LOCUS       sequence-300764-        6708 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 ORF7b.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6708)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6708)
  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-m4252935
FEATURES             Location/Qualifiers
     source          1..6708
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             9..29
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             30..47
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    116..237
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(127..146)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     181..200
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      352..398
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      428..564
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    609..680
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(906..1494)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(986..1005)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        1994..3203
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3040..3059
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3207..4235
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    4262..4310
                     /note="synthetic polyadenylation signal"
     CDS             join(4576..4644,4645..5436)
                     /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(4794..4813)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5449..5585
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5737..5808
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5813..5859)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5877..5925
                     /note="synthetic polyadenylation signal"
     enhancer        6006..6309
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6311..6513
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6464..6484
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 gtcacgccga aaacctttac tttcagtcgc accatcacca tcaccattaa ggatccgcag
       61 gcctctgcta gcttgactga ctgagataca gcgtaccttc agctcacaga catgataaga
      121 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
      181 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
      241 aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa
      301 agcaagtaaa acctctacaa atgtggtatt ggcccatctc tatcggtatc gtagcataac
      361 cccttggggc ctctaaacgg gtcttgaggg gttttttgtg cccctcgggc cggattgcta
      421 tctaccggca ttggcgcaga aaaaaatgcc tgatgcgacg ctgcgcgtct tatactccca
      481 catatgccag attcagcaac ggatacggct tccccaactt gcccacttcc atacgtgtcc
      541 tccttaccag aaatttatcc ttaaggtcgt cagctatcct gcaggcgatc tctcgatttc
      601 gatcaagaca ttcctttaat ggtcttttct ggacaccact aggggtcaga agtagttcat
      661 caaactttct tccctcccta atctcattgg ttaccttggg ctatcgaaac ttaattaacc
      721 agtcaagtca gctacttggc gagatcgact tgtctgggtt tcgactacgc tcagaattgc
      781 gtcagtcaag ttcgatctgg tccttgctat tgcacccgtt ctccgattac gagtttcatt
      841 taaatcatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
      901 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
      961 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     1021 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     1081 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     1141 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     1201 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     1261 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     1321 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     1381 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     1441 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     1501 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     1561 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     1621 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     1681 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc atttaaattt
     1741 ccgaactctc caaggccctc gtcggaaaat cttcaaacct ttcgtccgat ccatcttgca
     1801 ggctacctct cgaacgaact atcgcaagtc tcttggccgg ccttgcgcct tggctattgc
     1861 ttggcagcgc ctatcgccag gtattactcc aatcccgaat atccgagatc gggatcaccc
     1921 gagagaagtt caacctacat cctcaatccc gatctatccg agatccgagg aatatcgaaa
     1981 tcggggcgcg cctggcctcc gcgccgggtt ttggcgcctc ccgcgggcgc ccccctcgtc
     2041 acggcgagcg ctgccacgtc agacgaaggg cgcaggagcg tcctgatcct tccgcccgga
     2101 cgctcaggac agcggcccgc tgctcataag actcggcctt agaaccccag tatcagcaga
     2161 aggacatttt aggacgggac ttgggtgact ctagggcact ggttttcttt ccagagagcg
     2221 gaacaggcga ggaaaagtag tcccttctcg gcgattctgc ggagggatct ccgtggggcg
     2281 gtgaacgccg atgattatat aaggacgcgc cgggtgtggc acagctagtt ccgtcgcagc
     2341 cgggatttgg gtcgcggttc ttgtttgtgg atcgctgtga tcgtcacttg gtgagtagcg
     2401 ggctgctggg ctggccgggg ctttcgtggc cgccgggccg ctcggtggga cggaagcgtg
     2461 tggagagacc gccaagggct gtagtctggg tccgcgagca aggttgccct gaactggggg
     2521 ttggggggag cgcagcaaaa tggcggctgt tcccgagtct tgaatggaag acgcttgtga
