LOCUS       sequence-300419-        6786 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 6786)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6786)
  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-m4252992
FEATURES             Location/Qualifiers
     source          1..6786
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    25..146
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(62..81)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     116..135
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        complement(535..737)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(564..584)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        739..1042
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    1123..1171
                     /note="synthetic polyadenylation signal"
     terminator      1189..1235
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    1240..1311
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(1463..1599)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(join(1612..2403,2404..2472))
                     /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     2235..2254
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    2738..2786
                     /note="synthetic polyadenylation signal"
     CDS             complement(2813..3841)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     promoter        complement(3845..5054)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(3989..4008)
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     rep_origin      5554..6142
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6043..6062
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    6368..6439
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6484..6620)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     terminator      complement(6650..6696)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa
       61 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
      121 ttgtccaaac tcatcaatgt atcttatcat gtctgtgagc tgaaggtacg ctgtatctca
      181 gtcagtcaag ctagcagagg cctgcggatc cttataaggt tgccctgttg tccagcattt
      241 cttcacaaag cttgtttatg tctacagcac cctgcatgga aagcaaaaca gaaagtagtg
      301 aaaccatttt ttcaaaggct tcagtagtat ctttagctaa gagaatgtca ttgtgtaact
      361 ggacacattg agcccacaat ttagatgatg attctactct gagttgttgc aaaactgaga
      421 gtaagactac tgatgtgcac tttacatctg acattttcat ggtggcggcc gctggcgggt
      481 gtgtcgagtg gatggtagga tcgacaaaga tctgatctga cggttcacta aaccagctct
      541 gcttatatag acctcccacc gtacacgcct accgcccatt tgcgtcaatg gggcggagtt
      601 gttacgacat tttggaaagt cccgttgatt ttggtgccaa aacaaactcc cattgacgtc
      661 aatggggtgg agacttggaa atccccgtga gtcaaaccgc tatccacgcc cattgatgta
      721 ctgccaaaac cgcatcagca tggtaatagc gatgactaat acgtagatgt actgccaagt
      781 aggaaagtcc cataaggtca tgtactgggc ataatgccag gcgggccatt taccgtcatt
      841 gacgtcaata gggggcgtac ttggcatatg atacacttga tgtactgcca agtgggcagt
      901 ttaccgtaaa tactccaccc attgacgtca atggaaagtc cctattggcg ttactatggg
      961 aacatacgtc attattgacg tcaatgggcg ggggtcgttg ggcggtcagc caggcgggcc
     1021 atttaccgta agttatgtaa cgcggaactc catatatggg ctatgaacta atgaccccgt
     1081 aattgattac tagatctggc tacactcgat cagcagttag ataataaaat cgctatccat
     1141 cgaagatgga tgtgtgttgg ttttttgtgt gtgtaacgca acgattgata gcataacccc
     1201 ttggggcctc taaacgggtc ttgaggggtt ttttgaattg agaggtagct gaagctgctg
     1261 ttatgaccac tagagggaag aagatacctg tggagctaat ggtccaagat gtcgggagcc
     1321 gcgatcgctg ccaaaaaggt aagctgacga tagctgacga gattcgatgc ccgacgagca
     1381 aatgagcgcc acggctgttc gcgtaagctc gaaacgacgg agattcgata gagccagagt
     1441 cgagtttaaa cgattcggga tcttaaggat aaatttctgg taaggaggac acgtatggaa
     1501 gtgggcaagt tggggaagcc gtatccgttg ctgaatctgg catatgtggg agtataagac
     1561 gcgcagcgtc gcatcaggca tttttttctg cgccaatgca cctagaatcg gttaccaatg
     1621 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     1681 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     1741 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     1801 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctatcaa
     1861 ctgttgccgg gaagctagag taagtagttc gccagttaat agtttacgca aggttgttgc
     1921 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     1981 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     2041 cttcggtcct ccaatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     2101 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     2161 tgaatactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     2221 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     2281 aaagcgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     2341 gtaacccact cgcgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     2401 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     2461 ttgaatactc atgcgaaagc ctcctgctga tcctgtctct tgatagatgt ttgcaaaagc
     2521 taggactcga ttatacgttg accgatcaga ctatcaatat ctaggtttaa acagatcggt
     2581 acgctgccgt gaccaggctt gagtacaata tctgacgatt ggacttcctg ggtcccaagc
