LOCUS       sequence-300736-        6720 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 ORF10 protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6720)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6720)
  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-m4252918
FEATURES             Location/Qualifiers
     source          1..6720
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    87..158
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(163..209)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    227..275
                     /note="synthetic polyadenylation signal"
     enhancer        356..659
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        661..863
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     814..834
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      932..941
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1052..1072
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1073..1117
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     polyA_signal    1186..1307
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1197..1216)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1251..1270
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1422..1468
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1498..1634
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1679..1750
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1976..2564)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2056..2075)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3064..4273
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4110..4129
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4277..5305
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5332..5380
                     /note="synthetic polyadenylation signal"
     CDS             join(5646..5714,5715..6506)
                     /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(5864..5883)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6519..6655
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
ORIGIN
        1 cagccgtggc gctcatttgc tcgtcgggca tcgaatctcg tcagctatcg tcagcttacc
       61 tttttggcag cgatcgcggc tcccgacatc ttggaccatt agctccacag gtatcttctt
      121 ccctctagtg gtcataacag cagcttcagc tacctctcaa ttcaaaaaac ccctcaagac
      181 ccgtttagag gccccaaggg gttatgctat caatcgttgc gttacacaca caaaaaacca
      241 acacacatcc atcttcgatg gatagcgatt ttattatcta actgctgatc gagtgtagcc
      301 agatctagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
      361 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
      421 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
      481 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
      541 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
      601 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgc
      661 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
      721 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
      781 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
      841 gggaggtcta tataagcaga gctggtttag tgaaccgtca gatcagatct ttgtcgatcc
      901 taccatccac tcgacacacc cgccagcggc cgccaccatg gggtacataa atgttttcgc
      961 ctttccattc acgatctatt cactcctttt atgtaggatg aactctcgca attacattgc
     1021 tcaggtagac gtcgtgaact ttaacctaac tgaaaacctt tactttcagt cgaaagaaac
     1081 cgcggcagct aagttcgaga ggcagcacat ggacagttaa ggatccgcag gcctctgcta
     1141 gcttgactga ctgagataca gcgtaccttc agctcacaga catgataaga tacattgatg
     1201 agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg
     1261 atgctattgc tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt
     1321 gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa
     1381 acctctacaa atgtggtatt ggcccatctc tatcggtatc gtagcataac cccttggggc
     1441 ctctaaacgg gtcttgaggg gttttttgtg cccctcgggc cggattgcta tctaccggca
     1501 ttggcgcaga aaaaaatgcc tgatgcgacg ctgcgcgtct tatactccca catatgccag
     1561 attcagcaac ggatacggct tccccaactt gcccacttcc atacgtgtcc tccttaccag
     1621 aaatttatcc ttaaggtcgt cagctatcct gcaggcgatc tctcgatttc gatcaagaca
     1681 ttcctttaat ggtcttttct ggacaccact aggggtcaga agtagttcat caaactttct
     1741 tccctcccta atctcattgg ttaccttggg ctatcgaaac ttaattaacc agtcaagtca
     1801 gctacttggc gagatcgact tgtctgggtt tcgactacgc tcagaattgc gtcagtcaag
     1861 ttcgatctgg tccttgctat tgcacccgtt ctccgattac gagtttcatt taaatcatgt
     1921 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     1981 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     2041 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     2101 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     2161 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     2221 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     2281 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     2341 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     2401 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     2461 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     2521 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     2581 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     2641 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     2701 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     2761 ctatctcagc gatctgtcta tttcgttcat ccatagttgc atttaaattt ccgaactctc