     2581 ggcgggctgt gaggtcgttg aaacaaggtg gggggcatgg tgggcggcaa gaacccaagg
     2641 tcttgagccc ttcgctaatg cgggaaagct cttattcggg tgagatgggc tgggcaccat
     2701 ctggggaccc tgacgtgaag tttgtcactg actggagaac tcggtttgtc gtctgttgcg
     2761 ggggcggcag ttatggcggt gccgttgggc agtgcacccg tacctttggg agcgcgcgcc
     2821 ctcgtcgtgt cgtgacgtca cccgttctgt tggcttataa tgcagggtgg ggccaccagc
     2881 cggtaggtgt gcggtaggct tttctccgtc gcaggacgca gggttcgggc ctagggtagg
     2941 ctctcctgaa tcgacaggcg ccggacctct ggtgagggga gggataagtg aggcgtcagt
     3001 ttctttggtc ggttttatgt acctatcttc ttaagtagct gaagctccgg ttttgaacta
     3061 tgcgctcggg gttggcgagt gtgttttgtg aagtttttta ggcacctttt gaaatgtaat
     3121 catttgggtc aatatgtaat tttcagtgtt agacttgtaa attgtccgct aaattctggc
     3181 cgtttttggc ttttttgtta gacaacatgg gtaaaaagcc tgaactcacc gcgacgtctg
     3241 tcgagaagtt tctgatcgaa aagttcgaca gcgtctccga cctgatgcag ctctcggagg
     3301 gcgaagaatc tcgtgctttc agcttcgatg taggagggcg tggatatgtc ctgcgggtaa
     3361 atagctgcgc cgatggtttc tacaaagatc gttatgttta tcggcacttt gcatccgccg
     3421 cgctcccgat tccggaagtg cttgacattg gggagttcag cgagagcctg acctattgca
     3481 tctcccgccg tgcacagggt gtcacgttgc aagacctgcc tgaaaccgaa ctgcccgctg
     3541 ttctgcagcc ggtcgcggag gcaatggatg ccatcgctgc cgccgatctt agccagacga
     3601 gcgggttcgg cccattcgga ccgcaaggaa tcggtcaata cactacatgg cgtgatttca
     3661 tatgcgcgat tgctgatccc catgtgtatc actggcaaac tgtgatggac gacaccgtca
     3721 gtgcgtccgt cgcgcaggct ctcgatgagc tgatgctttg ggccgaggac tgccccgaag
     3781 tccggcacct cgtgcacgcg gatttcggct ccaacaatgt cctgacggac aatggccgca
     3841 taacagcggt cattgactgg agcgaggcga tgttcgggga ttcccaatac gaggtcgcca
     3901 acatcttctt ctggaggccg tggttggctt gtatggagca gcagacgcgc tacttcgagc
     3961 ggaggcatcc ggagcttgca ggatcgcccc ggctccgggc gtatatgctc cgcattggtc
     4021 ttgaccaact ctatcagagc ttggttgacg gcaatttcga tgatgcagct tgggcgcagg
     4081 gtcgatgcga cgcaatcgtc cgatccggag ccgggactgt cgggcgtaca caaatcgccc
     4141 gcagaagcgc cgccgtctgg accgatggct gtgtagaagt actcgccgat agtggaaacc
     4201 gacgccccag cactcgtccg agggcaaagg aataagctag tatgtaagcc tagtcttaga
     4261 taataaaatc gctatccatc gaagatggat gtgtgttggt tttttgtgtg tgtaacgcta
     4321 ggcgcgcctg gtgtaccgag aacgatcctc tcagtgcgag tctcgacgat ccatatcgtt
     4381 gcttggcagt cagccagtcg gaatccagct tgggacccag gaagtccaat cgtcagatat
     4441 tgtactcaag cctggtcacg gcagcgtacc gatctgttta aacctagata ttgatagtct
     4501 gatcggtcaa cgtataatcg agtcctagct tttgcaaaca tctatcaaga gacaggatca
     4561 gcaggaggct ttcgcatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     4621 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     4681 gatcagttgg gtgcgcgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     4741 gagagttttc gccccgaaga acgctttcca atgatgagca cttttaaagt tctgctatgt
     4801 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     4861 tctcagaatg acttggttga gtattcacca gtcacagaaa agcatcttac ggatggcatg
     4921 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     4981 cttctgacaa cgattggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     5041 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     5101 cgtgacacca cgatgcctgt agcaatggca acaaccttgc gtaaactatt aactggcgaa
     5161 ctacttactc tagcttcccg gcaacagttg atagactgga tggaggcgga taaagttgca
     5221 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     5281 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     5341 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     5401 gctgagatag gtgcctcact gattaagcat tggtaaccga ttctaggtgc attggcgcag
     5461 aaaaaaatgc ctgatgcgac gctgcgcgtc ttatactccc acatatgcca gattcagcaa
     5521 cggatacggc ttccccaact tgcccacttc catacgtgtc ctccttacca gaaatttatc
     5581 cttaagatcc cgaatcgttt aaactcgact ctggctctat cgaatctccg tcgtttcgag
     5641 cttacgcgaa cagccgtggc gctcatttgc tcgtcgggca tcgaatctcg tcagctatcg
     5701 tcagcttacc tttttggcag cgatcgcggc tcccgacatc ttggaccatt agctccacag
     5761 gtatcttctt ccctctagtg gtcataacag cagcttcagc tacctctcaa ttcaaaaaac
     5821 ccctcaagac ccgtttagag gccccaaggg gttatgctat caatcgttgc gttacacaca
     5881 caaaaaacca acacacatcc atcttcgatg gatagcgatt ttattatcta actgctgatc
     5941 gagtgtagcc agatctagta atcaattacg gggtcattag ttcatagccc atatatggag
     6001 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
     6061 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
     6121 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
     6181 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
     6241 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
     6301 attaccatgc tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
     6361 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
     6421 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
     6481 cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca gatcagatct
     6541 ttgtcgatcc taccatccac tcgacacacc cgccagcggc cgccaccatg atagaactat
     6601 cgctcatcga cttttatctg tgcttcttgg ctttcctgct ttttctggtg ctcattatgc
     6661 taatcatttt ttggttcagt ctggagttac aagatcataa tgagacat
//
