LOCUS       sequence-313567-        6263 bp    DNA     circular SYN 15-APR-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 6263)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6263)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pGBW-m4252860
FEATURES             Location/Qualifiers
     source          1..6263
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        306..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             712..732
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             733..750
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    803..924
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(814..833)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     868..887
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1039..1085
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1115..1251
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1296..1367
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1593..2181)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1673..1692)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2681..3890
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3727..3746
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3894..4493
                     /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(3894..3913)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4390..4410
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4520..4568
                     /note="synthetic polyadenylation signal"
     CDS             join(4834..4902,4903..5694)
                     /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(5052..5071)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5707..5843
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5995..6066
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6071..6117)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6135..6183
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catgctgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatca gatctttgtc
      541 gatcctacca tccactcgac acacccgcca gcggccgcca ccatgattga actttcatta
      601 attgacttct atttgtgctt tttagccttt ctgctattcc ttgttttaat tatgcttatt
      661 atcttttggt tctcacttga actgcaagat cataatgaaa cttgtcacgc cgaaaacctt
      721 tactttcagt cgcaccatca ccatcaccat taagctagct tgactgactg agatacagcg
      781 taccttcagc tcacagacat gataagatac attgatgagt ttggacaaac cacaactaga
      841 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
      901 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt
      961 cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtattggc
     1021 ccatctctat cggtatcgta gcataacccc ttggggcctc taaacgggtc ttgaggggtt
     1081 ttttgtgccc ctcgggccgg attgctatct accggcattg gcgcagaaaa aaatgcctga
     1141 tgcgacgctg cgcgtcttat actcccacat atgccagatt cagcaacgga tacggcttcc
     1201 ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa tttatcctta aggtcgtcag
     1261 ctatcctgca ggcgatctct cgatttcgat caagacattc ctttaatggt cttttctgga
     1321 caccactagg ggtcagaagt agttcatcaa actttcttcc ctccctaatc tcattggtta
     1381 ccttgggcta tcgaaactta attaaccagt caagtcagct acttggcgag atcgacttgt
     1441 ctgggtttcg actacgctca gaattgcgtc agtcaagttc gatctggtcc ttgctattgc
     1501 acccgttctc cgattacgag tttcatttaa atcatgtgag caaaaggcca gcaaaaggcc
     1561 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     1621 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     1681 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     1741 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     1801 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     1861 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     1921 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     1981 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     2041 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     2101 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     2161 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     2221 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     2281 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     2341 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     2401 cgttcatcca tagttgcatt taaatttccg aactctccaa ggccctcgtc ggaaaatctt
     2461 caaacctttc gtccgatcca tcttgcaggc tacctctcga acgaactatc gcaagtctct
     2521 tggccggcct tgcgccttgg ctattgcttg gcagcgccta tcgccaggta ttactccaat
     2581 cccgaatatc cgagatcggg atcacccgag agaagttcaa cctacatcct caatcccgat