     2821 caaggccctc gtcggaaaat cttcaaacct ttcgtccgat ccatcttgca ggctacctct
     2881 cgaacgaact atcgcaagtc tcttggccgg ccttgcgcct tggctattgc ttggcagcgc
     2941 ctatcgccag gtattactcc aatcccgaat atccgagatc gggatcaccc gagagaagtt
     3001 caacctacat cctcaatccc gatctatccg agatccgagg aatatcgaaa tcggggcgcg
     3061 cctggcctcc gcgccgggtt ttggcgcctc ccgcgggcgc ccccctcgtc acggcgagcg
     3121 ctgccacgtc agacgaaggg cgcaggagcg tcctgatcct tccgcccgga cgctcaggac
     3181 agcggcccgc tgctcataag actcggcctt agaaccccag tatcagcaga aggacatttt
     3241 aggacgggac ttgggtgact ctagggcact ggttttcttt ccagagagcg gaacaggcga
     3301 ggaaaagtag tcccttctcg gcgattctgc ggagggatct ccgtggggcg gtgaacgccg
     3361 atgattatat aaggacgcgc cgggtgtggc acagctagtt ccgtcgcagc cgggatttgg
     3421 gtcgcggttc ttgtttgtgg atcgctgtga tcgtcacttg gtgagtagcg ggctgctggg
     3481 ctggccgggg ctttcgtggc cgccgggccg ctcggtggga cggaagcgtg tggagagacc
     3541 gccaagggct gtagtctggg tccgcgagca aggttgccct gaactggggg ttggggggag
     3601 cgcagcaaaa tggcggctgt tcccgagtct tgaatggaag acgcttgtga ggcgggctgt
     3661 gaggtcgttg aaacaaggtg gggggcatgg tgggcggcaa gaacccaagg tcttgagccc
     3721 ttcgctaatg cgggaaagct cttattcggg tgagatgggc tgggcaccat ctggggaccc
     3781 tgacgtgaag tttgtcactg actggagaac tcggtttgtc gtctgttgcg ggggcggcag
     3841 ttatggcggt gccgttgggc agtgcacccg tacctttggg agcgcgcgcc ctcgtcgtgt
     3901 cgtgacgtca cccgttctgt tggcttataa tgcagggtgg ggccaccagc cggtaggtgt
     3961 gcggtaggct tttctccgtc gcaggacgca gggttcgggc ctagggtagg ctctcctgaa
     4021 tcgacaggcg ccggacctct ggtgagggga gggataagtg aggcgtcagt ttctttggtc
     4081 ggttttatgt acctatcttc ttaagtagct gaagctccgg ttttgaacta tgcgctcggg
     4141 gttggcgagt gtgttttgtg aagtttttta ggcacctttt gaaatgtaat catttgggtc
     4201 aatatgtaat tttcagtgtt agacttgtaa attgtccgct aaattctggc cgtttttggc
     4261 ttttttgtta gacaacatgg gtaaaaagcc tgaactcacc gcgacgtctg tcgagaagtt
     4321 tctgatcgaa aagttcgaca gcgtctccga cctgatgcag ctctcggagg gcgaagaatc
     4381 tcgtgctttc agcttcgatg taggagggcg tggatatgtc ctgcgggtaa atagctgcgc
     4441 cgatggtttc tacaaagatc gttatgttta tcggcacttt gcatccgccg cgctcccgat
     4501 tccggaagtg cttgacattg gggagttcag cgagagcctg acctattgca tctcccgccg
     4561 tgcacagggt gtcacgttgc aagacctgcc tgaaaccgaa ctgcccgctg ttctgcagcc
     4621 ggtcgcggag gcaatggatg ccatcgctgc cgccgatctt agccagacga gcgggttcgg
     4681 cccattcgga ccgcaaggaa tcggtcaata cactacatgg cgtgatttca tatgcgcgat
     4741 tgctgatccc catgtgtatc actggcaaac tgtgatggac gacaccgtca gtgcgtccgt
     4801 cgcgcaggct ctcgatgagc tgatgctttg ggccgaggac tgccccgaag tccggcacct
     4861 cgtgcacgcg gatttcggct ccaacaatgt cctgacggac aatggccgca taacagcggt
     4921 cattgactgg agcgaggcga tgttcgggga ttcccaatac gaggtcgcca acatcttctt
     4981 ctggaggccg tggttggctt gtatggagca gcagacgcgc tacttcgagc ggaggcatcc
     5041 ggagcttgca ggatcgcccc ggctccgggc gtatatgctc cgcattggtc ttgaccaact
     5101 ctatcagagc ttggttgacg gcaatttcga tgatgcagct tgggcgcagg gtcgatgcga
     5161 cgcaatcgtc cgatccggag ccgggactgt cgggcgtaca caaatcgccc gcagaagcgc
     5221 cgccgtctgg accgatggct gtgtagaagt actcgccgat agtggaaacc gacgccccag
     5281 cactcgtccg agggcaaagg aataagctag tatgtaagcc tagtcttaga taataaaatc
     5341 gctatccatc gaagatggat gtgtgttggt tttttgtgtg tgtaacgcta ggcgcgcctg
     5401 gtgtaccgag aacgatcctc tcagtgcgag tctcgacgat ccatatcgtt gcttggcagt
     5461 cagccagtcg gaatccagct tgggacccag gaagtccaat cgtcagatat tgtactcaag
     5521 cctggtcacg gcagcgtacc gatctgttta aacctagata ttgatagtct gatcggtcaa
     5581 cgtataatcg agtcctagct tttgcaaaca tctatcaaga gacaggatca gcaggaggct
     5641 ttcgcatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     5701 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     5761 gtgcgcgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     5821 gccccgaaga acgctttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     5881 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     5941 acttggttga gtattcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     6001 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     6061 cgattggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     6121 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     6181 cgatgcctgt agcaatggca acaaccttgc gtaaactatt aactggcgaa ctacttactc
     6241 tagcttcccg gcaacagttg atagactgga tggaggcgga taaagttgca ggaccacttc
     6301 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     6361 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     6421 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     6481 gtgcctcact gattaagcat tggtaaccga ttctaggtgc attggcgcag aaaaaaatgc
     6541 ctgatgcgac gctgcgcgtc ttatactccc acatatgcca gattcagcaa cggatacggc
     6601 ttccccaact tgcccacttc catacgtgtc ctccttacca gaaatttatc cttaagatcc
     6661 cgaatcgttt aaactcgact ctggctctat cgaatctccg tcgtttcgag cttacgcgaa
//
