LOCUS       sequence-300450-        6224 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 6224)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6224)
  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-m4252973
FEATURES             Location/Qualifiers
     source          1..6224
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      171..217
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      247..383
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    428..499
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(725..1313)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(805..824)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        1813..3022
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     2859..2878
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3026..3625
                     /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(3026..3045)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3522..3542
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    3652..3700
                     /note="synthetic polyadenylation signal"
     CDS             join(3966..4034,4035..4826)
                     /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(4184..4203)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4839..4975
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5127..5198
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5203..5249)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5267..5315
                     /note="synthetic polyadenylation signal"
     enhancer        5396..5699
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5701..5903
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5854..5874
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     polyA_signal    join(6159..6224,1..56)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6170..6189)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     join(6224,1..19)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca
       61 acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa
      121 gcaagtaaaa cctctacaaa tgtggtattg gcccatctct atcggtatcg tagcataacc
      181 ccttggggcc tctaaacggg tcttgagggg ttttttgtgc ccctcgggcc ggattgctat
      241 ctaccggcat tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt atactcccac
      301 atatgccaga ttcagcaacg gatacggctt ccccaacttg cccacttcca tacgtgtcct
      361 ccttaccaga aatttatcct taaggtcgtc agctatcctg caggcgatct ctcgatttcg
      421 atcaagacat tcctttaatg gtcttttctg gacaccacta ggggtcagaa gtagttcatc
      481 aaactttctt ccctccctaa tctcattggt taccttgggc tatcgaaact taattaacca
      541 gtcaagtcag ctacttggcg agatcgactt gtctgggttt cgactacgct cagaattgcg
      601 tcagtcaagt tcgatctggt ccttgctatt gcacccgttc tccgattacg agtttcattt
      661 aaatcatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
      721 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
      781 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
      841 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
      901 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
      961 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     1021 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     1081 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     1141 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     1201 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     1261 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     1321 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     1381 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     1441 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     1501 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgca tttaaatttc
     1561 cgaactctcc aaggccctcg tcggaaaatc ttcaaacctt tcgtccgatc catcttgcag
     1621 gctacctctc gaacgaacta tcgcaagtct cttggccggc cttgcgcctt ggctattgct
     1681 tggcagcgcc tatcgccagg tattactcca atcccgaata tccgagatcg ggatcacccg
     1741 agagaagttc aacctacatc ctcaatcccg atctatccga gatccgagga atatcgaaat
     1801 cggggcgcgc ctggcctccg cgccgggttt tggcgcctcc cgcgggcgcc cccctcgtca
     1861 cggcgagcgc tgccacgtca gacgaagggc gcaggagcgt cctgatcctt ccgcccggac
     1921 gctcaggaca gcggcccgct gctcataaga ctcggcctta gaaccccagt atcagcagaa
     1981 ggacatttta ggacgggact tgggtgactc tagggcactg gttttctttc cagagagcgg
     2041 aacaggcgag gaaaagtagt cccttctcgg cgattctgcg gagggatctc cgtggggcgg
     2101 tgaacgccga tgattatata aggacgcgcc gggtgtggca cagctagttc cgtcgcagcc
     2161 gggatttggg tcgcggttct tgtttgtgga tcgctgtgat cgtcacttgg tgagtagcgg
     2221 gctgctgggc tggccggggc tttcgtggcc gccgggccgc tcggtgggac ggaagcgtgt
     2281 ggagagaccg ccaagggctg tagtctgggt ccgcgagcaa ggttgccctg aactgggggt
