LOCUS       sequence-300418-        8888 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 RNA-dependent RNA 
            polymerase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8888)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 8888)
  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-m4252975
FEATURES             Location/Qualifiers
     source          1..8888
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    85..156
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(161..207)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    225..273
                     /note="synthetic polyadenylation signal"
     enhancer        354..657
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        659..861
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     812..832
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      930..939
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3781..3902
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3792..3811)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3846..3865
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      4017..4063
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      4093..4229
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4274..4345
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(4571..5159)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4651..4670)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        5659..6868
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     6705..6724
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             6872..7471
                     /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(6872..6891)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7368..7388
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    7498..7546
                     /note="synthetic polyadenylation signal"
     CDS             join(7812..7880,7881..8672)
                     /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(8030..8049)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      8685..8821
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
ORIGIN
        1 gccgtggcgc tcatttgctc gtcgggcatc gaatctcgtc agctatcgtc agcttacctt
       61 tttggcagcg atcgcggctc ccgacatctt ggaccattag ctccacaggt atcttcttcc
      121 ctctagtggt cataacagca gcttcagcta cctctcaatt caaaaaaccc ctcaagaccc
      181 gtttagaggc cccaaggggt tatgctatca atcgttgcgt tacacacaca aaaaaccaac
      241 acacatccat cttcgatgga tagcgatttt attatctaac tgctgatcga gtgtagccag
      301 atctagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
      361 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
      421 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
      481 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
      541 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
      601 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatgctg
      661 atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca
      721 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt
      781 ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg
      841 gaggtctata taagcagagc tggtttagtg aaccgtcaga tcagatcttt gtcgatccta
      901 ccatccactc gacacacccg ccagcggccg ccaccatggc tgatgcacaa tcgtttttaa
      961 accgggtttg cggtgtaagt gcagcccgtc ttacaccgtg cggcacaggc actagtactg
     1021 atgtcgtata cagggctttt gacatctaca atgataaagt agctggtttt gctaaattcc
     1081 taaaaactaa ttgttgtcgc ttccaagaaa aggacgaaga tgacaattta attgattctt
     1141 actttgtagt taagagacac actttctcta actaccaaca tgaagaaaca atttataatt
     1201 tacttaagga ttgtccagct gttgctaaac atgacttctt taagtttaga atagacggtg
     1261 acatggtacc acatatatca cgtcaacgtc ttactaaata cacaatggca gacctcgtct
     1321 atgctttaag gcattttgat gaaggtaatt gtgacacatt aaaagaaata cttgtcacat
     1381 acaattgttg tgatgatgat tatttcaata aaaaggactg gtatgatttt gtagaaaacc
     1441 cagatatatt acgcgtatac gccaacttag gtgaacgtgt acgccaagct ttgttaaaaa
     1501 cagtacaatt ctgtgatgcc atgcgaaatg ctggtattgt tggtgtactg acattagata
     1561 atcaagatct caatggtaac tggtatgatt tcggtgattt catacaaacc acgccaggta
     1621 gtggagttcc tgttgtagat tcttattatt cattgttaat gcctatatta accttgacca
     1681 gggctttaac tgcagagtca catgttgaca ctgacttaac aaagccttac attaagtggg
     1741 atttgttaaa atatgacttc acggaagaga ggttaaaact ctttgaccgt tattttaaat
