LOCUS       sequence-297956-        6042 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 6042)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6042)
  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-m4133320
FEATURES             Location/Qualifiers
     source          1..6042
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        366..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      637..646
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2449..2469
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             2470..2496
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    2549..2670
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2560..2579)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2614..2633
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2785..2831
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2861..2997
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3042..3113
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(3339..3927)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3419..3438)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(4673..4741,4742..5533)
                     /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(4891..4910)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5546..5682
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5834..5905
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5910..5956)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5974..6022
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 tagccagatc tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catgctgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatca gatctttgtc
      601 gatcctacca tccactcgac acacccgcca gcggccgcca ccatggcggt gggggcttgc
      661 gtactatgta acagccagac gtcattgcgc tgcggtgcat gtataagaag gccgtttctg
      721 tgttgcaaat gctgctacga tcatgttatt tcgacttctc acaagcttgt gctcagtgtc
      781 aatccttatg tgtgtaatgc ccccggatgc gacgtcacag atgttaccca actgtacctg
      841 ggcggcatgt cctattattg taagagccac aaaccaccaa tctccttccc cctgtgcgcc
      901 aacggacagg tgtttgggct ctacaaaaac acatgtgtgg gatctgacaa tgtcaccgac
      961 ttcaacgcta tcgccacatg cgattggact aatgcaggcg actacatttt agccaacacc
     1021 tgtaccgaac ggctgaagtt atttgcggct gagactctaa aggcaactga ggaaaccttc
     1081 aaactttcat atggtatcgc cacggtacgt gaggtgttgt ctgatcgaga actgcatctc
     1141 agttgggagg tcgggaagcc tagacctccc cttaatcgaa attacgtgtt cacaggttat
     1201 agggttacca agaacagcaa agtccagatc ggcgaatata cttttgagaa gggagattac
     1261 ggtgacgccg tagtgtatcg tggcacaaca acatacaaac tgaatgttgg ggattacttc
     1321 gtgctgacgt cccacaccgt tatgccgctc tcagctccaa ctttggttcc ccaggaacat
     1381 tacgtgcgga ttactggact ctatccaacg ctaaacataa gcgacgagtt ttccagcaac
     1441 gtggctaatt accaaaaggt cggcatgcag aagtactcca cccttcaggg tcctcctggg
     1501 actggcaaaa gtcacttcgc catcggactg gcactttact atcccagtgc gcgcattgta
     1561 tataccgcct gctcacatgc ggcagtggac gccctgtgtg agaaggcttt aaagtatctg
     1621 ccaatagaca aatgctctcg cattatccct gccagggcca gagtggagtg ctttgataag
     1681 tttaaagtga actcgaccct cgaacagtat gtcttctgca cagttaacgc attgcccgaa
     1741 acgaccgccg atatagttgt gttcgacgaa atctcaatgg ctaccaatta tgatctctcc
     1801 gtcgtcaatg cacggctgcg ggccaaacac tacgtctaca tcggggaccc cgcgcagtta
     1861 ccagcaccgc gaaccctact gactaagggc acacttgagc cagagtactt caacagcgta
     1921 tgtagactga tgaaaaccat tggccctgat atgttcctgg gaacatgtcg gaggtgcccc
     1981 gctgaaatag tggacaccgt ttcagcactt gtgtacgaca ataagctcaa agcacacaag
     2041 gataaatccg cacaatgttt taagatgttt tacaaagggg ttattaccca tgatgtgtct
     2101 agcgctatca atcgccctca gattggcgtg gtgagagagt ttttgacgcg aaatcccgcc
     2161 tggcgaaagg ccgtattcat cagtccttac aactcgcaaa atgccgtcgc tagtaagatt
     2221 ttaggactcc ctactcagac agttgatagc tctcaaggtt ctgaatacga ttatgtgatt
     2281 tttactcaga ctaccgaaac ggcacattct tgtaacgtca accgttttaa cgtggccatt
     2341 acaagggcca aggttggcat cttgtgtatc atgtcagacc gggacctgta tgataaactg
     2401 caattcacat ctttggagat accgcgccgc aatgtcgcta cccttcagga aaacctttac
     2461 tttcagtcgt acccctatga cgtaccagat tatgcgtaag ctagcttgac tgactgagat
