LOCUS       sequence-314731-        8430 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 8430)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 8430)
  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-m4133371
FEATURES             Location/Qualifiers
     source          1..8430
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        306..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      577..586
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2383..2403
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             2407..2472
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    2541..2662
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2552..2571)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2606..2625
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2777..2823
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2853..2989
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3034..3105
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(3331..3919)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3411..3430)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        4419..5628
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     5465..5484
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             5632..6660
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    6687..6735
                     /note="synthetic polyadenylation signal"
     CDS             join(7001..7069,7070..7861)
                     /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(7219..7238)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      7874..8010
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    8162..8233
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(8238..8284)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    8302..8350
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catgctgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatca gatctttgtc
      541 gatcctacca tccactcgac acacccgcca gcggccgcca ccatggttgg ggcttgtgtt
      601 ctttgcaatt cacagacttc attaagatgt ggtgcttgca tacgtagacc attcttatgt
      661 tgtaaatgct gttacgacca tgtcatatca acatcacata aattagtctt gtctgttaat
      721 ccgtatgttt gcaatgctcc aggttgtgat gtcacagatg tgactcaact ttacttagga
      781 ggtatgagct attattgtaa atcacataaa ccacccatta gttttccatt gtgtgctaat
      841 ggacaagttt ttggtttata taaaaataca tgtgttggta gcgataatgt tactgacttt
      901 aatgcaattg caacatgtga ctggacaaat gctggtgatt acattttagc taacacctgt
      961 actgaaagac tcaagctttt tgcagcagaa acgctcaaag ctactgagga gacatttaaa
     1021 ctgtcttatg gtattgctac tgtacgtgaa gtgctgtctg acagagaatt acatctttca
     1081 tgggaagttg gtaaacctag accaccactt aaccgaaatt atgtctttac tggttatcgt
     1141 gtaactaaaa acagtaaagt acaaatagga gagtacacct ttgaaaaagg tgactatggt
     1201 gatgctgttg tttaccgagg tacaacaact tacaaattaa atgttggtga ttattttgtg
     1261 ctgacatcac atacagtaat gccattaagt gcacctacac tagtgccaca agagcactat
     1321 gttagaatta ctggcttata cccaacactc aatatctcag atgagttttc tagcaatgtt
     1381 gcaaattatc aaaaggttgg tatgcaaaag tattctacac tccagggacc acctggtact
     1441 ggtaagagtc attttgctat tggcctagct ctctactacc cttctgctcg catagtgtat
     1501 acagcttgct ctcatgccgc tgttgatgca ctatgtgaga aggcattaaa atatttgcct
     1561 atagataaat gtagtagaat tatacctgca cgtgctcgtg tagagtgttt tgataaattc
     1621 aaagtgaatt caacattaga acagtatgtc ttttgtactg taaatgcatt gcctgagacg
     1681 acagcagata tagttgtctt tgatgaaatt tcaatggcca caaattatga tttgagtgtt
     1741 gtcaatgcca gattacgtgc taagcactat gtgtacattg gcgaccctgc tcaattacct
     1801 gcaccacgca cattgctaac taagggcaca ctagaaccag aatatttcaa ttcagtgtgt
     1861 agacttatga aaactatagg tccagacatg ttcctcggaa cttgtcggcg ttgtcctgct
     1921 gaaattgttg acactgtgag tgctttggtt tatgataata agcttaaagc acataaagac
     1981 aaatcagctc aatgctttaa aatgttttat aagggtgtta tcacgcatga tgtttcatct
