LOCUS       sequence-300614-        6290 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 ORF7b.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6290)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6290)
  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-m4252856
FEATURES             Location/Qualifiers
     source          1..6290
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      241..829
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     730..749
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    1055..1126
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(1171..1307)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     terminator      complement(1337..1383)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    1498..1619
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1535..1554)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1589..1608
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(1672..1716)
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             complement(1717..1737)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     promoter        complement(1941..2143)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1970..1990)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2145..2448
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    2529..2577
                     /note="synthetic polyadenylation signal"
     terminator      2595..2641
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    2646..2717
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(2869..3005)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(join(3018..3809,3810..3878))
                     /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     3641..3660
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    4144..4192
                     /note="synthetic polyadenylation signal"
     CDS             complement(4219..4818)
                     /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(4302..4322)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     4799..4818
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(4822..6031)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(4966..4985)
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
ORIGIN
        1 aaatgcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
       61 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
      121 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
      181 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
      241 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
      301 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      361 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      421 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      481 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      541 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      601 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      661 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      721 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      781 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      841 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatga tttaaatgaa
      901 actcgtaatc ggagaacggg tgcaatagca aggaccagat cgaacttgac tgacgcaatt
      961 ctgagcgtag tcgaaaccca gacaagtcga tctcgccaag tagctgactt gactggttaa
     1021 ttaagtttcg atagcccaag gtaaccaatg agattaggga gggaagaaag tttgatgaac
     1081 tacttctgac ccctagtggt gtccagaaaa gaccattaaa ggaatgtctt gatcgaaatc
     1141 gagagatcgc ctgcaggata gctgacgacc ttaaggataa atttctggta aggaggacac
     1201 gtatggaagt gggcaagttg gggaagccgt atccgttgct gaatctggca tatgtgggag
     1261 tataagacgc gcagcgtcgc atcaggcatt tttttctgcg ccaatgccgg tagatagcaa
     1321 tccggcccga ggggcacaaa aaacccctca agacccgttt agaggcccca aggggttatg
     1381 ctacgatacc gatagagatg ggccaatacc acatttgtag aggttttact tgctttaaaa
     1441 aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac
     1501 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     1561 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     1621 catgtctgtg agctgaaggt acgctgtatc tcagtcagtc aagctagctt aactgtccat
     1681 gtgctgcctc tcgaacttag ctgccgcggt ttctttcgac tgaaagtaaa ggttttcggc
     1741 gtgacatgtc tcattatgat cttgtaactc cagactgaac caaaaaatga ttagcataat
     1801 gagcaccaga aaaagcagga aagccaagaa gcacagataa aagtcgatga gcgatagttc
     1861 tatcatggtg gcggccgctg gcgggtgtgt cgagtggatg gtaggatcga caaagatctg
     1921 atctgacggt tcactaaacc agctctgctt atatagacct cccaccgtac acgcctaccg
     1981 cccatttgcg tcaatggggc ggagttgtta cgacattttg gaaagtcccg ttgattttgg
     2041 tgccaaaaca aactcccatt gacgtcaatg gggtggagac ttggaaatcc ccgtgagtca
     2101 aaccgctatc cacgcccatt gatgtactgc caaaaccgca tcagcatggt aatagcgatg
     2161 actaatacgt agatgtactg ccaagtagga aagtcccata aggtcatgta ctgggcataa
     2221 tgccaggcgg gccatttacc gtcattgacg tcaatagggg gcgtacttgg catatgatac