     2521 acagcgtacc ttcagctcac agacatgata agatacattg atgagtttgg acaaaccaca
     2581 actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt
     2641 gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt
     2701 caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt
     2761 attggcccat ctctatcggt atcgtagcat aaccccttgg ggcctctaaa cgggtcttga
     2821 ggggtttttt gtgcccctcg ggccggattg ctatctaccg gcattggcgc agaaaaaaat
     2881 gcctgatgcg acgctgcgcg tcttatactc ccacatatgc cagattcagc aacggatacg
     2941 gcttccccaa cttgcccact tccatacgtg tcctccttac cagaaattta tccttaaggt
     3001 cgtcagctat cctgcaggcg atctctcgat ttcgatcaag acattccttt aatggtcttt
     3061 tctggacacc actaggggtc agaagtagtt catcaaactt tcttccctcc ctaatctcat
     3121 tggttacctt gggctatcga aacttaatta accagtcaag tcagctactt ggcgagatcg
     3181 acttgtctgg gtttcgacta cgctcagaat tgcgtcagtc aagttcgatc tggtccttgc
     3241 tattgcaccc gttctccgat tacgagtttc atttaaatca tgtgagcaaa aggccagcaa
     3301 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     3361 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     3421 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     3481 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     3541 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     3601 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     3661 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     3721 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     3781 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     3841 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     3901 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     3961 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     4021 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     4081 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     4141 ctatttcgtt catccatagt tgcatttaaa tttccgaact ctccaaggcc ctcgtcggaa
     4201 aatcttcaaa cctttcgtcc gatccatctt gcaggctacc tctcgaacga actatcgcaa
     4261 gtctcttggc cggccttgcg ccttggctat tgcttggcag cgcctatcgc caggtattac
     4321 tccaatcccg aatatccgag atcgggatca cccgagagaa gttcaaccta catcctcaat
     4381 cccgatctat ccgagatccg aggaatatcg aaatcggggc gcgcctggtg taccgagaac
     4441 gatcctctca gtgcgagtct cgacgatcca tatcgttgct tggcagtcag ccagtcggaa
     4501 tccagcttgg gacccaggaa gtccaatcgt cagatattgt actcaagcct ggtcacggca
     4561 gcgtaccgat ctgtttaaac ctagatattg atagtctgat cggtcaacgt ataatcgagt
     4621 cctagctttt gcaaacatct atcaagagac aggatcagca ggaggctttc gcatgagtat
     4681 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     4741 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cgcgagtggg
     4801 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     4861 ctttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     4921 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     4981 ttcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     5041 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga ttggaggacc
     5101 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     5161 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     5221 aatggcaaca accttgcgta aactattaac tggcgaacta cttactctag cttcccggca
     5281 acagttgata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     5341 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     5401 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     5461 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     5521 taagcattgg taaccgattc taggtgcatt ggcgcagaaa aaaatgcctg atgcgacgct
     5581 gcgcgtctta tactcccaca tatgccagat tcagcaacgg atacggcttc cccaacttgc
     5641 ccacttccat acgtgtcctc cttaccagaa atttatcctt aagatcccga atcgtttaaa
     5701 ctcgactctg gctctatcga atctccgtcg tttcgagctt acgcgaacag ccgtggcgct
     5761 catttgctcg tcgggcatcg aatctcgtca gctatcgtca gcttaccttt ttggcagcga
     5821 tcgcggctcc cgacatcttg gaccattagc tccacaggta tcttcttccc tctagtggtc
     5881 ataacagcag cttcagctac ctctcaattc aaaaaacccc tcaagacccg tttagaggcc
     5941 ccaaggggtt atgctatcaa tcgttgcgtt acacacacaa aaaaccaaca cacatccatc
     6001 ttcgatggat agcgatttta ttatctaact gctgatcgag tg
//