     2341 tggggggagc gcagcaaaat ggcggctgtt cccgagtctt gaatggaaga cgcttgtgag
     2401 gcgggctgtg aggtcgttga aacaaggtgg ggggcatggt gggcggcaag aacccaaggt
     2461 cttgagccct tcgctaatgc gggaaagctc ttattcgggt gagatgggct gggcaccatc
     2521 tggggaccct gacgtgaagt ttgtcactga ctggagaact cggtttgtcg tctgttgcgg
     2581 gggcggcagt tatggcggtg ccgttgggca gtgcacccgt acctttggga gcgcgcgccc
     2641 tcgtcgtgtc gtgacgtcac ccgttctgtt ggcttataat gcagggtggg gccaccagcc
     2701 ggtaggtgtg cggtaggctt ttctccgtcg caggacgcag ggttcgggcc tagggtaggc
     2761 tctcctgaat cgacaggcgc cggacctctg gtgaggggag ggataagtga ggcgtcagtt
     2821 tctttggtcg gttttatgta cctatcttct taagtagctg aagctccggt tttgaactat
     2881 gcgctcgggg ttggcgagtg tgttttgtga agttttttag gcaccttttg aaatgtaatc
     2941 atttgggtca atatgtaatt ttcagtgtta gacttgtaaa ttgtccgcta aattctggcc
     3001 gtttttggct tttttgttag acaacatgac cgagtacaag cccacggtgc gcctcgccac
     3061 ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc
     3121 cacgcgccac accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact
     3181 cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc
     3241 ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg
     3301 cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga tggagggcct
     3361 cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc
     3421 cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggccgccga
     3481 gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga
     3541 gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg
     3601 catgacccgc aagcccggtg cctaggctag tatgtaagcc tagtcttaga taataaaatc
     3661 gctatccatc gaagatggat gtgtgttggt tttttgtgtg tgtaacgcta ggcgcgcctg
     3721 gtgtaccgag aacgatcctc tcagtgcgag tctcgacgat ccatatcgtt gcttggcagt
     3781 cagccagtcg gaatccagct tgggacccag gaagtccaat cgtcagatat tgtactcaag
     3841 cctggtcacg gcagcgtacc gatctgttta aacctagata ttgatagtct gatcggtcaa
     3901 cgtataatcg agtcctagct tttgcaaaca tctatcaaga gacaggatca gcaggaggct
     3961 ttcgcatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     4021 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     4081 gtgcgcgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     4141 gccccgaaga acgctttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     4201 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     4261 acttggttga gtattcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     4321 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     4381 cgattggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     4441 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     4501 cgatgcctgt agcaatggca acaaccttgc gtaaactatt aactggcgaa ctacttactc
     4561 tagcttcccg gcaacagttg atagactgga tggaggcgga taaagttgca ggaccacttc
     4621 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     4681 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     4741 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     4801 gtgcctcact gattaagcat tggtaaccga ttctaggtgc attggcgcag aaaaaaatgc
     4861 ctgatgcgac gctgcgcgtc ttatactccc acatatgcca gattcagcaa cggatacggc
     4921 ttccccaact tgcccacttc catacgtgtc ctccttacca gaaatttatc cttaagatcc
     4981 cgaatcgttt aaactcgact ctggctctat cgaatctccg tcgtttcgag cttacgcgaa
     5041 cagccgtggc gctcatttgc tcgtcgggca tcgaatctcg tcagctatcg tcagcttacc
     5101 tttttggcag cgatcgcggc tcccgacatc ttggaccatt agctccacag gtatcttctt
     5161 ccctctagtg gtcataacag cagcttcagc tacctctcaa ttcaaaaaac ccctcaagac
     5221 ccgtttagag gccccaaggg gttatgctat caatcgttgc gttacacaca caaaaaacca
     5281 acacacatcc atcttcgatg gatagcgatt ttattatcta actgctgatc gagtgtagcc
     5341 agatctagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
     5401 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
     5461 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
     5521 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
     5581 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
     5641 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgc
     5701 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
     5761 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
     5821 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
     5881 gggaggtcta tataagcaga gctggtttag tgaaccgtca gatcagatct ttgtcgatcc
     5941 taccatccac tcgacacacc cgccagcggc cgccaccatg attgaacttt cattaattga
     6001 cttctatttg tgctttttag cctttctgct attccttgtt ttaattatgc ttattatctt
     6061 ttggttctca cttgaactgc aagatcataa tgaaacttgt cacgcctaag ctagcttgac
     6121 tgactgagat acagcgtacc ttcagctcac agacatgata agatacattg atgagtttgg
     6181 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtg
//