     2041 gcaattaaca ggccacaaat aggcgtggta agagaattcc ttacacgtaa ccctgcttgg
     2101 agaaaagctg tctttatttc accttataat tcacagaatg ctgtagcctc aaagattttg
     2161 ggactaccaa ctcaaactgt tgattcatca cagggctcag aatatgacta tgtcatattc
     2221 actcaaacca ctgaaacagc tcactcttgt aatgtaaaca gatttaatgt tgctattacc
     2281 agagcaaaag taggcatact ttgcataatg tctgatagag acctttatga caagttgcaa
     2341 tttacaagtc ttgaaattcc acgtaggaat gtggcaactt tagaaaacct ttactttcag
     2401 tcgatggatt ataaggacca cgatggagac tacaaagatc atgacattga ttacaaggac
     2461 gacgatgaca aataaggatc cgcaggcctc tgctagcttg actgactgag atacagcgta
     2521 ccttcagctc acagacatga taagatacat tgatgagttt ggacaaacca caactagaat
     2581 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     2641 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     2701 gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtattggccc
     2761 atctctatcg gtatcgtagc ataacccctt ggggcctcta aacgggtctt gaggggtttt
     2821 ttgtgcccct cgggccggat tgctatctac cggcattggc gcagaaaaaa atgcctgatg
     2881 cgacgctgcg cgtcttatac tcccacatat gccagattca gcaacggata cggcttcccc
     2941 aacttgccca cttccatacg tgtcctcctt accagaaatt tatccttaag gtcgtcagct
     3001 atcctgcagg cgatctctcg atttcgatca agacattcct ttaatggtct tttctggaca
     3061 ccactagggg tcagaagtag ttcatcaaac tttcttccct ccctaatctc attggttacc
     3121 ttgggctatc gaaacttaat taaccagtca agtcagctac ttggcgagat cgacttgtct
     3181 gggtttcgac tacgctcaga attgcgtcag tcaagttcga tctggtcctt gctattgcac
     3241 ccgttctccg attacgagtt tcatttaaat catgtgagca aaaggccagc aaaaggccag
     3301 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     3361 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     3421 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     3481 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     3541 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     3601 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     3661 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     3721 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     3781 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     3841 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     3901 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     3961 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     4021 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     4081 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     4141 ttcatccata gttgcattta aatttccgaa ctctccaagg ccctcgtcgg aaaatcttca
     4201 aacctttcgt ccgatccatc ttgcaggcta cctctcgaac gaactatcgc aagtctcttg
     4261 gccggccttg cgccttggct attgcttggc agcgcctatc gccaggtatt actccaatcc
     4321 cgaatatccg agatcgggat cacccgagag aagttcaacc tacatcctca atcccgatct
     4381 atccgagatc cgaggaatat cgaaatcggg gcgcgcctgg cctccgcgcc gggttttggc
     4441 gcctcccgcg ggcgcccccc tcgtcacggc gagcgctgcc acgtcagacg aagggcgcag
     4501 gagcgtcctg atccttccgc ccggacgctc aggacagcgg cccgctgctc ataagactcg
     4561 gccttagaac cccagtatca gcagaaggac attttaggac gggacttggg tgactctagg
     4621 gcactggttt tctttccaga gagcggaaca ggcgaggaaa agtagtccct tctcggcgat
     4681 tctgcggagg gatctccgtg gggcggtgaa cgccgatgat tatataagga cgcgccgggt
     4741 gtggcacagc tagttccgtc gcagccggga tttgggtcgc ggttcttgtt tgtggatcgc
     4801 tgtgatcgtc acttggtgag tagcgggctg ctgggctggc cggggctttc gtggccgccg
     4861 ggccgctcgg tgggacggaa gcgtgtggag agaccgccaa gggctgtagt ctgggtccgc
     4921 gagcaaggtt gccctgaact gggggttggg gggagcgcag caaaatggcg gctgttcccg
     4981 agtcttgaat ggaagacgct tgtgaggcgg gctgtgaggt cgttgaaaca aggtgggggg
     5041 catggtgggc ggcaagaacc caaggtcttg agcccttcgc taatgcggga aagctcttat
     5101 tcgggtgaga tgggctgggc accatctggg gaccctgacg tgaagtttgt cactgactgg
     5161 agaactcggt ttgtcgtctg ttgcgggggc ggcagttatg gcggtgccgt tgggcagtgc
     5221 acccgtacct ttgggagcgc gcgccctcgt cgtgtcgtga cgtcacccgt tctgttggct
