LOCUS       sequence-299519-        4470 bp    DNA     circular SYN 15-DEC-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 4470)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4470)
  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-m4133693
FEATURES             Location/Qualifiers
     source          1..4470
                     /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..286
                     /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    339..460
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(350..369)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     404..423
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      575..621
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      651..787
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    832..903
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1129..1717)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1209..1228)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(2463..2531,2532..3323)
                     /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(2681..2700)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3336..3472
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3624..3695
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(3700..3746)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    3764..3812
                     /note="synthetic polyadenylation signal"
     enhancer        3893..4196
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4198..4400
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4351..4371
                     /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 ttcagtcgaa gaggcgatgg aaaaagaatt ttatcgcggt
      241 gagtgcagcc aacagattca aaaagatatc ctcgtctggt gccctctaag ctagcttgac
      301 tgactgagat acagcgtacc ttcagctcac agacatgata agatacattg atgagtttgg
      361 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
      421 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
      481 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta
      541 caaatgtggt attggcccat ctctatcggt atcgtagcat aaccccttgg ggcctctaaa
      601 cgggtcttga ggggtttttt gtgcccctcg ggccggattg ctatctaccg gcattggcgc
      661 agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc ccacatatgc cagattcagc
      721 aacggatacg gcttccccaa cttgcccact tccatacgtg tcctccttac cagaaattta
      781 tccttaaggt cgtcagctat cctgcaggcg atctctcgat ttcgatcaag acattccttt
      841 aatggtcttt tctggacacc actaggggtc agaagtagtt catcaaactt tcttccctcc
      901 ctaatctcat tggttacctt gggctatcga aacttaatta accagtcaag tcagctactt
      961 ggcgagatcg acttgtctgg gtttcgacta cgctcagaat tgcgtcagtc aagttcgatc
     1021 tggtccttgc tattgcaccc gttctccgat tacgagtttc atttaaatca tgtgagcaaa
     1081 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     1141 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     1201 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     1261 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     1321 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     1381 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     1441 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     1501 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     1561 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     1621 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     1681 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     1741 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     1801 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     1861 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     1921 agcgatctgt ctatttcgtt catccatagt tgcatttaaa tttccgaact ctccaaggcc
     1981 ctcgtcggaa aatcttcaaa cctttcgtcc gatccatctt gcaggctacc tctcgaacga
     2041 actatcgcaa gtctcttggc cggccttgcg ccttggctat tgcttggcag cgcctatcgc
     2101 caggtattac tccaatcccg aatatccgag atcgggatca cccgagagaa gttcaaccta
     2161 catcctcaat cccgatctat ccgagatccg aggaatatcg aaatcggggc gcgcctggtg
     2221 taccgagaac gatcctctca gtgcgagtct cgacgatcca tatcgttgct tggcagtcag
     2281 ccagtcggaa tccagcttgg gacccaggaa gtccaatcgt cagatattgt actcaagcct
     2341 ggtcacggca gcgtaccgat ctgtttaaac ctagatattg atagtctgat cggtcaacgt
     2401 ataatcgagt cctagctttt gcaaacatct atcaagagac aggatcagca ggaggctttc
     2461 gcatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     2521 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     2581 cgcgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     2641 ccgaagaacg ctttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     2701 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     2761 tggttgagta ttcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     2821 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     2881 ttggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     2941 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     3001 tgcctgtagc aatggcaaca accttgcgta aactattaac tggcgaacta cttactctag
     3061 cttcccggca acagttgata gactggatgg aggcggataa agttgcagga ccacttctgc
     3121 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     3181 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     3241 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     3301 cctcactgat taagcattgg taaccgattc taggtgcatt ggcgcagaaa aaaatgcctg
     3361 atgcgacgct gcgcgtctta tactcccaca tatgccagat tcagcaacgg atacggcttc
     3421 cccaacttgc ccacttccat acgtgtcctc cttaccagaa atttatcctt aagatcccga
     3481 atcgtttaaa ctcgactctg gctctatcga atctccgtcg tttcgagctt acgcgaacag
     3541 ccgtggcgct catttgctcg tcgggcatcg aatctcgtca gctatcgtca gcttaccttt
     3601 ttggcagcga tcgcggctcc cgacatcttg gaccattagc tccacaggta tcttcttccc
     3661 tctagtggtc ataacagcag cttcagctac ctctcaattc aaaaaacccc tcaagacccg
     3721 tttagaggcc ccaaggggtt atgctatcaa tcgttgcgtt acacacacaa aaaaccaaca
     3781 cacatccatc ttcgatggat agcgatttta ttatctaact gctgatcgag tgtagccaga
     3841 tctagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
     3901 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
     3961 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
     4021 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
     4081 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
     4141 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatgctga
     4201 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
     4261 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
     4321 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
     4381 aggtctatat aagcagagct ggtttagtga accgtcagat cagatctttg tcgatcctac
     4441 catccactcg acacacccgc cagcggccgc
//
