LOCUS       sequence-307287-        8388 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 helicase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8388)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 8388)
  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-m4133829
FEATURES             Location/Qualifiers
     source          1..8388
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        81..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..588
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     539..559
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      657..666
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2463..2483
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             2484..2510
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    2579..2700
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2590..2609)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2644..2663
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2815..2861
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2891..3027
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3072..3143
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(3369..3957)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3449..3468)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        4457..5666
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     5503..5522
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             5670..6698
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    6725..6773
                     /note="synthetic polyadenylation signal"
     CDS             join(7039..7107,7108..7899)
                     /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(7257..7276)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      7912..8048
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    8200..8271
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(8276..8322)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    8340..8388
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 atctaactgc tgatcgagtg tagccagatc tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catgctgatg cggttttggc agtacatcaa tgggcgtgga
      421 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
      481 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
      541 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctgg tttagtgaac
      601 cgtcagatca gatctttgtc gatcctacca tccactcgac acacccgcca gcggccgcca
      661 ccatggttgg ggcttgtgtt ctttgcaatt cacagacttc attaagatgt ggtgcttgca
      721 tacgtagacc attcttatgt tgtaaatgct gttacgacca tgtcatatca acatcacata
      781 aattagtctt gtctgttaat ccgtatgttt gcaatgctcc aggttgtgat gtcacagatg
      841 tgactcaact ttacttagga ggtatgagct attattgtaa atcacataaa ccacccatta
      901 gttttccatt gtgtgctaat ggacaagttt ttggtttata taaaaataca tgtgttggta
      961 gcgataatgt tactgacttt aatgcaattg caacatgtga ctggacaaat gctggtgatt
     1021 acattttagc taacacctgt actgaaagac tcaagctttt tgcagcagaa acgctcaaag
     1081 ctactgagga gacatttaaa ctgtcttatg gtattgctac tgtacgtgaa gtgctgtctg
     1141 acagagaatt acatctttca tgggaagttg gtaaacctag accaccactt aaccgaaatt
     1201 atgtctttac tggttatcgt gtaactaaaa acagtaaagt acaaatagga gagtacacct
     1261 ttgaaaaagg tgactatggt gatgctgttg tttaccgagg tacaacaact tacaaattaa
     1321 atgttggtga ttattttgtg ctgacatcac atacagtaat gccattaagt gcacctacac
     1381 tagtgccaca agagcactat gttagaatta ctggcttata cccaacactc aatatctcag
     1441 atgagttttc tagcaatgtt gcaaattatc aaaaggttgg tatgcaaaag tattctacac
     1501 tccagggacc acctggtact ggtaagagtc attttgctat tggcctagct ctctactacc
     1561 cttctgctcg catagtgtat acagcttgct ctcatgccgc tgttgatgca ctatgtgaga
     1621 aggcattaaa atatttgcct atagataaat gtagtagaat tatacctgca cgtgctcgtg
     1681 tagagtgttt tgataaattc aaagtgaatt caacattaga acagtatgtc ttttgtactg
     1741 taaatgcatt gcctgagacg acagcagata tagttgtctt tgatgaaatt tcaatggcca
     1801 caaattatga tttgagtgtt gtcaatgcca gattacgtgc taagcactat gtgtacattg
