LOCUS       sequence-301668-        6323 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 6323)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6323)
  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-m4252780
FEATURES             Location/Qualifiers
     source          1..6323
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    74..145
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(150..196)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    214..262
                     /note="synthetic polyadenylation signal"
     enhancer        343..646
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        648..850
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     801..821
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1054..1074
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1075..1152
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     polyA_signal    1205..1326
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1216..1235)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1270..1289
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1441..1487
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1517..1653
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1698..1769
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1995..2583)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2075..2094)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3083..4292
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4129..4148
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4296..4895
                     /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(4296..4315)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4792..4812
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4922..4970
                     /note="synthetic polyadenylation signal"
     CDS             join(5236..5304,5305..6096)
                     /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(5454..5473)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6109..6245
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
ORIGIN
        1 catttgctcg tcgggcatcg aatctcgtca gctatcgtca gcttaccttt ttggcagcga
       61 tcgcggctcc cgacatcttg gaccattagc tccacaggta tcttcttccc tctagtggtc
      121 ataacagcag cttcagctac ctctcaattc aaaaaacccc tcaagacccg tttagaggcc
      181 ccaaggggtt atgctatcaa tcgttgcgtt acacacacaa aaaaccaaca cacatccatc
      241 ttcgatggat agcgatttta ttatctaact gctgatcgag tgtagccaga tctagtaatc
      301 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
      361 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
      421 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
      481 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
      541 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
      601 tcctacttgg cagtacatct acgtattagt catcgctatt accatgctga tgcggttttg
      661 gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
      721 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
      781 taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
      841 aagcagagct ggtttagtga accgtcagat cagatctttg tcgatcctac catccactcg
      901 acacacccgc cagcggccgc caccatgatt gaactttcat taattgactt ctatttgtgc
      961 tttttagcct ttctgctatt ccttgtttta attatgctta ttatcttttg gttctcactt
     1021 gaactgcaag atcataatga aacttgtcac gccgaaaacc tttactttca gtcgaagagg
     1081 cgatggaaaa agaattttat cgcggtgagt gcagccaaca gattcaaaaa gatatcctcg
     1141 tctggtgccc tctaagctag cttgactgac tgagatacag cgtaccttca gctcacagac
     1201 atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc
     1261 tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa
     1321 caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag
     1381 gttttttaaa gcaagtaaaa cctctacaaa tgtggtattg gcccatctct atcggtatcg
     1441 tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgtgc ccctcgggcc
     1501 ggattgctat ctaccggcat tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt
     1561 atactcccac atatgccaga ttcagcaacg gatacggctt ccccaacttg cccacttcca
     1621 tacgtgtcct ccttaccaga aatttatcct taaggtcgtc agctatcctg caggcgatct
     1681 ctcgatttcg atcaagacat tcctttaatg gtcttttctg gacaccacta ggggtcagaa
     1741 gtagttcatc aaactttctt ccctccctaa tctcattggt taccttgggc tatcgaaact
     1801 taattaacca gtcaagtcag ctacttggcg agatcgactt gtctgggttt cgactacgct
     1861 cagaattgcg tcagtcaagt tcgatctggt ccttgctatt gcacccgttc tccgattacg
