LOCUS       sequence-300438-        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-m4252980
FEATURES             Location/Qualifiers
     source          1..6224
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    217..288
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(293..339)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    357..405
                     /note="synthetic polyadenylation signal"
     enhancer        486..789
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        791..993
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     944..964
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     polyA_signal    1249..1370
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1260..1279)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1314..1333
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1485..1531
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1561..1697
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1742..1813
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2039..2627)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2119..2138)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3127..4336
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4173..4192
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4340..4939
                     /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(4340..4359)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4836..4856
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4966..5014
                     /note="synthetic polyadenylation signal"
     CDS             join(5280..5348,5349..6140)
                     /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(5498..5517)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      join(6153..6224,1..65)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
ORIGIN
        1 cggatacggc ttccccaact tgcccacttc catacgtgtc ctccttacca gaaatttatc
       61 cttaagatcc cgaatcgttt aaactcgact ctggctctat cgaatctccg tcgtttcgag
      121 cttacgcgaa cagccgtggc gctcatttgc tcgtcgggca tcgaatctcg tcagctatcg
      181 tcagcttacc tttttggcag cgatcgcggc tcccgacatc ttggaccatt agctccacag
      241 gtatcttctt ccctctagtg gtcataacag cagcttcagc tacctctcaa ttcaaaaaac
      301 ccctcaagac ccgtttagag gccccaaggg gttatgctat caatcgttgc gttacacaca
      361 caaaaaacca acacacatcc atcttcgatg gatagcgatt ttattatcta actgctgatc
      421 gagtgtagcc agatctagta atcaattacg gggtcattag ttcatagccc atatatggag
      481 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      541 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      601 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      661 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      721 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      781 attaccatgc tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      841 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      901 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      961 cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca gatcagatct
     1021 ttgtcgatcc taccatccac tcgacacacc cgccagcggc cgccaccatg atagaactat
     1081 cgctcatcga cttttatctg tgcttcttgg ctttcctgct ttttctggtg ctcattatgc
     1141 taatcatttt ttggttcagt ctggagttac aagatcataa tgagacatgt cacgcctaag
     1201 ctagcttgac tgactgagat acagcgtacc ttcagctcac agacatgata agatacattg
     1261 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
     1321 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
     1381 attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt
     1441 aaaacctcta caaatgtggt attggcccat ctctatcggt atcgtagcat aaccccttgg
     1501 ggcctctaaa cgggtcttga ggggtttttt gtgcccctcg ggccggattg ctatctaccg
     1561 gcattggcgc agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc ccacatatgc
     1621 cagattcagc aacggatacg gcttccccaa cttgcccact tccatacgtg tcctccttac
     1681 cagaaattta tccttaaggt cgtcagctat cctgcaggcg atctctcgat ttcgatcaag
     1741 acattccttt aatggtcttt tctggacacc actaggggtc agaagtagtt catcaaactt
     1801 tcttccctcc ctaatctcat tggttacctt gggctatcga aacttaatta accagtcaag
     1861 tcagctactt ggcgagatcg acttgtctgg gtttcgacta cgctcagaat tgcgtcagtc
     1921 aagttcgatc tggtccttgc tattgcaccc gttctccgat tacgagtttc atttaaatca
