LOCUS       sequence-300599-        4383 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 4383)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4383)
  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-m4133176
FEATURES             Location/Qualifiers
     source          1..4383
                     /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"
     CDS             1065..1085
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1086..1130
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     polyA_signal    1183..1304
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1194..1213)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1248..1267
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1419..1465
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1495..1631
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1676..1747
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1973..2561)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2053..2072)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3307..3375,3376..4167)
                     /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(3525..3544)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4180..4316
                     /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 ccaccatgat agaactatcg ctcatcgact
      961 tttatctgtg cttcttggct ttcctgcttt ttctggtgct cattatgcta atcatttttt
     1021 ggttcagtct ggagttacaa gatcataatg agacatgtca cgccgaaaac ctttactttc
     1081 agtcgaaaga aaccgcggca gctaagttcg agaggcagca catggacagt taagctagct
     1141 tgactgactg agatacagcg taccttcagc tcacagacat gataagatac attgatgagt
     1201 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg
     1261 ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca
     1321 ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc
     1381 tctacaaatg tggtattggc ccatctctat cggtatcgta gcataacccc ttggggcctc
     1441 taaacgggtc ttgaggggtt ttttgtgccc ctcgggccgg attgctatct accggcattg
     1501 gcgcagaaaa aaatgcctga tgcgacgctg cgcgtcttat actcccacat atgccagatt
     1561 cagcaacgga tacggcttcc ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa
     1621 tttatcctta aggtcgtcag ctatcctgca ggcgatctct cgatttcgat caagacattc
     1681 ctttaatggt cttttctgga caccactagg ggtcagaagt agttcatcaa actttcttcc
     1741 ctccctaatc tcattggtta ccttgggcta tcgaaactta attaaccagt caagtcagct
     1801 acttggcgag atcgacttgt ctgggtttcg actacgctca gaattgcgtc agtcaagttc
     1861 gatctggtcc ttgctattgc acccgttctc cgattacgag tttcatttaa atcatgtgag
     1921 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     1981 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     2041 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     2101 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     2161 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     2221 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     2281 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     2341 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     2401 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     2461 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     2521 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     2581 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     2641 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     2701 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     2761 tctcagcgat ctgtctattt cgttcatcca tagttgcatt taaatttccg aactctccaa
     2821 ggccctcgtc ggaaaatctt caaacctttc gtccgatcca tcttgcaggc tacctctcga
     2881 acgaactatc gcaagtctct tggccggcct tgcgccttgg ctattgcttg gcagcgccta
     2941 tcgccaggta ttactccaat cccgaatatc cgagatcggg atcacccgag agaagttcaa
     3001 cctacatcct caatcccgat ctatccgaga tccgaggaat atcgaaatcg gggcgcgcct
     3061 ggtgtaccga gaacgatcct ctcagtgcga gtctcgacga tccatatcgt tgcttggcag
     3121 tcagccagtc ggaatccagc ttgggaccca ggaagtccaa tcgtcagata ttgtactcaa
     3181 gcctggtcac ggcagcgtac cgatctgttt aaacctagat attgatagtc tgatcggtca
     3241 acgtataatc gagtcctagc ttttgcaaac atctatcaag agacaggatc agcaggaggc
     3301 tttcgcatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     3361 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     3421 ggtgcgcgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     3481 cgccccgaag aacgctttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     3541 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     3601 gacttggttg agtattcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     3661 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     3721 acgattggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     3781 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     3841 acgatgcctg tagcaatggc aacaaccttg cgtaaactat taactggcga actacttact
     3901 ctagcttccc ggcaacagtt gatagactgg atggaggcgg ataaagttgc aggaccactt
     3961 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     4021 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     4081 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     4141 ggtgcctcac tgattaagca ttggtaaccg attctaggtg cattggcgca gaaaaaaatg
     4201 cctgatgcga cgctgcgcgt cttatactcc cacatatgcc agattcagca acggatacgg
     4261 cttccccaac ttgcccactt ccatacgtgt cctccttacc agaaatttat ccttaagatc
     4321 ccgaatcgtt taaactcgac tctggctcta tcgaatctcc gtcgtttcga gcttacgcga
     4381 aca
//
