LOCUS       sequence-311766-        6051 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 6051)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6051)
  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-m4134462
FEATURES             Location/Qualifiers
     source          1..6051
                     /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             2404..2445
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     polyA_signal    2498..2619
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2509..2528)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2563..2582
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2734..2780
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2810..2946
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2991..3062
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(3288..3876)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3368..3387)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(4622..4690,4691..5482)
                     /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(4840..4859)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5495..5631
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5783..5854
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5859..5905)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5923..5971
                     /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 tcggggaaac ctatacccaa tccacttctc ggcttagatt ctacctaagc tagcttgact
     2461 gactgagata cagcgtacct tcagctcaca gacatgataa gatacattga tgagtttgga
     2521 caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt
     2581 gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat
     2641 tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac
     2701 aaatgtggta ttggcccatc tctatcggta tcgtagcata accccttggg gcctctaaac
     2761 gggtcttgag gggttttttg tgcccctcgg gccggattgc tatctaccgg cattggcgca
     2821 gaaaaaaatg cctgatgcga cgctgcgcgt cttatactcc cacatatgcc agattcagca
     2881 acggatacgg cttccccaac ttgcccactt ccatacgtgt cctccttacc agaaatttat
     2941 ccttaaggtc gtcagctatc ctgcaggcga tctctcgatt tcgatcaaga cattccttta
     3001 atggtctttt ctggacacca ctaggggtca gaagtagttc atcaaacttt cttccctccc
     3061 taatctcatt ggttaccttg ggctatcgaa acttaattaa ccagtcaagt cagctacttg
     3121 gcgagatcga cttgtctggg tttcgactac gctcagaatt gcgtcagtca agttcgatct
     3181 ggtccttgct attgcacccg ttctccgatt acgagtttca tttaaatcat gtgagcaaaa
     3241 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     3301 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     3361 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     3421 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     3481 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     3541 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     3601 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     3661 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     3721 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     3781 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     3841 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     3901 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     3961 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     4021 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     4081 gcgatctgtc tatttcgttc atccatagtt gcatttaaat ttccgaactc tccaaggccc
     4141 tcgtcggaaa atcttcaaac ctttcgtccg atccatcttg caggctacct ctcgaacgaa
     4201 ctatcgcaag tctcttggcc ggccttgcgc cttggctatt gcttggcagc gcctatcgcc
     4261 aggtattact ccaatcccga atatccgaga tcgggatcac ccgagagaag ttcaacctac
     4321 atcctcaatc ccgatctatc cgagatccga ggaatatcga aatcggggcg cgcctggtgt
     4381 accgagaacg atcctctcag tgcgagtctc gacgatccat atcgttgctt ggcagtcagc
     4441 cagtcggaat ccagcttggg acccaggaag tccaatcgtc agatattgta ctcaagcctg
     4501 gtcacggcag cgtaccgatc tgtttaaacc tagatattga tagtctgatc ggtcaacgta
     4561 taatcgagtc ctagcttttg caaacatcta tcaagagaca ggatcagcag gaggctttcg
     4621 catgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     4681 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     4741 gcgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     4801 cgaagaacgc tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     4861 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     4921 ggttgagtat tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     4981 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     5041 tggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     5101 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     5161 gcctgtagca atggcaacaa ccttgcgtaa actattaact ggcgaactac ttactctagc
     5221 ttcccggcaa cagttgatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     5281 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     5341 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     5401 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     5461 ctcactgatt aagcattggt aaccgattct aggtgcattg gcgcagaaaa aaatgcctga
     5521 tgcgacgctg cgcgtcttat actcccacat atgccagatt cagcaacgga tacggcttcc
     5581 ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa tttatcctta agatcccgaa
     5641 tcgtttaaac tcgactctgg ctctatcgaa tctccgtcgt ttcgagctta cgcgaacagc
     5701 cgtggcgctc atttgctcgt cgggcatcga atctcgtcag ctatcgtcag cttacctttt
     5761 tggcagcgat cgcggctccc gacatcttgg accattagct ccacaggtat cttcttccct
     5821 ctagtggtca taacagcagc ttcagctacc tctcaattca aaaaacccct caagacccgt
     5881 ttagaggccc caaggggtta tgctatcaat cgttgcgtta cacacacaaa aaaccaacac
     5941 acatccatct tcgatggata gcgattttat tatctaactg ctgatcgagt gtagccagat
     6001 ctagtaatca attacggggt cattagttca tagcccatat atggagttcc g
//