     1981 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     2041 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     2101 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     2161 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     2221 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     2281 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     2341 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     2401 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     2461 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     2521 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     2581 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     2641 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     2701 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     2761 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     2821 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcatttaaa tttccgaact
     2881 ctccaaggcc ctcgtcggaa aatcttcaaa cctttcgtcc gatccatctt gcaggctacc
     2941 tctcgaacga actatcgcaa gtctcttggc cggccttgcg ccttggctat tgcttggcag
     3001 cgcctatcgc caggtattac tccaatcccg aatatccgag atcgggatca cccgagagaa
     3061 gttcaaccta catcctcaat cccgatctat ccgagatccg aggaatatcg aaatcggggc
     3121 gcgcctggcc tccgcgccgg gttttggcgc ctcccgcggg cgcccccctc gtcacggcga
     3181 gcgctgccac gtcagacgaa gggcgcagga gcgtcctgat ccttccgccc ggacgctcag
     3241 gacagcggcc cgctgctcat aagactcggc cttagaaccc cagtatcagc agaaggacat
     3301 tttaggacgg gacttgggtg actctagggc actggttttc tttccagaga gcggaacagg
     3361 cgaggaaaag tagtcccttc tcggcgattc tgcggaggga tctccgtggg gcggtgaacg
     3421 ccgatgatta tataaggacg cgccgggtgt ggcacagcta gttccgtcgc agccgggatt
     3481 tgggtcgcgg ttcttgtttg tggatcgctg tgatcgtcac ttggtgagta gcgggctgct
     3541 gggctggccg gggctttcgt ggccgccggg ccgctcggtg ggacggaagc gtgtggagag
     3601 accgccaagg gctgtagtct gggtccgcga gcaaggttgc cctgaactgg gggttggggg
     3661 gagcgcagca aaatggcggc tgttcccgag tcttgaatgg aagacgcttg tgaggcgggc
     3721 tgtgaggtcg ttgaaacaag gtggggggca tggtgggcgg caagaaccca aggtcttgag
     3781 cccttcgcta atgcgggaaa gctcttattc gggtgagatg ggctgggcac catctgggga
     3841 ccctgacgtg aagtttgtca ctgactggag aactcggttt gtcgtctgtt gcgggggcgg
     3901 cagttatggc ggtgccgttg ggcagtgcac ccgtaccttt gggagcgcgc gccctcgtcg
     3961 tgtcgtgacg tcacccgttc tgttggctta taatgcaggg tggggccacc agccggtagg
     4021 tgtgcggtag gcttttctcc gtcgcaggac gcagggttcg ggcctagggt aggctctcct
     4081 gaatcgacag gcgccggacc tctggtgagg ggagggataa gtgaggcgtc agtttctttg
     4141 gtcggtttta tgtacctatc ttcttaagta gctgaagctc cggttttgaa ctatgcgctc
     4201 ggggttggcg agtgtgtttt gtgaagtttt ttaggcacct tttgaaatgt aatcatttgg
     4261 gtcaatatgt aattttcagt gttagacttg taaattgtcc gctaaattct ggccgttttt
     4321 ggcttttttg ttagacaaca tgaccgagta caagcccacg gtgcgcctcg ccacccgcga
     4381 cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg
     4441 ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct
     4501 cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc
     4561 ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg
     4621 catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggagg gcctcctggc
     4681 gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca
     4741 ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggccg ccgagcgcgc
     4801 cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct
     4861 cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac
     4921 ccgcaagccc ggtgcctagg ctagtatgta agcctagtct tagataataa aatcgctatc
     4981 catcgaagat ggatgtgtgt tggttttttg tgtgtgtaac gctaggcgcg cctggtgtac
     5041 cgagaacgat cctctcagtg cgagtctcga cgatccatat cgttgcttgg cagtcagcca
     5101 gtcggaatcc agcttgggac ccaggaagtc caatcgtcag atattgtact caagcctggt
     5161 cacggcagcg taccgatctg tttaaaccta gatattgata gtctgatcgg tcaacgtata
     5221 atcgagtcct agcttttgca aacatctatc aagagacagg atcagcagga ggctttcgca
     5281 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     5341 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcgc
     5401 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     5461 aagaacgctt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     5521 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     5581 ttgagtattc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     5641 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgattg
     5701 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     5761 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     5821 ctgtagcaat ggcaacaacc ttgcgtaaac tattaactgg cgaactactt actctagctt
     5881 cccggcaaca gttgatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     5941 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     6001 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     6061 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     6121 cactgattaa gcattggtaa ccgattctag gtgcattggc gcagaaaaaa atgcctgatg
     6181 cgacgctgcg cgtcttatac tcccacatat gccagattca gcaa
//