     2641 tggattccga ctggctgact gccaagcaac gatatggatc gtcgagactc gcactgagag
     2701 gatcgttctc ggtacaccag gcgcgcctag cgttacacac acaaaaaacc aacacacatc
     2761 catcttcgat ggatagcgat tttattatct aagactaggc ttacatacta gcttattcct
     2821 ttgccctcgg acgagtgctg gggcgtcggt ttccactatc ggcgagtact tctacacagc
     2881 catcggtcca gacggcggcg cttctgcggg cgatttgtgt acgcccgaca gtcccggctc
     2941 cggatcggac gattgcgtcg catcgaccct gcgcccaagc tgcatcatcg aaattgccgt
     3001 caaccaagct ctgatagagt tggtcaagac caatgcggag catatacgcc cggagccggg
     3061 gcgatcctgc aagctccgga tgcctccgct cgaagtagcg cgtctgctgc tccatacaag
     3121 ccaaccacgg cctccagaag aagatgttgg cgacctcgta ttgggaatcc ccgaacatcg
     3181 cctcgctcca gtcaatgacc gctgttatgc ggccattgtc cgtcaggaca ttgttggagc
     3241 cgaaatccgc gtgcacgagg tgccggactt cggggcagtc ctcggcccaa agcatcagct
     3301 catcgagagc ctgcgcgacg gacgcactga cggtgtcgtc catcacagtt tgccagtgat
     3361 acacatgggg atcagcaatc gcgcatatga aatcacgcca tgtagtgtat tgaccgattc
     3421 cttgcggtcc gaatgggccg aacccgctcg tctggctaag atcggcggca gcgatggcat
     3481 ccattgcctc cgcgaccggc tgcagaacag cgggcagttc ggtttcaggc aggtcttgca
     3541 acgtgacacc ctgtgcacgg cgggagatgc aataggtcag gctctcgctg aactccccaa
     3601 tgtcaagcac ttccggaatc gggagcgcgg cggatgcaaa gtgccgataa acataacgat
     3661 ctttgtagaa accatcggcg cagctattta cccgcaggac atatccacgc cctcctacat
     3721 cgaagctgaa agcacgagat tcttcgccct ccgagagctg catcaggtcg gagacgctgt
     3781 cgaacttttc gatcagaaac ttctcgacag acgtcgcggt gagttcaggc tttttaccca
     3841 tgttgtctaa caaaaaagcc aaaaacggcc agaatttagc ggacaattta caagtctaac
     3901 actgaaaatt acatattgac ccaaatgatt acatttcaaa aggtgcctaa aaaacttcac
     3961 aaaacacact cgccaacccc gagcgcatag ttcaaaaccg gagcttcagc tacttaagaa
     4021 gataggtaca taaaaccgac caaagaaact gacgcctcac ttatccctcc cctcaccaga
     4081 ggtccggcgc ctgtcgattc aggagagcct accctaggcc cgaaccctgc gtcctgcgac
     4141 ggagaaaagc ctaccgcaca cctaccggct ggtggcccca ccctgcatta taagccaaca
     4201 gaacgggtga cgtcacgaca cgacgagggc gcgcgctccc aaaggtacgg gtgcactgcc
     4261 caacggcacc gccataactg ccgcccccgc aacagacgac aaaccgagtt ctccagtcag
     4321 tgacaaactt cacgtcaggg tccccagatg gtgcccagcc catctcaccc gaataagagc
     4381 tttcccgcat tagcgaaggg ctcaagacct tgggttcttg ccgcccacca tgccccccac
     4441 cttgtttcaa cgacctcaca gcccgcctca caagcgtctt ccattcaaga ctcgggaaca
     4501 gccgccattt tgctgcgctc cccccaaccc ccagttcagg gcaaccttgc tcgcggaccc
     4561 agactacagc ccttggcggt ctctccacac gcttccgtcc caccgagcgg cccggcggcc
     4621 acgaaagccc cggccagccc agcagcccgc tactcaccaa gtgacgatca cagcgatcca
     4681 caaacaagaa ccgcgaccca aatcccggct gcgacggaac tagctgtgcc acacccggcg
     4741 cgtccttata taatcatcgg cgttcaccgc cccacggaga tccctccgca gaatcgccga
     4801 gaagggacta cttttcctcg cctgttccgc tctctggaaa gaaaaccagt gccctagagt
     4861 cacccaagtc ccgtcctaaa atgtccttct gctgatactg gggttctaag gccgagtctt
     4921 atgagcagcg ggccgctgtc ctgagcgtcc gggcggaagg atcaggacgc tcctgcgccc
     4981 ttcgtctgac gtggcagcgc tcgccgtgac gaggggggcg cccgcgggag gcgccaaaac
     5041 ccggcgcgga ggccaggcgc gccccgattt cgatattcct cggatctcgg atagatcggg
     5101 attgaggatg taggttgaac ttctctcggg tgatcccgat ctcggatatt cgggattgga
     5161 gtaatacctg gcgataggcg ctgccaagca atagccaagg cgcaaggccg gccaagagac
     5221 ttgcgatagt tcgttcgaga ggtagcctgc aagatggatc ggacgaaagg tttgaagatt
     5281 ttccgacgag ggccttggag agttcggaaa tttaaatgca actatggatg aacgaaatag
     5341 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     5401 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     5461 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     5521 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     5581 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     5641 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     5701 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     5761 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     5821 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     5881 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     5941 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     6001 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     6061 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     6121 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     6181 ttgctggcct tttgctcaca tgatttaaat gaaactcgta atcggagaac gggtgcaata
     6241 gcaaggacca gatcgaactt gactgacgca attctgagcg tagtcgaaac ccagacaagt
     6301 cgatctcgcc aagtagctga cttgactggt taattaagtt tcgatagccc aaggtaacca
     6361 atgagattag ggagggaaga aagtttgatg aactacttct gacccctagt ggtgtccaga
     6421 aaagaccatt aaaggaatgt cttgatcgaa atcgagagat cgcctgcagg atagctgacg
     6481 accttaagga taaatttctg gtaaggagga cacgtatgga agtgggcaag ttggggaagc
     6541 cgtatccgtt gctgaatctg gcatatgtgg gagtataaga cgcgcagcgt cgcatcaggc
     6601 atttttttct gcgccaatgc cggtagatag caatccggcc cgaggggcac aaaaaacccc
     6661 tcaagacccg tttagaggcc ccaaggggtt atgctacgat accgatagag atgggccaat
     6721 accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg
     6781 aaacat
//
