LOCUS       sequence-307366-        6762 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 6762)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6762)
  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-m4252950
FEATURES             Location/Qualifiers
     source          1..6762
                     /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    295..416
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(306..325)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     360..379
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      531..577
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      607..743
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    788..859
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1085..1673)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1165..1184)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2173..3382
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3219..3238
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3386..4414
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    4441..4489
                     /note="synthetic polyadenylation signal"
     CDS             join(4755..4823,4824..5615)
                     /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(4973..4992)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5628..5764
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5916..5987
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5992..6038)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6056..6104
                     /note="synthetic polyadenylation signal"
     enhancer        6185..6488
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6490..6692
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6643..6663
                     /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 gatccgcagg
      241 cctctgctag cttgactgac tgagatacag cgtaccttca gctcacagac atgataagat
      301 acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg
      361 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca
      421 acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa
      481 gcaagtaaaa cctctacaaa tgtggtattg gcccatctct atcggtatcg tagcataacc
      541 ccttggggcc tctaaacggg tcttgagggg ttttttgtgc ccctcgggcc ggattgctat
      601 ctaccggcat tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt atactcccac
      661 atatgccaga ttcagcaacg gatacggctt ccccaacttg cccacttcca tacgtgtcct
      721 ccttaccaga aatttatcct taaggtcgtc agctatcctg caggcgatct ctcgatttcg
      781 atcaagacat tcctttaatg gtcttttctg gacaccacta ggggtcagaa gtagttcatc
      841 aaactttctt ccctccctaa tctcattggt taccttgggc tatcgaaact taattaacca
      901 gtcaagtcag ctacttggcg agatcgactt gtctgggttt cgactacgct cagaattgcg
      961 tcagtcaagt tcgatctggt ccttgctatt gcacccgttc tccgattacg agtttcattt
     1021 aaatcatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     1081 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     1141 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     1201 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     1261 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     1321 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     1381 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     1441 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     1501 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     1561 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     1621 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     1681 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     1741 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     1801 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     1861 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgca tttaaatttc
     1921 cgaactctcc aaggccctcg tcggaaaatc ttcaaacctt tcgtccgatc catcttgcag
     1981 gctacctctc gaacgaacta tcgcaagtct cttggccggc cttgcgcctt ggctattgct
     2041 tggcagcgcc tatcgccagg tattactcca atcccgaata tccgagatcg ggatcacccg
     2101 agagaagttc aacctacatc ctcaatcccg atctatccga gatccgagga atatcgaaat
     2161 cggggcgcgc ctggcctccg cgccgggttt tggcgcctcc cgcgggcgcc cccctcgtca
     2221 cggcgagcgc tgccacgtca gacgaagggc gcaggagcgt cctgatcctt ccgcccggac
     2281 gctcaggaca gcggcccgct gctcataaga ctcggcctta gaaccccagt atcagcagaa
     2341 ggacatttta ggacgggact tgggtgactc tagggcactg gttttctttc cagagagcgg
     2401 aacaggcgag gaaaagtagt cccttctcgg cgattctgcg gagggatctc cgtggggcgg
     2461 tgaacgccga tgattatata aggacgcgcc gggtgtggca cagctagttc cgtcgcagcc
     2521 gggatttggg tcgcggttct tgtttgtgga tcgctgtgat cgtcacttgg tgagtagcgg
     2581 gctgctgggc tggccggggc tttcgtggcc gccgggccgc tcggtgggac ggaagcgtgt
     2641 ggagagaccg ccaagggctg tagtctgggt ccgcgagcaa ggttgccctg aactgggggt
     2701 tggggggagc gcagcaaaat ggcggctgtt cccgagtctt gaatggaaga cgcttgtgag
