LOCUS       sequence-295248-        7481 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 leader protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7481)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7481)
  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-m4134162
FEATURES             Location/Qualifiers
     source          1..7481
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             542..562
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             911..928
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1021..1142
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1032..1051)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1086..1105
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1257..1303
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1333..1469
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1514..1585
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1811..2399)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1891..1910)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2899..4108
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3945..3964
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4112..5140
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5167..5215
                     /note="synthetic polyadenylation signal"
     CDS             join(5481..5549,5550..6341)
                     /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(5699..5718)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6354..6490
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6635..6706
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6711..6757)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6775..6823
                     /note="synthetic polyadenylation signal"
     enhancer        6904..7207
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7209..7411
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7362..7382
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(7480..7481,1..8)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 caccatggag agccttgtcc ctggtttcaa cgagaaaaca cacgtccaac tcagtttgcc
       61 tgttttacag gttcgcgacg tgctcgtacg tggctttgga gactccgtgg aggaggtctt
      121 atcagaggca cgtcaacatc ttaaagatgg cacttgtggc ttagtagaag ttgaaaaagg
      181 cgttttgcct caacttgaac agccctatgt gttcatcaaa cgttcggatg ctcgaactgc
      241 acctcatggt catgttatgg ttgagctggt agcagaactc gaaggcattc agtacggtcg
      301 tagtggtgag acacttggtg tccttgtccc tcatgtgggc gaaataccag tggcttaccg
      361 caaggttctt cttcgtaaga acggtaataa aggagctggt ggccatagtt acggcgccga
      421 tctaaagtca tttgacttag gcgacgagct tggcactgat ccttatgaag attttcaaga
      481 aaactggaac actaaacata gcagtggtgt tacccgtgaa ctcatgcgtg agcttaacgg
      541 agaaaacctt tactttcagt cgatgagtga caaaatcatt catctaaccg atgactcctt
      601 tgatacagac gtgttgaagg ccgatggtgc gatactggta gatttctggg ctgaatggtg
      661 tggcccgtgc aagatgatcg cacccattct ggacgagatc gccgacgagt atcaaggaaa
      721 attaactgtc gctaagctta atatagatca gaacccaggg acggcaccta aatacggcat
      781 caggggaata cctaccctcc tgctctttaa gaatggggaa gttgccgcga caaaagtggg
      841 agccctgtct aagggccagc taaaagagtt cttggacgct aacctggcag gttcaggctc
      901 ggggcacatg caccatcacc atcaccatag cagcgggctc gtgccacgcg gctaaggatc
      961 cgcaggcctc tgctagcttg actgactgag atacagcgta ccttcagctc acagacatga
     1021 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
     1081 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     1141 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt
     1201 tttaaagcaa gtaaaacctc tacaaatgtg gtattggccc atctctatcg gtatcgtagc
     1261 ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgtgcccct cgggccggat
