LOCUS       sequence-295966-        7139 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 7139)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7139)
  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-m4133552
FEATURES             Location/Qualifiers
     source          1..7139
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             545..565
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             566..610
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     polyA_signal    679..800
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(690..709)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     744..763
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      915..961
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      991..1127
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1172..1243
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1469..2057)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1549..1568)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2557..3766
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3603..3622
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3770..4798
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    4825..4873
                     /note="synthetic polyadenylation signal"
     CDS             join(5139..5207,5208..5999)
                     /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(5357..5376)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6012..6148
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6293..6364
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6369..6415)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6433..6481
                     /note="synthetic polyadenylation signal"
     enhancer        6562..6865
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6867..7069
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7020..7040
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(7138..7139,1..8)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 caccatggag tctctcgttc ccggctttaa tgaaaaaact catgtccagt tatccttgcc
       61 agtgcttcaa gtaagggatg tgctggtgcg gggtttcgga gacagtgtag aggaagtcct
      121 gtcagaggct agacagcacc tgaaggatgg gacctgtggc ttagtggaag ttgagaaggg
      181 agtcctcccg cagctggaac aaccttatgt gtttataaaa cgcagcgacg cgcgaacagc
      241 acctcacggg catgtaatgg tggagcttgt cgccgagttg gagggcatcc agtacggaag
      301 atcgggtgaa acgctgggag ttctagtgcc ccacgttggc gaaattccgg tggcctaccg
      361 taaggttttg ctcaggaaaa acggaaataa gggggctggt ggccatagct atggggcaga
      421 cctgaagtct ttcgatctgg gcgacgagct gggcaccgac ccatatgagg atttccagga
      481 aaactggaac acaaaacaca gctccggggt aactcgcgag ctaatgcgag aattgaatgg
      541 aggggaaaac ctttactttc agtcgaaaga aaccgcggca gctaagttcg agaggcagca
      601 catggacagt taaggatccg caggcctctg ctagcttgac tgactgagat acagcgtacc
      661 ttcagctcac agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
      721 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
      781 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg
      841 gggaggtgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt attggcccat
      901 ctctatcggt atcgtagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt
      961 gtgcccctcg ggccggattg ctatctaccg gcattggcgc agaaaaaaat gcctgatgcg
     1021 acgctgcgcg tcttatactc ccacatatgc cagattcagc aacggatacg gcttccccaa
     1081 cttgcccact tccatacgtg tcctccttac cagaaattta tccttaaggt cgtcagctat
     1141 cctgcaggcg atctctcgat ttcgatcaag acattccttt aatggtcttt tctggacacc
     1201 actaggggtc agaagtagtt catcaaactt tcttccctcc ctaatctcat tggttacctt
     1261 gggctatcga aacttaatta accagtcaag tcagctactt ggcgagatcg acttgtctgg
     1321 gtttcgacta cgctcagaat tgcgtcagtc aagttcgatc tggtccttgc tattgcaccc
     1381 gttctccgat tacgagtttc atttaaatca tgtgagcaaa aggccagcaa aaggccagga
     1441 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     1501 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     1561 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     1621 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     1681 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     1741 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     1801 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     1861 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     1921 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     1981 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     2041 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     2101 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     2161 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     2221 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     2281 catccatagt tgcatttaaa tttccgaact ctccaaggcc ctcgtcggaa aatcttcaaa
     2341 cctttcgtcc gatccatctt gcaggctacc tctcgaacga actatcgcaa gtctcttggc
     2401 cggccttgcg ccttggctat tgcttggcag cgcctatcgc caggtattac tccaatcccg
     2461 aatatccgag atcgggatca cccgagagaa gttcaaccta catcctcaat cccgatctat
     2521 ccgagatccg aggaatatcg aaatcggggc gcgcctggcc tccgcgccgg gttttggcgc
     2581 ctcccgcggg cgcccccctc gtcacggcga gcgctgccac gtcagacgaa gggcgcagga
     2641 gcgtcctgat ccttccgccc ggacgctcag gacagcggcc cgctgctcat aagactcggc
     2701 cttagaaccc cagtatcagc agaaggacat tttaggacgg gacttgggtg actctagggc
