LOCUS       sequence-297502-        4380 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 4380)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4380)
  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-m4133670
FEATURES             Location/Qualifiers
     source          1..4380
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             134..154
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             155..196
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     polyA_signal    249..370
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(260..279)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     314..333
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      485..531
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      561..697
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    742..813
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1039..1627)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1119..1138)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(2373..2441,2442..3233)
                     /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(2591..2610)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3246..3382
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3534..3605
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(3610..3656)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    3674..3722
                     /note="synthetic polyadenylation signal"
     enhancer        3803..4106
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4108..4310
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4261..4281
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgatt gaactttcat taattgactt ctatttgtgc tttttagcct ttctgctatt
       61 ccttgtttta attatgctta ttatcttttg gttctcactt gaactgcaag atcataatga
      121 aacttgtcac gccgaaaacc tttactttca gtcggggaaa cctataccca atccacttct
      181 cggcttagat tctacctaag ctagcttgac tgactgagat acagcgtacc ttcagctcac
      241 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
      301 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
      361 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg
      421 ggaggttttt taaagcaagt aaaacctcta caaatgtggt attggcccat ctctatcggt
      481 atcgtagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gtgcccctcg
      541 ggccggattg ctatctaccg gcattggcgc agaaaaaaat gcctgatgcg acgctgcgcg
      601 tcttatactc ccacatatgc cagattcagc aacggatacg gcttccccaa cttgcccact
      661 tccatacgtg tcctccttac cagaaattta tccttaaggt cgtcagctat cctgcaggcg
      721 atctctcgat ttcgatcaag acattccttt aatggtcttt tctggacacc actaggggtc
      781 agaagtagtt catcaaactt tcttccctcc ctaatctcat tggttacctt gggctatcga
      841 aacttaatta accagtcaag tcagctactt ggcgagatcg acttgtctgg gtttcgacta
      901 cgctcagaat tgcgtcagtc aagttcgatc tggtccttgc tattgcaccc gttctccgat
      961 tacgagtttc atttaaatca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     1021 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     1081 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     1141 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     1201 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     1261 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     1321 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     1381 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     1441 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     1501 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     1561 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     1621 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     1681 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     1741 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     1801 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     1861 tgcatttaaa tttccgaact ctccaaggcc ctcgtcggaa aatcttcaaa cctttcgtcc
     1921 gatccatctt gcaggctacc tctcgaacga actatcgcaa gtctcttggc cggccttgcg
     1981 ccttggctat tgcttggcag cgcctatcgc caggtattac tccaatcccg aatatccgag
     2041 atcgggatca cccgagagaa gttcaaccta catcctcaat cccgatctat ccgagatccg
     2101 aggaatatcg aaatcggggc gcgcctggtg taccgagaac gatcctctca gtgcgagtct
     2161 cgacgatcca tatcgttgct tggcagtcag ccagtcggaa tccagcttgg gacccaggaa
     2221 gtccaatcgt cagatattgt actcaagcct ggtcacggca gcgtaccgat ctgtttaaac
     2281 ctagatattg atagtctgat cggtcaacgt ataatcgagt cctagctttt gcaaacatct
     2341 atcaagagac aggatcagca ggaggctttc gcatgagtat tcaacatttc cgtgtcgccc
     2401 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     2461 aagtaaaaga tgctgaagat cagttgggtg cgcgagtggg ttacatcgaa ctggatctca
     2521 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ctttccaatg atgagcactt
     2581 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     2641 gtcgccgcat acactattct cagaatgact tggttgagta ttcaccagtc acagaaaagc
     2701 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     2761 acactgcggc caacttactt ctgacaacga ttggaggacc gaaggagcta accgcttttt
     2821 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     2881 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca accttgcgta
     2941 aactattaac tggcgaacta cttactctag cttcccggca acagttgata gactggatgg
     3001 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     3061 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     3121 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     3181 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taaccgattc
     3241 taggtgcatt ggcgcagaaa aaaatgcctg atgcgacgct gcgcgtctta tactcccaca
     3301 tatgccagat tcagcaacgg atacggcttc cccaacttgc ccacttccat acgtgtcctc
     3361 cttaccagaa atttatcctt aagatcccga atcgtttaaa ctcgactctg gctctatcga
     3421 atctccgtcg tttcgagctt acgcgaacag ccgtggcgct catttgctcg tcgggcatcg
     3481 aatctcgtca gctatcgtca gcttaccttt ttggcagcga tcgcggctcc cgacatcttg
     3541 gaccattagc tccacaggta tcttcttccc tctagtggtc ataacagcag cttcagctac
     3601 ctctcaattc aaaaaacccc tcaagacccg tttagaggcc ccaaggggtt atgctatcaa
     3661 tcgttgcgtt acacacacaa aaaaccaaca cacatccatc ttcgatggat agcgatttta
     3721 ttatctaact gctgatcgag tgtagccaga tctagtaatc aattacgggg tcattagttc
     3781 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
     3841 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
     3901 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
     3961 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
     4021 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
     4081 acgtattagt catcgctatt accatgctga tgcggttttg gcagtacatc aatgggcgtg
     4141 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
     4201 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
     4261 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga
     4321 accgtcagat cagatctttg tcgatcctac catccactcg acacacccgc cagcggccgc
//