     2761 gcgggctgtg aggtcgttga aacaaggtgg ggggcatggt gggcggcaag aacccaaggt
     2821 cttgagccct tcgctaatgc gggaaagctc ttattcgggt gagatgggct gggcaccatc
     2881 tggggaccct gacgtgaagt ttgtcactga ctggagaact cggtttgtcg tctgttgcgg
     2941 gggcggcagt tatggcggtg ccgttgggca gtgcacccgt acctttggga gcgcgcgccc
     3001 tcgtcgtgtc gtgacgtcac ccgttctgtt ggcttataat gcagggtggg gccaccagcc
     3061 ggtaggtgtg cggtaggctt ttctccgtcg caggacgcag ggttcgggcc tagggtaggc
     3121 tctcctgaat cgacaggcgc cggacctctg gtgaggggag ggataagtga ggcgtcagtt
     3181 tctttggtcg gttttatgta cctatcttct taagtagctg aagctccggt tttgaactat
     3241 gcgctcgggg ttggcgagtg tgttttgtga agttttttag gcaccttttg aaatgtaatc
     3301 atttgggtca atatgtaatt ttcagtgtta gacttgtaaa ttgtccgcta aattctggcc
     3361 gtttttggct tttttgttag acaacatggg taaaaagcct gaactcaccg cgacgtctgt
     3421 cgagaagttt ctgatcgaaa agttcgacag cgtctccgac ctgatgcagc tctcggaggg
     3481 cgaagaatct cgtgctttca gcttcgatgt aggagggcgt ggatatgtcc tgcgggtaaa
     3541 tagctgcgcc gatggtttct acaaagatcg ttatgtttat cggcactttg catccgccgc
     3601 gctcccgatt ccggaagtgc ttgacattgg ggagttcagc gagagcctga cctattgcat
     3661 ctcccgccgt gcacagggtg tcacgttgca agacctgcct gaaaccgaac tgcccgctgt
     3721 tctgcagccg gtcgcggagg caatggatgc catcgctgcc gccgatctta gccagacgag
     3781 cgggttcggc ccattcggac cgcaaggaat cggtcaatac actacatggc gtgatttcat
     3841 atgcgcgatt gctgatcccc atgtgtatca ctggcaaact gtgatggacg acaccgtcag
     3901 tgcgtccgtc gcgcaggctc tcgatgagct gatgctttgg gccgaggact gccccgaagt
     3961 ccggcacctc gtgcacgcgg atttcggctc caacaatgtc ctgacggaca atggccgcat
     4021 aacagcggtc attgactgga gcgaggcgat gttcggggat tcccaatacg aggtcgccaa
     4081 catcttcttc tggaggccgt ggttggcttg tatggagcag cagacgcgct acttcgagcg
     4141 gaggcatccg gagcttgcag gatcgccccg gctccgggcg tatatgctcc gcattggtct
     4201 tgaccaactc tatcagagct tggttgacgg caatttcgat gatgcagctt gggcgcaggg
     4261 tcgatgcgac gcaatcgtcc gatccggagc cgggactgtc gggcgtacac aaatcgcccg
     4321 cagaagcgcc gccgtctgga ccgatggctg tgtagaagta ctcgccgata gtggaaaccg
     4381 acgccccagc actcgtccga gggcaaagga ataagctagt atgtaagcct agtcttagat
     4441 aataaaatcg ctatccatcg aagatggatg tgtgttggtt ttttgtgtgt gtaacgctag
     4501 gcgcgcctgg tgtaccgaga acgatcctct cagtgcgagt ctcgacgatc catatcgttg
     4561 cttggcagtc agccagtcgg aatccagctt gggacccagg aagtccaatc gtcagatatt
     4621 gtactcaagc ctggtcacgg cagcgtaccg atctgtttaa acctagatat tgatagtctg
     4681 atcggtcaac gtataatcga gtcctagctt ttgcaaacat ctatcaagag acaggatcag
     4741 caggaggctt tcgcatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     4801 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     4861 atcagttggg tgcgcgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     4921 agagttttcg ccccgaagaa cgctttccaa tgatgagcac ttttaaagtt ctgctatgtg
     4981 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     5041 ctcagaatga cttggttgag tattcaccag tcacagaaaa gcatcttacg gatggcatga
     5101 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     5161 ttctgacaac gattggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     5221 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     5281 gtgacaccac gatgcctgta gcaatggcaa caaccttgcg taaactatta actggcgaac
     5341 tacttactct agcttcccgg caacagttga tagactggat ggaggcggat aaagttgcag
     5401 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     5461 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     5521 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     5581 ctgagatagg tgcctcactg attaagcatt ggtaaccgat tctaggtgca ttggcgcaga
     5641 aaaaaatgcc tgatgcgacg ctgcgcgtct tatactccca catatgccag attcagcaac
     5701 ggatacggct tccccaactt gcccacttcc atacgtgtcc tccttaccag aaatttatcc
     5761 ttaagatccc gaatcgttta aactcgactc tggctctatc gaatctccgt cgtttcgagc
     5821 ttacgcgaac agccgtggcg ctcatttgct cgtcgggcat cgaatctcgt cagctatcgt
     5881 cagcttacct ttttggcagc gatcgcggct cccgacatct tggaccatta gctccacagg
     5941 tatcttcttc cctctagtgg tcataacagc agcttcagct acctctcaat tcaaaaaacc
     6001 cctcaagacc cgtttagagg ccccaagggg ttatgctatc aatcgttgcg ttacacacac
     6061 aaaaaaccaa cacacatcca tcttcgatgg atagcgattt tattatctaa ctgctgatcg
     6121 agtgtagcca gatctagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
     6181 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
     6241 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
     6301 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
     6361 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
     6421 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
     6481 ttaccatgct gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
     6541 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
     6601 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
     6661 gtgtacggtg ggaggtctat ataagcagag ctggtttagt gaaccgtcag atcagatctt
     6721 tgtcgatcct accatccact cgacacaccc gccagcggcc gc
//