     1921 agtttcattt aaatcatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     1981 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     2041 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     2101 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     2161 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     2221 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     2281 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     2341 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     2401 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     2461 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     2521 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     2581 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     2641 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     2701 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     2761 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgca
     2821 tttaaatttc cgaactctcc aaggccctcg tcggaaaatc ttcaaacctt tcgtccgatc
     2881 catcttgcag gctacctctc gaacgaacta tcgcaagtct cttggccggc cttgcgcctt
     2941 ggctattgct tggcagcgcc tatcgccagg tattactcca atcccgaata tccgagatcg
     3001 ggatcacccg agagaagttc aacctacatc ctcaatcccg atctatccga gatccgagga
     3061 atatcgaaat cggggcgcgc ctggcctccg cgccgggttt tggcgcctcc cgcgggcgcc
     3121 cccctcgtca cggcgagcgc tgccacgtca gacgaagggc gcaggagcgt cctgatcctt
     3181 ccgcccggac gctcaggaca gcggcccgct gctcataaga ctcggcctta gaaccccagt
     3241 atcagcagaa ggacatttta ggacgggact tgggtgactc tagggcactg gttttctttc
     3301 cagagagcgg aacaggcgag gaaaagtagt cccttctcgg cgattctgcg gagggatctc
     3361 cgtggggcgg tgaacgccga tgattatata aggacgcgcc gggtgtggca cagctagttc
     3421 cgtcgcagcc gggatttggg tcgcggttct tgtttgtgga tcgctgtgat cgtcacttgg
     3481 tgagtagcgg gctgctgggc tggccggggc tttcgtggcc gccgggccgc tcggtgggac
     3541 ggaagcgtgt ggagagaccg ccaagggctg tagtctgggt ccgcgagcaa ggttgccctg
     3601 aactgggggt tggggggagc gcagcaaaat ggcggctgtt cccgagtctt gaatggaaga
     3661 cgcttgtgag gcgggctgtg aggtcgttga aacaaggtgg ggggcatggt gggcggcaag
     3721 aacccaaggt cttgagccct tcgctaatgc gggaaagctc ttattcgggt gagatgggct
     3781 gggcaccatc tggggaccct gacgtgaagt ttgtcactga ctggagaact cggtttgtcg
     3841 tctgttgcgg gggcggcagt tatggcggtg ccgttgggca gtgcacccgt acctttggga
     3901 gcgcgcgccc tcgtcgtgtc gtgacgtcac ccgttctgtt ggcttataat gcagggtggg
     3961 gccaccagcc ggtaggtgtg cggtaggctt ttctccgtcg caggacgcag ggttcgggcc
     4021 tagggtaggc tctcctgaat cgacaggcgc cggacctctg gtgaggggag ggataagtga
     4081 ggcgtcagtt tctttggtcg gttttatgta cctatcttct taagtagctg aagctccggt
     4141 tttgaactat gcgctcgggg ttggcgagtg tgttttgtga agttttttag gcaccttttg
     4201 aaatgtaatc atttgggtca atatgtaatt ttcagtgtta gacttgtaaa ttgtccgcta
     4261 aattctggcc gtttttggct tttttgttag acaacatgac cgagtacaag cccacggtgc
     4321 gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg
     4381 actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg gtcaccgagc
     4441 tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg
     4501 acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg
     4561 ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga
     4621 tggagggcct cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg
     4681 gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg
     4741 aggccgccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc
     4801 ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc
     4861 gcacctggtg catgacccgc aagcccggtg cctaggctag tatgtaagcc tagtcttaga
     4921 taataaaatc gctatccatc gaagatggat gtgtgttggt tttttgtgtg tgtaacgcta
     4981 ggcgcgcctg gtgtaccgag aacgatcctc tcagtgcgag tctcgacgat ccatatcgtt
     5041 gcttggcagt cagccagtcg gaatccagct tgggacccag gaagtccaat cgtcagatat
     5101 tgtactcaag cctggtcacg gcagcgtacc gatctgttta aacctagata ttgatagtct
     5161 gatcggtcaa cgtataatcg agtcctagct tttgcaaaca tctatcaaga gacaggatca
     5221 gcaggaggct ttcgcatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     5281 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     5341 gatcagttgg gtgcgcgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     5401 gagagttttc gccccgaaga acgctttcca atgatgagca cttttaaagt tctgctatgt
     5461 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     5521 tctcagaatg acttggttga gtattcacca gtcacagaaa agcatcttac ggatggcatg
     5581 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     5641 cttctgacaa cgattggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     5701 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     5761 cgtgacacca cgatgcctgt agcaatggca acaaccttgc gtaaactatt aactggcgaa
     5821 ctacttactc tagcttcccg gcaacagttg atagactgga tggaggcgga taaagttgca
     5881 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     5941 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     6001 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     6061 gctgagatag gtgcctcact gattaagcat tggtaaccga ttctaggtgc attggcgcag
     6121 aaaaaaatgc ctgatgcgac gctgcgcgtc ttatactccc acatatgcca gattcagcaa
     6181 cggatacggc ttccccaact tgcccacttc catacgtgtc ctccttacca gaaatttatc
     6241 cttaagatcc cgaatcgttt aaactcgact ctggctctat cgaatctccg tcgtttcgag
     6301 cttacgcgaa cagccgtggc gct
//