     1321 tgctatctac cggcattggc gcagaaaaaa atgcctgatg cgacgctgcg cgtcttatac
     1381 tcccacatat gccagattca gcaacggata cggcttcccc aacttgccca cttccatacg
     1441 tgtcctcctt accagaaatt tatccttaag gtcgtcagct atcctgcagg cgatctctcg
     1501 atttcgatca agacattcct ttaatggtct tttctggaca ccactagggg tcagaagtag
     1561 ttcatcaaac tttcttccct ccctaatctc attggttacc ttgggctatc gaaacttaat
     1621 taaccagtca agtcagctac ttggcgagat cgacttgtct gggtttcgac tacgctcaga
     1681 attgcgtcag tcaagttcga tctggtcctt gctattgcac ccgttctccg attacgagtt
     1741 tcatttaaat catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     1801 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     1861 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     1921 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     1981 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     2041 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     2101 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     2161 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     2221 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     2281 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     2341 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     2401 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     2461 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     2521 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     2581 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcattta
     2641 aatttccgaa ctctccaagg ccctcgtcgg aaaatcttca aacctttcgt ccgatccatc
     2701 ttgcaggcta cctctcgaac gaactatcgc aagtctcttg gccggccttg cgccttggct
     2761 attgcttggc agcgcctatc gccaggtatt actccaatcc cgaatatccg agatcgggat
     2821 cacccgagag aagttcaacc tacatcctca atcccgatct atccgagatc cgaggaatat
     2881 cgaaatcggg gcgcgcctgg cctccgcgcc gggttttggc gcctcccgcg ggcgcccccc
     2941 tcgtcacggc gagcgctgcc acgtcagacg aagggcgcag gagcgtcctg atccttccgc
     3001 ccggacgctc aggacagcgg cccgctgctc ataagactcg gccttagaac cccagtatca
     3061 gcagaaggac attttaggac gggacttggg tgactctagg gcactggttt tctttccaga
     3121 gagcggaaca ggcgaggaaa agtagtccct tctcggcgat tctgcggagg gatctccgtg
     3181 gggcggtgaa cgccgatgat tatataagga cgcgccgggt gtggcacagc tagttccgtc
     3241 gcagccggga tttgggtcgc ggttcttgtt tgtggatcgc tgtgatcgtc acttggtgag
     3301 tagcgggctg ctgggctggc cggggctttc gtggccgccg ggccgctcgg tgggacggaa
     3361 gcgtgtggag agaccgccaa gggctgtagt ctgggtccgc gagcaaggtt gccctgaact
     3421 gggggttggg gggagcgcag caaaatggcg gctgttcccg agtcttgaat ggaagacgct
     3481 tgtgaggcgg gctgtgaggt cgttgaaaca aggtgggggg catggtgggc ggcaagaacc
     3541 caaggtcttg agcccttcgc taatgcggga aagctcttat tcgggtgaga tgggctgggc
     3601 accatctggg gaccctgacg tgaagtttgt cactgactgg agaactcggt ttgtcgtctg
     3661 ttgcgggggc ggcagttatg gcggtgccgt tgggcagtgc acccgtacct ttgggagcgc
     3721 gcgccctcgt cgtgtcgtga cgtcacccgt tctgttggct tataatgcag ggtggggcca
     3781 cctgccggta ggtgtgcggt aggcttttct ccgtcgcagg acgcagggtt cgggcctagg
     3841 gtaggctctc ctgaatcgac aggcgccgga cctctggtga ggggagggat aagtgaggcg
     3901 tcagtttctt tggtcggttt tatgtaccta tcttcttaag tagctgaagc tccggttttg
     3961 aactatgcgc tcggggttgg cgagtgtgtt ttgtgaagtt ttttaggcac cttttgaaat
     4021 gtaatcattt gggtcaatat gtaattttca gtgttagact tgtaaattgt ccgctaaatt
     4081 ctggccgttt ttggcttttt tgttagacaa catgggtaaa aagcctgaac tcaccgcgac
     4141 gtctgtcgag aagtttctga tcgaaaagtt cgacagcgtc tccgacctga tgcagctctc
     4201 ggagggcgaa gaatctcgtg ctttcagctt cgatgtagga gggcgtggat atgtcctgcg
     4261 ggtaaatagc tgcgccgatg gtttctacaa agatcgttat gtttatcggc actttgcatc
     4321 cgccgcgctc ccgattccgg aagtgcttga cattggggag ttcagcgaga gcctgaccta
     4381 ttgcatctcc cgccgtgcac agggtgtcac gttgcaagac ctgcctgaaa ccgaactgcc