     1801 attgggatca gacataccac ccaaattgtg ttaactgttt ggatgacaga tgcattctgc
     1861 attgtgcaaa ctttaatgtt ttattctcta cagtgttccc acctacaagt tttggaccac
     1921 tagtgagaaa aatatttgtt gatggtgttc catttgtagt ttcaactgga taccacttca
     1981 gagagctagg tgttgtacat aatcaggatg taaacttaca tagctctaga cttagtttta
     2041 aggaattact tgtgtatgct gctgaccctg ctatgcacgc tgcttctggt aatctattac
     2101 tagataaacg cactacgtgc ttttcagtag ctgcacttac taacaatgtt gcttttcaaa
     2161 ctgtcaaacc cggtaatttt aacaaagact tctatgactt tgctgtgtct aagggtttct
     2221 ttaaggaagg aagttctgtt gaattaaaac acttcttctt tgctcaggat ggtaatgctg
     2281 ctatcagcga ttatgactac tatcgttata atctaccaac aatgtgtgat atcagacaac
     2341 tactatttgt agttgaagtt gttgataagt actttgattg ttacgatggt ggctgtatta
     2401 atgctaacca agtcatcgtc aacaacctag acaaatcagc tggttttcca tttaataaat
     2461 ggggtaaggc tagactttat tatgattcaa tgagttatga ggatcaagat gcacttttcg
     2521 catatacaaa acgtaatgtc atccctacta taactcaaat gaatcttaag tatgccatta
     2581 gtgcaaagaa tagagctcgc accgtagctg gtgtctctat ctgtagtact atgaccaata
     2641 gacagtttca tcaaaaatta ttgaaatcaa tagccgccac tagaggagct actgtagtaa
     2701 ttggaacaag caaattctat ggtggttggc acaacatgtt aaaaactgtt tatagtgatg
     2761 tagaaaaccc tcaccttatg ggttgggatt atcctaaatg tgatagagcc atgcctaaca
     2821 tgcttagaat tatggcctca cttgttcttg ctcgcaaaca tacaacgtgt tgtagcttgt
     2881 cacaccgttt ctatagatta gctaatgagt gtgctcaagt attgagtgaa atggtcatgt
     2941 gtggcggttc actatatgtt aaaccaggtg gaacctcatc aggagatgcc acaactgctt
     3001 atgctaatag tgtttttaac atttgtcaag ctgtcacggc caatgttaat gcacttttat
     3061 ctactgatgg taacaaaatt gccgataagt atgtccgcaa tttacaacac agactttatg
     3121 agtgtctcta tagaaataga gatgttgaca cagactttgt gaatgagttt tacgcatatt
     3181 tgcgtaaaca tttctcaatg atgatactct ctgacgatgc tgttgtgtgt ttcaatagca
     3241 cttatgcatc tcaaggtcta gtggctagca taaagaactt taagtcagtt ctttattatc
     3301 aaaacaatgt ttttatgtct gaagcaaaat gttggactga gactgacctt actaaaggac
     3361 ctcatgaatt ttgctctcaa catacaatgc tagttaaaca gggtgatgat tatgtgtacc
     3421 ttccttaccc agatccatca agaatcctag gggccggctg ttttgtagat gatatcgtaa
     3481 aaacagatgg tacacttatg attgaacggt tcgtgtcttt agctatagat gcttacccac
     3541 ttactaaaca tcctaatcag gagtatgctg atgtctttca tttgtactta caatacataa
     3601 gaaagctaca tgatgagtta acaggacaca tgttagacat gtattctgtt atgcttacta
     3661 atgataacac ttcaaggtat tgggaacctg agttttatga ggctatgtac acaccgcata
     3721 cagtcttata agctagcttg actgactgag atacagcgta ccttcagctc acagacatga
     3781 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
     3841 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     3901 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt
     3961 tttaaagcaa gtaaaacctc tacaaatgtg gtattggccc atctctatcg gtatcgtagc
     4021 ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgtgcccct cgggccggat
     4081 tgctatctac cggcattggc gcagaaaaaa atgcctgatg cgacgctgcg cgtcttatac
     4141 tcccacatat gccagattca gcaacggata cggcttcccc aacttgccca cttccatacg
     4201 tgtcctcctt accagaaatt tatccttaag gtcgtcagct atcctgcagg cgatctctcg
     4261 atttcgatca agacattcct ttaatggtct tttctggaca ccactagggg tcagaagtag
     4321 ttcatcaaac tttcttccct ccctaatctc attggttacc ttgggctatc gaaacttaat
     4381 taaccagtca agtcagctac ttggcgagat cgacttgtct gggtttcgac tacgctcaga
     4441 attgcgtcag tcaagttcga tctggtcctt gctattgcac ccgttctccg attacgagtt
     4501 tcatttaaat catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     4561 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     4621 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     4681 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     4741 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     4801 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     4861 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     4921 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     4981 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     5041 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     5101 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     5161 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     5221 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     5281 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     5341 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcattta
