LOCUS       sequence-301684-        7080 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 7080)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7080)
  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-m4252878
FEATURES             Location/Qualifiers
     source          1..7080
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        474..1683
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     1520..1539
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             1687..2715
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    2742..2790
                     /note="synthetic polyadenylation signal"
     CDS             join(3056..3124,3125..3916)
                     /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(3274..3293)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3929..4065
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4217..4288
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4293..4339)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4357..4405
                     /note="synthetic polyadenylation signal"
     enhancer        4486..4789
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4791..4993
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4944..4964
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             5197..5217
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             5566..5583
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    5676..5797
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5687..5706)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5741..5760
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      5912..5958
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      5988..6124
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6169..6240
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(6466..7054)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6546..6565)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
       61 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
      121 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
      181 gatctgtcta tttcgttcat ccatagttgc atttaaattt ccgaactctc caaggccctc
      241 gtcggaaaat cttcaaacct ttcgtccgat ccatcttgca ggctacctct cgaacgaact
      301 atcgcaagtc tcttggccgg ccttgcgcct tggctattgc ttggcagcgc ctatcgccag
      361 gtattactcc aatcccgaat atccgagatc gggatcaccc gagagaagtt caacctacat
      421 cctcaatccc gatctatccg agatccgagg aatatcgaaa tcggggcgcg cctggcctcc
      481 gcgccgggtt ttggcgcctc ccgcgggcgc ccccctcgtc acggcgagcg ctgccacgtc
      541 agacgaaggg cgcaggagcg tcctgatcct tccgcccgga cgctcaggac agcggcccgc
      601 tgctcataag actcggcctt agaaccccag tatcagcaga aggacatttt aggacgggac
      661 ttgggtgact ctagggcact ggttttcttt ccagagagcg gaacaggcga ggaaaagtag
      721 tcccttctcg gcgattctgc ggagggatct ccgtggggcg gtgaacgccg atgattatat
      781 aaggacgcgc cgggtgtggc acagctagtt ccgtcgcagc cgggatttgg gtcgcggttc
      841 ttgtttgtgg atcgctgtga tcgtcacttg gtgagtagcg ggctgctggg ctggccgggg
      901 ctttcgtggc cgccgggccg ctcggtggga cggaagcgtg tggagagacc gccaagggct
      961 gtagtctggg tccgcgagca aggttgccct gaactggggg ttggggggag cgcagcaaaa
     1021 tggcggctgt tcccgagtct tgaatggaag acgcttgtga ggcgggctgt gaggtcgttg
     1081 aaacaaggtg gggggcatgg tgggcggcaa gaacccaagg tcttgagccc ttcgctaatg
     1141 cgggaaagct cttattcggg tgagatgggc tgggcaccat ctggggaccc tgacgtgaag
     1201 tttgtcactg actggagaac tcggtttgtc gtctgttgcg ggggcggcag ttatggcggt
     1261 gccgttgggc agtgcacccg tacctttggg agcgcgcgcc ctcgtcgtgt cgtgacgtca
     1321 cccgttctgt tggcttataa tgcagggtgg ggccaccagc cggtaggtgt gcggtaggct
     1381 tttctccgtc gcaggacgca gggttcgggc ctagggtagg ctctcctgaa tcgacaggcg
     1441 ccggacctct ggtgagggga gggataagtg aggcgtcagt ttctttggtc ggttttatgt
     1501 acctatcttc ttaagtagct gaagctccgg ttttgaacta tgcgctcggg gttggcgagt
     1561 gtgttttgtg aagtttttta ggcacctttt gaaatgtaat catttgggtc aatatgtaat
     1621 tttcagtgtt agacttgtaa attgtccgct aaattctggc cgtttttggc ttttttgtta
     1681 gacaacatgg gtaaaaagcc tgaactcacc gcgacgtctg tcgagaagtt tctgatcgaa
     1741 aagttcgaca gcgtctccga cctgatgcag ctctcggagg gcgaagaatc tcgtgctttc
     1801 agcttcgatg taggagggcg tggatatgtc ctgcgggtaa atagctgcgc cgatggtttc
     1861 tacaaagatc gttatgttta tcggcacttt gcatccgccg cgctcccgat tccggaagtg
     1921 cttgacattg gggagttcag cgagagcctg acctattgca tctcccgccg tgcacagggt
     1981 gtcacgttgc aagacctgcc tgaaaccgaa ctgcccgctg ttctgcagcc ggtcgcggag
     2041 gcaatggatg ccatcgctgc cgccgatctt agccagacga gcgggttcgg cccattcgga
     2101 ccgcaaggaa tcggtcaata cactacatgg cgtgatttca tatgcgcgat tgctgatccc
     2161 catgtgtatc actggcaaac tgtgatggac gacaccgtca gtgcgtccgt cgcgcaggct
     2221 ctcgatgagc tgatgctttg ggccgaggac tgccccgaag tccggcacct cgtgcacgcg
     2281 gatttcggct ccaacaatgt cctgacggac aatggccgca taacagcggt cattgactgg
     2341 agcgaggcga tgttcgggga ttcccaatac gaggtcgcca acatcttctt ctggaggccg
     2401 tggttggctt gtatggagca gcagacgcgc tacttcgagc ggaggcatcc ggagcttgca
     2461 ggatcgcccc ggctccgggc gtatatgctc cgcattggtc ttgaccaact ctatcagagc
     2521 ttggttgacg gcaatttcga tgatgcagct tgggcgcagg gtcgatgcga cgcaatcgtc
     2581 cgatccggag ccgggactgt cgggcgtaca caaatcgccc gcagaagcgc cgccgtctgg
