LOCUS       sequence-369839-        4592 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 4592)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4592)
  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-m4134102
FEATURES             Location/Qualifiers
     source          1..4592
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     408..427
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             575..1174
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(575..594)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1071..1091
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    1201..1249
                     /note="synthetic polyadenylation signal"
     CDS             join(1515..1583,1584..2375)
                     /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(1733..1752)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      2388..2524
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2569..2640
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2866..3454)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2946..2965)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(3954..4592,1..571)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
ORIGIN
        1 acccaaggtc ttgagccctt cgctaatgcg ggaaagctct tattcgggtg agatgggctg
       61 ggcaccatct ggggaccctg acgtgaagtt tgtcactgac tggagaactc ggtttgtcgt
      121 ctgttgcggg ggcggcagtt atggcggtgc cgttgggcag tgcacccgta cctttgggag
      181 cgcgcgccct cgtcgtgtcg tgacgtcacc cgttctgttg gcttataatg cagggtgggg
      241 ccacctgccg gtaggtgtgc ggtaggcttt tctccgtcgc aggacgcagg gttcgggcct
      301 agggtaggct ctcctgaatc gacaggcgcc ggacctctgg tgaggggagg gataagtgag
      361 gcgtcagttt ctttggtcgg ttttatgtac ctatcttctt aagtagctga agctccggtt
      421 ttgaactatg cgctcggggt tggcgagtgt gttttgtgaa gttttttagg caccttttga
      481 aatgtaatca tttgggtcaa tatgtaattt tcagtgttag acttgtaaat tgtccgctaa
      541 attctggccg tttttggctt ttttgttaga caacatgacc gagtacaagc ccacggtgcg
      601 cctcgccacc cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga
      661 ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct
      721 gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga
      781 cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc
      841 cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat
      901 ggagggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg
      961 cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga
     1021 ggccgccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc
     1081 cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg
     1141 cacctggtgc atgacccgca agcccggtgc ctaggctagt atgtaagcct agtcttagat
     1201 aataaaatcg ctatccatcg aagatggatg tgtgttggtt ttttgtgtgt gtaacgctag
     1261 gcgcgcctgg tgtaccgaga acgatcctct cagtgcgagt ctcgacgatc catatcgttg
     1321 cttggcagtc agccagtcgg aatccagctt gggacccagg aagtccaatc gtcagatatt
     1381 gtactcaagc ctggtcacgg cagcgtaccg atctgtttaa acctagatat tgatagtctg
     1441 atcggtcaac gtataatcga gtcctagctt ttgcaaacat ctatcaagag acaggatcag
     1501 caggaggctt tcgcatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     1561 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     1621 atcagttggg tgcgcgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     1681 agagttttcg ccccgaagaa cgctttccaa tgatgagcac ttttaaagtt ctgctatgtg
     1741 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     1801 ctcagaatga cttggttgag tattcaccag tcacagaaaa gcatcttacg gatggcatga
     1861 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     1921 ttctgacaac gattggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     1981 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     2041 gtgacaccac gatgcctgta gcaatggcaa caaccttgcg taaactatta actggcgaac
     2101 tacttactct agcttcccgg caacagttga tagactggat ggaggcggat aaagttgcag
     2161 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     2221 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     2281 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     2341 ctgagatagg tgcctcactg attaagcatt ggtaaccgat tctaggtgca ttggcgcaga
     2401 aaaaaatgcc tgatgcgacg ctgcgcgtct tatactccca catatgccag attcagcaac
     2461 ggatacggct tccccaactt gcccacttcc atacgtgtcc tccttaccag aaatttatcc
     2521 ttaaggtcgt cagctatcct gcaggcgatc tctcgatttc gatcaagaca ttcctttaat
     2581 ggtcttttct ggacaccact aggggtcaga agtagttcat caaactttct tccctcccta
     2641 atctcattgg ttaccttggg ctatcgaaac ttaattaacc agtcaagtca gctacttggc
     2701 gagatcgact tgtctgggtt tcgactacgc tcagaattgc gtcagtcaag ttcgatctgg
     2761 tccttgctat tgcacccgtt ctccgattac gagtttcatt taaatcatgt gagcaaaagg
     2821 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     2881 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     2941 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     3001 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     3061 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     3121 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     3181 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     3241 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     3301 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     3361 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     3421 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     3481 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     3541 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     3601 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     3661 gatctgtcta tttcgttcat ccatagttgc atttaaattt ccgaactctc caaggccctc
     3721 gtcggaaaat cttcaaacct ttcgtccgat ccatcttgca ggctacctct cgaacgaact
     3781 atcgcaagtc tcttggccgg ccttgcgcct tggctattgc ttggcagcgc ctatcgccag
     3841 gtattactcc aatcccgaat atccgagatc gggatcaccc gagagaagtt caacctacat
     3901 cctcaatccc gatctatccg agatccgagg aatatcgaaa tcggggcgcg cctggcctcc
     3961 gcgccgggtt ttggcgcctc ccgcgggcgc ccccctcgtc acggcgagcg ctgccacgtc
     4021 agacgaaggg cgcaggagcg tcctgatcct tccgcccgga cgctcaggac agcggcccgc
     4081 tgctcataag actcggcctt agaaccccag tatcagcaga aggacatttt aggacgggac
     4141 ttgggtgact ctagggcact ggttttcttt ccagagagcg gaacaggcga ggaaaagtag
     4201 tcccttctcg gcgattctgc ggagggatct ccgtggggcg gtgaacgccg atgattatat
     4261 aaggacgcgc cgggtgtggc acagctagtt ccgtcgcagc cgggatttgg gtcgcggttc
     4321 ttgtttgtgg atcgctgtga tcgtcacttg gtgagtagcg ggctgctggg ctggccgggg
     4381 ctttcgtggc cgccgggccg ctcggtggga cggaagcgtg tggagagacc gccaagggct
     4441 gtagtctggg tccgcgagca aggttgccct gaactggggg ttggggggag cgcagcaaaa
     4501 tggcggctgt tcccgagtct tgaatggaag acgcttgtga ggcgggctgt gaggtcgttg
     4561 aaacaaggtg gggggcatgg tgggcggcaa ga
//
