LOCUS       sequence-300711-        6753 bp    DNA     circular SYN 15-APR-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 6753)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6753)
  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-m4252934
FEATURES             Location/Qualifiers
     source          1..6753
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      220..808
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     709..728
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    1034..1105
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(1150..1286)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     terminator      complement(1316..1362)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    1477..1598
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1514..1533)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1568..1587
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(1667..1744)
                     /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(1745..1765)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     regulatory      1876..1885
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1954..2156)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1983..2003)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2158..2461
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    2542..2590
                     /note="synthetic polyadenylation signal"
     terminator      2608..2654
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    2659..2730
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(2882..3018)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(join(3031..3822,3823..3891))
                     /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     3654..3673
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    4157..4205
                     /note="synthetic polyadenylation signal"
     CDS             complement(4232..5260)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     promoter        complement(5264..6473)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(5408..5427)
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
ORIGIN
        1 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
       61 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
      121 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
      181 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
      241 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
      301 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
      361 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
      421 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
      481 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
      541 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
      601 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
      661 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
      721 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
      781 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
      841 ggccttttgc tggccttttg ctcacatgat ttaaatgaaa ctcgtaatcg gagaacgggt
      901 gcaatagcaa ggaccagatc gaacttgact gacgcaattc tgagcgtagt cgaaacccag
      961 acaagtcgat ctcgccaagt agctgacttg actggttaat taagtttcga tagcccaagg
     1021 taaccaatga gattagggag ggaagaaagt ttgatgaact acttctgacc cctagtggtg
     1081 tccagaaaag accattaaag gaatgtcttg atcgaaatcg agagatcgcc tgcaggatag
     1141 ctgacgacct taaggataaa tttctggtaa ggaggacacg tatggaagtg ggcaagttgg
     1201 ggaagccgta tccgttgctg aatctggcat atgtgggagt ataagacgcg cagcgtcgca
     1261 tcaggcattt ttttctgcgc caatgccggt agatagcaat ccggcccgag gggcacaaaa
     1321 aacccctcaa gacccgttta gaggccccaa ggggttatgc tacgataccg atagagatgg
     1381 gccaatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg
     1441 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat
     1501 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     1561 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtga gctgaaggta
     1621 cgctgtatct cagtcagtca agctagcaga ggcctgcgga tccttagagg gcaccagacg
     1681 aggatatctt tttgaatctg ttggctgcac tcaccgcgat aaaattcttt ttccatcgcc
     1741 tcttcgactg aaagtaaagg ttttctgtga gattaaagtt aactacatct acttgtgcta
     1801 tgtagttacg agaattcatt ctgcacaaga gtagactata tatcgtaaac ggaaaagcga
     1861 aaacgtttat atagcccatg gtggcggccg ctggcgggtg tgtcgagtgg atggtaggat
     1921 cgacaaagat ctgatctgac ggttcactaa accagctctg cttatataga cctcccaccg
     1981 tacacgccta ccgcccattt gcgtcaatgg ggcggagttg ttacgacatt ttggaaagtc
     2041 ccgttgattt tggtgccaaa acaaactccc attgacgtca atggggtgga gacttggaaa
     2101 tccccgtgag tcaaaccgct atccacgccc attgatgtac tgccaaaacc gcatcagcat
     2161 ggtaatagcg atgactaata cgtagatgta ctgccaagta ggaaagtccc ataaggtcat
     2221 gtactgggca taatgccagg cgggccattt accgtcattg acgtcaatag ggggcgtact
     2281 tggcatatga tacacttgat gtactgccaa gtgggcagtt taccgtaaat actccaccca
     2341 ttgacgtcaa tggaaagtcc ctattggcgt tactatggga acatacgtca ttattgacgt
     2401 caatgggcgg gggtcgttgg gcggtcagcc aggcgggcca tttaccgtaa gttatgtaac
     2461 gcggaactcc atatatgggc tatgaactaa tgaccccgta attgattact agatctggct
     2521 acactcgatc agcagttaga taataaaatc gctatccatc gaagatggat gtgtgttggt
