LOCUS       sequence-300705-        6377 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 6377)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6377)
  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-m4252837
FEATURES             Location/Qualifiers
     source          1..6377
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      51..97
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    102..173
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(325..461)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(join(474..1265,1266..1334))
                     /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     1097..1116
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    1600..1648
                     /note="synthetic polyadenylation signal"
     CDS             complement(1675..2274)
                     /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(1758..1778)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     2255..2274
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(2278..3487)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(2422..2441)
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     rep_origin      3987..4575
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4476..4495
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    4801..4872
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4917..5053)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     terminator      complement(5083..5129)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5244..5365
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5281..5300)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5335..5354
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(5418..5495)
                     /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"
     CDS             complement(5496..5516)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     promoter        complement(5774..5976)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(5803..5823)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        5978..6281
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    join(6362..6377,1..33)
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 atcgaagatg gatgtgtgtt ggttttttgt gtgtgtaacg caacgattga tagcataacc
       61 ccttggggcc tctaaacggg tcttgagggg ttttttgaat tgagaggtag ctgaagctgc
      121 tgttatgacc actagaggga agaagatacc tgtggagcta atggtccaag atgtcgggag
      181 ccgcgatcgc tgccaaaaag gtaagctgac gatagctgac gagattcgat gcccgacgag
      241 caaatgagcg ccacggctgt tcgcgtaagc tcgaaacgac ggagattcga tagagccaga
      301 gtcgagttta aacgattcgg gatcttaagg ataaatttct ggtaaggagg acacgtatgg
      361 aagtgggcaa gttggggaag ccgtatccgt tgctgaatct ggcatatgtg ggagtataag
      421 acgcgcagcg tcgcatcagg catttttttc tgcgccaatg cacctagaat cggttaccaa
      481 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
      541 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
      601 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
      661 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatc
      721 aactgttgcc gggaagctag agtaagtagt tcgccagtta atagtttacg caaggttgtt
      781 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
      841 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
      901 tccttcggtc ctccaatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
      961 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     1021 ggtgaatact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     1081 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     1141 ggaaagcgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     1201 atgtaaccca ctcgcgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     1261 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     1321 tgttgaatac tcatgcgaaa gcctcctgct gatcctgtct cttgatagat gtttgcaaaa
     1381 gctaggactc gattatacgt tgaccgatca gactatcaat atctaggttt aaacagatcg
     1441 gtacgctgcc gtgaccaggc ttgagtacaa tatctgacga ttggacttcc tgggtcccaa
     1501 gctggattcc gactggctga ctgccaagca acgatatgga tcgtcgagac tcgcactgag
     1561 aggatcgttc tcggtacacc aggcgcgcct agcgttacac acacaaaaaa ccaacacaca
     1621 tccatcttcg atggatagcg attttattat ctaagactag gcttacatac tagcctaggc
     1681 accgggcttg cgggtcatgc accaggtgcg cggtccttcg ggcacctcga cgtcggcggt
     1741 gacggtgaag ccgagccgct cgtagaaggg gaggttgcgg ggcgcggagg tctccaggaa
     1801 ggcgggcacc ccggcgcgct cggcggcctc cactccgggg agcacgacgg cgctgcccag
     1861 acccttgccc tggtggtcgg gcgagacgcc gacggtggcc aggaaccacg cgggctcctt
     1921 gggccggtgc ggcgccagga ggccctccat ctgttgctgc gcggccagcc gggaaccgct
     1981 caactcggcc atgcgcgggc cgatctcggc gaacaccgcc cccgcttcga cgctctccgg
     2041 cgtggtccag accgccaccg cggcgccgtc gtccgcgacc cacaccttgc cgatgtcgag
     2101 cccgacgcgc gtgaggaaga gttcttgcag ctcggtgacc cgctcgatgt ggcggtccgg
     2161 atcgacggtg tggcgcgtgg cggggtagtc ggcgaacgcg gcggcgaggg tgcgtacggc
     2221 cctggggacg tcgtcgcggg tggcgaggcg caccgtgggc ttgtactcgg tcatgttgtc