     2761 actggttttc tttccagaga gcggaacagg cgaggaaaag tagtcccttc tcggcgattc
     2821 tgcggaggga tctccgtggg gcggtgaacg ccgatgatta tataaggacg cgccgggtgt
     2881 ggcacagcta gttccgtcgc agccgggatt tgggtcgcgg ttcttgtttg tggatcgctg
     2941 tgatcgtcac ttggtgagta gcgggctgct gggctggccg gggctttcgt ggccgccggg
     3001 ccgctcggtg ggacggaagc gtgtggagag accgccaagg gctgtagtct gggtccgcga
     3061 gcaaggttgc cctgaactgg gggttggggg gagcgcagca aaatggcggc tgttcccgag
     3121 tcttgaatgg aagacgcttg tgaggcgggc tgtgaggtcg ttgaaacaag gtggggggca
     3181 tggtgggcgg caagaaccca aggtcttgag cccttcgcta atgcgggaaa gctcttattc
     3241 gggtgagatg ggctgggcac catctgggga ccctgacgtg aagtttgtca ctgactggag
     3301 aactcggttt gtcgtctgtt gcgggggcgg cagttatggc ggtgccgttg ggcagtgcac
     3361 ccgtaccttt gggagcgcgc gccctcgtcg tgtcgtgacg tcacccgttc tgttggctta
     3421 taatgcaggg tggggccacc tgccggtagg tgtgcggtag gcttttctcc gtcgcaggac
     3481 gcagggttcg ggcctagggt aggctctcct gaatcgacag gcgccggacc tctggtgagg
     3541 ggagggataa gtgaggcgtc agtttctttg gtcggtttta tgtacctatc ttcttaagta
     3601 gctgaagctc cggttttgaa ctatgcgctc ggggttggcg agtgtgtttt gtgaagtttt
     3661 ttaggcacct tttgaaatgt aatcatttgg gtcaatatgt aattttcagt gttagacttg
     3721 taaattgtcc gctaaattct ggccgttttt ggcttttttg ttagacaaca tgggtaaaaa
     3781 gcctgaactc accgcgacgt ctgtcgagaa gtttctgatc gaaaagttcg acagcgtctc
     3841 cgacctgatg cagctctcgg agggcgaaga atctcgtgct ttcagcttcg atgtaggagg
     3901 gcgtggatat gtcctgcggg taaatagctg cgccgatggt ttctacaaag atcgttatgt
     3961 ttatcggcac tttgcatccg ccgcgctccc gattccggaa gtgcttgaca ttggggagtt
     4021 cagcgagagc ctgacctatt gcatctcccg ccgtgcacag ggtgtcacgt tgcaagacct
     4081 gcctgaaacc gaactgcccg ctgttctgca gccggtcgcg gaggcaatgg atgccatcgc
     4141 tgccgccgat cttagccaga cgagcgggtt cggcccattc ggaccgcaag gaatcggtca
     4201 atacactaca tggcgtgatt tcatatgcgc gattgctgat ccccatgtgt atcactggca
     4261 aactgtgatg gacgacaccg tcagtgcgtc cgtcgcgcag gctctcgatg agctgatgct
     4321 ttgggccgag gactgccccg aagtccggca cctcgtgcac gcggatttcg gctccaacaa
     4381 tgtcctgacg gacaatggcc gcataacagc ggtcattgac tggagcgagg cgatgttcgg
     4441 ggattcccaa tacgaggtcg ccaacatctt cttctggagg ccgtggttgg cttgtatgga
     4501 gcagcagacg cgctacttcg agcggaggca tccggagctt gcaggatcgc cccggctccg
     4561 ggcgtatatg ctccgcattg gtcttgacca actctatcag agcttggttg acggcaattt
     4621 cgatgatgca gcttgggcgc agggtcgatg cgacgcaatc gtccgatccg gagccgggac
     4681 tgtcgggcgt acacaaatcg cccgcagaag cgccgccgtc tggaccgatg gctgtgtaga
     4741 agtactcgcc gatagtggaa accgacgccc cagcactcgt ccgagggcaa aggaataagc
     4801 tagtatgtaa gcctagtctt agataataaa atcgctatcc atcgaagatg gatgtgtgtt
     4861 ggttttttgt gtgtgtaacg ctaggcgcgc ctggtgtacc gagaacgatc ctctcagtgc
     4921 gagtctcgac gatccatatc gttgcttggc agtcagccag tcggaatcca gcttgggacc
     4981 caggaagtcc aatcgtcaga tattgtactc aagcctggtc acggcagcgt accgatctgt
     5041 ttaaacctag atattgatag tctgatcggt caacgtataa tcgagtccta gcttttgcaa
     5101 acatctatca agagacagga tcagcaggag gctttcgcat gagtattcaa catttccgtg
     5161 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     5221 tggtgaaagt aaaagatgct gaagatcagt tgggtgcgcg agtgggttac atcgaactgg
     5281 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgcttt ccaatgatga
     5341 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     5401 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtattca ccagtcacag
     5461 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     5521 gtgataacac tgcggccaac ttacttctga caacgattgg aggaccgaag gagctaaccg
     5581 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     5641 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacct
     5701 tgcgtaaact attaactggc gaactactta ctctagcttc ccggcaacag ttgatagact
     5761 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     5821 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     5881 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     5941 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     6001 cgattctagg tgcattggcg cagaaaaaaa tgcctgatgc gacgctgcgc gtcttatact
     6061 cccacatatg ccagattcag caacggatac ggcttcccca acttgcccac ttccatacgt
     6121 gtcctcctta ccagaaattt atccttaaga tcgtttaaac tcgactctgg ctctatcgaa
     6181 tctccgtcgt ttcgagctta cgcgaacagc cgtggcgctc atttgctcgt cgggcatcga
     6241 atctcgtcag ctatcgtcag cttacctttt tggcagcgat cgcggctccc gacatcttgg
     6301 accattagct ccacaggtat cttcttccct ctagtggtca taacagcagc ttcagctacc
     6361 tctcaattca aaaaacccct caagacccgt ttagaggccc caaggggtta tgctatcaat
     6421 cgttgcgtta cacacacaaa aaaccaacac acatccatct tcgatggata gcgattttat
     6481 tatctaactg ctgatcgagt gtagccagat ctagtaatca attacggggt cattagttca
     6541 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
     6601 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
     6661 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
     6721 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
     6781 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
     6841 cgtattagtc atcgctatta ccatgctgat gcggttttgg cagtacatca atgggcgtgg
     6901 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
     6961 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
     7021 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa
     7081 ccgtcagatc agatctttgt cgatcctacc atccactcga cacacccgcc agcggccgc
//