     1861 gcgaccctgc tcaattacct gcaccacgca cattgctaac taagggcaca ctagaaccag
     1921 aatatttcaa ttcagtgtgt agacttatga aaactatagg tccagacatg ttcctcggaa
     1981 cttgtcggcg ttgtcctgct gaaattgttg acactgtgag tgctttggtt tatgataata
     2041 agcttaaagc acataaagac aaatcagctc aatgctttaa aatgttttat aagggtgtta
     2101 tcacgcatga tgtttcatct gcaattaaca ggccacaaat aggcgtggta agagaattcc
     2161 ttacacgtaa ccctgcttgg agaaaagctg tctttatttc accttataat tcacagaatg
     2221 ctgtagcctc aaagattttg ggactaccaa ctcaaactgt tgattcatca cagggctcag
     2281 aatatgacta tgtcatattc actcaaacca ctgaaacagc tcactcttgt aatgtaaaca
     2341 gatttaatgt tgctattacc agagcaaaag taggcatact ttgcataatg tctgatagag
     2401 acctttatga caagttgcaa tttacaagtc ttgaaattcc acgtaggaat gtggcaactt
     2461 tagaaaacct ttactttcag tcgtacccct atgacgtacc agattatgcg taaggatccg
     2521 caggcctctg ctagcttgac tgactgagat acagcgtacc ttcagctcac agacatgata
     2581 agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt
     2641 tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt
     2701 aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt
     2761 taaagcaagt aaaacctcta caaatgtggt attggcccat ctctatcggt atcgtagcat
     2821 aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gtgcccctcg ggccggattg
     2881 ctatctaccg gcattggcgc agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc
     2941 ccacatatgc cagattcagc aacggatacg gcttccccaa cttgcccact tccatacgtg
     3001 tcctccttac cagaaattta tccttaaggt cgtcagctat cctgcaggcg atctctcgat
     3061 ttcgatcaag acattccttt aatggtcttt tctggacacc actaggggtc agaagtagtt
     3121 catcaaactt tcttccctcc ctaatctcat tggttacctt gggctatcga aacttaatta
     3181 accagtcaag tcagctactt ggcgagatcg acttgtctgg gtttcgacta cgctcagaat
     3241 tgcgtcagtc aagttcgatc tggtccttgc tattgcaccc gttctccgat tacgagtttc
     3301 atttaaatca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     3361 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     3421 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     3481 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     3541 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     3601 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     3661 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     3721 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     3781 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     3841 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     3901 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     3961 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     4021 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     4081 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     4141 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcatttaaa
     4201 tttccgaact ctccaaggcc ctcgtcggaa aatcttcaaa cctttcgtcc gatccatctt
     4261 gcaggctacc tctcgaacga actatcgcaa gtctcttggc cggccttgcg ccttggctat
     4321 tgcttggcag cgcctatcgc caggtattac tccaatcccg aatatccgag atcgggatca
     4381 cccgagagaa gttcaaccta catcctcaat cccgatctat ccgagatccg aggaatatcg
     4441 aaatcggggc gcgcctggcc tccgcgccgg gttttggcgc ctcccgcggg cgcccccctc
     4501 gtcacggcga gcgctgccac gtcagacgaa gggcgcagga gcgtcctgat ccttccgccc
     4561 ggacgctcag gacagcggcc cgctgctcat aagactcggc cttagaaccc cagtatcagc
     4621 agaaggacat tttaggacgg gacttgggtg actctagggc actggttttc tttccagaga
     4681 gcggaacagg cgaggaaaag tagtcccttc tcggcgattc tgcggaggga tctccgtggg
     4741 gcggtgaacg ccgatgatta tataaggacg cgccgggtgt ggcacagcta gttccgtcgc
     4801 agccgggatt tgggtcgcgg ttcttgtttg tggatcgctg tgatcgtcac ttggtgagta
     4861 gcgggctgct gggctggccg gggctttcgt ggccgccggg ccgctcggtg ggacggaagc
     4921 gtgtggagag accgccaagg gctgtagtct gggtccgcga gcaaggttgc cctgaactgg
     4981 gggttggggg gagcgcagca aaatggcggc tgttcccgag tcttgaatgg aagacgcttg
     5041 tgaggcgggc tgtgaggtcg ttgaaacaag gtggggggca tggtgggcgg caagaaccca
     5101 aggtcttgag cccttcgcta atgcgggaaa gctcttattc gggtgagatg ggctgggcac