     2641 accgatggct gtgtagaagt actcgccgat agtggaaacc gacgccccag cactcgtccg
     2701 agggcaaagg aataagctag tatgtaagcc tagtcttaga taataaaatc gctatccatc
     2761 gaagatggat gtgtgttggt tttttgtgtg tgtaacgcta ggcgcgcctg gtgtaccgag
     2821 aacgatcctc tcagtgcgag tctcgacgat ccatatcgtt gcttggcagt cagccagtcg
     2881 gaatccagct tgggacccag gaagtccaat cgtcagatat tgtactcaag cctggtcacg
     2941 gcagcgtacc gatctgttta aacctagata ttgatagtct gatcggtcaa cgtataatcg
     3001 agtcctagct tttgcaaaca tctatcaaga gacaggatca gcaggaggct ttcgcatgag
     3061 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     3121 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcgcgagt
     3181 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     3241 acgctttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     3301 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     3361 gtattcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     3421 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgattggagg
     3481 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     3541 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     3601 agcaatggca acaaccttgc gtaaactatt aactggcgaa ctacttactc tagcttcccg
     3661 gcaacagttg atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     3721 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     3781 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     3841 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     3901 gattaagcat tggtaaccga ttctaggtgc attggcgcag aaaaaaatgc ctgatgcgac
     3961 gctgcgcgtc ttatactccc acatatgcca gattcagcaa cggatacggc ttccccaact
     4021 tgcccacttc catacgtgtc ctccttacca gaaatttatc cttaagatcc cgaatcgttt
     4081 aaactcgact ctggctctat cgaatctccg tcgtttcgag cttacgcgaa cagccgtggc
     4141 gctcatttgc tcgtcgggca tcgaatctcg tcagctatcg tcagcttacc tttttggcag
     4201 cgatcgcggc tcccgacatc ttggaccatt agctccacag gtatcttctt ccctctagtg
     4261 gtcataacag cagcttcagc tacctctcaa ttcaaaaaac ccctcaagac ccgtttagag
     4321 gccccaaggg gttatgctat caatcgttgc gttacacaca caaaaaacca acacacatcc
     4381 atcttcgatg gatagcgatt ttattatcta actgctgatc gagtgtagcc agatctagta
     4441 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
     4501 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
     4561 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
     4621 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
     4681 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
     4741 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgc tgatgcggtt
     4801 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
     4861 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
     4921 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
     4981 tataagcaga gctggtttag tgaaccgtca gatcagatct ttgtcgatcc taccatccac
     5041 tcgacacacc cgccagcggc cgccaccatg attgaacttt cattaattga cttctatttg
     5101 tgctttttag cctttctgct attccttgtt ttaattatgc ttattatctt ttggttctca
     5161 cttgaactgc aagatcataa tgaaacttgt cacgccgaaa acctttactt tcagtcgatg
     5221 agtgacaaaa tcattcatct aaccgatgac tcctttgata cagacgtgtt gaaggccgat
     5281 ggtgcgatac tggtagattt ctgggctgaa tggtgtggcc cgtgcaagat gatcgcaccc
     5341 attctggacg agatcgccga cgagtatcaa ggaaaattaa ctgtcgctaa gcttaatata
     5401 gatcagaacc cagggacggc acctaaatac ggcatcaggg gaatacctac cctcctgctc
     5461 tttaagaatg gggaagttgc cgcgacaaaa gtgggagccc tgtctaaggg ccagctaaaa
     5521 gagttcttgg acgctaacct ggcaggttca ggctcggggc acatgcacca tcaccatcac
     5581 catagcagcg ggctcgtgcc acgcggctaa ggatccgcag gcctctgcta gcttgactga
     5641 ctgagataca gcgtaccttc agctcacaga catgataaga tacattgatg agtttggaca
     5701 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     5761 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
     5821 tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa
     5881 atgtggtatt ggcccatctc tatcggtatc gtagcataac cccttggggc ctctaaacgg
     5941 gtcttgaggg gttttttgtg cccctcgggc cggattgcta tctaccggca ttggcgcaga
     6001 aaaaaatgcc tgatgcgacg ctgcgcgtct tatactccca catatgccag attcagcaac
     6061 ggatacggct tccccaactt gcccacttcc atacgtgtcc tccttaccag aaatttatcc
     6121 ttaaggtcgt cagctatcct gcaggcgatc tctcgatttc gatcaagaca ttcctttaat
     6181 ggtcttttct ggacaccact aggggtcaga agtagttcat caaactttct tccctcccta
     6241 atctcattgg ttaccttggg ctatcgaaac ttaattaacc agtcaagtca gctacttggc
     6301 gagatcgact tgtctgggtt tcgactacgc tcagaattgc gtcagtcaag ttcgatctgg
     6361 tccttgctat tgcacccgtt ctccgattac gagtttcatt taaatcatgt gagcaaaagg
     6421 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     6481 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     6541 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     6601 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     6661 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     6721 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     6781 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     6841 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     6901 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     6961 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     7021 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
//