     5281 tataatgcag ggtggggcca cctgccggta ggtgtgcggt aggcttttct ccgtcgcagg
     5341 acgcagggtt cgggcctagg gtaggctctc ctgaatcgac aggcgccgga cctctggtga
     5401 ggggagggat aagtgaggcg tcagtttctt tggtcggttt tatgtaccta tcttcttaag
     5461 tagctgaagc tccggttttg aactatgcgc tcggggttgg cgagtgtgtt ttgtgaagtt
     5521 ttttaggcac cttttgaaat gtaatcattt gggtcaatat gtaattttca gtgttagact
     5581 tgtaaattgt ccgctaaatt ctggccgttt ttggcttttt tgttagacaa catgggtaaa
     5641 aagcctgaac tcaccgcgac gtctgtcgag aagtttctga tcgaaaagtt cgacagcgtc
     5701 tccgacctga tgcagctctc ggagggcgaa gaatctcgtg ctttcagctt cgatgtagga
     5761 gggcgtggat atgtcctgcg ggtaaatagc tgcgccgatg gtttctacaa agatcgttat
     5821 gtttatcggc actttgcatc cgccgcgctc ccgattccgg aagtgcttga cattggggag
     5881 ttcagcgaga gcctgaccta ttgcatctcc cgccgtgcac agggtgtcac gttgcaagac
     5941 ctgcctgaaa ccgaactgcc cgctgttctg cagccggtcg cggaggcaat ggatgccatc
     6001 gctgccgccg atcttagcca gacgagcggg ttcggcccat tcggaccgca aggaatcggt
     6061 caatacacta catggcgtga tttcatatgc gcgattgctg atccccatgt gtatcactgg
     6121 caaactgtga tggacgacac cgtcagtgcg tccgtcgcgc aggctctcga tgagctgatg
     6181 ctttgggccg aggactgccc cgaagtccgg cacctcgtgc acgcggattt cggctccaac
     6241 aatgtcctga cggacaatgg ccgcataaca gcggtcattg actggagcga ggcgatgttc
     6301 ggggattccc aatacgaggt cgccaacatc ttcttctgga ggccgtggtt ggcttgtatg
     6361 gagcagcaga cgcgctactt cgagcggagg catccggagc ttgcaggatc gccccggctc
     6421 cgggcgtata tgctccgcat tggtcttgac caactctatc agagcttggt tgacggcaat
     6481 ttcgatgatg cagcttgggc gcagggtcga tgcgacgcaa tcgtccgatc cggagccggg
     6541 actgtcgggc gtacacaaat cgcccgcaga agcgccgccg tctggaccga tggctgtgta
     6601 gaagtactcg ccgatagtgg aaaccgacgc cccagcactc gtccgagggc aaaggaataa
     6661 gctagtatgt aagcctagtc ttagataata aaatcgctat ccatcgaaga tggatgtgtg
     6721 ttggtttttt gtgtgtgtaa cgctaggcgc gcctggtgta ccgagaacga tcctctcagt
     6781 gcgagtctcg acgatccata tcgttgcttg gcagtcagcc agtcggaatc cagcttggga
     6841 cccaggaagt ccaatcgtca gatattgtac tcaagcctgg tcacggcagc gtaccgatct
     6901 gtttaaacct agatattgat agtctgatcg gtcaacgtat aatcgagtcc tagcttttgc
     6961 aaacatctat caagagacag gatcagcagg aggctttcgc atgagtattc aacatttccg
     7021 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     7081 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgcg cgagtgggtt acatcgaact
     7141 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgct ttccaatgat
     7201 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     7261 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtatt caccagtcac
     7321 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     7381 gagtgataac actgcggcca acttacttct gacaacgatt ggaggaccga aggagctaac
     7441 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     7501 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     7561 cttgcgtaaa ctattaactg gcgaactact tactctagct tcccggcaac agttgataga
     7621 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     7681 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     7741 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     7801 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     7861 accgattcta ggtgcattgg cgcagaaaaa aatgcctgat gcgacgctgc gcgtcttata
     7921 ctcccacata tgccagattc agcaacggat acggcttccc caacttgccc acttccatac
     7981 gtgtcctcct taccagaaat ttatccttaa gatcccgaat cgtttaaact cgactctggc
     8041 tctatcgaat ctccgtcgtt tcgagcttac gcgaacagcc gtggcgctca tttgctcgtc
     8101 gggcatcgaa tctcgtcagc tatcgtcagc ttaccttttt ggcagcgatc gcggctcccg
     8161 acatcttgga ccattagctc cacaggtatc ttcttccctc tagtggtcat aacagcagct
     8221 tcagctacct ctcaattcaa aaaacccctc aagacccgtt tagaggcccc aaggggttat
     8281 gctatcaatc gttgcgttac acacacaaaa aaccaacaca catccatctt cgatggatag
     8341 cgattttatt atctaactgc tgatcgagtg tagccagatc tagtaatcaa ttacggggtc
     8401 attagttcat agcccatata tggagttccg
//
