctgttatca gtgcttcaac agctgcgggt
     1141 ggagagttcg tcaaagctct gggcccagtg cgtgcagctg cacaatgata tattgctcgc
     1201 gaaagatact acggaagctt tcgagaagat ggtcagcttg ctatctgttc tactgagcat
     1261 gcaaggagca gtagacatca acaaactttg cgaagagatg ttagacaacc gagccaccct
     1321 gcaataagga tccgcaggcc tctgctagct tgactgactg agatacagcg taccttcagc
     1381 tcacagacat gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa
     1441 aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct
     1501 gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg
     1561 tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtattggc ccatctctat
     1621 cggtatcgta gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgtgccc
     1681 ctcgggccgg attgctatct accggcattg gcgcagaaaa aaatgcctga tgcgacgctg
     1741 cgcgtcttat actcccacat atgccagatt cagcaacgga tacggcttcc ccaacttgcc
     1801 cacttccata cgtgtcctcc ttaccagaaa tttatcctta aggtcgtcag ctatcctgca
     1861 ggcgatctct cgatttcgat caagacattc ctttaatggt cttttctgga caccactagg
     1921 ggtcagaagt agttcatcaa actttcttcc ctccctaatc tcattggtta ccttgggcta
     1981 tcgaaactta attaaccagt caagtcagct acttggcgag atcgacttgt ctgggtttcg
     2041 actacgctca gaattgcgtc agtcaagttc gatctggtcc ttgctattgc acccgttctc
     2101 cgattacgag tttcatttaa atcatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     2161 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     2221 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     2281 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     2341 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     2401 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     2461 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     2521 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     2581 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     2641 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     2701 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     2761 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     2821 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     2881 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     2941 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     3001 tagttgcatt taaatttccg aactctccaa ggccctcgtc ggaaaatctt caaacctttc
     3061 gtccgatcca tcttgcaggc tacctctcga acgaactatc gcaagtctct tggccggcct
     3121 tgcgccttgg ctattgcttg gcagcgccta tcgccaggta ttactccaat cccgaatatc
     3181 cgagatcggg atcacccgag agaagttcaa cctacatcct caatcccgat ctatccgaga
     3241 tccgaggaat atcgaaatcg gggcgcgcct ggcctccgcg ccgggttttg gcgcctcccg
     3301 cgggcgcccc cctcgtcacg gcgagcgctg ccacgtcaga cgaagggcgc aggagcgtcc
     3361 tgatccttcc gcccggacgc tcaggacagc ggcccgctgc tcataagact cggccttaga
     3421 accccagtat cagcagaagg acattttagg acgggacttg ggtgactcta gggcactggt
     3481 tttctttcca gagagcggaa caggcgagga aaagtagtcc cttctcggcg attctgcgga
     3541 gggatctccg tggggcggtg aacgccgatg attatataag gacgcgccgg gtgtggcaca
     3601 gctagttccg tcgcagccgg gatttgggtc gcggttcttg tttgtggatc gctgtgatcg
     3661 tcacttggtg agtagcgggc tgctgggctg gccggggctt tcgtggccgc cgggccgctc
     3721 ggtgggacgg aagcgtgtgg agagaccgcc aagggctgta gtctgggtcc gcgagcaagg
     3781 ttgccctgaa ctgggggttg gggggagcgc agcaaaatgg cggctgttcc cgagtcttga
     3841 atggaagacg cttgtgaggc gggctgtgag gtcgttgaaa caaggtgggg ggcatggtgg
     3901 gcggcaagaa cccaaggtct tgagcccttc gctaatgcgg gaaagctctt attcgggtga
     3961 gatgggctgg gcaccatctg gggaccctga cgtgaagttt gtcactgact ggagaactcg