     2641 ctatccgaga tccgaggaat atcgaaatcg gggcgcgcct ggcctccgcg ccgggttttg
     2701 gcgcctcccg cgggcgcccc cctcgtcacg gcgagcgctg ccacgtcaga cgaagggcgc
     2761 aggagcgtcc tgatccttcc gcccggacgc tcaggacagc ggcccgctgc tcataagact
     2821 cggccttaga accccagtat cagcagaagg acattttagg acgggacttg ggtgactcta
     2881 gggcactggt tttctttcca gagagcggaa caggcgagga aaagtagtcc cttctcggcg
     2941 attctgcgga gggatctccg tggggcggtg aacgccgatg attatataag gacgcgccgg
     3001 gtgtggcaca gctagttccg tcgcagccgg gatttgggtc gcggttcttg tttgtggatc
     3061 gctgtgatcg tcacttggtg agtagcgggc tgctgggctg gccggggctt tcgtggccgc
     3121 cgggccgctc ggtgggacgg aagcgtgtgg agagaccgcc aagggctgta gtctgggtcc
     3181 gcgagcaagg ttgccctgaa ctgggggttg gggggagcgc agcaaaatgg cggctgttcc
     3241 cgagtcttga atggaagacg cttgtgaggc gggctgtgag gtcgttgaaa caaggtgggg
     3301 ggcatggtgg gcggcaagaa cccaaggtct tgagcccttc gctaatgcgg gaaagctctt
     3361 attcgggtga gatgggctgg gcaccatctg gggaccctga cgtgaagttt gtcactgact
     3421 ggagaactcg gtttgtcgtc tgttgcgggg gcggcagtta tggcggtgcc gttgggcagt
     3481 gcacccgtac ctttgggagc gcgcgccctc gtcgtgtcgt gacgtcaccc gttctgttgg
     3541 cttataatgc agggtggggc caccagccgg taggtgtgcg gtaggctttt ctccgtcgca
     3601 ggacgcaggg ttcgggccta gggtaggctc tcctgaatcg acaggcgccg gacctctggt
     3661 gaggggaggg ataagtgagg cgtcagtttc tttggtcggt tttatgtacc tatcttctta
     3721 agtagctgaa gctccggttt tgaactatgc gctcggggtt ggcgagtgtg ttttgtgaag
     3781 ttttttaggc accttttgaa atgtaatcat ttgggtcaat atgtaatttt cagtgttaga
     3841 cttgtaaatt gtccgctaaa ttctggccgt ttttggcttt tttgttagac aacatgaccg
     3901 agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc
     3961 tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg gaccgccaca
     4021 tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca
     4081 aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg
     4141 aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc
     4201 tggccgcgca gcaacagatg gagggcctcc tggcgccgca ccggcccaag gagcccgcgt
     4261 ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg
     4321 tcgtgctccc cggagtggag gccgccgagc gcgccggggt gcccgccttc ctggagacct
     4381 ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg
     4441 aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc taggctagta
     4501 tgtaagccta gtcttagata ataaaatcgc tatccatcga agatggatgt gtgttggttt
     4561 tttgtgtgtg taacgctagg cgcgcctggt gtaccgagaa cgatcctctc agtgcgagtc
     4621 tcgacgatcc atatcgttgc ttggcagtca gccagtcgga atccagcttg ggacccagga
     4681 agtccaatcg tcagatattg tactcaagcc tggtcacggc agcgtaccga tctgtttaaa
     4741 cctagatatt gatagtctga tcggtcaacg tataatcgag tcctagcttt tgcaaacatc
     4801 tatcaagaga caggatcagc aggaggcttt cgcatgagta ttcaacattt ccgtgtcgcc
     4861 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     4921 aaagtaaaag atgctgaaga tcagttgggt gcgcgagtgg gttacatcga actggatctc
     4981 aacagcggta agatccttga gagttttcgc cccgaagaac gctttccaat gatgagcact
     5041 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     5101 ggtcgccgca tacactattc tcagaatgac ttggttgagt attcaccagt cacagaaaag
     5161 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     5221 aacactgcgg ccaacttact tctgacaacg attggaggac cgaaggagct aaccgctttt
     5281 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     5341 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aaccttgcgt
     5401 aaactattaa ctggcgaact acttactcta gcttcccggc aacagttgat agactggatg
     5461 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     5521 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     5581 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     5641 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaaccgatt
     5701 ctaggtgcat tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt atactcccac
     5761 atatgccaga ttcagcaacg gatacggctt ccccaacttg cccacttcca tacgtgtcct
     5821 ccttaccaga aatttatcct taagatcccg aatcgtttaa actcgactct ggctctatcg
     5881 aatctccgtc gtttcgagct tacgcgaaca gccgtggcgc tcatttgctc gtcgggcatc
     5941 gaatctcgtc agctatcgtc agcttacctt tttggcagcg atcgcggctc ccgacatctt
     6001 ggaccattag ctccacaggt atcttcttcc ctctagtggt cataacagca gcttcagcta
     6061 cctctcaatt caaaaaaccc ctcaagaccc gtttagaggc cccaaggggt tatgctatca
     6121 atcgttgcgt tacacacaca aaaaaccaac acacatccat cttcgatgga tagcgatttt
     6181 attatctaac tgctgatcga gtgtagccag atctagtaat caattacggg gtcattagtt
     6241 catagcccat atatggagtt ccg
//