     5161 catctgggga ccctgacgtg aagtttgtca ctgactggag aactcggttt gtcgtctgtt
     5221 gcgggggcgg cagttatggc ggtgccgttg ggcagtgcac ccgtaccttt gggagcgcgc
     5281 gccctcgtcg tgtcgtgacg tcacccgttc tgttggctta taatgcaggg tggggccacc
     5341 tgccggtagg tgtgcggtag gcttttctcc gtcgcaggac gcagggttcg ggcctagggt
     5401 aggctctcct gaatcgacag gcgccggacc tctggtgagg ggagggataa gtgaggcgtc
     5461 agtttctttg gtcggtttta tgtacctatc ttcttaagta gctgaagctc cggttttgaa
     5521 ctatgcgctc ggggttggcg agtgtgtttt gtgaagtttt ttaggcacct tttgaaatgt
     5581 aatcatttgg gtcaatatgt aattttcagt gttagacttg taaattgtcc gctaaattct
     5641 ggccgttttt ggcttttttg ttagacaaca tgggtaaaaa gcctgaactc accgcgacgt
     5701 ctgtcgagaa gtttctgatc gaaaagttcg acagcgtctc cgacctgatg cagctctcgg
     5761 agggcgaaga atctcgtgct ttcagcttcg atgtaggagg gcgtggatat gtcctgcggg
     5821 taaatagctg cgccgatggt ttctacaaag atcgttatgt ttatcggcac tttgcatccg
     5881 ccgcgctccc gattccggaa gtgcttgaca ttggggagtt cagcgagagc ctgacctatt
     5941 gcatctcccg ccgtgcacag ggtgtcacgt tgcaagacct gcctgaaacc gaactgcccg
     6001 ctgttctgca gccggtcgcg gaggcaatgg atgccatcgc tgccgccgat cttagccaga
     6061 cgagcgggtt cggcccattc ggaccgcaag gaatcggtca atacactaca tggcgtgatt
     6121 tcatatgcgc gattgctgat ccccatgtgt atcactggca aactgtgatg gacgacaccg
     6181 tcagtgcgtc cgtcgcgcag gctctcgatg agctgatgct ttgggccgag gactgccccg
     6241 aagtccggca cctcgtgcac gcggatttcg gctccaacaa tgtcctgacg gacaatggcc
     6301 gcataacagc ggtcattgac tggagcgagg cgatgttcgg ggattcccaa tacgaggtcg
     6361 ccaacatctt cttctggagg ccgtggttgg cttgtatgga gcagcagacg cgctacttcg
     6421 agcggaggca tccggagctt gcaggatcgc cccggctccg ggcgtatatg ctccgcattg
     6481 gtcttgacca actctatcag agcttggttg acggcaattt cgatgatgca gcttgggcgc
     6541 agggtcgatg cgacgcaatc gtccgatccg gagccgggac tgtcgggcgt acacaaatcg
     6601 cccgcagaag cgccgccgtc tggaccgatg gctgtgtaga agtactcgcc gatagtggaa
     6661 accgacgccc cagcactcgt ccgagggcaa aggaataagc tagtatgtaa gcctagtctt
     6721 agataataaa atcgctatcc atcgaagatg gatgtgtgtt ggttttttgt gtgtgtaacg
     6781 ctaggcgcgc ctggtgtacc gagaacgatc ctctcagtgc gagtctcgac gatccatatc
     6841 gttgcttggc agtcagccag tcggaatcca gcttgggacc caggaagtcc aatcgtcaga
     6901 tattgtactc aagcctggtc acggcagcgt accgatctgt ttaaacctag atattgatag
     6961 tctgatcggt caacgtataa tcgagtccta gcttttgcaa acatctatca agagacagga
     7021 tcagcaggag gctttcgcat gagtattcaa catttccgtg tcgcccttat tccctttttt
     7081 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     7141 gaagatcagt tgggtgcgcg agtgggttac atcgaactgg atctcaacag cggtaagatc
     7201 cttgagagtt ttcgccccga agaacgcttt ccaatgatga gcacttttaa agttctgcta
     7261 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     7321 tattctcaga atgacttggt tgagtattca ccagtcacag aaaagcatct tacggatggc
     7381 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     7441 ttacttctga caacgattgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     7501 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     7561 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacct tgcgtaaact attaactggc
     7621 gaactactta ctctagcttc ccggcaacag ttgatagact ggatggaggc ggataaagtt
     7681 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     7741 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     7801 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     7861 atcgctgaga taggtgcctc actgattaag cattggtaac cgattctagg tgcattggcg
     7921 cagaaaaaaa tgcctgatgc gacgctgcgc gtcttatact cccacatatg ccagattcag
     7981 caacggatac ggcttcccca acttgcccac ttccatacgt gtcctcctta ccagaaattt
     8041 atccttaaga tcccgaatcg tttaaactcg actctggctc tatcgaatct ccgtcgtttc
     8101 gagcttacgc gaacagccgt ggcgctcatt tgctcgtcgg gcatcgaatc tcgtcagcta
     8161 tcgtcagctt acctttttgg cagcgatcgc ggctcccgac atcttggacc attagctcca
     8221 caggtatctt cttccctcta gtggtcataa cagcagcttc agctacctct caattcaaaa
     8281 aacccctcaa gacccgttta gaggccccaa ggggttatgc tatcaatcgt tgcgttacac
     8341 acacaaaaaa ccaacacaca tccatcttcg atggatagcg attttatt
//