     2581 tttttgtgtg tgtaacgcaa cgattgatag cataacccct tggggcctct aaacgggtct
     2641 tgaggggttt tttgaattga gaggtagctg aagctgctgt tatgaccact agagggaaga
     2701 agatacctgt ggagctaatg gtccaagatg tcgggagccg cgatcgctgc caaaaaggta
     2761 agctgacgat agctgacgag attcgatgcc cgacgagcaa atgagcgcca cggctgttcg
     2821 cgtaagctcg aaacgacgga gattcgatag agccagagtc gagtttaaac gattcgggat
     2881 cttaaggata aatttctggt aaggaggaca cgtatggaag tgggcaagtt ggggaagccg
     2941 tatccgttgc tgaatctggc atatgtggga gtataagacg cgcagcgtcg catcaggcat
     3001 ttttttctgc gccaatgcac ctagaatcgg ttaccaatgc ttaatcagtg aggcacctat
     3061 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     3121 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     3181 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     3241 tggtcctgca actttatccg cctccatcca gtctatcaac tgttgccggg aagctagagt
     3301 aagtagttcg ccagttaata gtttacgcaa ggttgttgcc attgctacag gcatcgtggt
     3361 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     3421 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc caatcgttgt
     3481 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     3541 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gaatactcaa ccaagtcatt
     3601 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     3661 cgcgccacat agcagaactt taaaagtgct catcattgga aagcgttctt cggggcgaaa
     3721 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gcgcacccaa
     3781 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     3841 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tgcgaaagcc
     3901 tcctgctgat cctgtctctt gatagatgtt tgcaaaagct aggactcgat tatacgttga
     3961 ccgatcagac tatcaatatc taggtttaaa cagatcggta cgctgccgtg accaggcttg
     4021 agtacaatat ctgacgattg gacttcctgg gtcccaagct ggattccgac tggctgactg
     4081 ccaagcaacg atatggatcg tcgagactcg cactgagagg atcgttctcg gtacaccagg
     4141 cgcgcctagc gttacacaca caaaaaacca acacacatcc atcttcgatg gatagcgatt
     4201 ttattatcta agactaggct tacatactag cttattcctt tgccctcgga cgagtgctgg
     4261 ggcgtcggtt tccactatcg gcgagtactt ctacacagcc atcggtccag acggcggcgc
     4321 ttctgcgggc gatttgtgta cgcccgacag tcccggctcc ggatcggacg attgcgtcgc
     4381 atcgaccctg cgcccaagct gcatcatcga aattgccgtc aaccaagctc tgatagagtt
     4441 ggtcaagacc aatgcggagc atatacgccc ggagccgggg cgatcctgca agctccggat
     4501 gcctccgctc gaagtagcgc gtctgctgct ccatacaagc caaccacggc ctccagaaga
     4561 agatgttggc gacctcgtat tgggaatccc cgaacatcgc ctcgctccag tcaatgaccg
     4621 ctgttatgcg gccattgtcc gtcaggacat tgttggagcc gaaatccgcg tgcacgaggt
     4681 gccggacttc ggggcagtcc tcggcccaaa gcatcagctc atcgagagcc tgcgcgacgg
     4741 acgcactgac ggtgtcgtcc atcacagttt gccagtgata cacatgggga tcagcaatcg
     4801 cgcatatgaa atcacgccat gtagtgtatt gaccgattcc ttgcggtccg aatgggccga
     4861 acccgctcgt ctggctaaga tcggcggcag cgatggcatc cattgcctcc gcgaccggct
     4921 gcagaacagc gggcagttcg gtttcaggca ggtcttgcaa cgtgacaccc tgtgcacggc
     4981 gggagatgca ataggtcagg ctctcgctga actccccaat gtcaagcact tccggaatcg
     5041 ggagcgcggc ggatgcaaag tgccgataaa cataacgatc tttgtagaaa ccatcggcgc
     5101 agctatttac ccgcaggaca tatccacgcc ctcctacatc gaagctgaaa gcacgagatt
     5161 cttcgccctc cgagagctgc atcaggtcgg agacgctgtc gaacttttcg atcagaaact
     5221 tctcgacaga cgtcgcggtg agttcaggct ttttacccat gttgtctaac aaaaaagcca
     5281 aaaacggcca gaatttagcg gacaatttac aagtctaaca ctgaaaatta catattgacc
     5341 caaatgatta catttcaaaa ggtgcctaaa aaacttcaca aaacacactc gccaaccccg
     5401 agcgcatagt tcaaaaccgg agcttcagct acttaagaag ataggtacat aaaaccgacc
     5461 aaagaaactg acgcctcact tatccctccc ctcaccagag gtccggcgcc tgtcgattca
     5521 ggagagccta ccctaggccc gaaccctgcg tcctgcgacg gagaaaagcc taccgcacac
     5581 ctaccggctg gtggccccac cctgcattat aagccaacag aacgggtgac gtcacgacac
     5641 gacgagggcg cgcgctccca aaggtacggg tgcactgccc aacggcaccg ccataactgc
     5701 cgcccccgca acagacgaca aaccgagttc tccagtcagt gacaaacttc acgtcagggt
     5761 ccccagatgg tgcccagccc atctcacccg aataagagct ttcccgcatt agcgaagggc
     5821 tcaagacctt gggttcttgc cgcccaccat gccccccacc ttgtttcaac gacctcacag
     5881 cccgcctcac aagcgtcttc cattcaagac tcgggaacag ccgccatttt gctgcgctcc
     5941 ccccaacccc cagttcaggg caaccttgct cgcggaccca gactacagcc cttggcggtc
     6001 tctccacacg cttccgtccc accgagcggc ccggcggcca cgaaagcccc ggccagccca
     6061 gcagcccgct actcaccaag tgacgatcac agcgatccac aaacaagaac cgcgacccaa
     6121 atcccggctg cgacggaact agctgtgcca cacccggcgc gtccttatat aatcatcggc
     6181 gttcaccgcc ccacggagat ccctccgcag aatcgccgag aagggactac ttttcctcgc
     6241 ctgttccgct ctctggaaag aaaaccagtg ccctagagtc acccaagtcc cgtcctaaaa
     6301 tgtccttctg ctgatactgg ggttctaagg ccgagtctta tgagcagcgg gccgctgtcc
     6361 tgagcgtccg ggcggaagga tcaggacgct cctgcgccct tcgtctgacg tggcagcgct
     6421 cgccgtgacg aggggggcgc ccgcgggagg cgccaaaacc cggcgcggag gccaggcgcg
     6481 ccccgatttc gatattcctc ggatctcgga tagatcggga ttgaggatgt aggttgaact
     6541 tctctcgggt gatcccgatc tcggatattc gggattggag taatacctgg cgataggcgc
     6601 tgccaagcaa tagccaaggc gcaaggccgg ccaagagact tgcgatagtt cgttcgagag
     6661 gtagcctgca agatggatcg gacgaaaggt ttgaagattt tccgacgagg gccttggaga
     6721 gttcggaaat ttaaatgcaa ctatggatga acg
//