     2281 taacaaaaaa gccaaaaacg gccagaattt agcggacaat ttacaagtct aacactgaaa
     2341 attacatatt gacccaaatg attacatttc aaaaggtgcc taaaaaactt cacaaaacac
     2401 actcgccaac cccgagcgca tagttcaaaa ccggagcttc agctacttaa gaagataggt
     2461 acataaaacc gaccaaagaa actgacgcct cacttatccc tcccctcacc agaggtccgg
     2521 cgcctgtcga ttcaggagag cctaccctag gcccgaaccc tgcgtcctgc gacggagaaa
     2581 agcctaccgc acacctaccg gctggtggcc ccaccctgca ttataagcca acagaacggg
     2641 tgacgtcacg acacgacgag ggcgcgcgct cccaaaggta cgggtgcact gcccaacggc
     2701 accgccataa ctgccgcccc cgcaacagac gacaaaccga gttctccagt cagtgacaaa
     2761 cttcacgtca gggtccccag atggtgccca gcccatctca cccgaataag agctttcccg
     2821 cattagcgaa gggctcaaga ccttgggttc ttgccgccca ccatgccccc caccttgttt
     2881 caacgacctc acagcccgcc tcacaagcgt cttccattca agactcggga acagccgcca
     2941 ttttgctgcg ctccccccaa cccccagttc agggcaacct tgctcgcgga cccagactac
     3001 agcccttggc ggtctctcca cacgcttccg tcccaccgag cggcccggcg gccacgaaag
     3061 ccccggccag cccagcagcc cgctactcac caagtgacga tcacagcgat ccacaaacaa
     3121 gaaccgcgac ccaaatcccg gctgcgacgg aactagctgt gccacacccg gcgcgtcctt
     3181 atataatcat cggcgttcac cgccccacgg agatccctcc gcagaatcgc cgagaaggga
     3241 ctacttttcc tcgcctgttc cgctctctgg aaagaaaacc agtgccctag agtcacccaa
     3301 gtcccgtcct aaaatgtcct tctgctgata ctggggttct aaggccgagt cttatgagca
     3361 gcgggccgct gtcctgagcg tccgggcgga aggatcagga cgctcctgcg cccttcgtct
     3421 gacgtggcag cgctcgccgt gacgaggggg gcgcccgcgg gaggcgccaa aacccggcgc
     3481 ggaggccagg cgcgccccga tttcgatatt cctcggatct cggatagatc gggattgagg
     3541 atgtaggttg aacttctctc gggtgatccc gatctcggat attcgggatt ggagtaatac
     3601 ctggcgatag gcgctgccaa gcaatagcca aggcgcaagg ccggccaaga gacttgcgat
     3661 agttcgttcg agaggtagcc tgcaagatgg atcggacgaa aggtttgaag attttccgac
     3721 gagggccttg gagagttcgg aaatttaaat gcaactatgg atgaacgaaa tagacagatc
     3781 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     3841 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     3901 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     3961 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     4021 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     4081 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     4141 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     4201 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     4261 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     4321 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     4381 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     4441 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     4501 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     4561 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     4621 ccttttgctc acatgattta aatgaaactc gtaatcggag aacgggtgca atagcaagga
     4681 ccagatcgaa cttgactgac gcaattctga gcgtagtcga aacccagaca agtcgatctc
     4741 gccaagtagc tgacttgact ggttaattaa gtttcgatag cccaaggtaa ccaatgagat
     4801 tagggaggga agaaagtttg atgaactact tctgacccct agtggtgtcc agaaaagacc
     4861 attaaaggaa tgtcttgatc gaaatcgaga gatcgcctgc aggatagctg acgaccttaa
     4921 ggataaattt ctggtaagga ggacacgtat ggaagtgggc aagttgggga agccgtatcc
     4981 gttgctgaat ctggcatatg tgggagtata agacgcgcag cgtcgcatca ggcatttttt
     5041 tctgcgccaa tgccggtaga tagcaatccg gcccgagggg cacaaaaaac ccctcaagac
     5101 ccgtttagag gccccaaggg gttatgctac gataccgata gagatgggcc aataccacat
     5161 ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata
     5221 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa
     5281 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     5341 tgtccaaact catcaatgta tcttatcatg tctgtgagct gaaggtacgc tgtatctcag
     5401 tcagtcaagc tagcttagag ggcaccagac gaggatatct ttttgaatct gttggctgca
     5461 ctcaccgcga taaaattctt tttccatcgc ctcttcgact gaaagtaaag gttttcgtca
     5521 atctccattg gctgttcctc atccagttgg gaatatttat tctcagttaa agacttacta
     5581 aggtttttta tgatgagatt aatgatgtag tccaggttcc aaatgctaac cttgaaggtt
     5641 cgcatgataa ttagcagtat ttctgcgatc gtgacctgaa aatccaccaa gtggaacatg
     5701 gtggcggccg ctggcgggtg tgtcgagtgg atggtaggat cgacaaagat ctgatctgac
     5761 ggttcactaa accagctctg cttatataga cctcccaccg tacacgccta ccgcccattt
     5821 gcgtcaatgg ggcggagttg ttacgacatt ttggaaagtc ccgttgattt tggtgccaaa
     5881 acaaactccc attgacgtca atggggtgga gacttggaaa tccccgtgag tcaaaccgct
     5941 atccacgccc attgatgtac tgccaaaacc gcatcagcat ggtaatagcg atgactaata
     6001 cgtagatgta ctgccaagta ggaaagtccc ataaggtcat gtactgggca taatgccagg
     6061 cgggccattt accgtcattg acgtcaatag ggggcgtact tggcatatga tacacttgat
     6121 gtactgccaa gtgggcagtt taccgtaaat actccaccca ttgacgtcaa tggaaagtcc
     6181 ctattggcgt tactatggga acatacgtca ttattgacgt caatgggcgg gggtcgttgg
     6241 gcggtcagcc aggcgggcca tttaccgtaa gttatgtaac gcggaactcc atatatgggc
     6301 tatgaactaa tgaccccgta attgattact agatctggct acactcgatc agcagttaga
     6361 taataaaatc gctatcc
//