     2281 acttgatgta ctgccaagtg ggcagtttac cgtaaatact ccacccattg acgtcaatgg
     2341 aaagtcccta ttggcgttac tatgggaaca tacgtcatta ttgacgtcaa tgggcggggg
     2401 tcgttgggcg gtcagccagg cgggccattt accgtaagtt atgtaacgcg gaactccata
     2461 tatgggctat gaactaatga ccccgtaatt gattactaga tctggctaca ctcgatcagc
     2521 agttagataa taaaatcgct atccatcgaa gatggatgtg tgttggtttt ttgtgtgtgt
     2581 aacgcaacga ttgatagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt
     2641 gaattgagag gtagctgaag ctgctgttat gaccactaga gggaagaaga tacctgtgga
     2701 gctaatggtc caagatgtcg ggagccgcga tcgctgccaa aaaggtaagc tgacgatagc
     2761 tgacgagatt cgatgcccga cgagcaaatg agcgccacgg ctgttcgcgt aagctcgaaa
     2821 cgacggagat tcgatagagc cagagtcgag tttaaacgat tcgggatctt aaggataaat
     2881 ttctggtaag gaggacacgt atggaagtgg gcaagttggg gaagccgtat ccgttgctga
     2941 atctggcata tgtgggagta taagacgcgc agcgtcgcat caggcatttt tttctgcgcc
     3001 aatgcaccta gaatcggtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     3061 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     3121 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     3181 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     3241 ttatccgcct ccatccagtc tatcaactgt tgccgggaag ctagagtaag tagttcgcca
     3301 gttaatagtt tacgcaaggt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     3361 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     3421 atgttgtgca aaaaagcggt tagctccttc ggtcctccaa tcgttgtcag aagtaagttg
     3481 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     3541 tccgtaagat gcttttctgt gactggtgaa tactcaacca agtcattctg agaatagtgt
     3601 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     3661 agaactttaa aagtgctcat cattggaaag cgttcttcgg ggcgaaaact ctcaaggatc
     3721 ttaccgctgt tgagatccag ttcgatgtaa cccactcgcg cacccaactg atcttcagca
     3781 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     3841 aagggaataa gggcgacacg gaaatgttga atactcatgc gaaagcctcc tgctgatcct
     3901 gtctcttgat agatgtttgc aaaagctagg actcgattat acgttgaccg atcagactat
     3961 caatatctag gtttaaacag atcggtacgc tgccgtgacc aggcttgagt acaatatctg
     4021 acgattggac ttcctgggtc ccaagctgga ttccgactgg ctgactgcca agcaacgata
     4081 tggatcgtcg agactcgcac tgagaggatc gttctcggta caccaggcgc gcctagcgtt
     4141 acacacacaa aaaaccaaca cacatccatc ttcgatggat agcgatttta ttatctaaga
     4201 ctaggcttac atactagcct aggcaccggg cttgcgggtc atgcaccagg tgcgcggtcc
     4261 ttcgggcacc tcgacgtcgg cggtgacggt gaagccgagc cgctcgtaga aggggaggtt
     4321 gcggggcgcg gaggtctcca ggaaggcggg caccccggcg cgctcggcgg cctccactcc
     4381 ggggagcacg acggcgctgc ccagaccctt gccctggtgg tcgggcgaga cgccgacggt
     4441 ggccaggaac cacgcgggct ccttgggccg gtgcggcgcc aggaggccct ccatctgttg
     4501 ctgcgcggcc agccgggaac cgctcaactc ggccatgcgc gggccgatct cggcgaacac
     4561 cgcccccgct tcgacgctct ccggcgtggt ccagaccgcc accgcggcgc cgtcgtccgc
     4621 gacccacacc ttgccgatgt cgagcccgac gcgcgtgagg aagagttctt gcagctcggt
     4681 gacccgctcg atgtggcggt ccggatcgac ggtgtggcgc gtggcggggt agtcggcgaa
     4741 cgcggcggcg agggtgcgta cggccctggg gacgtcgtcg cgggtggcga ggcgcaccgt
     4801 gggcttgtac tcggtcatgt tgtctaacaa aaaagccaaa aacggccaga atttagcgga
     4861 caatttacaa gtctaacact gaaaattaca tattgaccca aatgattaca tttcaaaagg
     4921 tgcctaaaaa acttcacaaa acacactcgc caaccccgag cgcatagttc aaaaccggag
     4981 cttcagctac ttaagaagat aggtacataa aaccgaccaa agaaactgac gcctcactta
     5041 tccctcccct caccagaggt ccggcgcctg tcgattcagg agagcctacc ctaggcccga
     5101 accctgcgtc ctgcgacgga gaaaagccta ccgcacacct accggctggt ggccccaccc
     5161 tgcattataa gccaacagaa cgggtgacgt cacgacacga cgagggcgcg cgctcccaaa
     5221 ggtacgggtg cactgcccaa cggcaccgcc ataactgccg cccccgcaac agacgacaaa
     5281 ccgagttctc cagtcagtga caaacttcac gtcagggtcc ccagatggtg cccagcccat
     5341 ctcacccgaa taagagcttt cccgcattag cgaagggctc aagaccttgg gttcttgccg
     5401 cccaccatgc cccccacctt gtttcaacga cctcacagcc cgcctcacaa gcgtcttcca
     5461 ttcaagactc gggaacagcc gccattttgc tgcgctcccc ccaaccccca gttcagggca
     5521 accttgctcg cggacccaga ctacagccct tggcggtctc tccacacgct tccgtcccac
     5581 cgagcggccc ggcggccacg aaagccccgg ccagcccagc agcccgctac tcaccaagtg
     5641 acgatcacag cgatccacaa acaagaaccg cgacccaaat cccggctgcg acggaactag
     5701 ctgtgccaca cccggcgcgt ccttatataa tcatcggcgt tcaccgcccc acggagatcc
     5761 ctccgcagaa tcgccgagaa gggactactt ttcctcgcct gttccgctct ctggaaagaa
     5821 aaccagtgcc ctagagtcac ccaagtcccg tcctaaaatg tccttctgct gatactgggg
     5881 ttctaaggcc gagtcttatg agcagcgggc cgctgtcctg agcgtccggg cggaaggatc
     5941 aggacgctcc tgcgcccttc gtctgacgtg gcagcgctcg ccgtgacgag gggggcgccc
     6001 gcgggaggcg ccaaaacccg gcgcggaggc caggcgcgcc ccgatttcga tattcctcgg
     6061 atctcggata gatcgggatt gaggatgtag gttgaacttc tctcgggtga tcccgatctc
     6121 ggatattcgg gattggagta atacctggcg ataggcgctg ccaagcaata gccaaggcgc
     6181 aaggccggcc aagagacttg cgatagttcg ttcgagaggt agcctgcaag atggatcgga
     6241 cgaaaggttt gaagattttc cgacgagggc cttggagagt tcggaaattt
//
