LOCUS       sequence-300713-        4461 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 4461)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4461)
  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-m4134238
FEATURES             Location/Qualifiers
     source          1..4461
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      285..873
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     774..793
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    1099..1170
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(1215..1351)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     terminator      complement(1381..1427)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    1542..1663
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1579..1598)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1633..1652
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(1716..1781)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             complement(1785..1805)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     promoter        complement(2063..2265)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2092..2112)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2267..2570
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    2651..2699
                     /note="synthetic polyadenylation signal"
     terminator      2717..2763
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    2768..2839
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(2991..3127)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(join(3140..3931,3932..4000))
                     /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     3763..3782
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gtttgaagat tttccgacga gggccttgga gagttcggaa atttaaatgc aactatggat
       61 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
      121 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
      181 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
      241 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
      301 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
      361 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
      421 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
      481 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
      541 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
      601 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
      661 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
      721 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
      781 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
      841 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
      901 ttcctggcct tttgctggcc ttttgctcac atgatttaaa tgaaactcgt aatcggagaa
      961 cgggtgcaat agcaaggacc agatcgaact tgactgacgc aattctgagc gtagtcgaaa
     1021 cccagacaag tcgatctcgc caagtagctg acttgactgg ttaattaagt ttcgatagcc
     1081 caaggtaacc aatgagatta gggagggaag aaagtttgat gaactacttc tgacccctag
     1141 tggtgtccag aaaagaccat taaaggaatg tcttgatcga aatcgagaga tcgcctgcag
     1201 gatagctgac gaccttaagg ataaatttct ggtaaggagg acacgtatgg aagtgggcaa
     1261 gttggggaag ccgtatccgt tgctgaatct ggcatatgtg ggagtataag acgcgcagcg
     1321 tcgcatcagg catttttttc tgcgccaatg ccggtagata gcaatccggc ccgaggggca
     1381 caaaaaaccc ctcaagaccc gtttagaggc cccaaggggt tatgctacga taccgataga
     1441 gatgggccaa taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc
     1501 ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt
     1561 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     1621 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtgagctga
     1681 aggtacgctg tatctcagtc agtcaagcta gcttatttgt catcgtcgtc cttgtaatca
     1741 atgtcatgat ctttgtagtc tccatcgtgg tccttataat ccatcgactg aaagtaaagg
     1801 ttttcgtcaa tctccattgg ctgttcctca tccagttggg aatatttatt ctcagttaaa
     1861 gacttactaa ggttttttat gatgagatta atgatgtagt ccaggttcca aatgctaacc
     1921 ttgaaggttc gcatgataat tagcagtatt tctgcgatcg tgacctgaaa atccaccaag
     1981 tggaacatgg tggcggccgc tggcgggtgt gtcgagtgga tggtaggatc gacaaagatc
     2041 tgatctgacg gttcactaaa ccagctctgc ttatatagac ctcccaccgt acacgcctac
     2101 cgcccatttg cgtcaatggg gcggagttgt tacgacattt tggaaagtcc cgttgatttt
     2161 ggtgccaaaa caaactccca ttgacgtcaa tggggtggag acttggaaat ccccgtgagt
     2221 caaaccgcta tccacgccca ttgatgtact gccaaaaccg catcagcatg gtaatagcga
     2281 tgactaatac gtagatgtac tgccaagtag gaaagtccca taaggtcatg tactgggcat
     2341 aatgccaggc gggccattta ccgtcattga cgtcaatagg gggcgtactt ggcatatgat
     2401 acacttgatg tactgccaag tgggcagttt accgtaaata ctccacccat tgacgtcaat
     2461 ggaaagtccc tattggcgtt actatgggaa catacgtcat tattgacgtc aatgggcggg
     2521 ggtcgttggg cggtcagcca ggcgggccat ttaccgtaag ttatgtaacg cggaactcca
     2581 tatatgggct atgaactaat gaccccgtaa ttgattacta gatctggcta cactcgatca
     2641 gcagttagat aataaaatcg ctatccatcg aagatggatg tgtgttggtt ttttgtgtgt
     2701 gtaacgcaac gattgatagc ataacccctt ggggcctcta aacgggtctt gaggggtttt
     2761 ttgaattgag aggtagctga agctgctgtt atgaccacta gagggaagaa gatacctgtg
     2821 gagctaatgg tccaagatgt cgggagccgc gatcgctgcc aaaaaggtaa gctgacgata
     2881 gctgacgaga ttcgatgccc gacgagcaaa tgagcgccac ggctgttcgc gtaagctcga
     2941 aacgacggag attcgataga gccagagtcg agtttaaacg attcgggatc ttaaggataa
     3001 atttctggta aggaggacac gtatggaagt gggcaagttg gggaagccgt atccgttgct
     3061 gaatctggca tatgtgggag tataagacgc gcagcgtcgc atcaggcatt tttttctgcg
     3121 ccaatgcacc tagaatcggt taccaatgct taatcagtga ggcacctatc tcagcgatct
     3181 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     3241 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     3301 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     3361 ctttatccgc ctccatccag tctatcaact gttgccggga agctagagta agtagttcgc
     3421 cagttaatag tttacgcaag gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     3481 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     3541 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc aatcgttgtc agaagtaagt
     3601 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     3661 catccgtaag atgcttttct gtgactggtg aatactcaac caagtcattc tgagaatagt
     3721 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     3781 gcagaacttt aaaagtgctc atcattggaa agcgttcttc ggggcgaaaa ctctcaagga
     3841 tcttaccgct gttgagatcc agttcgatgt aacccactcg cgcacccaac tgatcttcag
     3901 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     3961 aaaagggaat aagggcgaca cggaaatgtt gaatactcat gcgaaagcct cctgctgatc
     4021 ctgtctcttg atagatgttt gcaaaagcta ggactcgatt atacgttgac cgatcagact
     4081 atcaatatct aggtttaaac agatcggtac gctgccgtga ccaggcttga gtacaatatc
     4141 tgacgattgg acttcctggg tcccaagctg gattccgact ggctgactgc caagcaacga
     4201 tatggatcgt cgagactcgc actgagagga tcgttctcgg tacaccaggc gcgccccgat
     4261 ttcgatattc ctcggatctc ggatagatcg ggattgagga tgtaggttga acttctctcg
     4321 ggtgatcccg atctcggata ttcgggattg gagtaatacc tggcgatagg cgctgccaag
     4381 caatagccaa ggcgcaaggc cggccaagag acttgcgata gttcgttcga gaggtagcct
     4441 gcaagatgga tcggacgaaa g
//