     5401 aatttccgaa ctctccaagg ccctcgtcgg aaaatcttca aacctttcgt ccgatccatc
     5461 ttgcaggcta cctctcgaac gaactatcgc aagtctcttg gccggccttg cgccttggct
     5521 attgcttggc agcgcctatc gccaggtatt actccaatcc cgaatatccg agatcgggat
     5581 cacccgagag aagttcaacc tacatcctca atcccgatct atccgagatc cgaggaatat
     5641 cgaaatcggg gcgcgcctgg cctccgcgcc gggttttggc gcctcccgcg ggcgcccccc
     5701 tcgtcacggc gagcgctgcc acgtcagacg aagggcgcag gagcgtcctg atccttccgc
     5761 ccggacgctc aggacagcgg cccgctgctc ataagactcg gccttagaac cccagtatca
     5821 gcagaaggac attttaggac gggacttggg tgactctagg gcactggttt tctttccaga
     5881 gagcggaaca ggcgaggaaa agtagtccct tctcggcgat tctgcggagg gatctccgtg
     5941 gggcggtgaa cgccgatgat tatataagga cgcgccgggt gtggcacagc tagttccgtc
     6001 gcagccggga tttgggtcgc ggttcttgtt tgtggatcgc tgtgatcgtc acttggtgag
     6061 tagcgggctg ctgggctggc cggggctttc gtggccgccg ggccgctcgg tgggacggaa
     6121 gcgtgtggag agaccgccaa gggctgtagt ctgggtccgc gagcaaggtt gccctgaact
     6181 gggggttggg gggagcgcag caaaatggcg gctgttcccg agtcttgaat ggaagacgct
     6241 tgtgaggcgg gctgtgaggt cgttgaaaca aggtgggggg catggtgggc ggcaagaacc
     6301 caaggtcttg agcccttcgc taatgcggga aagctcttat tcgggtgaga tgggctgggc
     6361 accatctggg gaccctgacg tgaagtttgt cactgactgg agaactcggt ttgtcgtctg
     6421 ttgcgggggc ggcagttatg gcggtgccgt tgggcagtgc acccgtacct ttgggagcgc
     6481 gcgccctcgt cgtgtcgtga cgtcacccgt tctgttggct tataatgcag ggtggggcca
     6541 ccagccggta ggtgtgcggt aggcttttct ccgtcgcagg acgcagggtt cgggcctagg
     6601 gtaggctctc ctgaatcgac aggcgccgga cctctggtga ggggagggat aagtgaggcg
     6661 tcagtttctt tggtcggttt tatgtaccta tcttcttaag tagctgaagc tccggttttg
     6721 aactatgcgc tcggggttgg cgagtgtgtt ttgtgaagtt ttttaggcac cttttgaaat
     6781 gtaatcattt gggtcaatat gtaattttca gtgttagact tgtaaattgt ccgctaaatt
     6841 ctggccgttt ttggcttttt tgttagacaa catgaccgag tacaagccca cggtgcgcct
     6901 cgccacccgc gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta
     6961 ccccgccacg cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca
     7021 agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg
     7081 cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga
     7141 gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga
     7201 gggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt
     7261 ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc
     7321 cgccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt
     7381 ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac
     7441 ctggtgcatg acccgcaagc ccggtgccta ggctagtatg taagcctagt cttagataat
     7501 aaaatcgcta tccatcgaag atggatgtgt gttggttttt tgtgtgtgta acgctaggcg
     7561 cgcctggtgt accgagaacg atcctctcag tgcgagtctc gacgatccat atcgttgctt
     7621 ggcagtcagc cagtcggaat ccagcttggg acccaggaag tccaatcgtc agatattgta
     7681 ctcaagcctg gtcacggcag cgtaccgatc tgtttaaacc tagatattga tagtctgatc
     7741 ggtcaacgta taatcgagtc ctagcttttg caaacatcta tcaagagaca ggatcagcag
     7801 gaggctttcg catgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     7861 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     7921 agttgggtgc gcgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     7981 gttttcgccc cgaagaacgc tttccaatga tgagcacttt taaagttctg ctatgtggcg
     8041 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     8101 agaatgactt ggttgagtat tcaccagtca cagaaaagca tcttacggat ggcatgacag
     8161 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     8221 tgacaacgat tggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     8281 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     8341 acaccacgat gcctgtagca atggcaacaa ccttgcgtaa actattaact ggcgaactac
     8401 ttactctagc ttcccggcaa cagttgatag actggatgga ggcggataaa gttgcaggac
     8461 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     8521 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     8581 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     8641 agataggtgc ctcactgatt aagcattggt aaccgattct aggtgcattg gcgcagaaaa
     8701 aaatgcctga tgcgacgctg cgcgtcttat actcccacat atgccagatt cagcaacgga
     8761 tacggcttcc ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa tttatcctta
     8821 agatcccgaa tcgtttaaac tcgactctgg ctctatcgaa tctccgtcgt ttcgagctta
     8881 cgcgaaca
//