     4021 gtttgtcgtc tgttgcgggg gcggcagtta tggcggtgcc gttgggcagt gcacccgtac
     4081 ctttgggagc gcgcgccctc gtcgtgtcgt gacgtcaccc gttctgttgg cttataatgc
     4141 agggtggggc caccagccgg taggtgtgcg gtaggctttt ctccgtcgca ggacgcaggg
     4201 ttcgggccta gggtaggctc tcctgaatcg acaggcgccg gacctctggt gaggggaggg
     4261 ataagtgagg cgtcagtttc tttggtcggt tttatgtacc tatcttctta agtagctgaa
     4321 gctccggttt tgaactatgc gctcggggtt ggcgagtgtg ttttgtgaag ttttttaggc
     4381 accttttgaa atgtaatcat ttgggtcaat atgtaatttt cagtgttaga cttgtaaatt
     4441 gtccgctaaa ttctggccgt ttttggcttt tttgttagac aacatgggta aaaagcctga
     4501 actcaccgcg acgtctgtcg agaagtttct gatcgaaaag ttcgacagcg tctccgacct
     4561 gatgcagctc tcggagggcg aagaatctcg tgctttcagc ttcgatgtag gagggcgtgg
     4621 atatgtcctg cgggtaaata gctgcgccga tggtttctac aaagatcgtt atgtttatcg
     4681 gcactttgca tccgccgcgc tcccgattcc ggaagtgctt gacattgggg agttcagcga
     4741 gagcctgacc tattgcatct cccgccgtgc acagggtgtc acgttgcaag acctgcctga
     4801 aaccgaactg cccgctgttc tgcagccggt cgcggaggca atggatgcca tcgctgccgc
     4861 cgatcttagc cagacgagcg ggttcggccc attcggaccg caaggaatcg gtcaatacac
     4921 tacatggcgt gatttcatat gcgcgattgc tgatccccat gtgtatcact ggcaaactgt
     4981 gatggacgac accgtcagtg cgtccgtcgc gcaggctctc gatgagctga tgctttgggc
     5041 cgaggactgc cccgaagtcc ggcacctcgt gcacgcggat ttcggctcca acaatgtcct
     5101 gacggacaat ggccgcataa cagcggtcat tgactggagc gaggcgatgt tcggggattc
     5161 ccaatacgag gtcgccaaca tcttcttctg gaggccgtgg ttggcttgta tggagcagca
     5221 gacgcgctac ttcgagcgga ggcatccgga gcttgcagga tcgccccggc tccgggcgta
     5281 tatgctccgc attggtcttg accaactcta tcagagcttg gttgacggca atttcgatga
     5341 tgcagcttgg gcgcagggtc gatgcgacgc aatcgtccga tccggagccg ggactgtcgg
     5401 gcgtacacaa atcgcccgca gaagcgccgc cgtctggacc gatggctgtg tagaagtact
     5461 cgccgatagt ggaaaccgac gccccagcac tcgtccgagg gcaaaggaat aagctagtat
     5521 gtaagcctag tcttagataa taaaatcgct atccatcgaa gatggatgtg tgttggtttt
     5581 ttgtgtgtgt aacgctaggc gcgcctggtg taccgagaac gatcctctca gtgcgagtct
     5641 cgacgatcca tatcgttgct tggcagtcag ccagtcggaa tccagcttgg gacccaggaa
     5701 gtccaatcgt cagatattgt actcaagcct ggtcacggca gcgtaccgat ctgtttaaac
     5761 ctagatattg atagtctgat cggtcaacgt ataatcgagt cctagctttt gcaaacatct
     5821 atcaagagac aggatcagca ggaggctttc gcatgagtat tcaacatttc cgtgtcgccc
     5881 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     5941 aagtaaaaga tgctgaagat cagttgggtg cgcgagtggg ttacatcgaa ctggatctca
     6001 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ctttccaatg atgagcactt
     6061 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     6121 gtcgccgcat acactattct cagaatgact tggttgagta ttcaccagtc acagaaaagc
     6181 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     6241 acactgcggc caacttactt ctgacaacga ttggaggacc gaaggagcta accgcttttt
     6301 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     6361 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca accttgcgta
     6421 aactattaac tggcgaacta cttactctag cttcccggca acagttgata gactggatgg
     6481 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     6541 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     6601 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     6661 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taaccgattc
     6721 taggtgcatt ggcgcagaaa aaaatgcctg atgcgacgct gcgcgtctta tactcccaca
     6781 tatgccagat tc
//