     4441 cgctgttctg cagccggtcg cggaggcaat ggatgccatc gctgccgccg atcttagcca
     4501 gacgagcggg ttcggcccat tcggaccgca aggaatcggt caatacacta catggcgtga
     4561 tttcatatgc gcgattgctg atccccatgt gtatcactgg caaactgtga tggacgacac
     4621 cgtcagtgcg tccgtcgcgc aggctctcga tgagctgatg ctttgggccg aggactgccc
     4681 cgaagtccgg cacctcgtgc acgcggattt cggctccaac aatgtcctga cggacaatgg
     4741 ccgcataaca gcggtcattg actggagcga ggcgatgttc ggggattccc aatacgaggt
     4801 cgccaacatc ttcttctgga ggccgtggtt ggcttgtatg gagcagcaga cgcgctactt
     4861 cgagcggagg catccggagc ttgcaggatc gccccggctc cgggcgtata tgctccgcat
     4921 tggtcttgac caactctatc agagcttggt tgacggcaat ttcgatgatg cagcttgggc
     4981 gcagggtcga tgcgacgcaa tcgtccgatc cggagccggg actgtcgggc gtacacaaat
     5041 cgcccgcaga agcgccgccg tctggaccga tggctgtgta gaagtactcg ccgatagtgg
     5101 aaaccgacgc cccagcactc gtccgagggc aaaggaataa gctagtatgt aagcctagtc
     5161 ttagataata aaatcgctat ccatcgaaga tggatgtgtg ttggtttttt gtgtgtgtaa
     5221 cgctaggcgc gcctggtgta ccgagaacga tcctctcagt gcgagtctcg acgatccata
     5281 tcgttgcttg gcagtcagcc agtcggaatc cagcttggga cccaggaagt ccaatcgtca
     5341 gatattgtac tcaagcctgg tcacggcagc gtaccgatct gtttaaacct agatattgat
     5401 agtctgatcg gtcaacgtat aatcgagtcc tagcttttgc aaacatctat caagagacag
     5461 gatcagcagg aggctttcgc atgagtattc aacatttccg tgtcgccctt attccctttt
     5521 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     5581 ctgaagatca gttgggtgcg cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     5641 tccttgagag ttttcgcccc gaagaacgct ttccaatgat gagcactttt aaagttctgc
     5701 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     5761 actattctca gaatgacttg gttgagtatt caccagtcac agaaaagcat cttacggatg
     5821 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     5881 acttacttct gacaacgatt ggaggaccga aggagctaac cgcttttttg cacaacatgg
     5941 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     6001 acgagcgtga caccacgatg cctgtagcaa tggcaacaac cttgcgtaaa ctattaactg
     6061 gcgaactact tactctagct tcccggcaac agttgataga ctggatggag gcggataaag
     6121 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     6181 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     6241 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     6301 agatcgctga gataggtgcc tcactgatta agcattggta accgattcta ggtgcattgg
     6361 cgcagaaaaa aatgcctgat gcgacgctgc gcgtcttata ctcccacata tgccagattc
     6421 agcaacggat acggcttccc caacttgccc acttccatac gtgtcctcct taccagaaat
     6481 ttatccttaa gatcgtttaa actcgactct ggctctatcg aatctccgtc gtttcgagct
     6541 tacgcgaaca gccgtggcgc tcatttgctc gtcgggcatc gaatctcgtc agctatcgtc
     6601 agcttacctt tttggcagcg atcgcggctc ccgacatctt ggaccattag ctccacaggt
     6661 atcttcttcc ctctagtggt cataacagca gcttcagcta cctctcaatt caaaaaaccc
     6721 ctcaagaccc gtttagaggc cccaaggggt tatgctatca atcgttgcgt tacacacaca
     6781 aaaaaccaac acacatccat cttcgatgga tagcgatttt attatctaac tgctgatcga
     6841 gtgtagccag atctagtaat caattacggg gtcattagtt catagcccat atatggagtt
     6901 ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
     6961 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg
     7021 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
     7081 gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
     7141 gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
     7201 taccatgctg atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg
     7261 gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca
     7321 acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg
     7381 tgtacggtgg gaggtctata taagcagagc tggtttagtg aaccgtcaga tcagatcttt
     7441 gtcgatccta ccatccactc gacacacccg ccagcggccg c
//
