LOCUS       sequence-297397-        4410 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 ORF6 protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4410)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4410)
  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-m4133215
FEATURES             Location/Qualifiers
     source          1..4410
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             188..208
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             209..226
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    279..400
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(290..309)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     344..363
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      515..561
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      591..727
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    772..843
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1069..1657)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1149..1168)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(2403..2471,2472..3263)
                     /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(2621..2640)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3276..3412
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3564..3635
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(3640..3686)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    3704..3752
                     /note="synthetic polyadenylation signal"
     enhancer        3833..4136
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4138..4340
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4291..4311
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgttt catctcgttg actttcaggt tactatagca gagatattac taattattat
       61 gaggactttt aaagtttcca tttggaatct tgattacatc ataaacctca taattaaaaa
      121 tttatctaag tcactaactg agaataaata ttctcaatta gatgaagagc aaccaatgga
      181 gattgatgaa aacctttact ttcagtcgca ccatcaccat caccattaag ctagcttgac
      241 tgactgagat acagcgtacc ttcagctcac agacatgata agatacattg atgagtttgg
      301 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
      361 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
      421 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta
      481 caaatgtggt attggcccat ctctatcggt atcgtagcat aaccccttgg ggcctctaaa
      541 cgggtcttga ggggtttttt gtgcccctcg ggccggattg ctatctaccg gcattggcgc
      601 agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc ccacatatgc cagattcagc
      661 aacggatacg gcttccccaa cttgcccact tccatacgtg tcctccttac cagaaattta
      721 tccttaaggt cgtcagctat cctgcaggcg atctctcgat ttcgatcaag acattccttt
      781 aatggtcttt tctggacacc actaggggtc agaagtagtt catcaaactt tcttccctcc
      841 ctaatctcat tggttacctt gggctatcga aacttaatta accagtcaag tcagctactt
      901 ggcgagatcg acttgtctgg gtttcgacta cgctcagaat tgcgtcagtc aagttcgatc
      961 tggtccttgc tattgcaccc gttctccgat tacgagtttc atttaaatca tgtgagcaaa
     1021 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     1081 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     1141 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     1201 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     1261 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     1321 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     1381 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     1441 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     1501 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     1561 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     1621 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     1681 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     1741 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     1801 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     1861 agcgatctgt ctatttcgtt catccatagt tgcatttaaa tttccgaact ctccaaggcc
     1921 ctcgtcggaa aatcttcaaa cctttcgtcc gatccatctt gcaggctacc tctcgaacga
     1981 actatcgcaa gtctcttggc cggccttgcg ccttggctat tgcttggcag cgcctatcgc
     2041 caggtattac tccaatcccg aatatccgag atcgggatca cccgagagaa gttcaaccta
     2101 catcctcaat cccgatctat ccgagatccg aggaatatcg aaatcggggc gcgcctggtg
     2161 taccgagaac gatcctctca gtgcgagtct cgacgatcca tatcgttgct tggcagtcag
     2221 ccagtcggaa tccagcttgg gacccaggaa gtccaatcgt cagatattgt actcaagcct
     2281 ggtcacggca gcgtaccgat ctgtttaaac ctagatattg atagtctgat cggtcaacgt
     2341 ataatcgagt cctagctttt gcaaacatct atcaagagac aggatcagca ggaggctttc
     2401 gcatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     2461 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     2521 cgcgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     2581 ccgaagaacg ctttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     2641 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     2701 tggttgagta ttcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     2761 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     2821 ttggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     2881 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     2941 tgcctgtagc aatggcaaca accttgcgta aactattaac tggcgaacta cttactctag
     3001 cttcccggca acagttgata gactggatgg aggcggataa agttgcagga ccacttctgc
     3061 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     3121 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     3181 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     3241 cctcactgat taagcattgg taaccgattc taggtgcatt ggcgcagaaa aaaatgcctg
     3301 atgcgacgct gcgcgtctta tactcccaca tatgccagat tcagcaacgg atacggcttc
     3361 cccaacttgc ccacttccat acgtgtcctc cttaccagaa atttatcctt aagatcccga
     3421 atcgtttaaa ctcgactctg gctctatcga atctccgtcg tttcgagctt acgcgaacag
     3481 ccgtggcgct catttgctcg tcgggcatcg aatctcgtca gctatcgtca gcttaccttt
     3541 ttggcagcga tcgcggctcc cgacatcttg gaccattagc tccacaggta tcttcttccc
     3601 tctagtggtc ataacagcag cttcagctac ctctcaattc aaaaaacccc tcaagacccg
     3661 tttagaggcc ccaaggggtt atgctatcaa tcgttgcgtt acacacacaa aaaaccaaca
     3721 cacatccatc ttcgatggat agcgatttta ttatctaact gctgatcgag tgtagccaga
     3781 tctagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
     3841 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
     3901 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
     3961 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
     4021 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
     4081 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatgctga
     4141 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
     4201 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
     4261 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
     4321 aggtctatat aagcagagct ggtttagtga accgtcagat cagatctttg tcgatcctac
     4381 catccactcg acacacccgc cagcggccgc
//
